Amina Zorlak-Čavčić

This review proposes an integrative mechanistic model that brings together apoptosis, pyroptosis, ferroptosis, necroptosis, and necrosis within the broader context of heat-induced proteostasis disruption. Rather than depicting a strictly sequential or fully validated causal pathway, the model provides a conceptual framework through which interactions among cellular stress responses, endothelial dysfunction, heatstroke-induced coagulopathy (HIC), and multiple organ dysfunction syndrome (MODS) may be understood. Under extreme heat stress, widespread protein unfolding and chaperone saturation activate the unfolded protein response, disrupt endoplasmic (ER) calcium homeostasis, and couple through Ca²⁺-mediated injury to mitochondrial dysfunction and oxidative stress. These processes are reciprocally amplifying: reactive oxygen species (ROS) from failing mitochondria deepen proteostatic damage and potentiate necroptotic and ferroptotic signaling, while inflammation-driven immune activation further increases ROS production, closing a self-reinforcing cycle. The resulting engagement of multiple RCD pathways, apoptosis, pyroptosis, ferroptosis, and necroptosis, each with qualitatively distinct DAMP signatures, promotes inflammasome activation and a cytokine storm that destabilizes the endothelial glycocalyx, impairs barrier integrity, and triggers a pro-coagulant shift underlying HIC. We further examine biomarker candidates reflecting these mechanisms, evaluate emerging therapeutic strategies, ranging from HSP modulation and antioxidant therapy to endothelial-protective agents, mitophagy regulators, and nanomedicine, and outline the future role of multi-omics, machine learning, and regenerative approaches. By bringing together molecular, cellular, vascular, and systemic processes into a single cohesive model, this review proposes a conceptual foundation to guide diagnostic refinement, therapeutic innovation, and forensic interpretation in severe heat-related injury.

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.

Sex estimation is a fundamental component of biological profiling in forensic anthropology, particularly when skeletal remains are incomplete or fragmented. This study aimed to evaluate sex estimation of the cranial base using geometric morphometrics and to assess the predictive value of cranial base morphology for sex estimation. The study included 211 adult skulls (139 male, 72 female) from the Bosnian population. Each skull was digitized to generate 3D models, and 27 anatomical landmarks were recorded. Landmark coordinates were standardized using Generalized Procrustes Analysis, Principal Component Analysis, Discriminant Function Analysis with permutation testing, and regression of shape on centroid size. Statistically significant sex estimation was observed at both the form (shape and size) and shape levels. Classification accuracy based on cranial base form reached 92.81% for males and 86.11% for females. Shape-based classification, after removal of size effects, also showed high accuracy (90.65% for males and 81.94% for females). Regression analysis indicated that size contributed significantly but modestly to shape variation. The cranial base exhibits stable sexually dimorphic patterns and may represent a reliable anatomical region for sex estimation. These findings contribute to population-specific standards for the Bosnia and Herzegovina population and support the forensic applicability of 3D geometric morphometric approaches.

Background: Sudden death is an unexpected natural fatal event that may arise from both cardiac and noncardiac causes. Differentiating true primary cardiac pathology from incidental or secondary myocardial changes remains a major challenge in forensic evaluation, particularly in clearly noncardiac sudden deaths. Aim: The aim of this study was to assess histopathological changes in the heart in cases of sudden death of cardiac and noncardiac origin and to compare the distribution of these findings across different certified causes of death. Additionally, the study aimed to evaluate the frequency and potential forensic relevance of these cardiac alterations. Materials and Methods: A retrospective cross-sectional analysis was conducted on 78 medico-legal autopsy cases of sudden death examined at the Department of Forensic Medicine in Bosnia and Herzegovina (2016–2025). Paraffin-embedded cardiac tissue was evaluated histologically by three independent pathologists, while cases involving unnatural causes of death or significant autolysis were excluded. Results: Cardiac-origin deaths accounted for 76.9% of cases and were more common in males. Individuals who died from cardiac causes were significantly older than those with noncardiac causes (58.7 ± 19.2 vs. 45.7 ± 16.8 years; P = 0.012). Cardiac-origin deaths predominated in middle-aged and older adults, whereas noncardiac sudden deaths occurred more frequently in younger individuals. Myocardial infarction and scar tissue were significantly associated with cardiac-origin deaths ( P = 0.017 and P = 0.038). Hypertrophy and dispersed fibrosis increased progressively with age. Conclusion: Cardiac sudden deaths were more frequent and mainly associated with chronic myocardial changes rather than acute ischemia. Middle-aged individuals were the most vulnerable group, highlighting the need for improved cardiovascular risk assessment and prevention.

The aim of our research was to determine the progression of postmortem ocular changes, focusing on the retina and sclera, over a specified time period and to assess whether these changes are influenced by environmental or physiological factors. Twelve Wistar rats were divided into four experimental groups based on postmortem interval (PMI): immediate autopsy (control), 24 h, 48 h, and 72 h. The mechanism of death was asphyxia by hanging. Each eye sample was macroscopically and microscopically monitored for retinal and scleral changes depending on the PMI and eye exposure (open vs. closed). The most pronounced ocular changes were observed at the longest PMI, including complete loss of retinal stratification, degeneration of ganglion cells, and infiltration by mononuclear inflammatory cells. Similar degenerative changes were observed in the scleral epithelium, particularly in open eyes. This experimental model provides a foundational framework for studying postmortem ocular changes, particularly in the retina and sclera, and may enhance the accuracy of PMI estimation in forensic investigations.

Hypothermia is defined as a body core temperature below 35 °C andcan be caused by internal or external stress. Primary hypothermiais caused by excessive exposure to low environmental temperaturewithout any medical conditions prior to that. Secondary hypothermiais caused by alteration in thermoregulation by disease, trauma, surgery,drugs, or infections. The aim of the research is to investigate coretemperature values in rats subjected to specific water temperatures atfive different time points. It focuses on distinguishing between primaryand secondary hypothermia in these rats.The total 21 Wistar rats were divided into three experimental groupsas: Control group rats exposed only to hypothermic condition (n = 7);Alcohol + hypothermia (n = 7); and Benzodiazepines + hypothermia(n = 7). The temperature spots analyzed in the study were: normal coretemperature, core temperature during injection of 0,3 ml ketamine,temperature of immersion and the temperature at the onset ofhypothermia and temperature at the time of death.In our study the comparative analysis of body temperatures at varioustime points following submersion in water revealed significantdifferences among the study groups treated with either alcoholor benzodiazepines and the control group. Notable differenceswere observed in baseline temperature, post-anesthesia inductiontemperature, and immediate post-submersion temperature. Specifically,significant differences were discovered among the alcohol andbenzodiazepine groups (p < 0.001) and ranging from the alcohol andcontrol groups (p < 0.001). The analysis of survival times followinginduced hypothermia revealed a statistically significant differenceamong the three experimental groups (p = 0.04), though subsequentpost-hoc comparisons did not demonstrate significant differences inmean survival times.There is a difference in survival time between primary and secondaryhypothermia groups, depending on consumption and intoxication withalcohol or benzodiazepines. The analysis of survival times followinginduced hypothermia showed a statistically significant differenceamong the groups.