The Tomašica grave-site near Prijedor in the north of Bosnia is reported to be the largest primary mass grave discovered thus far relating to the 1992–95 war. A total of 275 complete bodies and 125 body parts were exhumed from it in 2013. Post mortem examinations of the victims showed that nearly all had died from gunshot injuries but an additional striking feature was the degree of preservation of many of the bodies, even 21 years on, with skin, soft tissues and internal organs still present in abundance and gross structures clearly identifiable. Histology was performed on 68 samples of soft tissue from a total 13 bodies, on both skin and internal organs, and the degree of preservation was assessed in terms of the ability to recognize microscopic structure. Further comparison was made with samples taken a month or so later (56 tissue samples from 9 bodies, all but one different from the first group), after the bodies had been covered in salt as a means of general preservation. Generally, at a microscopic level, skin and subcutaneous tissues were better preserved than internal organs, while tissues sampled at the time of autopsy were better preserved than those sampled weeks later.

Microscopic signs are not specific only to drowning, but to any other type of suffocation, the background of which is mechanical obstruction. What about cases where there are no soft tissues when he has a skeletal body? This pilot research aims to examine for the first time the attendance of diatoms in teeth and bones, with regard to the peripheral vascularization of bones and teeth, in the Bosnia River. The thirty rats were divided into groups: Group A (n = 6; autopsy—1 hour after death; cause of death-hanging); Group B (n = 6; autopsy—72 hours after death; cause of death-hanging); Group C (n = 6; autopsy—immediately after drowning; cause of death-drowning); Group D (n = 6; autopsy—24 hours after drowning; cause of death-drowning) and Group E (n = 6; autopsy—72 hours after drowning; cause of death-drowning). In Group D and Group E, diatoms were found in one rat per group. By optimizing the “Diatom Test” in experimental settings, it could potentially become a routine method in the future. This is just the initial research that leads us toward optimizing testing and taking samples in cases of uncleared etiology, when there are no preserved soft tissue structures for autopsy.

Hypothermia-related deaths present significant diagnostic challenges due to non-specific and often inconsistent autopsy findings. This study investigated the histological and immunohistochemical alterations associated with primary and secondary hypothermia in an experimental Rattus norvegicus model, focusing on the effects of benzodiazepine and alcohol ingestion. Twenty-one male rats were divided into three groups: control (K), benzodiazepine-treated (B), and alcohol-treated (A). After two weeks of substance administration, hypothermia was induced and multiple organ samples were analyzed. Histologically, renal tissue showed hydropic and vacuolar degeneration, congestion, and acute tubular injury across all groups, with no significant differences in E-cadherin expression. Lung samples revealed congestion, emphysema, and hemorrhage, with more pronounced vascular congestion in the alcohol and benzodiazepine groups. Cardiac tissue exhibited vacuolar degeneration and protein denaturation, particularly in substance-exposed animals. The spleen showed preserved architecture but increased erythrocyte infiltration and significantly elevated myeloperoxidase (MPO)-positive granulocytes in the intoxicated groups. Liver samples demonstrated congestion, focal necrosis, and subcapsular hemorrhage, especially in the alcohol group. Immunohistochemical analysis revealed statistically significant differences in MPO expression in both lung and spleen tissues, with the highest levels observed in the benzodiazepine group. Similarly, CK7 and CK20 expression in the gastroesophageal junction was significantly elevated in both alcohol- and benzodiazepine-treated animals compared to the controls. In contrast, E-cadherin expression in the kidney did not differ significantly among the groups. These findings suggest that specific histological and immunohistochemical patterns, particularly involving pulmonary, cardiac, hepatic, and splenic tissues, may help differentiate primary hypothermia from substance-related secondary hypothermia. The study underscores the value of integrating toxicological, histological, and molecular analyses to enhance the forensic assessment of hypothermia-related fatalities. Future research should aim to validate these markers in human autopsy series and explore additional molecular indicators to refine diagnostic accuracy in forensic pathology.

Introduction/Objective. The diffuse axonal injury has a very important place in clinical and forensic aspects of neurotraumatology. A special challenge is proving it in situations of short survival (less than two hours) after a craniocerebral injury. The aim of this study was to determine the efficacy of beta-amyloid precursor protein (ßAPP) immunohistochemical staining in postmortem diagnosis of axonal injuries in head injury survival shorter than two hours, its expression and distribution through the brain tissue of the deceased. Methods. 36 adult fatalities, both sexes, injured by acceleration-deceleration mechanisms were divided into two groups: died up to two hours and died more than two hours after the injury. Immunostaining of brain tissue samples (frontal parasagittal white mass, genu and splenium of the corpus callosum and rostral pons) was used to register ßAPP positivity. Data were processed by methods of descriptive and inferential nonparametric statistics, and p < 0.05 was considered statistically significant. Results. The ßAPP immunopositivity was shown in 88.9% of cases (82.3% of ≤ two hours group vs. 94.7% of > two hours group). βAPP expression was enhanced towards the posterior structures of the brain. The shortest survival period with detected βAPP immunopositivity was 20-25 minutes, in three cases. There was an association of βAPP expression in the brainstem and interhemispheric/paramesencephalic subarachnoid hemorrhage (p = 0.035). Conclusion. βAPP immunohistochemical staining is effective in proving diffuse axonal injury in casualties that survived less than half an hour. Interhemispheric/paramesencephalic subarachnoid hemorrhage may indicate a more severe form of axonal injury.

Postmortem biochemistry is a valuable tool in forensic investigations, providing insights into the tissue damage and organ dysfunction associated with death. This study aimed to identify biochemical markers that distinguish primary and secondary hypothermia. Twenty-one Wistar rats were allocated into three groups: the Control group (n = 7), which was exposed only to hypothermic conditions, the Alcohol + Hypothermia group (n = 7), and the Benzodiazepines + Hypothermia group (n = 7). The temperature metrics assessed included the normal core temperature, the post-ketamine (0.3 ml injection) core temperature, the immersion temperature, temperature at the onset of hypothermia, and temperature at death. Blood samples were collected from the thoracic aorta in EDTA vacuum tubes for biochemical analysis. The key biochemical parameters measured included the Total Protein (g/L), Albumin (g/L), Globulin (g/L), Albumin to Globulin Ratio, Alanine Aminotransferase (U/L), Alkaline Phosphatase (U/L), Cholesterol (mmol/L), Amylase (U/L), and Lipase (U/L), using an automated IDEXX (Netherlands) cell counter. Significant between-group differences were found for the total protein and globulin levels (p < 0.001 and p = 0.002, respectively), with post-hoc tests confirming differences between the alcohol and control, and benzodiazepine and control groups. The cholesterol levels were found to be significantly different through an omnibus test (p = 0.03), but post hoc tests did not confirm these differences on a statistically significant level. The amylase levels varied significantly across all groups (p < 0.001), with post hoc tests confirming significant differences among all pairs: alcohol vs. benzodiazepine (p = 0.002), alcohol vs. control (p = 0.003), and benzodiazepine vs. control (p < 0.001). The lipase levels showed significant differences in the omnibus test (p = 0.030), but there was no significance in the post hoc tests. Amylase emerged as the most significant parameter in our study, with reduced levels strongly associated with secondary hypothermia. These findings highlight the potential use of total protein, globulin, and amylase levels as biomarkers to differentiate between primary and secondary hypothermia in forensic contexts.