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.

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.

Background: Lyme borreliosis is a multisystemic infection caused by the spirochete Borrelia burgdorferi. Erythema migras is the main clinical marker of the disease. Objective: This study aimed was to investigate the frequency and clinical manifestations of European borreliosis on the skin, and to determine the significance of these findings for diagnosis and therapy. Methods: A retrospective-prospective clinical study of outpatients treated and monitored in a private clinic of an infectologist was conducted over nine years from to 2013-2021. The study was clinical, descriptive and analytical in nature. Results: In the investigated period, 509 (30.8%) patients with borreliosis symptoms were treated. EM in our patients occurred under the following conditions: a) ringed redness, b) redness of target cels and d) continuous round or oval redness of different sizes of individual redness, or multiple occurrences with primary dissemination. Skin changes with multiorgan chronic symptoms of borreliosis occurred in 67.7% of cases the including: walking redness of different shapes and sizes, pink borreliosis stretch marks, white borreliosis stretch marks, borreliosis palms and soles, psoriatic changes, Acrodermatitis chronica atrophicans, Scleroderma circumscripta-morphae, Erythema nodosum, Granuloma anulare and Lichen striatus et atrophicans. Of the 509 patients treated for borreliosis, 32.3% with multi-organ symptomatology had no skin changes. Conclusion: The skin manifestations of European borreliosis are multi-layered and Erythema migrans are basic, but not the only markers of the disease. ‘Pink borreliose stretch marks, “white borreliosis striae”, “borreliosis palms or soles”, and intermittent redness accompanied by itching are unique markers for the diagnosis of chronic borreliosis, if they are manifested.

INTRODUCTION Diatom tests are rarely used during autopsy to confirm drowning as the cause of death (COD) because of limitations of the current literature involving these techniques. Instead, experts rely on physical examination by the pathologist. Due to interpretive concerns regarding Diatom tests, they are often insufficient in establishing a diagnosis, but offer the potential to be an extremely useful diagnostic tool with further understanding. The aim of study is to optimize "Diatom Tests" for use in forensic medicine in Bosnia and Herzegovina. METHODS A randomized prospective experimental study was conducted, using albino Wistar rat models (Rattus norvegicus), at the Veterinary Facility, University of Sarajevo. Thirty-two adult albino rats, were used and distributed into groups as follows: Group A (6 deceased rats with COD other than drowning, but due to mechanical asphyxia, which were then submerged for 1 h after death); Group B (6 deceased rats with COD other than drowning, but due to mechanical asphyxia, which were then submerged for 72 h after death); Group C (6 rats that were immediately autopsied after drowning, with COD determined as drowning); Group D (6 rats that underwent a 48-hour postmortem period after drowning); Group E (COD: drowning, post-mortem 72hrs after death, remained submerged in water until PM). Live algological material was collected for the research of the systematics of algae from the Bosna river, Sarajevo, and transported to the University of Sarajevo (Department of Biology, Faculty of Science). Periphyllon, epiphyllon and epipelon were used to collect phytobenthos. The material was fixed with 4% formalin solution. Laboratory processing of diatoms was performed using the methods described by Hustedt (16). In the process of obtaining pure diatom valves, part of the material is digested with potassium permanganate (KMnO4), sulfuric acid (H2SO4), and oxalic acid (C2H2O4). In the next step, the cleaned diatom valves were mounted in Canadian balsam. A light microscope under 1000x magnification (Best Scope 2020) was used to evaluate and analyze the species. The identification of diatoms was performed using the reference of Cantonati et al (17). The nomenclature of diatom species was performed according to Guiry & Guiry's worldwide electronic internet database. RESULTS No diatoms were found in Groups A and B. However, Navicula sp. and Sellaphora sp. cf., were discovered during bone analysis of Group C where rats were immediately autopsied after drowning. Hantzschia amphioxus taxon was present in Group D, which underwent a 48-hour postmortem period after drowning and before samples were taken. In Groups C and D, where drowning was the COD, Diatoma vulgaris i Pinnularia major, Achnanthidium minutissimum i Melosira varians were present in the tooth samples. CONCLUSION Optimization of the "Diatom Test" method could potentially lead to its future use as a routine method within experimental settings. This experimental study is a starting point that guides forenscic medicine pracitioners towards the optimization of tests and sampling in cases of unexplained etiology, where preserved soft tissue structures is not available. In these cases, teeth and bones serve as accessible materials for diagnosing COD, alongside standardized nonspecific findings in the absence of organs for micro- and macroanalysis.

Ankylosing spondylitis is a serious ailment that affects people, and the first signs or symptoms usually occurr between the ages of 15 and 45. While the condition is mostly prevalent in men, women are not immune to this disease. This problem is diagnosed with a combination of clinical history and X-rays, pathology and HLAB27 test. The aim of this case study is to demonstrate how macroscopic and microscopic analysis can be used for identification of the disease from a forensic point of view. In April 2018, we exhumed 11 remains near the city Višegrad, twenty-five years after the last war. All the remains were completely skeletonized. The skeleton of a female was specific and shaped like a bamboo branch, with a partial knitting of vertebral bodies in the lumbar region of spine and with total knitting in the thoracic part. The spinous processes were completely knitted. Her son gave informations for verbal autopsy that she had trouble walking and doing normal activities during life. Samples for analysis and pathological diagnostics were used to determine the real bone condition for forensic purposes. To our best knowledge our case is first one in the literature which combines macroscopic and microscopic analysis of AK in exhumed skeletal remains after 25 years of death in modern era of Europe.

Heat-related biomarkers: Focus on the correlation of troponin I and 70 kDa heat shock protein Abstract Introduction: There is intensive research related to the forensic importance of biomarkers that would be the standard for postmortem damage to cardiomyocytes and the mechanism of the resulting damage. The aim of the research was to examinate the forensic-medical significance of serum levels of biomarkers as detectors of terminal hyperthermic damage to the myocardium. MATERIAL AND METHOD 40 laboratory animals were divided into groups: the first group was the control (n = 8) exposed to a physiological temperature of 37 °C, the second group was divided into two subgroups: antemortem (n = 8) and postmortem (n = 8), which included a exposure temperature of 41 °C and the third group was divided into two subgroups: antemortem (n = 8) and postmortem (n = 8), which included a exposure temperature of 44 °C. The concentration of cardiac TnI and Hsp70 was resoluted in serum by immunochemical enzyme-labeled immunoabsorption method. RESULTS A positive correlation was found between the temperature measured at the time of death and the serum values of cTnI (p = 0.02), in G41, and Hsp70 values did not significantly correlate with the core temperature in this group, p > 0.005. A positive correlation was significant between the concentration of Hsp 70 and the body temperature of rats in the group of rats with a fatal outcome was determined, p = 0.03. CONCLUSION Changes in the concentration of cTnI and Hsp70 in rat serum may indicate hyperthermic damage to the myocardium in the Wistar rat model of heat stroke.