Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation and destruction of cartilage, as well as by extra-articular manifestations. Rheumatoid nephropathy is a common complication of RA and its principal target is the renal corpuscle. Vitamin D and its analogs exert immunomodulatory actions throughout the body due to the widespread of their receptors. Our study aimed to compare the effects of cholecalciferol (vitamin D3) and alfacalcidol on renal corpuscle changes in pristane-induced RA model following a 28-day treatment, using geometric morphometrics. Forty female Wistar rats (190–210 g; 12–13 weeks old) were randomly assigned to four groups: the control (Cont) group (n = 10) received saline i.c., the PIA group (n = 10) was administered pristane i.c., PIA-ALF group (n = 10) was administered pristane i.c. and alfacalcidol orally, and the PIA-CH group (n = 10) was injected i.c. with pristane and received cholecalciferol orally. Pristane administration was used for RA induction. At the end of the experiment, the left kidneys were removed and processed by standard histological procedures for geometric morphometric analysis. Geometric morphometric analysis demonstrated that, compared with the control group, the architecture of the renal corpuscles was altered in the PIA (p < 0.0001) and PIA-CH (p = 0.0065) groups. In contrast, no statistically significant differences were observed in the PIA-ALF group (p = 0.3011). Geometric morphometric analysis demonstrated that alfacalcidol, but not cholecalciferol, exertedaprotective effect on the renal corpuscle architecture in pristane-induced rheumatoid arthritis in rats.

Abstract.

Background: Many deaths during heat waves stem not only from body overheating but also from heat stress, which can intensify pre-existing medical conditions, leading to fatal outcomes. Aim: This study aimed to investigate whether the intensity of pathological changes in the heart muscle and lung tissue of albino rats exposed to hyperthermia correlates with different water temperatures (WT) and to determine whether the histological structure of the myocardium and lungs varies. Methods: A sample of 21 albino rats was exposed to water temperatures of 37°C, 41°C, and 44°C. Temperature readings were recorded before immersion, immediately after immersion, at the point of reaching hyperthermia, at 20 minutes, and at the time of death. Tissue samples were collected from the dissected rats, fixed in 10% buffered formalin at room temperature, embedded in paraffin, sectioned into 4–5 μm slices, and stained using the hematoxylin-eosin method. Results: The severity of myocardial histopathological alterations increased with both higher temperatures and longer exposure durations WT. However, the progression of morphological alterations in cardiomyocytes was not markedly significant, likely due to the brief exposure time, which limited the visualization of subcellular alterations in Hematoxylin and Eosin staining-stained tissue. All lung samples from the seven rats exposed to the highest temperatures displayed bronchiolitis and acute bronchitis, along with early bronchopneumonia. Conclusion: While some organs exhibit greater tolerance to heat stroke than other organs, most organs show similar alterations characterized by capillary dilation, vascular pathway disruption, and extravasation. The extent of pathological changes in myocardial and lung tissue intensified with higher temperatures and longer exposure durations to elevated WT. However, the progression of morphological alterations in cardiomyocytes did not demonstrate marked significance, likely because of the brief exposure period. This short duration may limit the detection of subcellular changes when using hematoxylin-eosin staining.

Background: Primary hypothermia occurs when the body is exposed to extremely low temperatures in an environment with no underlying health conditions. Secondary hypothermia, on the other hand, results from disruptions in thermoregulation due to diseases, trauma, surgery, drugs, alcohol, or infections. Postmortem biochemistry has become a crucial factor in forensic examinations, offering valuable apprehension into tissue of and organ dysfunction associated with the process of dying. Aim: This research aims to explore various biochemical markers and their significance in distinguishing primary from secondary hypothermia. Methods: This study involved 21 Wistar rats, which were separated into three experimental groups: CG (n = 7), which were exposed only to hypothermic conditions; AHG (n = 7); and BHG (n = 7). We tested these parameters in each rat: glucose, urea, creatinine, blood urea nitrogen to creatinine ratio, phosphorus, calcium, sodium, potassium, sodium to potassium ratio, chloride, and calculated osmolality. Results: Distinct biochemical differences were noted between primary and secondary hypothermia. Glucose and creatinine levels exhibited significant variations (p < 0.001). Urea concentrations also manifested notable differences between the groups (p < 0.001). Phosphorus levels demonstrated significant differences (p = 0.004), with post hoc analyses revealing significant contrasts between the AHG and BHG (p = 0.014) and between the BHG and CG (p = 0.014). Potassium levels and the sodium-to-potassium ratio differed significantly (p < 0.001). Osmolality also varied significantly across experimental groups (p < 0.001), with post hoc tests confirming significant differences between the AHG and CG (p = 0.013) and between the BHG and CG (p = 0.002). Conclusion: The calculated osmolality exhibited significant variation among the different groups, indicating a notable impact of the substances on the biochemical profile related to hypothermia. This study focused on the effectiveness of biochemical markers in distinguishing primary hypothermia from secondary hypothermia.

Background: Different dietary components can affect hematological and biochemical profiles, potentially causing pathohistological changes in liver and kidney tissue. Aim: The animals in the experiment consumed various bakery and meat products, and ultimately, the potential effects on hematological, biochemical, and pathological parameters were evaluated. Methods: The study involved 24 clinically healthy adult rats, randomized into three groups of eight rats each, as follows: rats that consumed meat products (group M), rats that consumed bakery products (group H), and a control group that consumed conventional rodent food (group K) for 7 weeks. After 7 weeks, hematological and biochemical blood analyses were conducted along with pathohistological examinations of the liver and kidneys. Results: Significant differences (p < 0.05) were observed among groups for several hematological and biochemical parameters, including creatinine (CREA), urea, blood urea nitrogen /CREA, calcium, alanine transaminase, alkaline phosphatase, and lipase. Consuming meat products had a less favorable impact on the occurrence of kidney function disorders. Group H exhibited significant differences in leukocyte and platelet counts compared with groups M and K. Extreme echinocytosis was recorded in group M, whereas sideropenic anemia was prominent in group H. Analysis of the livers of rats in groups K and H did not show significant differences in the observed parameters (gamma-glutamyl transferase and total bilirubin), whereas group M had a significantly higher degree of hepatocyte degeneration and steatosis, and the observed infiltrate was also more pronounced, but not significantly. The kidneys of group M showed discrete alterations of the microstructure, i.e., slightly increased cellularity of renal corpuscles and hypertrophy of proximal nephrocyte, whereas the kidney tissue of group K had a regular appearance. Conclusion: Consuming meat products was associated with adverse liver and kidney changes, whereas bakery products led to sideropenic anemia and altered hematological values.

Abstract Multiple studies have shown the importance of adequate nutrition for animals and humans and its effect on overall health. Therefore, the aim of this study was to investigate the effects of different nutritional regimes on the intestinal health of rats by evaluating different morphological and morphometric characteristics of small intestines, with the emphasis on the villus height:crypt depth ratio (V:C). For the experimental study, 24 clinically healthy adult Wistar rats were used. The rats were randomly divided into 3 groups: the control group (group A) was fed with conventional food, the second group (group B) with bakery products, and the third group (group C) with meat products. Samples of the duodenum and jejunum were collected for detailed morphological and morphometric analysis. A significant increase in the duodenal villi height was reported in group B (661.59 µm) and C (602.83 µm) compared to the control group (475.34 µm). The crypt depth values in the jejunum were significantly higher in group B (191.41µm) and C (246.23 µm) compared with the control (145.14 µm). The jejunal V:C ratio was significantly lower in groups B and C. The study showed significant morphological changes in the intestinal parameters in rats fed predominantly with meat and bakery products. These findings could be applicable in both veterinary and human medicine, underlining the significance of consumed food on gut health.

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.

Microscopic signs indicative of drowning are not specific to drowning but also to any other form of suffocation where mechanical obstruction is involved. Our study aimed to evaluate both macroscopic and microscopic findings across different groups sharing a common mechanism of death but differing causes and to compare the diatom test with pathohistological examination.Twenty-nine adult Wistar rats, weighing within recommended ranges, were divided into four groups (L1-L4). The diatom test followed established guidelines for diatoms in water from the Bosna River. Microscopic examination revealed diatoms in the lungs of rats in L3 and L4 groups. Pathohistological findings showed varying degrees of changes including consolidation and inflammatory cell infiltration, dominated by lymphocytes and macrophages, with some samples also showing eosinophilic leukocytes.Significant differences were observed between animals whose cause of death was mechanical asphyxia (suffocatio) and those that were submersed for1 hour versus those that were submersed for 72 hours after death. Diatoms identified in group L4 samples 3, 4, and 5 included Navicula sp. (U3 and U6) and Ulnaria ulna (U4).Our findings suggest combining the diatom test with pathohistological analysis to support a drowning diagnosis. Further examination of other organs could enhance result reliability.

Introduction: Diabetes mellitus is associated with systemic complications, including the development of pulmonary injury, characterized mainly by excessive accumulation of extracellular matrix components and inflammatory cell infiltration in lung tissue. This process is driven by oxidative stress and chronic inflammation, both caused and exacerbated by hyperglycemia. N-acetylcysteine (NAC) and glycine, known for their antioxidant and anti-inflammatory effects, offer potential therapeutic benefits in mitigating diabetes-induced lung injury. Objective: The study aimed to investigate the effects of supplementation by either NAC or glycine or their combination on reducing lung injury in rats with type 1 diabetes Materials and methods: The study used 30 adult Wistar albino rats (10 weeks old, weighing between 180 g and 380 g). Six of them were used as controls, while 24 adult rats (10 weeks old, 180-380 g) with type 1 diabetes, induced through a single intraperitoneal injection of streptozotocin (STZ) at a dose of 55 mg/kg, were randomly assigned to four experimental groups: control (CTL), diabetic (Db), NAC treatment (diabetic+NAC), glycine treatment (diabetic+glycine), and combined NAC and glycine treatment (diabetic+NAC+glycine). NAC (100 mg/kg) and glycine (250 mg/kg) were administered orally for 12 weeks. At the end of the study, lung tissues were collected for histopathological examination. Qualitative, semi-quantitative, and stereological histological analysis was used to analyze structural changes in the lung tissue. Semi-quantitative scoring was carried out to evaluate the extent of inflammation, while stereological analysis was performed to determine the volume density of alveolar spaces and septal connective tissue. The semi-quantitative scoring included scores ranging from 0 (absent), 1 (minimal), 2 (mild), 3 (moderate), to 4 (severe). Results: Qualitative histological analysis revealed pronounced inflammation and fibrosis in the lungs of untreated diabetic rats, characterized by thickened alveolar septa and immune cell infiltration. Both treatments with NAC and glycine individually reduced inflammation and fibrosis compared to untreated diabetic rats. The greatest improvement was observed in the NAC+glycine group, where the alveolar structure appeared almost normal, with minimal inflammation. Semiquantitative analysis showed statistically significant differences in peribronchial and peribrochiolar infiltrates between the diabetic group (2.16±0.47) and the control group (0.33±0.21, p=0.026). The combination of NAC and glycine significantly reduced peribronchial and peribronchiolar infiltrates (0.33±0.33, p=0.026) compared to the diabetic group. Similarly, septal inflammatory infiltrates were significantly lower in the NAC+glycine group (1±0.36) compared to diabetic rats (3.33±0.33, p=0.004). Total airway inflammatory infiltration was also significantly reduced in the NAC+glycine group (1.33±0.33, p=0.002) compared to the diabetic group (5.5±0.5). Conclusion: As the combination of NAC and glycine demonstrated protective effects against lung inflammation and fibrosis in diabetic rats, a synergistic effect of NAC and glycine in mitigating pulmonary complications associated with type 1 diabetes may be suggested. These findings warrant further exploration of the combination for managing diabetic lung disease and potentially other fibrotic conditions.

Background: There is no specified diagnostic procedure that can help in determining the cause of death and the diagnosis of drowning because the pathohistological signs are almost identical and non-specified. Aim: Our study aims to recognize and prove diatom appearance in organs from a forensic aspect in Bosnia and Herzegovina, and to examine which is the more specific method in the diagnosis of drowning, the diatom test or the pathohistological finding. Methods: Rats of the recommended body weight were divided into four groups: G1 (n = 8; mechanism of death—asphyxia; cause of death—suffocation, submerged 1 hour after death); G2 (n = 8: mechanism of death-asphyxia; cause of death-suffocation, immersed 72 hours after death); G3 (n = 8: mechanism of death-asphyxia; cause of death-drowning, autopsy immediately after death), and G4 (n = 8: mechanism of death-asphyxia; cause of death-drowning, post mortem 24 hours after death). Results: During the diatom analysis, four species of diatoms, Diatoma vulgaris, Melosira varians, Epithemia adnata, and Cymbella sp, were successfully recovered from the stomach. Microscopic analysis did not detect diatoms in the kidneys and brains of rats, while the pathohistological changes were relatively uniform. Conclusion: Our results propose that the diatom test is a sustainable tool for supporting the diagnosis of drowning in the forensic pathology analysis of the cause of death. This experimental study is a starting point toward the optimization of tests and sampling in cases of unexplained etiology.

INTRODUCTION Hypothermia is defined as a body core temperature below 35 °C and can be caused by internal or external stress. Primary hypothermia is caused by excessive exposure to low environmental temperature without any medical conditions prior to that. Secondary hypothermia is caused by alteration in thermoregulation by disease, trauma, surgery, drugs, or infections. The aim of the research is to investigate core temperature values in rats subjected to specific water temperatures at five different time points. It focuses on distinguishing between primary and secondary hypothermia in these rats. METHODS The total 21 Wistar rats were divided into three experimental groups as: 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 core temperature, core temperature during injection of 0,3 ketamine, temperature of immersion and the temperature at the onset of hypothermia and temperature at the time of death. RESULTS In our study the comparative analysis of body temperatures at various time points following submersion in water revealed significant differences among the study groups treated with either alcohol or benzodiazepines and the control group. Notable differences were observed in baseline temperature, post-anesthesia induction temperature, and immediate post-submersion temperature. Specifically, significant differences were discovered among the alcohol and benzodiazepine groups (p < 0.001) and ranging from the alcohol and control groups (p < 0.001). The analysis of survival times following induced hypothermia revealed a statistically significant difference among the three experimental groups (p = 0.04), though subsequent post-hoc comparisons did not demonstrate significant differences in mean survival times. CONCLUSION There is a difference in survival time between primary and secondary hypothermia groups, depending on consumption and intoxication with alcohol or benzodiazepines. The analysis of survival times following induced hypothermia showed a statistically significant difference among the groups.

Commensal rats are highly adaptable to coexisting with human populations, often relying on human dwellings and immediate access to food, water, shelter and space. We present the case of an adult commensal brown rat, Rattus norvegicus, that was found in an urban area of Sarajevo (Bosnia and Herzegovina). Histopathological examination revealed numerous eggs of Calodium hepaticum in the liver parenchyma and coprological examination identified species of the class Cestoda (Rodentolepis spp., Hymenolepis diminuta) and the class Nematoda (Trichuris muris, Nippostrongylus brasiliensis and Strongyloides ratti). Haematological parameter analysis was conducted using the “Idexx ProCyte Dx” cell counter and biochemical parameters of the rat's serum were tested. Haematocrit, haemoglobin content and erythrocyte indices (MCV, MCH) were decreased and moderate quantities of spherocytes were present (4.95 %). The total number of leukocytes was within the physiological range, with minimally increased values of monocytes and neutrophils and decreased values of eosinophils. The results of the morphometric examinations were in the expected range for adult rats. This case report highlights the public health risk posed by commensal rats if systematic rodent control measures are not fully implemented. It should serve as a guide for future research on the population of commensal rats in the cities of Bosnia and Herzegovina, aimed at a detailed identification of parasitic zoonotic pathogens.

Objective: The aim of the study was to determine the possible impact of the total daily amount of skim milk on the level of bilirubin and liver enzymes through regression analysis. Materials and Methods: The study included 63 Holstein-Friesian cows. They were formed in 3 groups, based on the amount of daily milk production. Peripheral blood was punctured, through which the activities of total bilirubin were analyzed (μmol/L), as well as liver enzymes: alanine aminotransferase – ALT (U/L), aspartate aminotransferase – AST (U/L), lactate dehydrogenase – LDH (U/L) and alkaline phosphatase – ALP (U/L). Results: The lowest concentration of total bilirubin in blood plasma was recorded in the group of cows that have the lowest daily milk production (1.295 ± 0.255 µmol/L), and highest concentration is in cows that produce the most milk (1.855 ± 0.159 µmol/L), but intergroup differences are not significant. Regression analysis found a statistically significant relationship between the amount of produced daily milk and the concentration of total bilirubin (R2=0.132, p=0.0050.05). Conclusion: The activities of bilirubin and liver enzymes in the examined cows were in physiological balance. This indicates that the cows on the farm are raised in modern and good zootechnical and feeding conditions. In such conditions, dairy cows are able to maintain blood composition and homeostatic integrity within physiological limits and adequate reproductive and productive capacity.

Listeria is a food-borne pathogen that poses a great threat to human health. Severe symptoms followed by a high mortality rate are mainly caused by L. monocytogenes. Despite the zero-tolerance policy, the genus Listeria is still present in raw and ready-to-eat food. There are no recent studies regarding contamination of food with Listeria species in Bosnia and Herzegovina. This research is focused on obtaining basic information on the level of food contamination with this alarming pathogen. In this study, a total of 238 samples were analysed, where 105 samples were various types of raw food and 133 samples were ready-to-eat foods, and 18 samples tested positive for Listeria species. After isolation and detection, the confirmation of positive isolates of Listeria spp. was conducted using the biochemical kit API ® Listeria .