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: 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.

Background Biological mesh derived from porcine small intestinal submucosa (SIS) has a higher porosity and is more hydrophilic than tissue derived from bovine and cow dermal tissues. Therefore, we believe SIS mesh will lead to a milder inflammatory reaction than other, polypropylene and polypropylene-polydioxanone meshes, fewer adhesions, and less mesh shrinkage. Methods Ninety rats were divided randomly into three groups: in group 1, polypropylene mesh was implanted; in group 2, polypropylene-polydioxanone; and in group 3, the SIS mesh. The meshes were fixed intra-abdominally, in the upper part of the abdomen. Ten animals from each group were sacrificed on days 7, 28, and 60 after the implantation. Relaparotomy was performed, with a left paramedial incision and the adhesions formed were assessed according to the Surgical Membrane Study Group (SMSG) score, along with the percentage of shrinkage of the mesh, and any inflammation. Results There were no differences in terms of inflammatory reaction or the formation of adhesions between the meshes tested on the 7th day after implantation. However, the shrinkage of the SIS mesh was more expressed. On days 28 and 60, the SIS mesh caused less inflammatory reaction and formation of adhesions in relation to the other meshes tested. On day 60, there was no significant difference in the size of the meshes. Conclusion This study confirmed that, despite conflicting views on biological mesh, SIS mesh results in less inflammatory reaction, less adhesion formation, and a lesser degree of shrinkage, and can take its place in hernia repair.

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.

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.

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.

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.

Attacks by dogs, primarily stray dogs, are a very common problem faced by both underdeveloped and developed local goverments. The damage caused by attacks from stray dogs has multiple, and often long-lasting, harmful consequences for their victims, which are also reflected on members of their families. This manuscript provides a veterinary-epidemiological definition of the concept of dogs, as well as their legal definition in the context of „dangerous things“, and the psychological aspect that is regularly manifested in victims. Analyzing the legislation of Bosnia and Herzegovina, with a comparative presentation of the regulations of the EU member states, systematic overviews and answers are given, which are important for the improvement of existing regulations and harmonization of law and jurisprudence.