Bone and Tooth: Substrates for determining Drowning: A new diagnostic procedure in forensic medicine Practice?

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.

Geometric and Morphometric Analysis of the Auditory Ossicles in the Red Fox (Vulpes vulpes).

The use of carnivores as experimental models in auditory biology has led to a significant improvement regarding our understanding of the structure and function of the ear. Considering that data regarding the anatomy of the middle ear in the red fox are rare, this study aimed to describe the morphological and morphometrical features of the auditory ossicles in the red fox, as well as to provide their shape characteristics by geometric analysis. Nine adult red foxes were used in the study. The malleus, incus and stapes were extracted from the middle ear, prepared, photographed and measured using the software. For the geometric analysis, 19 landmarks were used. Following Principal Component Analysis (PCA), PC1 was found to explain approximately half of all variance (incus: 49.97%; malleus: 49.93%; stapes: 58.49%). The study demonstrated the similar anatomical organization of the auditory ossicles in line with important morphometric and basic geometric data, which can contribute to this field and add a useful perspective to the literature.

Heat-related biomarkers: Focus on the correlation of troponin I and 70 kDa heat shock protein.

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.

CT advantages of potential use of polymer plastic clips in neurocranium.

Aim Clips in neurosurgery are made of titanium alloys, which reduce artifacts on computed tomography (CT). The radiological advantage of plastic clips on the CT image was demonstrated when they were placed in an inter-hemispherical position at an angle of 90º. The aim of this study was to investigate the behaviour of the clip placed at different angles. Methods Sixty heads of domestic pigs were divided into two groups, in group 1 a titanium clip was placed to the interhemispheric position at an angle of 90º, 45º, 0º, ten heads for each angle. In group 2 a plastic clip was placed in the same way. CT scan of the brain was performed for each angle. The size of the density and possible artifact were measured on CT. Results The size of the titanium clip ranged from 17.05 mm at an angle of 0º in the axial plane to 91.47 mm at an angle of 0º in the sagittal plane. The average size of the plastic clip ranged from 6.4 mm at an angle of 0º in the axial plane to 23.22 mm in an angle of 90º in the sagittal plane. Artifacts were observed only in the titanium clip. Conclusion Plastic clips have shown radiological advantages over titanium clips in the CT image. The average density size of the plastic clip in all planes and all angles was smaller than the titanium clip.

Tissue Reaction and the Formation of Adhesion after the Use of DS Clip in Laparoscopic Appendectomy.

BACKGROUND AND OBJECTIVES: Titanium DS clips are made to secure the appendiceal stump during laparoscopic appendectomy. The assumption is that they behave like stapler clips in the body, being made from titanium. However, the construction and weight of DS clips differs from staplers. Their biocompatibility may have implications for their clinical use. METHODS: One hundred and twenty rats were randomized into four experimental groups: in the first group the appendiceal stump was secured by Vicryl® ligature, in the second by linear staplers, in the third by Hem-o-lok® plastic clips, and in the fourth by DS titanium clips. Ten animals from each group were sacrificed 7, 28, and 60 days post-surgery. Histopathological data and adhesion formation were assessed. RESULTS: On postoperative day 7, a statistically significant difference was found in the occurrence of inflammation between the Hem-o-lok® and stapler groups. The adhesion score was significantly higher in the Vycril® and Hem-o-lok® in comparison to the DS and stapler groups. On postoperative day 28, a statistically significant difference was found in the occurrence of inflammation between the Vycril® and stapler groups. The adhesion score was significantly higher in the Vycril® than in the DS and stapler groups. On postoperative day 60, there were no statistical differences between any of the groups. The adhesion score was significantly higher in the Hem-o-lok® group in comparison to the DS and stapler groups. CONCLUSION: Staplers and DS clips have advantages over other methods of securing the appendiceal stump, because of their high biocompatibility and lower adhesion score.

Dog bites and attacks on athletes: lack of effective prevention mechanisms.

Athletes who train in public places in urban and rural areas are just as attacked and injured by dogs of known owners as they are by dogs with no owners, in a relatively equal proportion. The largest number of bites occurs in the summer, what makes up half of all bites, just when sports activity is most pronounced. Athletes who are most often exposed to potential attacks and bites are cyclists, long-distance athletes, marathon runners, recreational athletes, etc. both during training and competitions. Off-road cyclists are at a significantly higher risk of dog attacks because cycling takes place off-road, that is, away from urban areas. Dog attacks can adversely affect the psycho-physical readiness of athletes. In Bosnia and Herzegovina there have been no cases of injuries to athletes recorded by competent medical institutions or umbrella sports associations. It is necessary to work on more efficient administration (registration and recording of attacks and bites of dogs). It is of utmost importance to educate athletes on dog behaviour, the reasons for their aggressive behaviour and causal mechanisms of dog attacks as well as the first aid education, what can have a great impact on reducing further complications.

Interdisciplinary aspects of possible negative effects of dogs on humans in Bosnia and Herzegovina.

The coexistence of humans and dogs, in addition to all positive effects, can result in negative effects on human health. A particular risk is posed by a population of stray dogs, that is, dogs without owners and veterinary supervision. A contact with dogs in addition to bites, carries the risk of viral, bacterial and parasitic zoonoses, and can also cause psychological trauma. Children, the elderly and pregnant women are the categories most susceptible to the negative effects of dogs. The aim of the paper was to make an interdisciplinary analysis of the negative effects of dogs on humans. Dog bites cause wounds and dysfunction of damaged tissue, and often lead to various infections. The risks of rabies and tetanus are particularly significant if proper and timely treatment is not performed. Ongoing training for dog owners can significantly reduce the number of bites inflicted by owned dogs, but stray dogs remain a serious social problem and pose potential health risks of some zoonosis. Timely and adequate management of bite wounds and the use of rabies-post-exposure prophylaxis as well as psycho-therapy, where indicated, significantly reduce possible adverse health effects for patients who have been bitten by dogs.

Morphological Changes in Blood Cells After Implantation of Titanium and Plastic Clips in the Neurocranium - Experimental Study on Dogs.

INTRODUCTION: Various studies confirm the biocompatibility and efficacy of clips for certain target tissues, but without any comparative analysis of hematological parameters. Therefore, we conducted a study to assess the possible association of the implantation of titanium and plastic clips in the neurocranium with possible morphological changes in the blood cells of experimental animals. MATERIALS AND METHODS: As a control, the peripheral blood smears were taken before surgery from 12 adult dogs that were divided into two experimental groups. After placing titanium and plastic clips in the neurocranium, the peripheral blood of the first group was analyzed on the seventh postoperative day, while the peripheral blood of the second group was analyzed on the sixtieth day. By microscopy of the blood smears, the following parameters were analyzed: the presence of poikilocytosis of the red blood cells, degenerative changes in the leukocytes and leukogram. RESULTS: There were no statistically significant differences between the mean values of the groups. Monocytosis was detected (first group 22.83 % and second 16.30 %), as well as neutropenia (46.80 %, in the second group). Degenerative changes to neutrophils and the occurrence of atypical lymphocytes were observed in the second experimental group (60th postoperative day). CONCLUSION: A mild adverse effect from the biomaterials present in the neurocranium of dogs was detected, affecting the majority of leukocytic cells. A chronic recurrent inflammatory process was caused by the presence of the plastic and titanium clips in the brain tissue. No adverse effect of biomaterials on erythrocytes in the neurocranium was detected in the dogs studied. Further studies are necessary to explain the occurrence of degenerative changes in the neutrophils and lymphocytes.

Biocompatibility of Plastic Clip in Neurocranium - Experimental Study on Dogs.

AIM: A potential advantage of the use of the plastic clips in neurosurgery is their property of causing fewer artifacts than titanium clips as assessed by computed tomography and magnetic resonance scans. The biocompatibility of plastic clips was demonstrated in the peritoneal cavity, but their behavior in the neurocranium is not known. MATERIAL AND METHODS: Twelve aggressive stray dogs designated for euthanasia were taken for this experimental study. The animals were divided into two groups. In all cases, after anesthesia, a craniotomy was performed, and after opening the dura, a proximal part titanium clip was placed on the isolated superficial Sylvian vein (a permanent Yasargil FT 746 T clip at a 90° angle, while a plastic Hem-o-lok clip ML was placed on another part of the vein). The first group of animals was sacrificed on the 7 < sup > th < /sup > postoperative day and the second group on the 60 < sup > th < /sup > postoperative day. Samples of tissue around the clips were taken for a histopathological evaluation. RESULTS: The plastic clip caused a more intensive tissue reaction than the titanium clip on the 7 < sup > th < /sup > postoperative day, but there was no statistical difference. Even on the 60 < sup > th < /sup > postoperative day there was no significant difference in tissue reaction between the titanium and plastic clips. CONCLUSION: These preliminary results confirm the possibility for the use of plastic clips in neurosurgery. Before their use in human neurosurgery, further studies are needed to investigate the long-term effects of the presence of plastic clips in the neurocranium, as well as studies of the aneurysmal model.

Formation of adhesion after intraperitoneal application of TiMesh: experimental study on a rodent model.

BACKGROUND: After laparoscopic repair of an incisive hernia, intraperitoneal prosthetic mesh, as a foreign material, is a strong stimulus for the development of adhesion, which may be the cause of serious complications. This experimental study compared three different meshes and their ability to prevent the formation of adhesion and shrinkage. METHODS: Ninety rats were divided randomly into three groups: in Group 1 Proceed mesh was implanted, in Group 2 Ultrapro mesh was implanted, and in Group 3 TiMesh was implanted. Mesh samples were fixed as an intraabdominal mesh in the upper part of the abdomen. Ten animals from each group were sacrificed on days 7, 28 and 60 post-surgery. After opening the abdomen, the formation of adhesion was assessed according to the Surgical Membrane Study Group (SMSG) score, the percentage of shrinkage of the mesh was established and inflammatory reaction scored. RESULTS: The SMSG score for adhesion was statistically significantly higher on all the postoperative days in the Proceed and Ultrapro mesh groups than in the TiMesh group which caused milder inflammatory reaction on 60th day than others meshes. The size of the mesh after 7 days was statistically significantly smaller in the Proceed and Ultrapro groups than in the TiMesh group, but after 60 days it was statistically significantly larger than in the TiMesh group. CONCLUSION: The least formation of adhesion was noted in the TiMesh group, in which the highest level of shrinkage was noticed after 28 and 60 days. TiMesh has advantages over the other meshes studied, but a larger size mesh may be recommended for intraperitoneal application.

Biocompatibility and adhesion formation of different endoloop ligatures in securing the base of the appendix.

BACKGROUND AND OBJECTIVES: The common technique used in securing the base of the appendix is Endoloop ligature (Ethicon, Somerville, NJ, USA). Vicryl (polyglactin 910) (Ethicon) and polydioxanone (PDS) (Ethicon) Endoloop ligatures can be used. There are potential benefits of the use of PDS Plus (Ethicon) Endoloop ligature. However, the use of different materials may vary in terms of inflammation, foreign-body reaction, rate of infection in the surgical area, or rate of adhesion formation. An ideal suture would induce minimal inflammatory response and adhesion formation. METHODS: Ninety rats were randomized into 3 groups: group I, in which appendectomy was performed with Vicryl ligature; group II, in which appendectomy was performed with PDS ligature; and group III, in which appendectomy was performed with PDS Plus ligature. The animals were killed on the seventh, 28th, and 60th days after surgery. The secured stump was used for histopathologic and immunohistochemistry analysis, as well as evaluation of the formation of adhesions. RESULTS: Mild and moderate inflammation was more frequent in the PDS and PDS Plus groups than in the Vicryl group on the seventh postoperative day. There were no significant differences in the degree of inflammation on the 28th and 60th postoperative days. The lowest degree of postoperative adhesions was observed in the PDS group. CONCLUSION: Milder postoperative inflammatory changes and a lower degree of postoperative adhesions were seen in the PDS ligature group, suggesting that this could be the standard Endoloop used to secure the base of the appendix.

Tissue reaction to absorbable endoloop, nonabsorbable titanium staples, and polymer Hem-o-lok clip after laparoscopic appendectomy.

BACKGROUND AND OBJECTIVES: The standard technique for securing the base of the appendix during laparoscopic appendectomy is by absorbable endoloop ligature, although clinical reports favor the use of the stapler. Nonabsorbable Hem-o-lok clips have been shown to be an alternative technique to this. However, it is currently not clear whether nonabsorbable clips have any effects on the intestine or promote infection in the surgical area. MATERIALS AND METHODS: Sixty Wistar albino rats were randomized into 3 treatment groups: group I (n=20) the base of the appendix was secured by endoloop 2-0 ligature; group II (n=20) dissection of the appendix was performed by a 45-mm thick stapler; and group III (n=20) the base of the appendix was secured by a Hem-o-lok plastic clip. The animals were sacrificed on the 14th and 28th days after surgery. The secured stump was used for histopathological examination. RESULTS: There were no significant differences in histopathological changes observed on the 14th postoperative day between the groups. On the 28th postoperative day, it was proved that mild and moderate inflammation is more frequent in the endoloop and Hem-o-lok groups than in the stapler group. Reaction to a foreign body is more frequent in the endoloop than in stapler and Hem-o-lok groups. CONCLUSION: The mildest postoperative inflammatory changes were seen in the stapler group, followed by the Hem-o-lok group. However, because of the price of the plastic clip and the simplicity of its application, its use is still favored during laparoscopic appendectomy.