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.

Qualitative Histologic Assessment vs. Geomorphometric Analysis of Nerve Fiber Shape after the Intraneural Application of Liposomal Bupivacaine / Evaluación Histológica Cualitativa Versus Análisis Geomorfométrico de la Forma de la Fibra Nerviosa Después de la Aplicación Intraneural de Bupivacaína Liposomal

SUMMARY: The preserved form of all components of the nerve fiber is a prerequisite for the proper conduction of the nerve impulse. various factors can change the shape of nerve fibers. In everyday practice, qualitative histological analysis is the gold standard for detecting changes in shape. Geometric morphometry is an innovative method that objectively enables the assessment of changes in nerve fibers' shape after local anesthetics action. A total of sixty sciatic nerves were used as material, which was intraneural injected with saline solution in the control group (n=30), and a solution of 1.33 % liposomal bupivacaine (n=30) in the test group. After the animals were sacrificed, nerve samples were taken and histological preparations were made. The preparations were first described and examined using a qualitative histological method, after which digital images were made. The images were entered into the MorphoJ program and processed using the method of geometric morphometry. Qualitative histological examination revealed no differences in nerve fibers after intraneurally applied physiological solution and liposomal bupivacaine. Using the method of geometric morphometry, a statistically significant change in the shape of axons was found after intraneurally applied saline solution and liposomal bupivacaine (p=0.0059). No significant differences in histological changes were found after the qualitative histological analysis of nerve fiber cross-section preparations. A statistically significant change in the shape of nerve fiber axons was observed after geometric morphometric analysis of digital images after intraneural application of saline and liposomal bupivacaine.

La forma conservada de todos los componentes de la fibra nerviosa es un requisito previo para la conducción correcta del impulso nervioso. Varios factores pueden cambiar la forma de las fibras nerviosas. En la práctica diaria, el análisis histológico cualitativo es el estándar de oro para detectar cambios de forma. La morfometría geométrica es un método innovador que permite evaluar objetivamente los cambios en la forma de las fibras nerviosas después de la acción de los anestésicos locales. Se utilizó como material un total de sesenta nervios ciáticos, que se inyectaron intraneuralmente con solución salina en el grupo control (n=30), y una solución de bupivacaína liposomal al 1,33 % (n=30) en el grupo de prueba. Después de sacrificados los animales, se tomaron muestras de nervios y se realizaron preparaciones histológicas. Primero se describieron y examinaron las preparaciones utilizando un método histológico cualitativo, después de lo cual se tomaron imágenes digitales. Las imágenes fueron ingresadas al programa MorphoJ y procesadas mediante el método de morfometría geométrica. El examen histológico cualitativo no reveló diferencias en las fibras nerviosas después de la aplicación intraneural de solución fisiológica y bupivacaína liposomal. Usando el método de morfometría geométrica, se encontró un cambio estadísticamente significativo en la forma de los axones después de la aplicación intraneural de solución salina y bupivacaína liposomal (p = 0,0059). No se encontraron diferencias significativas en los cambios histológicos después del análisis histológico cualitativo de las preparaciones de secciones transversales de fibras nerviosas. Se observó un cambio estadísticamente significativo en la forma de los axones de las fibras nerviosas después del análisis de morfometría geométrica de imágenes digitales después de la aplicación intraneural de solución salina y bupivacaína liposomal.

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.

A Geometric Morphometrics Approach for Sex Estimation Based on the Orbital Region of Human Skulls from Bosnian Population.

Background: Understanding the anatomy and morphological variability of the orbital region is of great importance in clinical practice, forensic medicine, and biological anthropology. Several methods are used to estimate sex based on the skeleton or parts of the skeleton: classic methods and the geometric morphometric method. The objective of this research was to analyse sex estimation of the orbital region on a sample of skulls from a Bosnian population using the geometric morphometric method. Materials and Methods: The research was conducted on three-dimensional models of 211 human adult skulls (139 males and 72 females) from the Osteological Collection at the Faculty of Medicine in Sarajevo. The skulls were recorded using a laser scanner to obtain skull 3D models. We marked 12 landmarks on each model to analyse sexual dimorphism. Landmarks were marked using the program Landmark Editor. After marking the landmarks, we used the MorphoJ program to analyse the morphological variability between male and female orbital regions. Results: After Procrustes superimposition, generating a covariant matrix, and introducing sex as a variable for classification, a discriminant functional analysis (DFA) was applied which determined the estimation for males with 86.33% accuracy and for females with 88.89% based on the form of the orbital region. The results of regression analysis showed that the size of the orbital region has a statistically significant effect on its shape's sexual dimorphism. After excluding the influence of size and providing DFA, we concluded that sex estimation was possible with 82.01% accuracy for males and 80.55% accuracy for females based on the shape of the orbital region in the examined sample. Conclusion: Sex estimation based on the orbital region was possible with more than 80% accuracy for both sexes, which is a high percentage of correct estimation. Therefore, we recommend using the orbital region of the skull for sex estimation.

Sex estimation based on foramen magnum: a three-dimensional geometric morphometrics approach / Estimación del sexo basado en el foramen magno: un enfoque de morfometría geométrica tridimensional

SUMMARY: The foramen magnum is an important topographic opening which connects cranial cavity and spinal canal. The analysis of the bone material established that there are differences in the shape of the foramen magnum between individuals. The aim of this study was to determine sex based on shape and size of foramen magnum using geometric morphometrics method. A study was performed on three-dimensional models (3D models) of 214 human skulls of known sex and known age (141 male skulls and 73 female skulls). The skulls are located at the museum of Medical Faculty, University of Sarajevo. Skulls belong to Bosnian population from the mid-twentieth century. All examined skulls were scanned with a laser scanner to obtain their 3D models. On 3D models of the examined skulls, four landmarks were marked on foramen magnum. Analysis of sex determination was performed using the MorphoJ program. Results of this study showed that there are sex differences in the shape and size of the foramen magnum. Sex determination based on the shape and size of the foramen magnum was showed 65.25 % accuracy for male and 63.01 % accuracy for female using geometric morphometrics method. Examination of the effect of size of foramen magnum on sexual dimorphism of shape of foramen magnum showed a statistically significant effect. Sex determination based just on the shape of foramen magnum using geometric morphometrics method was possible with 62.41 % accuracy for male and 58.90 % accuracy for female on examined sample. Sex differences on shape and size of foramen magnum were found using geometric morphometrics method on three-dimensional models of the examined skulls. The percentage of accuracy was higher for male based on the shape and size of the foramen magnum than for female.

El foramen magno es una importante abertura topográfica que conecta la cavidad craneal y el canal espinal. El análisis del material óseo estableció que existen diferencias en la forma del foramen magno entre individuos. El objetivo de este estudio fue determinar el sexo en función de la forma y el tamaño del foramen magno utilizando morfometría geométrica. El estudio se realizó en modelos tridimensionales (modelos 3D) de 214 cráneos humanos de sexo y edad conocidos (141 cráneos masculinos y 73 cráneos femeninos). Los cráneos se encuentran en el museo de la Facultad de Medicina de la Universidad de Sarajevo. Los cráneos pertenecen a población bosnia de mediados del siglo XX. Todos los cráneos examinados fueron escaneados con un escáner láser para obtener sus modelos 3D. En los modelos 3D de los cráneos examinados, se marcaron cuatro puntos de referencia en el foramen magno. El análisis de determinación de sexo se realizó utilizando el programa MorphoJ. Los resultados de este estudio mostraron que existen diferencias de sexo en la forma y el tamaño del foramen magno. La determinación del sexo basada en la forma y el tamaño del foramen magno mostró una precisión del 65,25 % para los hombres y del 63,01 % para las mujeres utilizando morfometría geométrica. El examen del efecto del tamaño del foramen magno sobre el dimorfismo sexual de la forma del foramen magno mostró un efecto estadísticamente significativo. La determinación del sexo basada solo en la forma del foramen magno utilizando morfometría geométrica fue posible con una precisión del 62,41 % para los hombres y del 58,90 % para las mujeres en la muestra examinada. Se encontraron diferencias de sexo en la forma y el tamaño del foramen magno utilizando morfometría geométrica en modelos tridimensionales de los cráneos examinados. El porcentaje de precisión fue mayor para los hombres en función de la forma y el tamaño del foramen magno que para las mujeres.

Morphometric analysis of clinically significant parameters of the main trunk of the left coronary artery.

Aim To determine the value of angles between the left coronary artery main trunk (LMT) and its branches, the anterior interventricular branch (LAD) and the circumflex branch (CX), and their possible relationship with the LMT length. Methods A total of 29 cadaveric hearts were used. The left coronary artery and its branches were dissected. The hearts were then classified according to the number of branches. The LMT length was measured with a digital gauge, and the LAD-CX angle, LMTLAD angle and LMT-CX angle with a manual goniometer. Results The average value of the LMT length was 9.0 mm (6.0-13.5). In 20 (68.97%) samples, the LMT was divided into two terminal branches. There was no statistically significant difference (p=0.321) in LMT length between the hearts with a bifurcation and without it. The average value of the LAD-CX angle was 89.0° (74.5-93.0), with a statistically significant difference (p=0.020) comparing to hearts with trifurcation. The mean value of the LMT-LAD angle was 30.83±9.23° and it was significantly lower (p=0.006) in the group of hearts with bifurcation compared to the group with trifurcation of the main trunk. Conclusion The LMT length shows great variability and is not related to the LAD-CX, LMT-LAD or the LMT-CX angle. Knowledge of the left coronary variation is essential in order to avoid misinterpretation of arteriogram.

Assessment of Work Ability of Patients with COPD in Relation to the Workplace.

Context: Chronic obstructive pulmonary disease (COPD) is one of the most common chronic lung diseases and is an important cause of mortality and morbidity in the world. Chronic obstructive pulmonary disease is a heterogeneous and multisystemic disease. Aims of the study was to assess workability of patients with COPD in relation to the workplace. Settings and Design: Prospective study. Methods and Material: The study was conducted on 150 patients with COPD. Each patient was examined by an occupational medicine specialist and a standardized COPD Questionnaire was completed. A physical examination was performed with special emphasis on auscultatory findings on the lungs. Spirometric testing was performed. Using the statistical methods, the results of the parameters of temporary (number of lost working days) and permanent incapacity (exercised right to some kind of disability) of the examined group were analyzed. Statistical Analysis Used: Descriptive statistics. SPSS 10.0. Results of the study showed that out of 150 patients, 48.67% had preserved working capacity for jobs. There are mostly jobs without special working conditions. Out of 150 patients, 51.33% had reduced working capacity for jobs. Conclusions: These are mostly workplaces with special working conditions, where respondents are exposed to the harmfulness of the workplace. Reduced working capacity was found in 51.33% of examined simple.

Qualitative and quantitative characteristics of the passive compartment underneath the myocardial bridge comprising the blood vessel adventitia and the perivascular connective tissue / Características cualitativas y cuantitativas del compartimento pasivo debajo del puente miocárdico que comprende la adventicia del vaso sanguíneo y el tejido conectivo perivascular

SUMMARY: Myocardial bridges are inborn anomalies frequently found in authopsies. Although tipically clinically silent, they are occasionally associated with severe clinical manifestations, e.g. myocardial ischemia or even sudden death. The pathophysiology and risk factors for these manifestations have not yet been completely elucidated. The connective tissue underneath the bridge has been considered as one of the factors the symptoms depend on. Thus, the aim of this research was to determine the histological characteristics of the connective tissue lying underneath the myocardial bridge and to contribute to a better understanding of the protective effects this passive compartment might have in prevention of severe clinical manifestations of myocardial bridging. The study was carried out on twenty hearts with myocardial bridges. Length of the bridge was determined using a precise electronic caliper. Sections of the myocardial bridges with the underlying connective tissue were obtained and prepared for qualitative and quantitative analysis. The connective tissue underneath the bridges was composed of adipose tissue and loose connective tissue in different ratios. The tissue underneath thin bridges was predominantly composed of adipose tissue, while loose connective tissue was the dominant component under thick bridges. The myocardial bridges had an average thickness of 0,98 ± 0.44 mm and an average length of 15,25±5,65 mm. We found a strong positive correlation between the myocardial bridge thickness and length (r = 0,860, p = 0,0001). The thickness of the passive connective tissue compartment under the myocardial bridges was 0,58±0,22 mm, and there was no correlation between this parameter and the myocardial bridge thickness (r = -0,011; p = 0,963). In the clinical evaluation of patients with these anomalies it is necessary to take into account independently the myocardial bridge thickness and length on one side and the thickness of the connective tissue lying underneath it on the other.

Los puentes miocárdicos son anomalías congénitas que se encuentran con frecuencia en las autopsias. Aunque típicamente éstos son clínicamente silenciosos, ocasionalmente se asocian con manifestaciones clínicas graves, como isquemia miocárdica o incluso muerte súbita. La fisiopatología y los factores de riesgo de estas manifestaciones aún no se han dilucidado por completo. El tejido conectivo debajo del puente se ha considerado como uno de los factores de los que dependen los síntomas. Por lo tanto, el objetivo de esta investigación fue determinar las características histológicas del tejido conectivo que se encuentra debajo del puente miocárdico y contribuir a una mejor comprensión de los efectos protectores que este compartimento pasivo podría tener en la prevención de manifestaciones clínicas graves de puente miocárdico. El estudio se llevó a cabo en veinte corazones con puentes miocárdicos. La longitud del puente se determinó utilizando un calibrador electrónico preciso. Se obtuvieron secciones de los puentes miocárdicos con el tejido conjuntivo subyacente y se prepararon para análisis cualitativo y cuantitativo. El tejido conectivo debajo de los puentes estaba compuesto de tejido adiposo y tejido conectivo laxo en diferentes proporciones. El tejido debajo de los puentes delgados estaba predominantemente compuesto de tejido adiposo, mientras que el tejido conectivo laxo era el componente dominante debajo de los puentes gruesos. Los puentes de miocardio tenían un espesor promedio de 0,98 ± 0,44 mm y una longitud promedio de 15,25 ± 5,65 mm. Encontramos una fuerte correlación positiva entre el grosor y la longitud del puente miocárdico (r = 0,860, p = 0,0001). El grosor del compartimiento de tejido conectivo pasivo debajo de los puentes miocárdicos era de 0,58±0,22 mm, y no hubo correlación entre este parámetro y el grosor del puente miocárdico (r = -0,011; p = 0,963). En la evaluación clínica de pacientes con estas anomalías es necesario tener en consideración de forma independiente el grosor y la longitud del puente de miocardio por un lado y el grosor del tejido conectivo que se encuentra debajo del mismo por el otro.

Model "P" in gender prediction based on the mastoid process.

Aim To determine the degree of accuracy in determining the sex of the skull based on classical morphometric analysis of the mastoid process. Morphometric analysis excluded a subjective approach expressed in osteoscopic analysis. Methods The study was conducted on a sample of 100 macerated skulls of known gender and age from the second half of the 20th century, including the Bosnian population. Of the 100 skulls, 50 (50%) were male and 50 (50%) were female. Male skulls were on average 60 (47-71) years old and female 57 (43-76) years old. At each mastoid process, 3 measurements were made: mastoid length, mastoid width (medio-lateral diameter) ML) and anteroposterior diameter (AP) of the mastoid process. Results Using the univariate method, we found a significant difference between length, width, AP diameter and size of mastoid processes as well as between gender (p <0.05). Multivariate binary logistic regression showed statistically significant differences in AP diameter of the mastoid process (p<0.05). Conclusion The created model "P" ("P"=exp [X]/1+exp [X]) for sex determination based on mastoid process showed sensitivity of 82% correct prediction for female skulls and 65% accurate prediction for male skulls. This discourse with respect of population standards grants most effective anthropological proof and as such may be suggested for forensic expertise based on human skull.