Left ventricular hypertrophy is associated with overexpression of HSP60, TLR2, and TLR4 in the myocardium.

Left ventricular hypertrophy is a common adaptive response to increased cardiac workload. Cardiomyocytes growth and increase in contractile force are conditioned by sufficient energy production, which implies appropriate mitochondrial function. The 60 kDa heat shock protein (HSP60) is a chaperone essential for mitochondrial proteostasis, but when translocates from mitochondria, it can also act as a potent inflammatory mediator binding to toll-like receptors (TLRs). In this study, we aimed to compare the expression pattern of HSP60, TLR2, and TLR4 in hypertrophic vs non-hypertrophic, normal human myocardium. We further examined whether HSP60 in situ binds to TLRs in hypertrophic myocardial tissue. In addition, expression of activated downstream targets of TLR 2/4 pathways was also evaluated.For this purpose, immunohistochemical expression analyses were performed on myocardial tissue samples obtained during the autopsy of human subjects in which left ventricular hypertrophy was the only cardiopathological finding and had died from sudden cardiac death, as well as from the subjects without any cardiac pathology, that died by unnatural death (accident or suicide). Double immunofluorescence was used to examine HSP60 translocation, while proximity ligation assay (PLA) was performed to assess HSP60 and TLRs interactions.Hypertrophic myocardium showed significantly higher expression of HSP60, TLR2, and TLR4 compared to normal myocardium. Furthermore, in hypertrophic cardiomyocytes, we found membrane translocation of HSP60 and signs of HSP60/TLR interactions.Conclusion: The obtained data point to an important supportive role of HSP60 in adaptive cardiomyocytes growth, while concomitant induction of TLR2 and TLR4 candidates HSP60-TLRs interactions as an early events during pathogenesis of secondary complications consequently to the left ventricular hypertrophy.

Alagille syndrome case report: implications for forensic pathology and anthropology.

This case report offers a multidisciplinary interpretation of the violent death of a 4-year-old girl suffering from Alagille syndrome who died after a low-height fall that resulted in temporal bone fracture and a large epidural hematoma. The article evidences the macroscopical and microscopical characteristics of the syndrome, focusing especially on the skeletal findings that emerged during autopsy. In the case report, distinction is made between a possible accidental or non-accidental nature of the injuries and the characteristics of the injury have been interpreted in the light of the existing data on Alagille syndrome. In conclusion, the death was documented as accidental since abnormalities in the skeletal system evidenced during autopsy have predisposed the death of the child albeit through a very mild head trauma. The case report evidences the importance of studying features of skull macro- and microstructure in patients with Alagille syndrome, which have been, until now, underreported in literature and which might contribute to fracture vulnerability in these patients. Although rare, Alagille syndrome is a condition that should be known to forensic medicine practitioners and whose features and peculiarities must be taken into consideration in pediatric autopsy and suspected child abuse cases.

Manual Snow Removal and Sudden Death.

The aim was to analyze the causes of sudden death in middle-aged and elderly men during manual snow removal. During snowy winter months in Zagreb, from January 2013 to January 2014, four males aged 52, 65, 72 and 81, died suddenly while manually removing snow. They were all autopsied. All of them have suffered from arterial hypertension and coronary heart disease, and one suffered from metabolic syndrome. The cause of death in two was probable malignant ventricular arrhythmia. In the third who fell down on the icy surface, consequences were cerebral contusion and neck vertebral luxation. In the fourth who fell down from the top of a 15 m tall building during snow removal, the cause of death were multiple injuries: fractures of both clavicles, ribs and vertebrae's Th5, Th6, hematothorax, cardiac contusion, hematopericardium, thoracic aorta rupture, contusions and ruptures of both lungs, rupture of the diaphragm, liver rupture, hematoperitoneum and cerebral edema. The estimated death rate in the City of Zagreb for males aged 30-64 years is 5.44/1,000,000 inhabitants, which is less than in those aged 65-85 years (40.03/1,000,000; p = 0.2269). Sudden strenuous physical effort due to manual snow removal in two non-trained persons, who have suffered from arterial hypertension and coronary heart disease, was the cause of sudden death. Manual snow removal is an important cause of sudden death, as it is a very arduous effort in non-adapted middle-aged and elderly persons.

Calpain expression in the brain cortex after traumatic brain injury.

Traumatic brain injury (TBI) is the leading cause of death and disability worldwide. Calpains, a family of cysteine proteases have been implicated in cells death following TBI. Using immunohistochemistry calpain expression was analyzed in post mortem brain tissue obtained from patients who died after TBI, and findings were compared with the brain tissue from patients who died from sudden cardiac arrest. In the injured cortex an increase in calpain expression was observed in all resident brain cells: neurons, glial and endothelial cells in comparison to the control group (all p < 0.001). Calpain expression was analyzed in different post-traumatic intervals, from day 0 until 10 days post-injury, in order to establish a time course of expression in the brain cortex after TBI. Expression was detected in the cortex 5 hours after the accident, peaked at 72 hours, and substantially reduced by 10 days after TBI. Calpain expression in the cortex significantly changed during the time from TBI to death (p < 0.001), and the most prominent expression was detected in the cortex 3 days after TBI. Our results indicate that prolonged calpain expression in resident brain cells (neurons, glial and endothelial cells) plays an important role in neuronal degeneration following TBI.

Disposal of the body after murder committed by juvenile offender.

Disposal of the body by setting in concrete or sealing with bricks represents relatively rare form of corpse hiding after crime. We report a unique case of juvenile offender who killed his father and subsequently concealed the body. The victim had been reported missing by members of his family but the body was not found until 12 months later. Hidden in concrete body was well preserved allowing not only identification of the victim but also determination of the cause of death.

Investigation of WWII/postwar mass burials in Croatia-The implementation of the Croatian model of searching for the imprisoned and missing persons.

The search for victims of World War Two (WWII) and the immediate aftermath period (postwar) in Croatia started together with the search for Homeland War victims in 1991. It continued through years, most often sporadically and in a non-homogenous way. It was just with the adoption of the Law on Research, Arrangement and Maintenance of Military Cemeteries, Cemeteries of Victims of WWII and Postwar Period in 2013 that the search became more structured and gained a formal governmental body responsible for the organization and supervision of the activities related to it. It was then that the well-established model of searching for Homeland War victims, based on many years of field work and research, started to be implemented in the search and analysis of WWII/postwar victims. The model represents a continuously growing and developing project which encompasses a wide variety of steps and procedures, from the investigation of alleged burial locations to the analysis and reburial of recovered mortal remains. From its implementation in 2016, it allowed the successful investigation of 1300 alleged burial locations, 484 field surveys, 42 exhumations and the recovery of remains of more than 1600 individuals. Besides, care for military cemeteries, marking of mass burial sites and arranging of international treaties are conducted in order to guarantee proper handling, relocation and repatriation of all those that perished during WWII and the postwar period.

Suicidal drowning in Southwestern Croatia: a 25-year review.

We designed a retrospective study to analyze suicidal drowning deaths occurring between 1981 and 2005. During the study period there were 134 cases of suicidal drowning accounting 10% of all suicides, and 31% of all drowning deaths. Of the 134 drowning suicides, 76 (57%) incidents occurred in the sea, 46 (34%) in water wells, 8 (6%) in rivers, and 4 (3%) in bathtubs. During the study period, cases of suicidal drowning were not identified in males and females under the age of 20. In females aged 65 years and older, drowning represents the most common type of suicide. Commonly, the victims were sober at the time of the incident. Cases of suicidal drowning committed in water wells are unexpectedly high, considering abundance of salt water.

Trauma related fat embolism syndrome in forensic practice.

The fat embolism syndrome (FES) in forensic practice is observed usually in cases of polytrauma related deaths. FES is rare, but serious complication after trauma. The most cases of post traumatic fat embolism are not fatal and it's very likely that many cases of mild fat embolism are overlooked. We describe a case of fat embolism syndrome in a young man after high energy injury. Upon concrete ceiling fell on his shoulders he had open fractures of both tibias and massive haematoma of the left hip joint. Transport immobilization of both legs was performed with admitting of analgesia during transportation to the hospital. Immediately after admission to hospital he underwent surgery. Several hours after osteosynthesis of both tibias, in ICU patient became hemodinamically and respiratory unstable. He developed global cyanosis and metabolic acidosis with significant decrease of oxygen pressure in the blood. Control chest radiograph showed typical "snow-storm" like pulmonary infiltrations. Deep bradicardy occurred followed by cardiopulmonary arrest. CPR was unsuccessful and the patient suddenly died within 12 hours from the incident. Histologic confirmation of fatty droplets the most commonly observed in the lung capillaries, brain capillaries or disseminated throughout the body remains diagnostic standard. We present one case of FES to emphasize the arising need of a quantitative analysis of both the size and localization of the fat emboli in order to grade the severity of FES and its relative contribution in pathophysiology of death. The postmortem diagnosis of fat embolism syndrome (FES), traditionally based on the histological demonstration of fat globules seems not to be enough, nowadays. A quantitative analysis of both the size and localization of the fat emboli has been discussed as reliable method of grading the pulmonary fat embolism in order to determine its relative functional contribution in death pathogenesis.

Immunohistochemical surfactant protein-A expression: fatal drowning vs. postmortem immersion.

The postmortem diagnosis of drowning continues to be one of the most difficult in forensic pathology because of unspecific autopsy findings. It must be always remembered that disposal of a victim body in water is not unknown in homicide. The most important physiological consequence in fatal drowning is hypoxemia. The air-liquid interface of alveoli and distal airways of the mature lung are lined with a thin layer of lung surfactant, composed of phospholipids, proteins and neutral lipids. Surfactant components are synthesized and/or incorporated into lung surfactant in alveolar type II cells, and secreted to form an airspace lining film. The composition and function of lung surfactant is disturbed in cases od acute lung injury (ALI) including drowning. Surfactant protein-A (SP-A) is the most abundant surfactant protein. Surfactant protein-A (SP-A) is secreted by type II alveolar cells and cells. It's immunohistochemical distribution is observed in two different pathways: a linear membranous staining and a granular intra-alveolar staining. We hypothesize the significance of immunohistochemical detection of SP-A and its help in determination of the time of death, and possibly distinguishing of death by immersion vs. postmortem immersion using the morphological analysis applied on SP-A immunohistochemical stained lung tissue samples. We also argue in favor of routine use of SP-A staining in selected forensic cases where pathogenesis includes mechanical asphyxia and lung pathology. Although some studies reached conclusions to define the mechanisms involved in pathogenesis of mechanical asphyxia and aspiration necessity of additional studies arose. The mechanism of the production of massive aggregates remains to be determined. In the mean time the detection of SP-A (immunohistochemical) as well as biochemical is potentially useful tool in the forensic practice with possible application in daily practice.

Pedestrian traffic fatalities in Southwestern Croatia.

Pedestrians are often considered the most vulnerable group of road users. The aim of our study was to analyze the forensic aspects of pedestrian traffic fatalities in Rijeka region. We analyzed pedestrian fatalities in a 5-year period (2002-2006), which included 44 fatally injured pedestrians examined at the Rijeka Institute of Forensic Medicine. The male:female ratio was 1:1. The median age was 57 (ranging from 2-95), with 66% aging over 60 and 89% aging over 40. The least of the accidents happened in July and during weekend. Almost 65% of the fatally injured pedestrians were sober (0.00 g/kg) and 24% had blood alcohol level of more than 1.50 g/kg. Our study showed that pedestrian fatalities in Rijeka region have specific forensic characteristics. These findings suggest the necessity of the specific approach and caution in planning of prevention measures for specific traffic fatalities, in this case pedestrian ones.

Traffic accidents with fatally injured drivers in Southwestern Croatia.

Traffic accidents represent a significant cause of death in Croatia, therefore being an important social and public-health problem. The aim of our study was to analyze the forensic characteristics of traffic accidents with fatally injured drivers in Southwestern Croatia. Study population included 277 fatally injured drivers, in a 10 year period (between 1994 and 2004). As for the gender, 92% were male and only 8% were female. The median age was 35 (13-86), with 53% of them aging 20-39, therefore being a part of the working population. Most of the accidents happened during the summertime (44%) and weekend (57%). Half of the fatally injured drivers (139) were sober (0.00 g/kg) and only 10% (28) had blood alcohol level of 0.00-0.5g/kg. Our results suggest that, as opposed to the current public opinion in Croatia, alcohol might not be the only crucial factor contributing to the drivers' mortality in Croatia. As for the prevention measures, other factors (drug intoxication, fatigue...) should also be taken into consideration and further studied.

The methods of committing and alcohol intoxication of suicides in Southwestern Croatia from 1996 to 2005.

Alcohol is the psychoactive substance most frequently associated with suicidal behavior. We hypothesized that the level of alcohol intoxication and the choice of specific methods of committing suicide are related. Suicides in the Southwestern Croatia in a 10-year period, especially regarding the method of committing and alcohol intoxication, were analyzed. Ten various methods of committing suicide were recorded. The average blood alcohol concentration at the moment of suicide was 0.68 g/kg with male, and 0.29 g/kg with female victims. The highest blood alcohol levels at the moment of suicide were recorded with suicides by explosive device (with the average blood alcohol concentration of 1.71 g/kg). This especially drastic method of suicide was rare in Croatia in a period prior to the Croatian Independence War (1991-1995), but its incidence significantly increased during the war and in a post-war period. These victims were not chronic alcoholics, but the excessive alcohol consumption occurred as a result of psychiatric disorders as a consequence of war stress they suffered.

Aspergilloma in a pulmonary abscess--the first manifestation of malignant bronchial obstruction.

We report a case of aspergilloma in a pulmonary abscess being the first manifestation of malignant bronchial obstruction. In our case, only the autopsy revealed that planocellular carcinoma was the real cause of aspergilloma. Malignant bronchial obstruction finally resulted in pulmonary abscess with secondary fungal colonization. The co-existence of these pathologies is uncommon, and the situation we report is extremely rare. Even in cases when there are no radiological, bronchoscopical or cytological signs of malignancy, as in our case, careful observation of these patients is necessary. Surgical approach should be considered whenever possible.

Primary non-Hodgkin's lymphoma of the humerus presenting as osteomyelitis.

We report a case of primary non-Hodgkin's lymphoma of the humerus that occurred in a 21-year-old male patient. A previously healthy man presented with the history of severe pain and a differential diagnosis from osteomyelitis to malignant lesion was suspected. A biopsy was performed and the final pathologic diagnosis was a diffuse large B-cell lymphoma. The patient was treated for lymphoma only with radiotherapy. To date, the patient remains tumor free 10 years after the onset of the disease.

Temporal Trends in Rainwater Tank Suicides in Rijeka, Croatia-A 30-year Study.

Although drowning in rainwater tanks is a generally rare phenomenon, this method for suicide has been observed in parts of Croatia. Review of autopsy records at the University of Rijeka, Croatia, was undertaken from 1987 to 2016 to examine this phenomenon. Of 469 drowning deaths, there were 35 suicides in rainwater tanks (7.5%). Overall drowning deaths showed no temporal trends. In contrast, suicidal drownings in rain water tanks showed a marked decline over the years, with 15 cases in 1987-1991, seven in 1992-1996, six in 1997-2001, six in 2002-2006, one in 2007-2011, and none in 2012-2016. Thus, suicidal drowning cases as a percentage of overall drownings dropped from 18% to zero (p<0.001). This study reports a very unusual method of suicide that may have occurred because of the ready availability of large water reservoirs in homes. The recent fall in numbers may relate to less reliance on domestic reservoirs of water.

Herpes simplex virus 1 miRNA sequence variations in latently infected human trigeminal ganglia.

Human herpes simplex virus 1 (HSV-1) expresses numerous miRNAs, the function of which is not well understood. Several qualitative and quantitative analyses of HSV-1 miRNAs have been performed on infected cells in culture and animal models, however, there is very limited knowledge of their expression in human samples. We sequenced small-RNA libraries of RNA derived from human trigeminal ganglia latently infected with HSV-1 and Varicella zoster virus (VZV) and detected only a small subset of HSV-1 miRNA. The most abundantly expressed miRNAs are miR-H2, miRNA that regulates the expression of immediate early gene ICP0, and miR-H3 and -H4, both miRNAs expressed antisense to the transcript encoding the major neurovirulence factor ICP34.5. The sequence of many HSV-1 miRNAs detected in human samples was different from the sequences deposited in miRBase, which might significantly affect targeted functional analyses.

Propane-associated autoerotic asphyxiation: accident or suicide?

Autoerotic fatalities refer to a death resulting from the participation in a sex-related activity designed to heighten sexual arousal. The overwhelming majority of these deaths are due to asphyxia, most often by hanging or ligature compression of the neck. Here we report a case of "atypical" autoerotic fatality in which death was not induced by ligature compression. Unusual death of a 24-year-old man, involving propane inhalation and plastic bag suffocation, is described. The special characteristics of propane gas as an asphyxiant agent are discussed, as well as its effect on the human body.

War and suicidal deaths by explosives in southwestern Croatia.

A total of 853 suicides, from which 29 were committed by explosives, were recorded in the examined region during three distinctly specific intervals-peace time (1986-1990), wartime (1991-1995), and the postwar time (1996-2000). Suicides caused by explosives, without a terrorist background, from 1986-2000 were reviewed. Out of 29 suicides by explosives committed in the examined period, the age and sex of suicide victims and alcohol intoxication at the time of suicide were analyzed. In the prewar period no suicides by explosives were recorded, in distinction from wartime and the postwar period when 20 and 9 such suicides were recorded, respectively. Alcohol testing was performed in 76% of the cases, showing the highest alcohol concentration in suicides' blood in wartime with an average value of 1.58 g/kg of absolute alcohol. As to the suicides' sex, 93% of the victims were male and 7% female. The paper emphasizes the drastically increased number of suicides committed by explosives in wartime.

Heart embolization with the Kirschner wire without cardiac tamponade.

The case of the heart embolization with the Kirschner wire that was used for shoulder trauma fixation, 2 years previously in a 67-year-old female, is reported. This case is unique; although embolization of foreign bodies to the heart is not a novel occurrence, heart embolization with non-broken Kirschner wire with a total length of 13.5 cm without cardiac tamponade was not described in medical literature so far.

Cell death and cell proliferation in mouse submandibular gland during early post-irradiation phase.

The effects of irradiation on different cell compartments in the submandibular gland were analyzed in adult C57BL/6 mice exposed to X-ray irradiation and followed up for 10 days. Apoptosis was quantified using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end labeling method (TUNEL). Cell proliferation was detected using immunohistochemistry for proliferating cell nuclear antigen (PCNA). Radiation-induced apoptosis occurred rapidly, reaching a maximum 3 days post-irradiation. The percentage of apoptotic cells increased with the irradiation dose. At day 1 post-irradiation, cell proliferation was significantly reduced in comparison to sham-irradiated controls. After post-irradiation arrest of the cell cycle, proliferation increased in all gland compartments, reaching a maximum at day 6 post-irradiation. The proliferation response corresponded to the dose of irradiation. We suggest that the reason for gland dysfunction could be the coexistence of high apoptotic and proliferative activity in the irradiated gland.