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.

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.

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.

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.

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.

The assessment of lens opacity postmortem and its implication in forensics.

Visual impairment, mostly due to cataracts, has been demonstrated to be an important factor associated with traffic accidents. Although vision screening is standard procedure during licensing in order to prevent motor vehicle accidents, an eye exam is not typically administered after an accident has already occurred. Postmortem assessment of lens opacity in victims of car accidents would provide helpful information for attesting to the liability of the parties in specific accidents, determining the circumstances of the accident, and developing preventive measures for both drivers and pedestrians alike. In this paper, we explore the use of different methods and their limitations for assessing lens opacity postmortem. We discuss the possible use and benefits of a simple, but as-yet untested method: retrobulbar translucency. The method would be based on the recording of shadows formed by opaque regions of the lens while the eye is illuminated from the back with a rigid source of light. The efficacy and objectivity of the method, its reproducibility, and the inter- and intra-observer error should be tested before implementing such a technique to be regularly used to determine lens opacity in cadavers.

Dive-related fatalities among tourist and local divers in the northern Croatian littoral (1980-2010).

BACKGROUND: The aim of the study was to retrospectively analyze diving fatalities occurring in Primorje-Gorski Kotar County (northern Croatian littoral), Croatia between 1980 and 2010 in order to identify differences between fatally injured tourist and resident divers, as well as temporal changes in the frequency of diver deaths. METHODS: Medico-legal and police reports of 47 consecutive fatal diving cases were reviewed to determine the frequency of death among divers in relation to year and month of death, age, sex, nationality, organization of diving, diving type, and health condition. RESULTS: The majority of victims were foreign citizens (59.6%) most of whom fell victim to scuba diving (70.4%). It was found that 79% of resident divers succumbed during free-diving. The number of diving fatalities increased significantly in the last three decades, especially among free-divers. Of the victims, 93% were males, usually belonging to younger age groups with tourist divers being significantly older than local divers. And 31.9% of divers, mostly tourists, showed signs of acute, chronic, or congenital pathological conditions. CONCLUSION: Fatally injured foreign divers differ from resident diver fatalities in diving method and age. Tourists are the group most at risk while scuba diving according to the Croatian sample. Occupational scuba divers and free-divers are the group most at risk among resident divers. This study is an important tool in uncovering the most common victims of diving and the related risk factors. It also highlights the problems present in the legal and medical monitoring of recreational divers and discusses possible pre-event, event, and post-event preventive actions that could lead to reduced mortality rates in divers.

Car-to-pedestrian accident with a unique decollement injury.

The authors present a unique case of decollement injury found on an 85-year-old victim that was run-over by a tank truck. While external examination evidenced multiple severe injuries, autopsy confirmed the preliminary findings and revealed also the presence of an extensive decollement that spread from the left hemithorax to a wound on the left ankle, through which parts of the small intestines and pertaining mesentery protruded. The article offers an interpretation of the injuries sustained by the victim, focusing on the most probable decollement mechanism. The forensic pathologist in this case could rely also on the valuable help of surveillance cameras of a nearby bank office that helped to better understand the events that brought to the fatal injuries. The authors concluded that the expulsion of the jejunum was produced by a combination of two movements: a forward passage that created the decollement and detachment of the jejunum and a backward movement that completed the expulsion of the jejunum through the open fracture of the ankle. This case report evidences the importance of the forensic pathologist and a correct and detailed investigation of injuries in reconstructing an accident, as well that of surveillance cameras as investigation tool in forensic cases.

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.

Introduction of calpain inhibitors in traumatic brain injury: a novel approach?

Traumatic brain injury (TBI) is a major cause of death and disability throughout the world. In recent years, researchers focused on the pathological significance of calcium accumulation in the brain after TBI. Neuronal calcium homeostasis disturbances may result in the activation of calpain a ubiquitous calcium-sensitive protease. The calpain family has a well-established causal role in neuronal cell death following acute brain injury: their activation has been observed to progressively increase after either contusive or diffuse brain trauma in animals, suggesting calpain to be a mediator of early neuronal damage. We hypothesize that pretreatment with the calpain inhibitors in population at objective risk (military soldiers' pre combat) in appropriate dose would open therapeutic time window expected to prevent and reduce extensive brain damage by providing optimal TBI neuroprotection. Additional therapeutic strategy for TBI, based on calpain modulating actions such as pretreatment with calpain inhibitors has been proposed. Since calpain overexpression has been well established in acute neuronal injury and further subsequent neurodegeneration, from a clinical viewpoint, we speculate that if this hypothesis proves correct pretreatment inhibitors introduction may become a therapeutic option for different brain pathologies to be approached and treated with.

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.

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.

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.

Toxicology and methods of committing suicide other than overdose.

Suicide represents a serious public health problem throughout the world. Toxicology plays a crucial role in the investigation of suicide. Psychoactive substances are recognized in the literature as the main suicide-generating stimuli. Ethanol is known to produce disinhibition and increased self-confidence. Other psychoactive substances can predominantly be central nervous system (CNS) stimulants or depressors. In cases of overdose as a method of suicide, the link between toxicology and the method of suicide is a matter of common sense and requires no additional explanation. On the other hand, in cases of non-overdose suicides this link is much more complex, and has not yet been extensively elucidated. We hypothesize a close relationship between toxicology and the choice of the method of committing suicide other than overdose. Negative findings may reflect either poor prescribed drug compliance in psychiatric patients, or suggest the role of other (non-toxicological) suicide-generating stimuli. On the other hand, positive findings influence the choice of the suicide method in a way that it depends on the prevalence of either stimulation or depression of the CNS, and consequent degree of behavioral aggression. Simplified, if the prevailing effect is CNS stimulation (with an increase in aggression), the method would be more drastic and more immediately fatal one, while with the CNS depression the method would be less immediately fatal and less drastic. There are only a few studies on the prevalence of psychoactive substances amongst completed suicides and even fewer studies have attempted to examine the relationship between substances used and the circumstances surrounding suicide. In evaluation of our hypothesis, we suggest thorough studies of toxicology and the choice of methods of committing suicides other than overdose. The scientific knowledge gained this way will eventually make toxicology a useful target in suicide prevention, especially amongst younger population.

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.

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.