HSV-1 miRNAs are post-transcriptionally edited in latently infected human ganglia.

IMPORTANCE: Herpes simplex virus 1 is an important human pathogen that has been intensively studied for many decades. Nevertheless, the molecular mechanisms regulating its establishment, maintenance, and reactivation from latency are poorly understood. Here, we show that HSV-1-encoded miR-H2 is post-transcriptionally edited in latently infected human tissues. Hyperediting of viral miRNAs increases the targeting potential of these miRNAs and may play an important role in regulating latency. We show that the edited miR-H2 can target ICP4, an essential viral protein. Interestingly, we found no evidence of hyperediting of its homolog, miR-H2, which is expressed by the closely related virus HSV-2. The discovery of post-translational modifications of viral miRNA in the latency phase suggests that these processes may also be important for other non-coding viral RNA in the latency phase, including the intron LAT, which in turn may be crucial for understanding the biology of this virus.

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.

"Selfie"-related electrocution.

Electrical injuries contribute a small but significant fraction to all burn trauma cases, and an even smaller proportion to those that are lethal. Accidental death rates among teenagers are the highest of all age-groups, and taking a "selfie" has been a practice that has, on rare occasions, led to such a death, often from electrocution. The case of a young man and his friend, who were electrocuted after climbing onto the roof of a parked railway wagon to take selfies is reported. One died at the scene, while the other survived. This case demonstrates that electrocution may occur with parked railway wagons, and in the young may be associated with selfie taking.

Homicide-suicide events in Southwestern Croatia, 1986-2009.

The aim of this study was to investigate the occurrence and characteristics of homicide-suicide events in Southwestern Croatia from 1986-2009 relying on autopsy reports of the Department of Forensic Medicine and Criminalistics, Rijeka and police records. A total of 17 cases involving 19 victims were identified. The perpetrators were most often men (82%), living in a spousal relationship with the victim. In 76.5% cases, suicide was committed immediately after homicide, and the same mean (most often an illegal firearm) was used for both. After examining the belongings of those involved in homicide-suicides, two factors differentiated our study from others, i.e. more victims and perpetrators were from the lower socioeconomic class and explosives were used in two homicide-suicide (12%) cases. The greater frequency of explosive used in homicide-suicide events may be a result of factors associated with the recent war fought in Croatia. These data prove the influence of sociological, historical and also political factors on the characteristics of a rare event such as homicide-suicide.

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.

Detection of Sars-Cov-2 antigens in thyroid gland showing histopathological features of subacute thyroiditis.

The clinical and laboratory findings of subacute thyroiditis have been repeatedly reported as being associated with acute Sars-Cov-2 infection and post-COVID-19 syndrome. The exact mechanisms and histopathological correlations underlying thyroid involvement remained unresolved, but current insights suggest either direct viral damage, systemic inflammatory reaction, or an autoimmune response as possible noxious effectors. Here we present findings of immunohistochemical/immunofluorescence detection of Sars-Cov-2 viral proteins (spike/S and nucleocapside proteins) in relation to histoarchitectonic changes of autoptic thyroid tissue obtained from patient who deceased from COVID-19.

Detection of SARS-CoV-2 Antigens in the AV-Node of a Cardiac Conduction System-A Case Report.

Mounting evidence indicates that new arrhythmic events frequently occur during and after coronavirus disease (COVID-19), posing additional mortality risk in older-aged and critically ill patients. However, the underlying mechanisms and cardio pathological substrates of COVID-related arrhythmias have not been clarified yet. Here, we report findings of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens and genes in the atrioventricular node (AV-node) of a cardiac conduction system, pointing to its direct infection as a possible arrhythmogenic factor.

Hepatoregenerative role of bone morphogenetic protein-9.

Bone morphogenetic protein-9 (BMP-9) is a member of the transforming growth factor beta (TGF-ß) superfamily of cytokines, which regulate cell growth and differentiation during embryogenesis. Apart of that, the hypoglycemic potential of BMP-9 is of great interest. It has been confirmed that BMP-9, like insulin, improves glycemia in diabetic mice and regulates directional glucose metabolism in hepatocytes; therefore it is proposed to be a candidate hepatic insulin-sensitizing substance (HISS). In liver fibrosis, due to the portocaval shunt, insulin bypasses the organ and the liver undergoes atrophy. Parenteral administration of insulin reverses atrophy by stimulating mitogenic activity of the hepatocytes. Because BMP-9 has a signaling pathway similar to other BMPs and insulin, it is to be expected that BMP-9 has a certain regenerative role in the liver, supporting the above-mentioned is evidence of BMP-9 expression in Dissè's spaces and BMP-7's mitogenic activity in mucosal cells. However, further studies are needed to confirm the possible regenerative role of BMP-9.

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.