Jože Balažic

The association between brain-derived neurotrophic factor polymorphism (BDNF Val66Met) and suicide

BACKGROUND Brain-derived neurotrophic factor (BDNF) mediates neural plasticity, mood, different behaviours, and stress response. A functional BDNF polymorphism (BDNF Val66Met) was reported to influence the effects of stressful life events or childhood adversity on depression and suicidal behaviour in various psychopathologies. The study evaluated the association between BDNF Val66Met variants and suicide, committed with violent or non-violent methods, in victims with or without stressful childhood experience. METHODS BDNF Val66Met polymorphism was genotyped on 560DNA samples from 359 suicide victims and 201 control subjects collected on autopsy from unrelated Caucasian subjects and subdivided according to gender, method of suicide, and influence of childhood adversity. RESULTS A similar frequency of BDNF Val66Met variants was found between all included suicide victims and the control groups, and also between the male groups. The frequency of the combined Met/Met and Met/Val genotypes and the homozygous Val/Val genotype was significantly different between the female suicide victims and female controls, between the female suicide victims who used violent suicide methods and female controls, and between all included suicide victims with or without stressful life events. The combined Met/Met and Met/Val genotypes contributed to this significance. LIMITATION A small group of suicide victims with available data on childhood adversity was studied. CONCLUSIONS The combined Met/Met and Met/Val genotypes of the BDNF Val66Met variant could be the risk factor for violent suicide in female subjects and for suicide in victims exposed to childhood trauma. These results confirm a major role of BDNF in increased vulnerability to suicide.

Promoter and functional polymorphisms of HTR2C and suicide victims

In Europe, the countries with the highest suicide rates form a so‐called J‐curve, which starts in Finland and extends down to Slovenia—a country with one of the worlds highest suicide rates. So far, the strongest association between suicide and genes has been shown for the serotonergic system. A functional polymorphism 68G>C (Cys23Ser) and a promoter polymorphism–995G>A of serotonin receptor 2C (HTR2C) have already been investigated, but no associations with suicide were determined. In the present study 334 suicide victims and 211 controls of Slovenian origin were genotyped for the above‐mentioned polymorphisms using standard methods. In the case of the polymorphism–995G>A no association with suicide was found. However, a significant association was observed between female suicide victims and polymorphism 68G>C. The significance remained when we combined alleles of female and male populations. An excess of GG genotype and allele G was observed. However, no statistically important differences were present when only males were analyzed. Haplotype analysis on female population showed marginal association of haplotype G‐C with suicide. The present study speaks for the plausible implication of the HTR2C in suicide susceptibility.

Frequency and number of resuscitation related rib and sternum fractures are higher than generally considered

AIM OF THE STUDY This study aimed to establish the incidence, number and location of CPR-related skeletal chest injuries (SCI) and to investigate the influence of age, gender, changes in resuscitation guidelines and technique of resuscitation. METHODS We analysed SCI in 2148 patients who had undergone resuscitation for non-traumatic cardiac arrest, as shown by autopsies performed at the Institute of Forensic Medicine in Ljubljana in the period 2004-2013. RESULTS External cardiac massage caused SCI in 86% of males and in 91% of females; sternum fractures occurred in 59% of males and 79% of females, rib fractures in 77% of males and 85% of females and sternocostal separations in 33% of males and 12% of females. The average number of all SCI per person was thus almost the same in males and females: 10.95 vs. 10.96. The percentage of patients injured and the number of SCI increased with age. Changes in resuscitation guidelines were also identified as a factor contributing to the incidence and number of SCI. No adverse effect of the use of LUCAS was found. CONCLUSION It is generally considered that at least 1/3 of resuscitated patients sustain rib fractures and at least 1/5 sustains sternum fractures. However, our study showed that these injuries are much more frequent and that increased compression rate and depth cause more SCI. Since in the period 2011-2013 accompanying severe injuries occurred in only 1.85% of cases, the resuscitation technique has not yet jeopardised patients safety, but further close monitoring is needed.

Sequence polymorphism of the mitochondrial DNA control region in the Slovenian population

The forensic application of mitochondrial DNA (mtDNA) typing requires large and regionally well-defined databases. To expand the database for forensic identification purposes in Slovenia, the mtDNA control region sequences of the hypervariable regions HVI and HVII were determined in a population of 129 maternally unrelated Slovenians, using a fluorescent-based capillary electrophoresis sequencing method. A total of 111 different haplotypes resulting from 124 polymorphic positions (80 polymorphic positions in HVI and 44 in HVII) were found. Of these, 101 mtDNA types were unique, 6 haplotypes were shared by 2 individuals, 1 haplotype by 3 individuals, 2 haplotypes by 4 individuals, and the most common haplotype was found in 5 individuals. The most frequent haplotypes in the Slovenian population ,263(G), 315.1(C) and 263(G), 309.1(C), 315.1(C) are also the most common in other European populations. The data support the concept that these haplotypes may represent a common European mtDNA sequence types. The sequence poymorphisms were compared to the databases of west Austria and central Italy and the HVI and HVII sequence matching probabilities within and between populations were calculated. It is 1.1–4.5 times more likely to find a sequence match in a random pair of Slovenians than in a random Slovenian-Italian pair and in a random Slovenian-Austrian pair. The length heteroplasmy in the homopolymeric C-stretch regions located at nucleotide positions 16184–16193 in HVI and at positions 303–315 in HVII was observed in 17% and 8% of individuals, respectively. A statistical estimate of the results for this population showed the random match probability and the genetic diversity of 1.16% and 0.996, respectively.

Alcoholic ketoacidiosis: a cause of sudden death of chronic alcoholics

In forensic medicine we often deal with cases of sudden death of chronic alcoholics from various aspects; above all we must establish the immediate cause of death. Most of the time the cause of death becomes clear from autopsies, histological, alcoholimetric and toxicological tests, but in a smaller number of cases it remains indeterminate even after these procedures. In our study we attempted to establish whether these cases of sudden death could be cases of alcoholic ketoacidosis. We studied 30 cases of sudden death of chronic alcoholics, for all of which autopsies, histological, alcoholimetric and toxicological tests had been performed, and for which we determined the β-hydroxybutyrate concentration in bodily fluid samples (blood, urine, cerebrospinal fluid and vitreous humour). In the group with indeterminate cause of death (seven cases) the average measured concentration of β-hydroxybutyrate in the blood, urine and vitreous humour was at least ten times higher than the concentrations in the group with clear causes of death, while the concentration of β-hydroxybutyrate in the cerebrospinal fluid did not differ significantly between the two groups.

Intracranial injuries by a screwdriver

Two patients with unusual intracranial foreign bodies are presented. Intracranial injuries produced by a piece of screwdriver as a wounding agent have not yet been reported in the literature. In one of our two cases we had to deal with homicide and in another with accidental self-inflicted injury. Both patients died, 5 and 14 days, respectively, after successful removal of the foreign body. In both cases death was due to brain ischaemia and the resulting oedema secondary to arterial injury.

Identical Human Papillomavirus (HPV) Genomic Variants Persist in Recurrent Respiratory Papillomatosis for up to 22 Years

Seventy initial and 125 follow-up tissue specimens of laryngeal papillomas, obtained from 70 patients who had had recurrent respiratory papillomatosis for from 1-22 years, were investigated for the presence of human papillomavirus (HPV) DNA and HPV E5a, LCR and/or full-length genomic variants. HPV-6 was found in 130/195, HPV-11 in 63/195, and HPV-6/HPV-11 in 2/195 samples. Within 67/70 (95.7%) patients, all follow-up HPV isolates genetically matched completely initial HPV isolate over the highly variable parts of the genome or over the entire genome. Frequent recurrence of laryngeal papillomas is a consequence of long-term persistence of the identical initial HPV genomic variant.

Parentage testing with 14 STR loci and population data for 5 STRs in the Slovenian population

Abstract In order to apply a set of 14 short tandem repeat (STR) loci in parentage testing, we performed a population genetic study on a sample of 260 unrelated people from the Slovenian population. Genotypes for the 14 STRs were determined using three multiplex polymerase chain reactions (PCR) and automated fluorescent detection. The allele frequencies of the STR loci D5S818, D13S317, D7S820, D8S1179 and D18S51 showed no deviation from the Hardy-Weinberg equilibrium and agreed well with other Caucasian populations. We resolved a series of 181 parentage disputes of which 29 were exclusions. In all cases, evidence for exclusion was obtained by at least 4 informative STRs out of the 14 loci analysed. The 14 loci combined comprise a highly discriminating test suitable for paternity and identity testing in the Slovenian population, with an average estimated mutation rate of 1.2×10–3, a combined calculated power of exclusion of 99.99974% and paternity index (PI) value of >106 in 72% of the inclusion cases and >105 in 91% of the inclusion cases.

Sudden death caused by laryngeal papillomatosis.

Laryngeal papillomatosis (LP) is the most frequent benign neoplasm of the larynx. Clinically it causes hoarseness and upper airway obstruction. Though the LP has the potential to endanger life by asphyxiation, this unfortunate outcome is extremely rare. We report the case of a 19-year-old female who suddenly died of asphyxiation caused by massive LP.

Amniotic fluid embolism with involvement of the brain, lungs, adrenal glands, and heart.

The case of a healthy 31-year-old woman in the 40th week of second pregnancy is presented. During preparation for an emergency caesarean section, she developed an amniotic fluid embolism (AFE) with unusual and unique features. The acute onset of disease with cardiorespiratory failure with hypotension, tachycardia, cyanosis, respiratory disturbances and loss of consciousness, suggested at first a pulmonary thromboembolism, but the appearance of convulsions led to the diagnosis of AFE. The patient died after 5 days due to an untreatable brain edema. At autopsy, AFE with the usually associated disseminated intravascular coagulation was found in the lungs, brain, left adrenal gland, kidneys, liver and heart. Eosinophilic inflammatory infiltrates were found in the lungs, hepatic portal fields and especially in the heart, suggesting a specific hypersensitivity reaction to fetal antigens. Moreover, intravascular accumulation of macrophages in the lungs also favored a non-specific immune reaction to amniotic fluid constituents.