Viability of human chondrocytes in an ex vivo model in relation to temperature and cartilage depth.

Chondrocytes in human articular cartilage remain viable post-mortem. It has however not been established yet how the storage temperature affects their survival, which is essential information when post-mortem cartilage is used for toxicologic studies. Our aim was to construct a simple model of explanted knee cartilage and to test the influences of time and temperature on the viability of chondrocytes in the ex vivo conditions. Osteochondral cylinders were procured from the cadaveric femoral condyles. The cylinders were embedded in water-tight rubber tubes, which formed separate chondral and osteal compartments. Tubes were filled with normal saline, without additives, to keep chondrocytes under close-to-normal conditions. The samples were divided into two groups stored at 4 degrees C and 35 degrees C, respectively. Three samples of each of these two groups were analysed at the time of removal, and then three and nine days later. Images of Live-Dead staining were scanned by a confocal laser microscope. Count of viable chondrocytes in four regions, from surface to bone, was obtained using image analysis software. The regression model revealed that the number of viable chondrocytes decreased every day by 19% and that an increase in temperature by 1 degree C decreased their viability by 5.8%. The temperature effect fell by 0.2 percentage points for every 100 microm from the surface to the bone. Herein we demonstrate that chondrocytes remain viable in the ex vivo model of human knee cartilage long enough to be able to serve as a model for toxicologic studies. Their viability is, however, significantly influenced by time and temperature.

The completed suicide as interplay of genes and environment.

The review will tackle the interplay of genetic and environmental risk factors behind the completed suicide. First, individual differences in suicidal behaviour in relation to heritability are presented followed by a brief discussion of genetic methods currently used to investigate candidate genes for the completed suicide. Further along the polygenetic, multi-factorial model of genetic proneness to suicidal behaviour is presented as interplay of genes and environment. Finally, the future implications in this quickly blossoming field of research are discussed.

Harmful alcohol use of those who died a violent death (the extended region of Ljubljana 1995-1999).

Consumption of alcohol increases the risk of dying a violent death. We wanted to establish a connection between harmful alcohol use and dying a violent death. We analyzed all such victims in the extended region of Ljubljana. The research included 1630 deceased, who were autopsied at the Forensic Institute of the Ljubljana, Faculty of Medicine in the period from 1995 to 1999. Presence of alcohol was established in 76.3% of the cases. From all included in the research, 38.2% of all work accident victims, 28.8% of all murder victims, 25.4% of suicides, 24.6% of victims involved in traffic accidents and 19.3% of those who died in accidents at home. 23.2% of all violent death victims had a concentration of alcohol above 1.5 g/kg; among those, victims of traffic accidents, suicides and accidents at home represent the largest part. The lowest values of alcohol in blood were found in those who died because of accidents at work. The highest values were found in males aged 35-44. The research confirmed that consumption of alcohol in Slovenia was strongly connected to violent deaths. The blood levels of alcohol of the victims are distinctively higher where there are practically no limitations of alcohol consumption and lower in the environment or activities where legal restrictions prohibit or at least explicitly limit harmful use of alcohol (working environment).

Expression of fibronectin suicidal in gunshot wounds.

Fibronectin has been proposed as the reliable marker of wound vitality and potent tool for age estimation in cases where survival time is at least few minutes. Cases of suicidal gunshot wounds were divided into two groups according to projectile energy and damage of the vital structures. Immunohistochemical reaction on fibronectin was evaluated with semiquantitative scale, which included sum of intensity and widespread of the reaction. Mann-Whitney test has shown statistical significant difference between groups (P< or =0.001).

Our experiences with fatal ecstasy abuse (two case reports).

Ecstasy is a psychostimulative drug (ab)used mostly by teenagers and young adults in discotheques and on the "rave" parties. Older adults ecstasy abusing cases are very rare. Among four cases of ecstasy abuse with fatal outcome noticed and examined in Slovenia, two were examined at our Institute of Forensic Medicine in Ljubljana. The first case was the accidental intoxication with 3,4 methylenedioxymethamphetamine (MDMA) on "rave" party, the second case was suicidal intoxication with combination of insulin and MDMA. Because of the increasing popularity of MDMA, it is important for all emergency physicians to be well educated in prompt recognition of MDMA intoxication symptoms. It is important that emergency physician carefully examines the death scene.

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.

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.

Human Y-specific STR haplotypes in a Slovenian population sample.

The allele distribution of the systems DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385 and YCAII were investigated in a sample of 121 unrelated males from Slovenia

Analysis of nine short tandem repeat (STR) loci in the Slovenian population.

A polymerase chain reaction (PCR)-based short tandem repeat (STR) system consisting of nine loci has recently been introduced in Slovenia for use in routine forensic identity testing. Fluorescently labelled PCR products were analysed using an ABI PRISM 310 Genetic Analyzer. The STR loci analysed exhibit between 6 and 14 observed alleles per locus and have a combined matching probability of 2.3 x 10(-10).

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.

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.