Forensic osteological investigations in Kosovo.

A team of Finnish forensic experts performed investigations of alleged mass graves in Kosovo under the mandate of the European Union (EU). Human skeletal remains from two locations were examined. The remains contained three almost complete skeletons, and individual bones and bone fragments, part of which were burned. Injuries, pathological changes, and findings for identification purposes were examined and documented using standard methods of forensic pathology and osteology. Gunshot injuries were found in some cases, but reliable determination of the cause and manner of death was not possible. A discrepancy arose between the number of victims reported in information received from the presiding district court, and results of the investigations. The estimation of the minimum number of victims was mostly acquired by DNA analysis.

Independent forensic autopsies in an armed conflict: investigation of the victims from Racak, Kosovo.

In January 1999, a team of Finnish forensic experts under the mandate of the European Union (EU forensic expert team, EU-FET) performed forensic investigations in a sovereign state, in Kosovo, the Federal Republic of Yugoslavia (FRY). The team served as a neutral participant in the forensic investigation of victims of an incident at Racak, which was receiving considerable international attention. The Finnish team performed forensic autopsies, monitored forensic autopsies performed by local experts and verified findings of earlier executed autopsies. The victims had sustained varying numbers of gunshot wounds, which were established to be the cause of death. The manner of death remained undetermined by the EU-FET, because the scene investigation and the chain of custody for the bodies from the site of the incident to the autopsy were impossible to verify by the team. The events at Racak were the first of those leading to charges by the International Criminal Tribunal for the former Yugoslavia (ICTY) against the highest authorities in power in the FRY for crimes against humanity and violations of the laws or customs of war.

Practical and legal aspects of forensic autopsy expert team operations.

Finnish forensic experts have investigated remains of the victims of alleged mass violence in the former Yugoslavia in 1996 under the mandate of the United Nations, and in 1998 and 1999 under the mandate of the European Union. The investigative documents later were surrendered to the International Criminal Tribunal for the former Yugoslavia (ICTY). The indictments issued by the ICTY include charges even against the highest authorities of the Federal Republic of Yugoslavia. This study describes the experience gained in organising forensic expert team operations in a foreign state by the Finnish team. The establishment and operation of a forensic expert team in a foreign state involve, among other things, legal issues, often related to differing legal systems. From an independent forensic expert team, great objectivity and self-constraint are expected. Moreover, a clear and sufficiently detailed agreement on the mandate of the team and on the possibility for unhindered and safe access of the experts to the alleged mass graves, as well as the assembling and briefing of the team members and provision of the necessary equipment should be completed before the beginning of the mission. Furthermore, the application of adequate and internationally recognised methods for processing and documenting the examination is essential for team's credibility.

Radiology in forensic expert team operations.

Radiological methods are widely used in forensic pathology. Their most common applications are in complementing human identification, particularly in investigations of mass disasters and decomposed bodies, and in searching for foreign material inside corpses. A team of Finnish forensic experts investigated human skeletal remains in Bosnia and Herzegovina (1996) and in Kosovo, the Federal Republic of Yugoslavia (1998). It also investigated more recently deceased victims in Kosovo (1999). In these investigations, the benefit of X-ray was in the detection of foreign material inside victims and their remains. For identification purposes, X-rays were mainly used to provide the best evidence possible of any pathological changes, physical characteristics, and injuries present.

Forensic investigations in Kosovo: experiences of the European Union Forensic Expert Team.

The international community has intervened ethnic hostilities in Kosovo, the Federal Republic of Yugoslavia. A team of Finnish forensic experts worked in December 1998 and January 1999 in Kosovo under the mandate of the European Union. The team investigated human remains from three locations of alleged ethnic violence. Concerning the events in Klecka, Volujak and Racak, the International Criminal Tribunal for the former Yugoslavia received documents of great gravity. The standard methods of forensic pathology were complemented with radiological, forensic odontological, physical anthropological and molecular genetic investigations.

Glycoprotein IIIa Pl(A) polymorphism associates with progression of coronary artery disease and with myocardial infarction in an autopsy series of middle-aged men who died suddenly.

Glycoprotein IIIa (GPIIIa) has a key role in the aggregation of thrombocytes, and it also mediates intimal hyperplasia after endothelial injuries; the possible association of the Pl(A1/A2) polymorphism of the gene for GPIIIa with coronary thrombosis and with the progression of coronary artery disease (CAD) is still to be confirmed. Therefore, the association of the Pl(A) polymorphism with the development of coronary atherosclerosis, coronary narrowing, and myocardial infarction (MI) was studied in a prospective, consecutive autopsy series of 300 middle-aged, white Finnish men (33 to 69 years) suffering sudden out-of-hospital or violent death. Coronary atherosclerosis was measured morphometrically and the coronary stenosis percentage determined from a cast rubber model of the coronary tree. We found a significant inverse relation (P=0.01) between the Pl(A2)-positive genotype and coronary artery stenosis. The frequency of possessing the Pl(A2) allele was significantly (odds ratio [OR] 0.45, 95% confidence interval [CI] 0.22 to 0.98) lower among men with >50% coronary stenosis (18.3%) than among those with <25% stenosis (32.9%). Although the Pl(A) polymorphism was not directly associated with MI, the Pl(A2) allele was present in 11 of the 22 men (50%) with MI and coronary thrombosis (OR 6.6, 95% CI 2.1 to 22.8) but in only 6 of the 47 (12.8%) with MI associated with severe stenosis in the absence of thrombosis. In line with this result, men possessing the Pl(A2) allele also had a larger area of fissured and ulcerated complicated lesions in their coronary arteries (P<0.05). The present results suggest that the Pl(A) polymorphism is involved in the development of CAD and MI. Men with the Pl(A2) allele may harbor more thin-walled, vulnerable coronary plaques, plaques prone to rupture, leading to massive, fatal thrombosis. In contrast, men homozygous for the Pl(A1) allele may more often show stable plaques and present with infarction caused by progressive coronary stenosis.

Age-dependent association of apolipoprotein E genotype with coronary and aortic atherosclerosis in middle-aged men: an autopsy study.

BACKGROUND: Apolipoprotein E (apoE) polymorphism is one of the genetic determinants of serum cholesterol values. The apoE epsilon4 allele has been associated with advanced coronary heart disease (CHD) diagnosed by angiography, but the role of the apoE genotype in atherosclerosis has not been confirmed at vessel-wall level, nor is any age-dependent effect of the apoE genotype on the development of CHD known. METHODS AND RESULTS: The right and left anterior descending coronary arteries (RCA and LAD) and the aorta from 700 male autopsy cases (Helsinki Sudden Death Study) in 1981-1982 and 1991-1992 (average age 53 years, range 33 to 70 years) were stained for fat, and all areas covered with fatty streaks, fibrotic plaques, and complicated lesions were measured. In the RCA and LAD, the apoE genotype was significantly associated with the area of total atherosclerotic lesions in men <53 years old but not with that in older men (P=0.0085 and P=0.041, respectively, for age-by-genotype interaction). Men <53 years old with the epsilon4/3 genotype showed 61% larger total atherosclerotic lesion area in the RCA (P=0.0027) and 26% larger area in the LAD (P=0.12) than did men with the epsilon3/3. The apoE epsilon4/3 was also associated with atherosclerotic lesions in the abdominal (P=0.014) and thoracic (P=0.12) aorta, but this effect, unlike that of the coronary arteries, was not age-related. CONCLUSIONS: In men, the apoE epsilon4 allele is a significant genetic risk factor for coronary atherosclerosis in early middle age. This suggests that at older age, other known risk factors of CHD play a more important role in the atherosclerotic process than apoE polymorphisms.

Genotyping of five short tandem repeat loci via triplex and duplex PCR.

We have developed a triplex PCR method for D3S1359, HumTH01 and HumTPO tetranucleotide loci and a duplex PCR method for HumFES/FPS and HumvWA31A tetranucleotide loci using high resolution polyacrylamide gel electrophoresis and silver staining. The methods were evaluated for paternity testing and individual identification and allele frequencies at these loci are reported for 189-3387 unrelated individuals in the Finnish population. The D3S1359 locus, especially, was found to be a highly informative locus. Seventeen alleles were found in the D3S1359 locus with a highest observed allele frequency of 0.199, a high exclusion power (PE) in paternity testing (0.78) and a high observed heterozygosity (0.89). The combined PE for these five loci was 0.99.

Moderate alcohol consumption and disorders of human spermatogenesis.

Environmental factors are suspected to be responsible in part for the deterioration in semen quality observed worldwide during the recent few decades. Alcohol might be one factor, considering the frequent changes in testicular function associated with heavy drinking. The dose-dependent effects of alcohol on human spermatogenesis are, however, not well known. We analyzed spermatogenesis and testicular tissue morphology of 195 men, aged 35-69 years, with computer-assisted microscopy in this autopsy study. The men were categorized into controls and four "consumption groups" according to the average daily alcohol consumption, which was determined on the basis of blind interviews with relatives and acquaintances. When the average daily alcohol consumption was 40 g or less, 59 (66%) of the 90 men showed normal spermatogenesis, whereas 31 (34%) had partial spermatogenic arrest (SA). Of the 31 men with average daily intake between 40 and 80 g, 17 (54%) showed normal spermatogenesis, 13 (42%) had partial or complete SA, and 1 man exhibited more severe testicular damage-"Sertoli cell only" (SCO) syndrome. Among men with daily intake between 80 and 160 g, only 13 of 35 men showed normal spermatogenesis (37%), 19 (54%) had partial or complete SA (odds ratio = 2.92), and 3 (9%) had the SCO syndrome (odds ratio = 16.85). The frequencies of spermatogenic disorders were similar in men drinking in excess of 160 g. Both SA and the SCO syndrome showed a clear dependence on daily dose; p < 0.0001 and p < 0.0004, respectively. We conclude that long-term average daily consumption of < 40 g of alcohol seems not to be associated with disorders of spermatogenesis. Consumption of moderate amounts of alcohol may affect semen quality more often than previously thought, whereas high alcohol consumption may even be associated with serious disorders of spermatogenesis.

Early perivenular fibrogenesis--precirrhotic lesions among moderate alcohol consumers and chronic alcoholics.

Cirrhosis is believed to be preceded by "precirrhotic" lesions indicating initial fibrogenesis in the perivenular area. We investigated three previously described markers of early perivenular fibrogenesis: the thickness of the rim of the terminal hepatic venule, perivenular fibrosis and perivenular fibronectin deposition. The frequencies of these features were evaluated and compared to long-term daily alcohol intake in autopsy series of 120 males comprising abstainers, moderate alcohol consumers and chronic heavy alcohol consumers. Thickening of the rim of the terminal hepatic venule showed no correlation to the long-term daily ethanol intake. In contrast, compared to abstainers, daily alcohol intake between 40 and 80 g for an average of 25 years was associated in an increased number of subjects with perivenular fibrosis (13.3% vs 38.9%, p < 0.05), and with perivenular fibronectin deposition (13.3% vs 44.4%, p < 0.025). Similarly, daily intake exceeding 80 g was associated in an increased number of subjects with perivenular fibrosis (56.1%, p < 0.001) and with perivenular fibronectin deposition (56.1%, p < 0.001). A daily intake exceeding 80 g (110 g/d vs 240 g/d) did not, however, further increase the occurrence of these lesions. A daily intake below 40 g of absolute alcohol was not associated with signs of early perivenular fibrogenesis. In this study, the frequencies of subjects with perivenular fibrosis and perivenular fibronectin deposition correlated with the amount of daily alcohol intake. A daily intake between 40 and 80 g was associated with approximately a three-fold and a daily intake exceeding 80 g with approximately a five-fold increase in the risk of these features. This could suggest that daily intake between 40 and 80 g represents a "threshold" level, beyond which the risk of alcoholic liver fibrosis increases significantly. The majority (50-70%) of chronic heavy alcohol consumers presented signs of early perivenular fibrogenesis, whereas cirrhosis was found in only 20% of heavy consumers. This could suggest that alcohol-induced fibrotic lesions of the liver may develop in the majority of people, but factors other than cumulative alcohol consumpation may have a critical impact on the progression of early liver fibrosis to cirrhosis.

Cytokeratin inclusions in alcoholic liver disease and their relation to the amount of alcohol intake.

In the present study, the frequency and the distribution pattern of immunoreactive hepatocytic cytokeratin (CK) inclusions was investigated using the monoclonal antibody (MAb) CAM 5.2 detecting CKs 8, 18 and 19. The CK antigenicity of the inclusions was confirmed on frozen sections with MAbs for the CKs 7, 8, 17, 18 and 19. The frequency of hepatocytic CK aggregates was compared to the presence of non-alcoholic and alcoholic liver disease (ALD) as well as to the average all-year daily ethanol intake of 195 consecutive males subjected to medico-legal autopsy. Hepatocytic CK inclusions were infrequent in patients with normal liver histology, portal fibrosis and/or portal inflammation. In ALD, however, inclusions were observed in 35.6% of patients with fatty liver, in 63.2% of patients with alcoholic hepatitis, in 30.8% of patients with bridging fibrosis and in 60.0% of patients with liver cirrhosis. In all specimens studied, the inclusions were reactive only for CKs 8 and 18, CKs 7, 17, and 19 being unreactive. The frequency of inclusion bodies increased, paralleling increasing average all-year daily alcohol consumption. Compared to non-drinkers, a daily intake of between 40 and 80 g of absolute alcohol was associated with a statistically significantly (relative risk, RR = 6.6) increased risk of formation of aggregates. Similarly, daily consumption exceeding 80 g was related to increased (RR = 6.0) frequency of CK aggregates. The frequency of full-blown "classical" Mallory bodies (MBs) was, however, increased (RR = 8.9) only in patients with a daily intake exceeding 160 g.(ABSTRACT TRUNCATED AT 250 WORDS)

Pleural plaques and exposure to mineral fibres in a male urban necropsy population.

OBJECTIVES: The study aimed to evaluate the risk of pleural plaques according to the degree of past exposure to asbestos, type of amphibole asbestos, and smoking, as well as to estimate the aetiologic fraction of asbestos as a cause of plaques among urban men. METHODS: The occurrence and extent of pleural plaques were recorded at necropsies of 288 urban men aged 33 to 69 years. The pulmonary concentration of asbestos and other mineral fibres was analysed with scanning electron microscopy. The probability of past exposure was estimated from the last occupation. RESULTS: Pleural plaques were detected in 58% of the cases and their frequency increased with age, probability of past occupational exposure to asbestos, pulmonary concentration of asbestos fibres, and smoking. The risk of both moderate and widespread plaques was raised among asbestos exposed cases, and the risk estimates were higher for widespread plaques than for moderate plaques. The age adjusted risk was higher for high concentrations of crocidolite/amosite fibres than for anthophyllite fibres. The aetiologic fraction of pulmonary concentration of asbestos fibres exceeding 0.1 million fibres/g was 43% for widespread plaques and 24% for all plaques. The median pulmonary concentrations of asbestos fibres were about threefold greater among cases with widespread plaques than among those without plaques. No increased risk of pleural plaques was associated with raised total concentrations of non-asbestos fibres. CONCLUSION: The occurrence of pleural plaques correlated closely with past exposure to asbestos. The risk was dependent on the intensity of exposure. Due to methodological difficulties in detecting past exposures to chrysotile and such low exposures that may still pose a risk of plaques, the aetiologic fractions calculated in the study probably underestimate the role of asbestos.

Lumbar disc degeneration and atherosclerosis of the abdominal aorta.

STUDY DESIGN: This study analyzed the relationship of disc degeneration to atherosclerotic changes in the abdominal aorta and stenosis of the ostia of the lumbar and middle sacral arteries, 86 plain radiographs of lumbosacral spines, and the corresponding abdominal aortas were evaluated in connection with routine autopsy. RESULTS: Disc degeneration, advanced atherosclerotic manifestations, and stenosis of the ostia of the lumbar and middle sacral arteries all increased with age (P < 0.001). After partial rank correlation analysis, keeping the effect of age constant, there was still a statistically significant association between the grade of disc degeneration and stenosis of the ostia of the arteries supplying the disc. The association was stronger at upper lumbar levels (0.001 < P < 0.01) than at lower ones (0.01 < P < 0.05). Furthermore, with complicated lesions in the abdominal aorta, disc degeneration at all lumbar levels increased. CONCLUSIONS: Atherosclerosis in the abdominal aorta and especially stenosis of the ostia of segmental arteries may play a part in lumbar disc degeneration.

Asbestos exposure and pulmonary fiber concentrations of 300 Finnish urban men.

OBJECTIVES: The aim of the study was to determine the pulmonary concentrations of mineral fibers in the Finnish male urban population and to evaluate the analysis of pulmonary fiber burden by scanning electron microscopy (SEM) as an indicator of past fiber exposure. METHODS: The pulmonary concentration of mineral fibers was determined by SEM and compared with occupational history for a series of 300 autopsies of urban men aged 33 to 69 years. RESULTS: The concentration of fibers (f) longer than 1 micron ranged from < 0.3 to 163.10(6) per gram of dry tissue (f.g-1). Asbestos fiber concentrations exceeding 1.10(6) f.g-1 were observed in 33% of the cases with probable occupational exposure to asbestos and 1% of the cases with unlikely occupational exposure. Even asbestos fiber concentrations of 0.3 to 1.10(6) f.g-1, especially of crocidolite-amosite fibers, were rare among the men with unlikely occupational exposure. Fiber concentrations exceeding or equaling 1.10(6) f.g-1 were 10 times more frequent among the men more than 60 years of age as compared with those less than 40 years of age. Inorganic fibers other than asbestos had a weaker correlation with occupational history and age. Smoking habits had no significant effect on the pulmonary fiber counts. CONCLUSIONS: Asbestos fiber concentrations exceeding 1.10(6) f.g-1 are highly indicative of past occupational exposure to asbestos. The distribution of fiber concentrations in the different age groups of this study indicated decreasing asbestos exposure in Finland since the 1970s.

Sudden and unexpected death of a 6-month-old baby with silent heart failure due to anomalous origin of the left coronary artery from the pulmonary artery.

A 6-month-old girl died suddenly without any previous symptoms of heart failure. Autopsy examination showed cardiomegaly (97 g) with a severely fibrotized myocardium. The left coronary artery was originating from the pulmonary artery. Histologically, the myocardium showed myocardial infarcts of different ages, as well as grossly thickened arterial branches due to increased flow in left-right shunt. We suggest that rare anomalies of the coronary arteries should be considered in the autopsies of suspected sudden infant death syndrome cases.

Radiographic demonstration of esophageal and tracheal fistulas at autopsy using a contrasting medium that vulcanizes at room temperature.

Esophageal and tracheal fistulas, which occur in 0.05% of medicolegal autopsies, were demonstrated in three cases by a postmortem radiographic technique using silicone rubber/lead oxide as a contrasting medium that vulcanizes at room temperature. In one 83-year-old male, a tracheoesophageal fistula was detected, which had developed after surgical repair of an esophageal rupture caused by a flexible fiberoptic endoscope. In a second case, carcinoma of the esophagus in a 78-year-old male had eroded the trachea and arcus of the aorta creating a fatal tracheoesophagoaortic fistula. In a third case, 55-year-old female developed a tracheobrachicephalic artery fistula as a result of an infiltrating cystic adenocarcinoma of the trachea, resulting in a fatal hemorrhage into the trachea. The results of this study indicate that diagnostic radiologic methods using a vulcanized contrasting medium are useful in supplementing normal dissection in autopsy cases with suspected fistulas of the esophagus or trachea.

Peripheral zonal hepatic necrosis caused by accidental ingestion of methyl ethyl ketone peroxide.

Fatal massive peripheral zonal hepatic necrosis developed in a 47-year-old man who accidentally ingested a solution of methyl ethyl ketone peroxide (MEKP) in dimethyl phthalate. Such solutions contain about 10% active oxygen. The clinical course was characterized by temporary cardiac arrest, abdominal burns, severe metabolic acidosis, rapid hepatic failure, rhabdomyolysis and respiratory insufficiency. A fatal outcome resulted 4 d afterwards from hepatic coma associated with blood coagulation disorders. Microscopical examination revealed massive periportal hepatic necrosis accompanied by atypical pseudoductular proliferation. The proliferating cells were probably of bile duct origin and exhibited atypia and mitoses. The pathogenetic mechanism may involve lipid peroxidation caused by free oxygen radicals derived from MEKP.

Lectin binding of subunits of placental and serum oxytocinase after electrophoresis and transblotting to nitrocellulose.

1. Oxytocinase enzymes were purified from maternal serum and human placenta, run by SDS-PAGE and transferred onto nitrocellulose. Both enzymes were homogeneous in protein staining with Mr of 145,000. 2. Both serum and placental oxytocinases bound concanavalin A (Con A), limax flavus agglutinin (LFA) and wheat germ agglutinin (WGA). The WGA-binding of the placental enzyme was more strongly inhibited by 0.2 M N-acetylglucosamine than that of the serum enzyme which may indicate a higher sialic acid content in the serum enzyme. 3. Neuraminidase treatment did not affect the binding of Con A but decreased the binding of WGA to serum and placental enzymes. Serum enzyme showed a pl 4.7 on isoelectric focusing.