[The origin, distribution and relocability of supravital hemorrhages]. / Vznik, distribuce a mobilita supravitálních hematomu.

The hemorrhage as a result of intravital bleeding is considered, from forensic point of view, as important sign of vital reaction of injury. However, in special cases it must be accepted that hemorrhage occurred after the death. The formation of supravital changes is evident, e.g., in organ donors whose blood circulation and pulmonary ventilation is kept after the brain death. The post-mortem origin of hemorrhages can also be seen in donors of eyeballs after enucleation made before the autopsy at Institute of Forensic Medicine and Toxicology. These hemorrhages are manifested after several hours when eyeballs were removed. Moreover, we observed the origin of hemorrhage in orbit which was caused nor by intravital bleeding neither by direct force. Its origin could not be explained nothing but postmortem propagation of the hemorrhage from the fracture of anterior fossa of the scull base. We did not find information about postmortem origin or relocation of hemorrhage of such extent in the literature. In the frame of knowledge about supravital reaction, this finding is of general importance with forensic impact. The documentation from the scene of death allowed correcting the appreciation of the mechanism of injury and traumatic process from the point of view of foreign culpability which should be considered in such case (e.g., a blow to the orbit with following fall under passing subway train).

[Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) as a cause of sudden death]. / Arytmogenní dysplazie/kardiomyopatie pravé srdecní komory jako prícina náhlé smrti.

Common cause of sudden death in children and teenagers are primary cardiomyopathies, mostly hypertrophic. The authors refer to arrhythmogenic right ventricular dysplasia/cardiomyopathy as a possible cause of sudden death. The disease is not limited to the right hearth only, the presence of fat and scar tissue in the in the hearth muscle is variable.

[Correlation among clinical and morphological findings in a diffuse axonal injury]. / Korelace klinických a morfologických nálezu u difuzního axonálního poranení.

In a man of 56 years--a victim of traffic accident, contusions in both temporal lobes of the brain and a small subdural and subarachnoid haemorrhage were diagnosed by CT and MR examination. According to the findings of petechial haemorrhages in the corpus callosum a diffuse axonal injury was clinically diagnosed. After 2 months the patient died. Excisions of corpus callosum and the brain-stem were examined by histological staining including Palmgren@s method and by immunohistochemical detection of neuron-specific enolase (NSE) in damaged axons. Clinical findings were compared with morphological findings, i.e. autopsy, histological and immunohistochemical examination. The possibilities of clinical diagnostics of diffuse axonal injury are discussed.

[Detection of myocardial and lung injury by psychotropic and narcotic agents by immunohistochemical detection of metalloproteinases]. / Prukaz poskození myokardu a plic psychotropními a omamnými látkami imunohistochemickou detekcí metaloproteináz.

Positive findings of intoxicant (narcotic) and psychotropic drugs (OPL) have been regularly recorded in clinical patients and deceased persons over the last years at the Institute of Forensic Medicine and Toxicology, 1st Medical faculty and General Teaching Hospital, Prague; stimulants and opioids represent the most frequent cause of death. Their misuse results in damage to various organs. In order to follow the development of pathological changes in the process of remodeling extracellular matrix directly in tissues, the methods of immunohistochemical detection of the matrix metalloproteinases in myocardium and lungs as well as fibrinogen in cardiomyocytes were selected for analysis in a group of 18 deceased individuals. In the intoxication with stimulants we usually demonstrated MMP 2 in the myocardium interstitium, MMP 9 being observed in two cases and MMP 1 in one case. The analysis of lungs always demonstrated MMP 1, especially in the lung interstitium and also on the surface of some alveoli, which accepted the appearance reaching up to "hyaline membranes" as well as in cellular elements of macrophage type and and the same was true for MMP2. Fibrinogen was not always demonstrated in cardiomyocytes. The detection of metalloproteinases was less prominent in the case of opioids. The demonstration of MMP explains well the evolution to more advanced pathomorphological changes, which have been found in myocardium and lungs of OPL users and fits to the nosological status of earlier phases of intoxications with these drugs.

[Immunohistochemical findings in prolonged cerebral hypoxia]. / Imunohistochemické nálezy pri protrahované mozkové hypoxii.

In the development of secondary changes of brain damage caused by injury, tumour and hypoxia of different origin with unconsciousness also the activity of the metalloproteinase matrix (MMP) may participate. Their activation occurs first in the vascular wall which they damage and it may be assumed that MMP potentiate thus secondary changes in the sense of interfering with the haematoencephalic barrier with development of vasogenic cerebral oedema. In the cells of the vascular wall and in glial elements already after two days MMP-2 and MMP-9 activity was proved. While in focal injuries the findings were in particular at the site of contusion foci, in prolonged hypoxia for various reasons these changes were diffuse.

[Time-related expression of adhesive proteins and other markers of age of injuries]. / K casovým relacím exprese adhezivních bílkovin a dalsích markeru stárí poranení.

UNLABELLED: In addition to evidence of the vital reaction it is important from the forensic aspect to assess the age of wounds, in particular in wounds with a short time of survival. To both can contribute detection of adhesive molecules, identified in recent years, possibly in combination with other markers of early stages of reparative inflammation. The submitted paper comprises the results of investigation of 465 skin wounds. The investigated samples were obtained from necroptic material, excision from wounds of treated patients and from experimental injuries in mice. Assessment of the age of injuries by means of endothelial adhesive molecules was made in paraffin sections after transfer into Varioclave and using the ABC techniqueref.. In human material a strong positive reaction of ICAM-1 was observed first after 1 and a half hours and latest after 3 and a half days, in P-selectin first after 3 mins., latest after 7 hours, in E-selectin first after 1 hour and latest after 17 days, in VCAM-1 first after 3 hours and latest 3 and a half days after the development of the injury. Expression of L-selectin was not typical for the injury. In skin injuries of mice positive immunohistochemical reactions were found as a rule sooner than in skin injuries of humans. Fibronectin was detected in paraffin sections from 70 skin wounds of dissected subjects immunohistochemically by the indirect immunoperoxidase reaction and by the use of the ABC technique labelled with alkaline phosphatase. Positive evidence was observed first after 5 minutes and latest after 8 hours. CONCLUSION: Detection of a rise of expression of ICAM-1, VCAM-1 and P- and E. selectin and the formation of basic netlike fibronectin structures improves the assessment of the age of wounds with a short survival period.

[Immunohistochemical detection of lung damage]. / Imunohistochemická detekce poskození plic.

Important risk factors associated with sudden infant deaths include in addition to the prone position also other exogenous factors which influence the respiratory function of the infant, in particular during the first half of his life. The negative action of maternal smoking during pregnancy damages foetal development and the child can suffer damage also from cigarette smoke in the environment. The investigation indicates possible toxic effects of activated matrixins by nitrogen oxides in pulmonary tissue caused by their presence in the environment.

[Polarographic determination of metals in firearm injuries]. / Polarografické stanovení kovu u strelných poranení.

There was tested the use of the polarographic determination of some metals (Pb, Cu, Fe, Sb) in the skin in deceased due to multiple firearm injuries. The authors present on a real case, in which the shots through the dress of victim penetrated, the possibility to determine entry and exit wound.

[Morphologic findings in chronic abuse of heroin and pervitine]. / Morfologické nálezy u chronického abúzu heroinu a pervitinu.

In a deceased 39-year-old man with a history of 10-year consumption of heroin and pervitin the authors made histological and immunohistochemical examinations of organs focused on detection of old and recent pathological changes. In the brain they detected oedema, venostasis, inflammatory infiltrates in the wall and surroundings of some vessels and hypoxic changes of neurons with a drop or disappearance of neuron-specific enolase. The myocardium was marked by oedema of the interstitium, focal diminution to disappearance of basophilia of myocyte nuclei with increased eosinophilia of some fibres and smaller and larger foci of fibrosis formed by mature and less mature connective tissue. Immunohistochemical examination revealed focal fibrinogen deposits in myocytes. A surprise was to a certain extent the finding of dispersely distributed caspase-8 in myocytes, caspase being one of the substances signalizing apoptosis. On microscopic examination of the lungs severe haemorrhagic oedema dominated. In the liver they found acute venostasis and chronic inflammatory changes with connective tissue proliferation in the portal areas. The finding in the kidneys suggested acute tubular nephrosis. The authors discuss the problem of direct toxicity of the ingested drugs, the influence of repeated states of hypoxia and infection.

[Terminology used in evaluating the severity of injuries for agencies involved with the penal system]. / Vymezení pojmu pouzívaných pri posuzování závaznosti poranení pro orgány cinné v trestním rízení.

The authors deal with medical and juristis terms concerning the evaluation of the grievousness of injuries for the police of the Czech Republic.

[Detection of metalloproteinases in pulmonary tissue after sudden and violent death]. / Prukaz metaloproteináz v plicní tkáni u náhlé a násilné smrti.

During early stages of chronic hypoxia under experimental conditions in the lungs alveolar macrophages are activated and in the latter an increased amount of matrix-metalloproteinase-collagenolytic enzymes is produced. Their presence was assessed in the present study in the lungs of infants who died suddenly (SIDS) and also in subjects who died from a violent death incl. acute and protracted external suffocation. The positive findings of these matrixins, in particular MMP-9 (gelatinase) were assessed in alveolar macrophages not only after protracted suffocation but also in the majority of sudden infant deaths. These findings supplement the mosaic of changes which indicate that sudden infant deaths are preceded by a period when the infant was exposed to hypoxia.

[Detection of caspase as a sign of tissue damage]. / Detekce kaspázy jako príznaku poskozené tkáne.

Caspases are cysteine proteases which participate in different stages of apoptosis. Apoptosis, cell death, programmed by its nucleus is associated also with a number of diseases and tissue damage. To this process increasing attention is paid also in the sphere of forensic medicine, in particular to make use of the diagnostic contribution in investigation of cause of death, vital reaction and time when the injury developed. In damaged tissues by immunohistochemical methods caspase activity was assessed. Caspase 8 activity (Flice) was detected in the heart muscle in congenital heart disease in a two-week old infant but also in ischaemia in a 58-year old woman with thrombosis of the coronary artery or in acute circulatory failure after an overdosage of pervitin in a 39-year-old drug addict. In the liver there was a positive finding in hepatocytes in chronic inflammatory changes caused by chronic alcohol abuse. An early caspase 8 reaction after an injury is suggested by positive findings in skeletal muscles of the neck and larynx in a 47-year-old man who was strangled. A positive reaction was proved only at the site of the line caused by the strangulation tool. However macroscopically nor by common histological staining the muscle tissue did not display any signs of injury at the mentioned sites. The results suggest that immunohistochemical evidence of caspase 8 can be used as a suitable complementary examination not only for detection of damaged tissue but also for evaluation of early stages of the vital reaction.

[Glycophorin in forensic medicine]. / Vyuzití glycophorinu v soudním lékarství.

Immunohistological evidence of glycophorin from paraffin sections of tissues fixed with formol, makes its possible to visualize membranes of preserved erythrocytes and fragments after their disintegration. This fact permits even after development of post-mortem changes to differentiate actual extravasation, manifested by a positive finding of glycophorin from mere infiltration of tissue by blood pigment after death, where glycophorin is lacking. The method is very sensitive and can reveal even very discrete haemorrhage. From different grades of erythrocyte disintegration in haematomas in the same subject sometimes conclusions can be drawn also on the different time of development of traumatic tissue changes.

[Immunohistochemical methods in the differential diagnosis of primary traumatic and subsequent secondary cerebral changes]. / Imunohistochemické metody v diferenciální diagnostice primárních úrazových a následných sekundárních zmen mozku.

In a 22-year-old man, driver of a personal motor vehicle, who died within 39 hours after a traffic injury, the authors made histological and immunohistochemical examinations of the brain focused on differentiation of primary traumatic and subsequent secondary changes. In haematomas the authors revealed the presence bi- and trivalent iron by Turnbull's and Perl's reaction as well as glycophorin by immunohistochemical reactions. White matter lesions were evaluated histologically by staining according to Palmgren and immunohistochemically by detection of neuron-specific enolase, beta-amyloid protein precursor and low molecular neurofilaments. Minor contusion foci in the corpus callosum and in the peripheral portion of the pons revealed the presence of extracellular bivalent iron and exceptionally also the presence of intracellular iron. Glycophorin was present not only in erythrocyte membranes but also in the form of lumps signalizing haemolysis. In the haematoma in the median portion of the pons neither iron nor free glycophorin were detected. At all investigated sites (subcortical areas of the white matter of the hemispheres, capsula interna, corpus callosum, pons Varolii) the authors detected numerous axonal deformities (oedema or formation of retraction spheroids) which revealed on immunohistochemical examination an intense reaction with antibodies in particular against neuron-specific enolase and beta-amyloid protein precursor, and to a smaller extent against low-molecular neurofilaments. The combination of the mentioned immunohistochemical examinations seemed a suitable method for differentiating primary cerebral injury (diffuse axonal injury and minor contusion foci in the corpus callosum and the margin of the pons) from secondary changes (haemorrhages in the median portion of the pons) which developed shortly before death as a manifestation of haemodynamic disorders associated with cerebral oedema).

[Determination of the ABH blood group system in the placenta]. / Príspevek k urcování krevnich skupinových vlastností systému A B H v placente

Blood group substances A, B, H were detect in placental tissue paraffin cuts (after 10% formaline fixation) by immunohistochemistry using indirect immunoperoxidase-two-layers technology or biotin-streptavidin complex signed by alkaline phosphates. Both fetal and maternal part of placenta are to be investigated which enables their group markers to be identified. Immunohistochemical detection of the A, B, H blood group substances in placenta can give conclusion about the blood group of fetus and of fetus and of mother as well.

[Detection of eosinophilic cationic protein in the lungs in sudden infant death]. / Prukaz eozinofilního kationického proteinu v plicích u náhlé smrti kojencu.

In a comparative study the authors detected presence of eosinophilic protein in infant's lung at sudden death. Degranulation of eosinophillic leucocytes and extracellular foci of this protein were found in a case of such sudden death. Discussion concerned possibility that the release of eosinophilic cationic protein could represent one of the triggering moments of infant sudden death.

[Immunohistochemical findings in the heart conduction system]. / Imunohistochemické nálezy na prevodním systému srdce.

Immunohistochemical investigation of heart conductive system showed that degenerative changes described by James (9) in some cells of the system had a nature of programmed death. Extinction of certain of number of cells of a reducted part of the system was found in membranous septum. Apoptotic antigen (21) could be proved in some destructed cells by using anti-Bax and anti-bcl-2 antibodies.

[Detection of free histamine and serotonin in the evaluation of the vital reaction in injuries]. / Prukaz volného histaminu a sérotoninu pri posuzování vitální reakce poranení.

Skin injury alive produces quick and distinct increase of free histamine and serotonin. A fluorimetric method for detection of these compounds in tissues has been the only one described in the literature up to now but it needed a time consuming extraction and big amount of primary material (about 3 g of skin devoid of subcutaneous fat tissue) as well. The authors succeeded in substituting the method and developed a voltametric detection of histamine and serotonin using the computerized ECO-TRIBO-Polarograph. It enabled to omit the extraction of skin compounds and to decreased the amount of primary tissue to a quarter. The detection of free histamine and serotonin levels is presumed to enable a more accurate determination of the time of injury, particulary in short interval before death; it presents, with a simultaneous of glycophorin, a convenient complementation of immunohistochemical investigation of fibronectin.

[Histochemistry and immunohistochemistry of the lung in sudden infant death]. / Histochemie a imunohistochemie plic u náhlé smrti kojencu.

Histochemical and immunohistochemical findings in respiratory tract of suddenly deceased infants were a bit different from those with violent death immediately after trauma. The bronchial wall inflammatory infiltration and foci of peribronchial lymphatic tissue did not show any substantial difference in both groups but findings in lung interstitium proper did. A regular finding was the presence of activated macrophages in interstitium and alveolar lumina and an increase of B and T lymphocytes in lung tissue as well. Lung findings in suddenly deceased infants do not have character of a shock lesion bud represents an initial phase of acute inflammation.