[Taxus baccata poisoning]. / Smrtelná otrava tisem cerveným.

Common yew poisoning occurs by per oral application of needles or extracts of needles usually. The determination of the cause of death is more difficult in the latter case. In our article, we advise of circumstances which could be helpful in diagnostic. In addition, we describe the substances contained in yew, their effect, importance and toxicological detection.

[3,5-dimethoxyfenol--marker intoxication with Taxus baccata]. / 3,5-dimethoxyfenol-marker intoxikace tisem cerveným.

Autopsy findings of fatal intoxication with yew (Taxus baccata) are nonspecific. A presence of plant residues in the digestive tract can signalize yew intoxication. If yew decoction is consumed, plant residues are not found. In such a case the intoxication can be signalized by the presence of 3,5-dimethoxyphenol in biological material. Authors of this article describe the proof and quantification of the 3,5-dimethoxyphenol in two cases of fatal intoxication with yew. In both cases the liquid/liquid extraction and solid phase extraction was used. Extracts obtained from the acidic and basic environment were analysed. Extracts from the acidic environment were methylated and the extracts from the basic environment were acetylated. The analyses were carried out by gas chromatography/mass spectrometry. In the blood of both intoxicated persons the presence of 3,5-dimethoxyphenol was proved and its concentration 82 ng/ml and 417 ng/ml was quantificated. In both cases the presence of 3,5-dimethoxyphenol was also proved in the gastric contents and urine.

Correlation of ultrastructural changes of endothelial cells and astrocytes occurring during blood brain barrier damage after traumatic brain injury with biochemical markers of BBB leakage and inflammatory response.

Focal cerebral contusion can be dynamic and expansive. It has been proved that subsequent expansive contusion is caused by brain parenchyma damage, especially BBB damage. We investigated a group of patients with traumatic brain injury. The patients (n=18) were divided into group I (n=7) of patients submitted to neurosurgery due to expansive contusion, and group II (n=11) of patients without surgery. Serum concentrations of NSE and S-100B protein were measured by electrochemiluminescence immunoassay, interleukin-6 (IL-6) was measured by chemiluminescent sequential immunometric assay and matrix metalloproteinases (MMP-9, MMP-2) were measured by immunoassays. Cortical biopsy specimens of brain were investigated by electron microscopy in patients with trauma brain injury submitted to neurosurgery. Biochemical investigation from first day up to third day after traumatic brain injury proved increased values of IL-6 (302.2+/-119.9 vs. 59.6+/-11.9 ng/l, p<0.02) and S-100B protein (3.064+/-1.064 vs. 0.649+/-0.182 microg/l, p<0.05) in patients with expansive lesion compared to patients without expansive contusion. Significantly higher levels of MMP-9 (150.4+/-28.46 vs. 74.11+/-13.16 ng/l, p<0.05) and of MMP-2 (814.5+/-126.3 vs. 523.1+/-25.28 ng/l, p<0.05) were found during first 3 days after admission in group I compared to group II. MMP-9 has also elevated in group II from lower values after admission (74.11+/-13.16 ng/l) up to high levels on the 10th day of hospitalization (225.1+/-49.35 ng/l). Ultrastructural investigation of endothelial cells and surrounded tissue revealed perivascular hemorrhage, increased pinocytic activity of endothelial cells, and cytotoxic edema of astroglial cells. Multivesical bodies were disclosed inside the endothelial cells. Higher levels of serum protein S-100B and IL-6 correlated with ultrastructural changes of endothelial cells, and with inflammatory response following TBI, respectively.

[Evaluation of relevance in concussion and damage of health by monitoring of neuron specific enolase and S-100b protein]. / Hodnocení závaznosti otresu mozku a poruchy zdraví pomocí monitorování hladin neuron specifické enolázy a proteinu S-100b v séru.

INTRODUCTION: Proteins released to the circulation from affected glial (neuron specific enolasis, NSE) or ganglial cells (S-100b protein) during traumatic brain injury might be used in diagnosis of traumatic brain injury in cases with negative finding on computer tomography scan (concussion) or in patients where the serious clinical status does not corresponde with mild changes on CT scan (diffuse axonal injury, DAI). Classification of DAI according Gennarelli considered the concussion as lower degree of DAI. MATERIALS AND METHOD: 15 patients were divided into group I of mild conccussion (n=3) with 1-day duration of hospitalisation, group II of serious concussion (n=4) with more days duration of hospitalisation with negative findings on CT scan and group III of patients with diagnosis of DAI (n=8). Blood samples were investigated by immunoanalysis for NSE and protein S-100b (Elecsys 2010, Roche). RESULTS: Values of NSE (16.30 +/- 2.33 vs. 110.48 +/- 34.99 vs. 24.07 +/- 6.29 microg/l), and protein S-100b (0.207 +/- 0.03 vs. 0.945 +/- 0.69 vs. 0.736 +/- 0.36 microg/l) overdrow the reference value in cases of group I, II, and III. We discuss the biomechanics of trauma and the blood brain barrier damage in comparison with values of NSE and S-100b protein. CONCLUSION: [corrected] We proved the significantly higher values of the NSE in group of serious concussion compared to group of DAI. We demonstrated that concussions in some cases lead to serious damage of health.

[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.

[Trauma of cervical spinal cord--an immunohistochemical study]. / Poranení krcní míchy--imunohistochemická studie.

The histologica and immunohistochemical examination of cervical spinal cord were used in the group of 11 deceased of violent death. The positivity of neuron-specific enolase (NSE) in axons of cervical spinal cord was examined, and findings in the group of deceased on craniocervical injury (7 cases) and the group of deceased without cranio-cervical trauma (4 cases) were compared. While a routine histological examination including Palmgren's staining showed no significant differences between bouth groups, we noticed an increased frequency of appearance of NSE deceased with craniocervical injury.

Diagnosis of traumatical and hypoxial changes of the CNS--immunohistochemical study.

The immunohistochemical detection of neuron-specific enolase and b-amyloid percursor protein were used in the group of deceased on craniocerebral injury and those who died of prolonged hypoxy without mechanical injury of the brain. Neuron-specific enolase (NSE) is produced by nerve cells and is a suitable marker for both the damage of neurons and axons. While undamaged nerve cells show immunoreactivity with the antibody anti-NSE, a significant decrease of this protein substance was noticed within two hours both in mechanical injury and in cases of prolonged hypoxy. We noticed the presence of NSE in damaged axons already several minutes after the injury whereas the hypoxy of brain without mechanical injury didn't show any or a very slight reaction of axons when examined with anti-NSE without topographic link to axonal lesion. b-amyloid percursor protein (b-APP) is a low molecular protein which the normal values of are not to be found in axons detected by standard immunohistochemistry. We noticed an increased frequency of appearance of this protein substance in axons changed by injury, while a reactive positivity to anti-body b-APP was to be found only rarely at the brain hypoxy without mechanical injury CNS.

Early and late fatal complications of inhalation injury.

Severe inhalation injury causes a substantial deterioration in the prognosis and increases the general mortality of patients with extensive burns. Recently, in particular due to the development of invasive monitoring of patients and effective treatment of acute burn shock, we encounter with increasing frequency patients who survive the acute stage, including complications such as ARDS, and reach the stage of late complications. The latter include tracheooesophageal fistulas that develop on the basis of pressure ulcers and chondromalacia, usually at the site of the balloon of the tracheostomic cannula, and the overproduction of fibrous tissue in the area of the airways which leads to the development of stenosis, pulmonary fibrosis and bronchiectasia. Frequently, different early and late complications combine.

[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.

[Differences in immunohistochemical findings in traumatic and hypoxic changes in the central nervous system]. / Rozdílnosti v imunohistochemických nálezech pri úrazových a hypoxických zmenách CNS.

The authors made in a group of deceased subjects with craniocerebral injuries and subjects with protracted hypoxia without mechanical brain injury immunohistochemical investigations of neuron-specific enolase and beta-amyloid protein precursor. Neuron-specific enolase (NSE) is produced by nerve cells and is a suitable marker of neuron as well as axon damage. While the bodies of intact nerve cells display immunoreactivity with the anti-NSE antibody, in damaged neurons already within two hours after injury a marked drop of this protein substance was observed after mechanical injury as well as after protracted hypoxia. In axons altered by injury the authors observed the presence of NSE already within several tens of minutes after injury while hypoxia of the brain without mechanical injury did not produce any or only a very weak reaction of axons on examination with anti-NSE without a topographic link to the axonal lesion. Beta-amyloid protein precursor (beta-APP) is a low molecular protein the normal values of which are not detectable in axons by standard immunochemistry. In axons altered by injury the authors observed an increased incidence of this protein substance while in cerebral hypoxia without mechanical injury of the CNS only in rare instances a positive reaction with anti-beta-APP antibody was found.

[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.

[Effect of autolysis on histochemical examinations of the central nervous system]. / Vliv autolýzy na imunohistochemické vysetrení CNS.

The objective of the study was to explain the effect of autolysis on immunohistochemical detection of neurone-specific enolase (NSE), beta-amyloid protein precursor (beta-APP) and ubiquitine in cerebral tissue. The examination was made in 6 deceased subjects without mechanical injury of the CNS and 6 subjects with a craniocerebral injury who survived from 6 hours to 3 days. In all deceased subjects the post-mortem examination was made within 24 hours after death. For immunohistochemical examination tissue excisions were taken from standard sites of the brain. The first tissue excisions were immersed into 10% formol after a post-mortem interval of 24 hours. The remaining tissue slices were subjected to autolysis at room temperature and gradually immersed into formol after 24-hour intervals, the longest post-mortem interval being 168 hours, i.e. 7 days. For visualization of the linked primary antibody the biotin-streptavidin system labelled with alkaline phosphatase was selected. In the group of 6 subjects who died after a craniocerebral injury in 4 instances axonal lesions were detected, i.e. axonal oedema or formation of retraction spheroids. The damaged axons were positive on examination with all investigated antibodies, whereby it was possible even after a 168-hour post-mortem interval to differentiate damaged and not damaged axons. In the group of 6 subjects without mechanical injury of the CNS in 5 instances axonal oedema was found, however, it was not positive with anti-NSE antibodies nor with anti-beta-APP. After the 24-hour post-mortem interval in this group in 3 instances ubiquitine positivity was found in axons but already after a post-mortem interval exceeding 2 days the axons were ubiquitine positive in all 6 subjects. Lumpy deposits of this substance could be detected in axons also beyond axonal structures.

[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.

Lethal burn trauma in children.

The aims of this retrospective study covering the years 1984-1998 were: 1. to survey burn injuries in children at the present time and 2. to compare the current results with the conclusions of an analogous study performed in the years 1964-1983. A decline in the occurrence of lethal burn wounds was found, as well as in burn shock as a direct cause of death. Children 1-4 years old continue to be the most frequent victims of fatal accidents. The most common cause of burn injury in this group remains scalding in the household.

[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).

[The effect of autolysis on the interpretation of results of immunohistochemical detection of myoglobin and fibrinogen in the myocardium]. / Vliv autolýzy na interpretaci výsledku imunohistochemického prukazu myoglobinu a fibrinogenu v myokardu.

Immunohistochemical detection of myoglobin and fibrinogen in the myocardium makes it possible to diagnose very early stages of ischaemic/hypoxic changes of the heart muscle. The authors demonstrate on the myocardium of a 58-years-old female patients who died suddenly with the finding of acute infarction of the anterior wall of the left ventricle with transmural rupture and cardiac tamponade the effect of autolysis on this examination. Tissue excision taken from the close vicinity of the rupture were subjected to autolysis at room temperature and immersed in formol for fixation within an interval of 24 hours. The control series of examinations was made in a similar way on the heart muscle of a 20-years-old woman who died from violent death, and ischaemic changes of the heart muscle were not anticipated. While in the early stages of autolysis it was possible to use combined immunohistochemical detection of myoglobin and fibrinogen, after a longer postmortal interval extensive artificial losses of myoglobin were observed, and the method of detection of fibrinogen deposits in damaged cardiomyocytes seemed more suitable. Even after a postmortal interval of 168 hours it was possible to differentiate reliably damaged myocytes with fibrinogen deposits from intact muscle fibres, where fibrinogen deposits were not observed.

[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 central nervous system in sudden infant death]. / Imunohistochemické nálezy na CNS u náhlé smrti kojencu.

In a group of 7 infants suddenly dying at the age up to 1 year a histological and immunohistochemical investigation was performed aimed to prove possible chronic hypoxic and latent traumatic changes. Presence of ubiquitin and neuron-specific enolase (NSE) was detected in axon and neurons. In six suddenly dying infants an immunohistochemical positivity was found with antibodies against ubiquitin in brain stem neurons. Antibodies against NSE showed positivity of the white matter was lacking in the detection both of ubiquitin and NSE. Among 12 persons dying of craniocerebral injury an intensive NSE immunopositivity was found in traumatized axons of 7 cases and ubiquitin positivity in 5 cases while brain stem neurons did not show any ubiquitin positivity. Bain stem neurons at sudden death were strongly NSE positive in 5 case, weakly positive in 3 cases and negative in 4 deceased. The combined proof of ubiquitin and NSE in neurons and axons at infant sudden death appears to he advantageous because it combines the detection of traumatic changes and chronic hypoxic lesions as well.