[Microglial activation and GFAP astroglial immunopositivity in drug abusers]. / Aktivace mikroglie a GFAP imunopozitivita astroglie u toxikomanu

INTRODUCTION: The neurotoxicity brought about by application of toxic and psychotropic substances is accompanied by an activation of astroglial and microglial cells in the brain. MATERIALS AND METHOD: We investigated clinically 42 patients addicted to psychotropic substances (hospitalised in the Motol Teaching Hospital). The NSE, S1OOB, and manganese concentrations in the blood were measured in the patients. In 14 deceased patients with drug evidence in the hair indicating a chronic abuse of addictive substances, the brain tissue glial cells were immunohistochemically labelled with antibody against CD68 and GFAP. RESULTS: In 8 hospitalised patients, there were increased NSE, S100B values in the blood (p < 0.05). Manganese in the blood was increased (3,03±1,9 µg/l, p < 0,05) in all patients. In deceased persons, the CD68 positivity of microglial cells and neurophagy have been proved. GFAP positive astroglial cells have been evidenced in the gray and white matter. CONCLUSION: The CD68 and GFAP positive glial cells in brain tissue can be a pathomorphological correlate of neurotoxicity in chronic abusers. The neurotoxicity can be monitored with NSE and S100B markers of damaged neuronal cells.

Immunohistochemistry and serum values of S-100B, glial fibrillary acidic protein, and hyperphosphorylated neurofilaments in brain injuries.

INTRODUCTION: Traumatic brain injury (TBI) triggers a series of reactions resulting in cytoskeletal-related changes varying between focal and diffuse injuries. METHODS: The patients (n=38) were divided into group of diffuse axonal injuries (DAI, n=10) and focal (n=28) injuries. Serum hyperphosphorylated neurofilaments (NF-H) and glial fibrillary acidic protein (GFAP) were measured by Biovendor immunoassay, and serum S-100B protein was measured by Cobas e411 (Roche) by immunoassay. Immunohistochemistry was performed with monoclonal antibodies (Chemicon, USA). RESULTS: The median serum S-100B concentration was higher in patients with focal mass lesions (1.72±0.4 µg/l vs. 0.37±0.1 µg/l, p<0,05) compared to patients with DAI during 10 days of hospitalisation. With respect to all patients, the highest peak of serum S-100B values (4.21±1.1 µg/l) and GFAP (8.58±2.4 µg/l) were found in expansive lesions. The median serum NF-H was higher in DAI compared to focal TBI (0.625±0.14 vs 0.139±0.02 ng/l, p<0.05) during all 10 days after admission. Further, immunohistochemical investigation, in deceased patients with DAI , using NF-H antibody proved positive varicose and waving axons, and retraction balls. Time-dependent profile of serum NF-H demonstrated the increase of values within 4th up to 10th day in both groups. Values ranged from 0.263 up to 1.325 ng/l in DAI, and from 0.103 up to 1.108 ng/l in focal injuries. Patients with expansive contusions had similar levels of serum NF-H as patients without expansive lesions. Immunohistochemistry of cytoskeletal proteins presented strong positive staining of vinculin, vimentin in vessels, GFAP, and S-100B in DAI compared to weak staining in expansive lesions. CONCLUSION: The time-profile kinetics of all markers may reflect different types of pathophysiological changes of the BBB or axonal damage in focal and diffuse injuries. KEYWORDS: brain contusions - diffuse axonal injury - S-100B protein - GFAP - hyperphosphorylated neurofilaments.

[Protein S100b in differential diagnosis of brain concussion and superficial scalp injury in inebriated patients]. / Protein S-100b v diferenciální diagnostice komoce mozkové a povrchního poranení hlavy u pacientu v ebriete

INTRODUCTION: Concussion cannot be differentiated from superficial scalp injury, especially in inebriated or uncooperative patients. This can have serious medical or forensic consequences. The aim of the study was to determine whether serum concentrations of S-100b in mild traumatic brain injury (MTBI) patients are significantly higher than those in patients with superficial scalp injury with scalp wound and alcohol intoxication. MATERIAL AND METHODS: A total of 50 patients with head injury, 25 with mild concussion without scalp wound and alcohol intoxication, 25 superficial scalp injury patients with scalp wound and clinical signs of inebriety. Neurological status and cranial CT scan were evaluated 60-120 minutes after injury in all the 50 patients to exclude focal cerebral injury or skull fracture. RESULTS: The serum levels of S-100b were significantly increased in patients with concussion (median 0.36 ± 0.15 µg/l ) in comparison with the group of patients with scalp injury and alcohol intoxication (median 0.09 ± 0.002 µg/l). All 50 patients had a normal cranial CT finding and neurological status. In all superficial scalp injury patients alcohol intoxication was confirmed (0.96 - 3.11s). CONCLUSION: We proved significantly higher values of S-100b in patients with brain concussion. Diagnostically decisive value of S-100b concentration in the serum was set at 146 µg/l and higher (94% sensitivity and 100% specificity). Alcohol intoxication (up to 3.11 s) and scalp wound seem to have had no crucial impact on serum S-100b level.

[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).

[Another mechanism of décollement]. / Jiný mechanismus décollement.

The cause of décollement is usually considered to be tangential brute blunt force impacting the body surface especially in case of hitting or running-over injury of the pedestrian's body by a car. The term rolling effect or rolling mechanism is used as well. The dissociation of tissue layers with other epiphenomenon occurs. The presented group of 152 décollement determined in 103 autopsy cases during the 4 years period comprises the observation of décollement of different etiology of the injuries (traffic accidents, falls from the high, compression of the torso); in the traffic accidents the occurrence in various participants of the traffic, not only in the case of the collision of the pedestrians with various traffic vehicles, but also in drivers of various traffic vehicles, and fellow-travelers as well. The topic, the localization, the content, the extent, and vital reaction and combination injuries were followed-up. According to the variability of the injury etiology, not restricted to the traditional conception of the décollement mechanism, it is obvious that the passed on rule by far is not covering the whole content of this concept. We didn't find any alternative interpretation of until now presented mechanism in the literature. The medical literature focuses mostly on the clinical aspect of this injury. In this paper, the biomechanics of the décollement origin also in case of the tissue compression by the pressure applied perpendicularly to the body surface, the dependences on physical properties of the actively or passively affecting object, the relevance of the ratio of the tissue structures compression in one direction and transversal dilation in other two directions according to the Poisson's constant, the question of tangential factor of the force in case of vertical falling on the horizontal plane, and biomechanical relations in case of body landing on an oblique surface are discussed. The mechanism of décollement is more complex as presented until now. The forensic interpretation of findings should reflex the above-mentioned facts.

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.

[Using of scanning electron microscopy for detection of gunshot residue]. / Pouzití skenovací elektronové mikroskopie pri detekci povýstrelových cástic.

Scanning electron microscope improves the possibility of investigation of surroundings near of gunshot wounds in forensic medicine, it is the next subsequent method for differentiating of area of entrance and exit wound, supplemental method for determination of firing distance, permit of detection (GSR) on the hand of shooter and ensured describing of samples and their stored. Detection of GSR provides many information about composition of bullet and primer. Authors are demonstrating the possibility of detection of GSR on experimental shooting to the krupon (pigs' skin) in different situation (such as in a room and in outside area) and using of different weapon (hand gun CZ No.75 and machine gun No.58).

[Biochemical and immunohistochemical markers of brain injury]. / Biochemické a imunohistochemické markery poskození mozku.

Proteins released to circulation from affected tissues during primary or secondary trauma brain injury might be used as serum markers of glial or ganglial cells damage (neuron specific enolasis and S100 B protein). Other markers of trauma can be proved as relatively specific of diffuse axonal injury by immunohistochemical detectoin (amyloid prekurzor protein, neuron specific enolasis, glial fibrilar acidic protein and superficial antigen receptor CD 68). Some markers are associated with blood brain barrier damage (matrix metaloproteinases (MMP-2, MMP-9) and synthase of nitric oxide (iNOS)). We aimed in our short communication on biomechanics of developed of trauma, primary or secondary kinds of trauma brain injury and use of trauma brain injury markers for clinical diagnostics and management of patients.

Inhalation trauma in burns, cytology and biochemical findings.

We have followed in a pilot study a group of patients for cytological and biochemical changes of lavage in the upper and lower respiratory system. Into the study patients with respiratory burns confirmed by bronchoscopy and skiagraphy were included. We divided patients according to the Lung Injury Scores (LIS). We obtained the values in our sample group between intubation and last swab (before extubation). Listed are the risk factors, probability of survival, and lung histology results are listed for patients who died.

[Findings in myocardial extracellular matrix in fatal poisoning with addictive substances]. / Nálezy v extracelulární matrix myokardu u zemrelých otrávených návykovou látkou.

It have been reported that chronic drug abuse induces changes in extracellular matrix (ECM) composition of myocardium. Matrix metaloproteinases (MMP) play important role in ECM remodelling and alteration in the ECM of myocardium. They are able to cleave ECM components and may contribute to progression of heart damage. Remodelling implies an alteration in the ECM and in the spatial orientation of cells and intercellular components. In this report we evaluated the extracellular matrix of myocardium of deceased drug abusers and controls. The myocardium of 5 toxicologically positive cases and 4 control cases were analysed by zymography of MMP, electrophoresis of collagens and histologicaly. Gelatinases (MMP-2 and MMP-9) and collagenase (MMP-1) activities were increased in drug abusers compare to the controls. In contrast, alpha chain of collagen type I after 160 hours collagen extraction, were not observed in control specimens. The activation of MMP proenzymes is one of the critical steps that lead to ECM breakdown. Our findings suggest that the myocardial collagen matrix is damaged in drug abusers.

[Objective evaluation of thoracic injuries and associated injuries using the Abbreviated Injury Scale and the New Injury Severity Score]. / Objektivizace poranení hrudníku a s nimi sdruzených poranení pomocí Abbreviated Injury Scale a New Injury Severity Score.

The authors focused attention on objectivization of the severity of chest injuries and associated injuries using the Abbreviated Injury Scale (AIS) and the New Injury Severity Score (NISS). They evaluated injuries detected on post-mortem examination in subjects who died from the sequelae of injuries in the Faculty Hospital Prague 10. The group comprised a total of 90 subjects regardless of sex aged 17-94 years who died in the hospital in 1996-2000. Traffic injuries accounted for 70% deaths, other blunt injuries for 27.8% and only 2.2% were penetrating injuries of the chest. The mean AIS value of the chest was 3.7, the mean value of NISS was 50.7. The NISS value declined in relation to age of the deceased patients and the period of survival. The period of survival varied from 30 mins. to 136 days. The mean NISS value on survival up to 5 hours after injury was 55.0. On comparison of our group we found that the results were consistent with those of American studies.

[Findings in bronchial lavage in patients with burns of the upper respiratory tract]. / Nálezy v bronchiálních lavázích u pacientu s popálením horních dýchacích cest.

The study is focused on cytologic and biochemical changes in bronchoalveolar lavage (BAL) at the group of patients from the Clinic of Burns Medicine at the University Hospital Královské Vinohrady in Prague. Only patients with inhalation trauma were involved in the study. The obtained data are from BAL measurements collected between intubation and extubation. There was the greater number of polymorphonuclears in the first two weeks in all patients. After 2-3 weeks there was a gradual increase in the number of macrophages at four patients and three patients had still the greater number of polymorphonuclears. There was a positive findings of PCNA at one patient and other markers (bcl-2, p53) were negative. The cytoflowmetry proved the increase in number of CD3 cells by 20% in one patient. The amount of proteins in the last BAL just before extubation was lower than 22 g/l. Serin proteinase activity was lower than 50 ncat/l at five patients and the ratio of MMP-8/TIMP-2 was higher than one in all analysed samples. There was no development of lung inflammation into chronic stadium. Two patients died.