Identification of the skeletal remains of the Czech communist regime crime victim, priest Josef Toufar.

Parish priest Josef Toufar died as a direct consequence of torture committed by Communist State Security Service agents, forcing him to confess that "miraculous" movement of crucifix above the main altar during the Holy Mass held in the Roman-Catholic church in Cíhost was staged by using a technical equipment. Josef Toufar was presumably buried in a mass grave at the cemetery in Prague-Dáblice under a false name Josef Zouhar. In 2013 the Czech Bishops' Conference grant an approval to begin the process of his beatification. However, the beatification required the exhumation and identification of the remains. In this case report, we describe the process of searching, exhumation, and the combined A-STR/Y-STR DNA analysis of remains of Pater Josef Toufar. His identification was feasible due to kinship analysis: buccal swabs of three family members (niece, grand-niece, and grand-nephew) were available for testing.

[Determination of AB0 blood group system from degraded blood stains on serological and molecular genetic level]. / Urcování krevního systému AB0 z tepelne degradovaných krevních stop na sérologické a molekulárne genetické urovni.

The AB0 blood group system typing remains one of the basic laboratory tasks in a forensic practice. However, problems arise when the analysed samples are seriously degraded. We took blood samples from six volunteers (three men, three women) and made blood stains on pieces of sterile cotton cloth. Blood stains were incubated at three different temperatures (22 degrees C, 37 degrees C, 56 degrees C) for various periods of time (1 day, 1 week, 14 days, 1 month, 3 months, 6 months, 1 year). For blood stains degraded at 22 degrees C we also analysed the samples after 3.5 hours of incubation. Moreover, we tried to determine the AB0 blood group system after thermal degradation at high temperature, accurately at 200 degrees C for 10 min. For the AB0 blood group system typing a Polymerase Chain Reaction method was used to amplify glycosyltransferase gene, when DNA had been isolated from artificially created blood stains, followed by their subsequent artificial thermal degradation. For serological AB0 typing the mixed agglutination and the Therkelsen method were used. The DNA analysis seemed to solve problems with seriously degraded blood stains but we found out that classical serological methods were even better in some cases.

[B0 aberrant genotype of AB0 blood group system]. / Aberantní genotyp B0 v krevním systému AB0.

The AB0 blood group system typing remains one of the basic laboratory tasks in a forensic practice. However, problems arise when the analysed samples are seriously degraded. The DNA analysis promised to solve this but an unexpected complication was encountered. For the AB0 blood group system typing a Polymerase Chain Reaction method was used to amplify glycosyltransferase gene, when DNA had been isolated from artificially created blood stains, followed by their subsequent artificial thermal degradation. In the B0 genotype an aberrant genotype was suprisingly found and its structure was confirmed by sequencing. This meant that a newly formed B00 (not the original BO) genotype was present. Such a finding, to our best knowledge, had not been observed yet and we were unable to find any references in the professional literature. The explanation of this result thus remains unclear.

[Findings in the detection of fibronectin in skin burns]. / Nekteré poznatky získané pri prukazu fibronectinu u popálenin kuze.

Occurrence of fibronectin was detected in paraffin sections of burn skin samples, excised from surviving patients, as well as skin samples removed from the death persons, who succumbed burn wounds. The other groups of samples, used for comparison with the previous one, consisted of the intact skin excisions and skin samples post-mortem exposed to thermic changes. Indirect immunoperoxidase reaction was the immunohistochemical method used in all examinations. In post-mortem burn skin samples there was a loss of fibronectin activity in the epidermo-dermal junction area as well as in the papillary dermis. In burn skin samples, excised from patients who only survived for very short time their (serious skin) burns, no evidence of any fibronectin activity was seen. Also in two other cases, when patients--due to their severe burns--survived for several hours only, there were no conspicuous differences in the intensity of fibronectin activity seen in comparison with features found in the intact skin samples. Fibronectin activity was, however, increased in all other burn skin samples, where the survival time of patients was from 30 minutes to 5 weeks and there were following differences in its intensity and also in its pattern of distribution. In the 1st-degree and in a superficial 2nd-degree skin burn wounds, fibronectin was also present in the epidermis. In the papillary dermis, fibronectin was distributed rather diffusely or in a spot-like pattern while in the reticular dermis, there was a tendency to form net-like structures among collagen fibers. In deeper 2nd-degree and in the 3rd-degree burn wounds, fibronectin was deposited in vicinity of blood vessels and skin appendages in a fibrillar pattern. In 6 out of 11 samples, where the survival time ranged from 7 to 21 days, fibroblasts were arranged among fine collagen fibers and some of these cells exhibited positive fibronectin activity on their surface. Numerous fibroblasts with finely scattered fibronectin spots and also a decrease of fibronectin activity were observed in more mature granulation tissue, present in burn skin samples where survival time of patients was five weeks.

[Effect of Stopangin mouth spray on blood alcohol levels measured by the Alcotest 7410 analyser made by Drager]. / Ovlivnení hodnoty hladiny alkoholu merené analyzátorem Alcotest 7410 fy Dräger sprayem Stopangin.

During a check-up of a driver by a police patrol in the Czech Republic by means of a breath analyzer of Dräger Co. an alcohol blood level of 0.50 g/kg was assessed. Later the driver reported that before the test he used at 30- and 5-minute intervals the drug Stopangin spray. In the Institute of Forensic Medicine the authors made an experiment which revealed that 22 minutes after administration of the drug the apparatus gave a negative result. Agreement with these conclusions was expressed also by a representative of Dräger Co. and the results were published as an expert opinion with a recommended procedure for the police of the Czech Republic in the South Moravian region.

Monoclonal antibody against DNA adducts with osmium structural probes.

Osmium tetroxide complexes with nitrogen ligands (Os,L) have been widely used as probes of the DNA structure. A monoclonal antibody OsBP7H8 against DNA adducts with Os,L was produced in mice. OsBP7H8 does not bind to proteins or total yeast RNA modified with Os,2,2'-bipyridine (bipy) nor to the unmodified nucleic acids and proteins. The antibody recognizes DNA modified with Os,bipy (DNA-Os,bipy) or with OsO4,1,10-phenanthroline (DNA-Os,phen) but it does not cross-react with oxidized DNA and with DNA adducts of osmium tetroxide complexes with other ligands (such as pyridine, TEMED and bathophenanthroline disulfonic acid). The affinity of OsBP7H8 to DNA-Os,phen is about five-fold higher as compared to DNA-Os,bipy. The antibody can be thus applied either for recognition of single-stranded and distorted regions in DNA (after DNA modification with Os,bipy) or for detection of both single-stranded and double-stranded DNAs (after DNA modification with Os,phen). A new simplified procedure for the dot-blot analysis is proposed, not requiring the purification of DNA-osmium adduct prior to its application to the membrane.