[Evidence of DNA from epithelial cells of the hand of the suspect found on the tool of the crime]. / Hinweis auf Täter-DNA durch die Untersuchung von Hautschuppen an Tatwerkzeugen.

A young woman was found unconscious in her flat strangulated with a children's scarf and was at first thought to be dead. The victim survived but is disabled for life. Neither bloodstaines nor sperm were found at the scene. Swebs of fleece were prepared from two fingers of a one-way glove which were found at the scene and from each end of the scarf in order to find some epithelial cells from the criminal. DNA was extracted and amplified by the polymerase-chain-reaction (PCR). The DNA-profile of the victim as well as that of the suspected apart living husband were found in these mixed stains. The suspect was severely incriminated by these findings and other evidence. Consequently the court convicted him to eleven years of inprisonment because of attempted manslaughter.

Frequency data for the STR loci HumFibra (FGA) and HumACTBP2 (SE33) in a population of Germans and Turks from South-West Germany.

Population studies were carried out on German and Turkish individuals from South-West Germany using the short tandem repeat (STR) systems HumFibra (n = 138 Turkish and 1161 German individuals) and HumACTBP2 (n = 202 Turkish and 1338 German individuals). After electrophoresis 19 alleles could be identified for HumFibra and 55 for HumACTBP2. No significant deviations from Hardy-Weinberg equilibrium were observed.

[Measuring precision in determining the length of restriction fragments within the scope of DNA analysis]. / Untersuchungen zur Messpräzision bei der Grössenbestimmung von Restriktions-Fragmentlängen im Rahmen der DNA-Analyse.

Because of the measurement error that is known from the length determination of restriction fragments during DNA analysis, it is necessary to determine the precision by which identical DNA can be evaluated. Besides the collection of frequency data from a certain population this is one of the basic prerequisites to calculate the likelihood ratio of an established polymorphism. By multiple measurement of restriction fragments from different persons indicated by the probes MS1, MS31, MS43A, g3 and YNH24 on different blots the within-laboratory variation was determined in the complete separation area from 1 to 19 kb. The values were compared to procedures used by other laboratories.

Safety of virus inactivated antithrombin III concentrate antithrombin III immuno (AT III).

In a prospective clinical trial the risk of infection after application of virus inactivated antithrombin III concentrate ANTITHROMBIN III IMMUNO (AT III) was investigated in patients undergoing cardiovascular surgery. The study was conducted according to the recommendations of the International Committee on Thrombosis and Hemostasis (ICTH), with the exception that most patients required additional blood products as well as AT III. Twenty-seven patients were eligible to test for the risk of acquiring hepatitis B. Twenty-six patients could be evaluated in terms of hepatitis NANB transmission considering ALT-levels whereas 20 patients could be tested for anti-HCV one year after surgery. Samples from 78 patients could be monitored for anti-HIV-1. None of these patients showed any signs of infection. AT III IMMUNO seems to be an antithrombin III concentrate with low or absent infectivity.

Safety of vapour heated prothrombin complex concentrate (PCC) Prothromplex S-TIM 4.

The risk of infection after application of vapour heated prothrombin complex concentrate PROTHROMPLEX S-TIM 4 (PCC) was investigated in patients undergoing cardiovascular surgery. The study was conducted according to the recommendations of the International Committee on Thrombosis and Hemostasis (ICTH) with the exception that most patients required other blood products in addition to PCC. Twenty-One patients were eligible to test for the risk of acquiring hepatitis NANB (ALT-levels) and samples from 12 patients were available that could be screened for anti-HCV. Twenty patients qualified for evaluation of the risk of developing hepatitis B, and 67 patients qualified to test for HIV-1-Infection. None of these patients showed any signs of infection. Vapour heating of prothrombin complex concentrate seems to lower the risk of transmitting viral diseases considerably.

Lung vascular injury after administration of viable hemolysin-forming Escherichia coli in isolated rabbit lungs.

Escherichia coli hemolysin, a transmembrane pore-forming exotoxin, is considered an important virulence factor. In the present study, the possible significance of hemolysin production was investigated in a model of septic lung failure through infusion of viable bacteria in isolated rabbit lungs; 10(4) to 10(7) E. coli/ml perfusate caused a dose- and time-dependent appearance of hemolysin, accompanied by release of potassium, thromboxane A2, and PGI2 into the perfusate. Concomitantly, marked pulmonary hypertension developed. Inhibitor studies suggested that the pressor response was predominantly mediated by pulmonary thromboxane generation. Administration of hemolysin-forming E. coli additionally caused a protracted, dose-dependent increase in the lung capillary filtration coefficient, followed by severe edema formation. The permeability increase was independent of lung prostanoid generation. An E. coli strain that releases an inactive form of hemolysin completely failed to provoke the described biophysical and biochemical responses. Preapplication of 2 x 10(8) human granulocytes was without effect in the present experimental model. We conclude that the hemolysin produced by low numbers of E. coli organisms can provoke thromboxane-mediated pulmonary hypertension and severe vascular leakage. E. coli hemolysin and, possibly, other related cytolysins may thus contribute directly to the pathogenesis of acute respiratory failure under conditions of sepsis or pneumonia.

Functional similarity between the haemolysins of Escherichia coli and Morganella morganii.

Haemolysin produced by a clinical isolate of Morganella morganii was examined for antigenic relatedness to the haemolysin of Escherichia coli and for similarities in mode of action. The M. morganii haemolysin migrated in SDS-PAGE as a single protein band with a slightly higher molecular weight than that of E. coli haemolysin. Several murine monoclonal antibodies against E. coli haemolysin cross-reacted with the M. morganii haemolysin in Western blots. Diminished haemolysis in the presence of osmotically-stabilising solutes indicated the formation of a pore by M. morganii haemolysin with an effective diameter of 1.5-3 nm. Results from dose-response experiments indicated that a single "hit" was sufficient for lysis of an erythrocyte. Detergent solubilisation of toxin-treated membranes led to recovery of bound toxin exclusively in monomeric form. M. morganii haemolysin was a potent leucocidin, that caused rapid leakage of ATP and death of human polymorphonuclear leucocytes. Under in-vitro conditions M. morganii haemolysin displayed similar leucocidal and haemolytic efficiency. The data demonstrate that M. morganii haemolysin shows functional properties virtually identical with those of E. coli haemolysin.

ABO and Lewis typing of secretion stains on nitrocellulose membranes using a new dot-blot-ELISA technique.

A new method for ABO and Lewis typing of body fluids is described. It combines the advantages of a good antigen binding to nitrocellulose membranes, the need of only very small amounts of stain material and the high sensitivity of an enzyme-linked immunosorbent assay for antigen detection. This is of special interest because conventional ABO and Lewis typing of secretion stains need relatively large stain dimensions. The method is very easy to handle, does not need any expensive equipment and gives a permanent record. Furthermore the high sensitivity offers the possibility of analyzing even sweat and urine stains without the need of concentrating these extracts.

[Gm/Km determination in sperm traces]. / Zur Gm/Km-Bestimmung an Spermaspuren.

In the forensic laboratories of the Federal Republic of Germany and West-Berlin 23 different semen stains and in our own laboratory 20 semen stains were typed in the gm/km-system doing 125 and 61 (own) test respectively. Examination was carried out by means of the haemagglutination method, which has been used successfully in typing bloodstains. Our critical assessment based on earlier experiences with semen stains was now confirmed and statistically evaluated: typing was successful in about 35-50% of the tests, but besides false-negative results, there was also a considerable percentage (4-10%) of false-positive ones. Therefore for the present it seems best to exclude the gm/km-typing of secretion stains from forensic investigations in order to avoid false incriminations or exonerations of suspects.

Potent leukocidal action of Escherichia coli hemolysin mediated by permeabilization of target cell membranes.

The contribution of Escherichia coli hemolysin (ECH) to bacterial virulence has been considered mainly in context with its hemolytic properties. We here report that this prevalent bacterial cytolysin is the most potent leukocidin known to date. Very low concentrations (approximately 1 ng/ml) of ECH evoke membrane permeability defects in PMN (2-10 x 10(6) cells/ml) leading to an efflux of cellular ATP and influx of propidium iodide. The attacked cells do not appear to repair the membrane lesions. Human serum albumin, high density and low density lipoprotein, and IgG together protect erythrocytes and platelets against attack by even high doses (5-25 micrograms/ml) of ECH. In contrast, PMN are still permeabilized by ECH at low doses (50-250 ng/ml) in the presence of these plasma inactivators. Thus, PMN become preferred targets for attack by ECH in human blood and protein-rich body fluids. Kinetic studies demonstrate that membrane permeabilization is a rapid process, ATP-release commencing within seconds after application of toxin to leukocytes. It is estimated that membrane permeabilization ensues upon binding of approximately 300 molecules ECH/PMN. This process is paralleled by granule exocytosis, and by loss of phagocytic killing capacity of the cells. The recognition that ECH directly counteracts a major immune defence mechanism of the human organism through its attack on granulocytes under physiological conditions sheds new light on its possible role and potential importance as a virulence factor of E. coli.

Quantitative study of the binding and hemolytic efficiency of Escherichia coli hemolysin.

Mono- and polyclonal antibodies were used to construct a sandwich enzyme-linked immunosorbent assay that permitted quantitation of Escherichia coli hemolysin in soluble and membrane-bound forms. Toxin concentrations of 4 to 14 micrograms/ml were measured in culture supernatants of E. coli LE 2001 at times of peak hemolytic activity. Quantitative studies on the binding of E. coli hemolysin to rabbit erythrocytes were conducted at 0 and 37 degrees C. At 37 degrees C, 85 to 95% of bindable toxin was cell bound after 60 min, and no saturability of binding was observed in the studied range of concentrations, which resulted in deposition of approximately 100 to 50,000 toxin molecules per cell. Binding was slower and less effective at 0 degrees C; however, hemolysis did occur at low temperature. The number of cell-bound toxin molecules required to generate a hemolytic lesion within 60 min was estimated to be approximately 100 molecules per cell at 37 degrees C and 800 to 1,000 molecules per cell at 0 degrees C. Upon prolonged incubation (5 to 20 h, 37 degrees C), the number of molecules evoking a functional lesion decreased to approximately 5 to 20 per cell. These results are compatible with the concept that E. coli hemolysin first adsorbs to the cell surface, with membrane insertion and pore formation following in a second step that may be temporally dissociated from that of binding. The data support the pore concept of toxin action by showing that attachment of a low and finite number of toxin molecules to an erythrocyte will ultimately generate a cytolytic lesion.

Isoelectric focusing of group specific component in semen and vaginal stains on ultrathin immobilized pH gradient gels.

Subtyping of the group specific component in secretions of semen and vaginal fluids is impossible with conventional detection systems. By means of a highly sensitive alkaline-phosphatase secondary antibody system the group specific component can reliably be detected in semen stains. Results with vaginal swabs were inconsistent and in saliva stains Gc activity could not be detected.

Alterations in lipid composition of liver cell plasma membranes during development.

Liver plasma membranes of Swiss-Webster CFW mice ranging in age from 5 to 165 days were isolated employing a two-phase dextran/polyethylene glycol polymer system. Electron microscopy revealed low mitochondrial and microsomal contamination. Lipids were quantitatively extracted and subjected to two dimensional thin layer chromatography. Individual lipids were eluted and phosphorus was measured by the Malachite green method. Cholesterol was quantitated by the micro Liberman-Burchard method and sphingolipids were quantitated fluorometrically with fluorescamine. During development, phosphatidylethanolamine levels decreased from 43.2% to 19.5% whereas phosphatidylcholine levels rose from 26% to 45.2%. The ration of phospholipid/cholesterol remained essentially constant during development; but the sphingolipid/cholesterol and sphingolipid/phospholipid ratios increased from 0.75 to 1.79 and 0.11 to 0.34, respectively.