Identification of platelet proteins separated by two-dimensional gel electrophoresis and analyzed by matrix assisted laser desorption/ionization-time of flight-mass spectrometry and detection of tyrosine-phosphorylated proteins.

Different search programs were compared to judge their particular efficiency in protein identification. We established a human blood platelet protein map and identified tyrosine-phosphorylated proteins. The cytosolic fraction of human blood platelets was separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and phosphorylated proteins were detected by Western blotting using anti-phosphotyrosine antibodies. Visualized protein spots were excised, digested with trypsin and analyzed by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The obtained mass fingerprint data sets have been analyzed using ProFound, MS-Fit and Mascot. For those protein spots with no significant search results MALDI post source decay (PSD) spectra have been acquired on the same sample. For automatic interpretation of these fragment ion spectra, the SEQUEST and Mascot algorithm were applied. Another approach for the identification of phosphorylated proteins is immunoprecipitation using an anti-phosphotyrosine antibody. A method for immunoprecipitation of tyrosine-phosphorylated peptides was optimized.

[HIV retrospective study of the German Red Cross blood donation service in Germany]. / HIV-Lookback-Studie der DRK-Blutspendedienste in der BRD.

OBJECTIVE: It was tried to retrospectively identify HIV infections in recipients of transfusions from donors who were tested HIV positive at a subsequent donation. These lookback data were traced back to answer the following questions: 1. How many transfusion recipients were infected before the start of the routine HIV testing in 1985? 2. How great is the risk of HIV infections from infected but not yet HIV antibody-positive donors? 3. Furthermore, the transfusion of HIV-infected transfusion recipients was traced back to the involved donor to establish causality. DESIGN: Retrospective ('lookback') study. SETTING: HIV Study Group of the Red Cross Blood Banks of the Federal Republic of Germany. PARTICIPANTS: Preceding donations of HIV antibody-positive repeat donors were traced back to the transfusion recipients in order to establish their HIV antibody status. In a second lookback study, HIV-infected transfusion recipients and their corresponding donors were investigated after they had been reported to the blood bank as infected by transfusion-associated HIV. INTERVENTIONS: None. RESULTS: Recipients of 156 respectively 133 transfusions from repeat donors found to be Western blot-positive were investigated from 1985 to 1987 and from 1987 to 1992, respectively. About 50% of the recipients had died. About 40% of the recipients could not be examined, because they either were not available for testing or refused to be tested or because it was impossible to clarify the fate of the blood products. 25 HIV recipients were identified from 1981 to 1985, when routine HIV testing began. Nine transfusion-associated HIV infections were identified from 1985 to 1992. 25 million units of blood were prepared during this period. CONCLUSIONS: The risk of HIV transmission by tested transfusions is extremely rare (in the order of 1:1 million). The second lookback study suggests that in more than 50% of the blood recipients in whom HIV infection was attributed to transfusion, a causal relationship to an infected donor could not be established.

Rh33 in two of three German siblings with D+ C+ c+ E- e+ red cells.

The proposita in a German family of three siblings has D+ C+ c+ E- e+ f+ Rh: -17,19,33,34 red cells with weak C, e, f, Rh19, and Rh34 and stronger-than-usual Rh33 expression. One sibling has D+ C+ c+ E- e+ f+ Rh:17,19,33,34 red cells with weak f and ordinary-strength Rh33, and the other sibling has D+ C+ c+ E- e+ f+ Rh:17,19,-33,34 red cells. In the absence of any further family members, the proposita's unusual phenotype suggests that she has an RoHar haplotype and a "new" Rh haplotype, provisionally named R1Lisa, that encodes Rh33, normal-strength D, weak C, weak or nondemonstrable e, Rh19, and Rh34, but not Rh17. Her Rh:33 sibling may have R1 and RoHar and her Rh:-33 sibling R1 and r haplotypes.

[Epidemiology of HIV in blood donors in West Germany]. / Epidemiologie des HIV bei Blutspendern in der Bundesrepublik Deutschland.

Of a total of 5.6 million blood donations by 1.6 million German Red Cross donors between July, 1985 and December, 1987 tested for HIV antibodies 210 were found to be positive in at least one Western blot. Calculated per quarter, the prevalence (for first-time donors) of 10 per 100,000 in 1985 decreased to 4 per 100,000 in 1986, while the incidence (for multiple donors) was a constant 2 per 100,000 for each quarter in 1987. This indicates that new HIV infections were rare in blood donors and there was no exponential increase. Further analysis revealed that over 90% of those with confirmed Western blot positive results were members of risk groups or their direct contacts. The age and sex distribution conformed to the known pattern for AIDS cases in the Federal Republic of Germany. The data demonstrate that there is at present no indication of a rapid spread of HIV infections transmitted heterosexually among blood donors.

[Prevalence of HIV antibodies in blood donors in the West Germany and West Berlin]. / Prävalenz von HIV-Antikörpern bei Blutspendern in der Bundesrepublik Deutschland und in West-Berlin.

Since 1985 all donor blood in the Federal Republic of Germany (FRG) and West Berlin (WB) have been routinely tested for HIV antibodies. The blood donor services of the German and Bavarian Red Cross have pooled their anti-HIV test data for the first year of operation (July 1985-June 1986) to obtain information on the prevalence of HIV antibodies in blood donors and in donated blood. Of 2.27 million units of blood, donated by 1.33 million donors, 218 were anti-HIV positive in the Western Blot Test, i.e. a prevalence rate of 10 per 100,000 donors. The prevalence was, as expected, higher in males than females. During the test period the anti-HIV prevalence fell, both as a total and in all subpopulations. In the second quarter of 1986 it had decreased to 6 per 100,000 donated units.