Comparison of fresh frozen plasma with a standardized serum protein solution following therapeutic plasma exchange in patients with autoimmune disease: a prospective controlled clinical trial.

The aim of the study was the comparison of the influence of fresh frozen plasma (FFP) (Freiburg, Germany) and Biseko, Biotest Pharma GmbH (Dreieich, Germany), as a plasma substitute (a standardized, virus inactivated human serum protein solution) on the coagulation factors, inhibitors, proteins, and complement factors in the plasma of autoimmune disease patients following membrane plasma separation. Patients (n = 24) with autoimmune disease were randomized to receive either FFP or Biseko for membrane plasma separation therapy. During each plasma exchange, 100% of the plasma volume was replaced by the respective substitute. Plasma exchange volume was performed once daily for 3 days. Target test parameters of the coagulation system were fibrinogen, fibrinopeptide A, factor VIII (FVIIIC), von Willebrand factor antigen (vWFAg), partial thromboplastin time (PTT), thromboplastin time (Quick value), and antithrombin (AT III). The immunoglobulins were IgG, IgA, and IgM and C-reactive protein (CRP). The thrombocytes were platelet factor 4 (PF4), and complement factors were C3 and C4. Biseko was well tolerated with 1 mild adverse drug reaction (ADR) (n = 1) while FFP gave rise to ADR on 7 occasions (n = 4). Statistically significant differences in the 2 groups were observed for fibrinogen, PTT, Quick value, and AT III. From the clinical point of view, all fluctuations and differences in parameter levels remained clinically silent. The differences had no clinical consequences. Reflecting on a potential decrease in the risk of infections in comparison to FFP therapy and the lower rate of adverse drug reactions, it is possible to postulate an advantage of Biseko for plasma exchange therapy.

Standardization of the PFA-100(R) platelet function test in 105 mmol/l buffered citrate: effect of gender, smoking, and oral contraceptives.

The PFA-100(R) (PFA) diagnostic system for the detection of platelet dysfunction was evaluated to determine reference ranges in a normal population. The PFA determines the primary haemostasis capacity (PHC) of anticoagulated whole blood, expressed by the system's closure time (CT). In this study the CT reference ranges were determined for blood samples collected in 105 mmol/l (3.2%) buffered citrate and the effect of gender, smoking, and use of oral contraceptives on reference ranges was assessed. Each of the 309 healthy blood donors from five blood centres was confirmed to have normal platelet function before inclusion in the study. Blood samples were tested in duplicate with both the collagen/epinephrine (Col/Epi) and collagen/ADP (Col/ADP) test cartridges. PFA reference ranges (90% central intervals of measured closure times) for both cartridge types were similar for all groups. Subgroup analysis showed that neither gender nor oral contraceptive usage had any effect on PHC. The 95% cut-off value for the Col/Epi CT was slightly higher for smokers than for non-smokers, an effect more pronounced in female than in male donors. However, the small difference did not justify establishment of specific reference ranges for smokers. Data from all included subjects were pooled to calculate the CT reference ranges for blood samples collected in 105 mmol/l buffered citrate (Col/Epi 82-150 s; Col/ADP 62-100 s). Normal levels of fibrinogen, as well as normal platelet counts and normal haematocrit levels, appeared not to influence the PHC. Because slight but significant differences of the reference ranges were observed between some of the participating sites, in-house confirmation of these reference range guidelines is recommended.

[Capture-R Ready-ID and DiaMed-ID for identification of erythrocyte bound antibodies after acid elution]. / Capture-R Ready-ID und DiaMed-ID zur Identifizierung erythrozytär gebundener Antikörper nach Säureelution.

The DiaMed-ID (D-ID) gel system is known to be a very sensitive and specific method for the detection of red cell antibodies. In various cases, we failed to find an antibody in the eluate in which red-cell-bound antibodies (IgG) where proven by a positive direct antiglobulin test (DAT). SPRCA (Capture-R, Ready Screen, Immucor; C-R) seems also to be very sensitive, in part due to the antihuman, IgG-coated indicator cells. Therefore, we compared 39 acid eluates from patients who had a positive DAT (monospecific rabbit antihuman IgG) both in the D-ID and in the C-R system. Patients (19 female, 20 male; mean age 62 years) were suspected either to have an autoimmune hemolytic anemia or an alloantibody. Identification of the antibodies was done with the system's own panel cells. Agglutination strength was scored from 1 to 4. Quantification of selected eluates was performed by titration, using the same cells in both systems. From 39 eluates, 31 were positive in the C-R and 25 in the D-ID. Nine eluates were negative in the C-R and 14 in the D-ID. If only eluates with a DAT reaction strength of 2 or lower were considered, obviously more negative results were found with the D-ID technique (p < 0.027) than with the C-R technique. In all eluates the degree of test reaction was stronger in the C-R system. However, titration endpoints of 6 quantified antibodies did not differ significantly. In 2 patients with slightly positive DAT, antibody typing was negative or not clear in the serum. In the corresponding eluates, an anti-K and an anti-JKa could be identified only by the C-R technique. In such instances we recommend to use the C-R technique to prevent transfusion complications.

[Advanced age--a risk factor in autologous blood donation?]. / Fortgeschrittenes Lebensalter--ein Risiko zur Eigenblutspende?

Tolerance of autologous blood donation was investigated in a patient group aged between 66 and 75 years (median = 70 years). Autologous blood donors between 18 and 65 years (median = 51 years) served as a control. A total of 38 patients were examined. Only blood donors which did not exceed ASA criteria II were accepted. Blood donation was performed weekly with a daily ferrum intake of 200 mg. Patients were divided into two groups with and without volume replacement, respectively. The parameters investigated were: blood pressure and pulse before and after the first and second autologous blood donation, circulatory response during 24 h after blood donation, and hemoglobin concentration before the first, second, and third donation. Regarding pulse and blood pressure, there was no statistical difference between the elder and younger patient group. Hemoglobin reduction from the first to the second donation was 1.1 g/dl in both groups and from the second to the third donation 1.1 g/dl in the younger group and 1.2 g/dl in the elder patient group. Again, no significant difference between both groups could be shown. None of the 38 patients showed negative side effects regarding the circulatory response during a time period of 24 h after blood donation.

[The gel centrifugation test: comparison of antibody identification at 37 degrees C with antibody identification at room temperature]. / Der Gelzentrifugationstest: Vergleich der Antikörpersuche bei 37 degrees C mit der Antikörpersuche bei Raumtemperatur.

The indirect antiglobulin test (antibody screening) (IAT) using the 'DiaMed-ID Micro Typing System' (ID system) was performed in two different ways: (1) incubation (Liss/Coombs version) at room temperature (RT) and (2) incubation (also Liss/Coombs version) at 37 degrees C. 106 antibody-containing sera were tested by IAT at RT and at 37 degrees C. The comparison of the results showed a high correlation between both methods: 49 of 106 antibodies were positive in the IAT at both RT and 37 degrees C, 51 antibodies were negative at both temperatures. Five antibodies were identified only in IAT performed at RT. 1/106 antibodies of the specificity anti-D (Rh prophylaxis) was not detected in the antibody screening at RT and in the conventional tube test. The antibody screening at RT using the ID System could be performed simultaneously with AB0 blood group typing without losing sensitivity and specificity. The modification of IAT using the ID System is more sensitive than the conventional tube test.

Two years serial lymphocytapheresis of progressive hyperleukocytosis in a patient with a non-Hodgkin's lymphoma.

A patient with non-Hodgkin's lymphoma was treated by ambulant lymphocytapheresis once a month over a period of 2 years and 8 months. The cytoreductive effect resulted in a decrease of white blood cells from 410/nl to 130/nl. The patient tolerated the procedure, which lasted 3-4 h, without any side effects. He could be kept in good condition by monthly apheresis. So lymphocytapheresis is a means to reduce hyperleukocytosis in patients with non-Hodgkin's lymphoma.

[Effects of an infusion pump with peristaltic mechanism of action on various blood components in vitro]. / Einflüsse einer Infusionspumpe mit peristaltischem Wirkprinzip auf verschiedene Blutkomponenten in vitro.

Apart from a slight hemolysis at very low flow rates, the peristaltic infusion pump IVAC 560 has no harmful effects on blood components such as packed red cells, whole blood, platelet concentrates and fresh frozen plasma. So it may be a convenient option to perform volume processed transfusions.

Detection of antibodies to hepatitis C virus in blood donors and their relationship to surrogate markers.

Stored serum samples from 7,179 nonselected blood donors were tested for anti-HCV using Ortho EIA first generation. Results were compared to data acquired by anti-HBc testing and ALT levels found in routine testing. 24 donors (0.33%) were repeatedly reactive with Ortho HCV EIA, 230 (3.20%) were anti-HBc-positive and 138 (1.92%) had raised ALT levels > or = 36 IU/l. A low correlation was found between HCV antibody screening with EIA and surrogate testing. When tested in addition with the Abbott HCV EIA, 20 of the 24 Ortho EIA-positive subjects showed a positive reaction. In the Abbott neutralization test 13 of these 20 (65%) were reactive. 8 (33.33%) of the 24 Ortho-EIA-positive donors were positive in the two-antigen-RIBA (first generation), 8 were indeterminate and 8 were nonreactive. The neutralization test and the RIBA can be used as supplementary tests fo further analyze HCV-EIA-positive specimens.

Influence of different surfactants on the separation of von Willebrand factor multimers by use of agarose gel electrophoresis and semi dry blotting technology.

The separation of vWF-multimers by use of sodium dodecyl sulfate (SDS) agarose gel electrophoresis and semi dry blotting instead of tank blotting procedure has been studied. We could demonstrate, that multimers in the range of highest molecular weights could not be visualized using this technique. Therefore we have investigated the influence of different types of anionic surfactants on the separation of vWF-multimers and could show that the hydrophilic/lipophilic balance (HLB) is an important value to characterize the capacity of a surfactant to separate vWF-multimers. The best separation quality was achieved with sodium dodecyl benzene sulfonate (SDBS). This surfactant is a superior alternative to SDS in separating vWF-multimers of the highest molecular weights.

Biosynthesis and secretion of M- and Z-type alpha 1-proteinase inhibitor by human monocytes. Effect of inhibitors of glycosylation and of oligosaccharide processing on secretion and function.

The biosynthesis and secretion of M-type and Z-type alpha 1-antitrypsin was studied in human monocytes. In monocytes of PiMM individuals alpha 1-antitrypsin represented 0.08% of the newly synthesized proteins and 0.44% of the secreted proteins. Two molecular forms of alpha 1-antitrypsin could be identified: a 51-kDa intracellular form, susceptible to endoglucosaminidase H, thus representing the high-mannose type precursor form and a 56-kDa form resistant to endoglucosaminidase H which was secreted into the medium. Inhibition of de novo glycosylation by tunicamycin impaired the secretion of M-type alpha 1-antitrypsin by about 75% whereas inhibition of oligosaccharide processing by the mannosidase II inhibitor swainsonine did not alter the secretion of M-type alpha 1-antitrypsin. alpha 1-Antitrypsin secreted by human monocytes was functionally active as measured by complex formation with porcine pancreatic elastase. Even unglycosylated alpha 1-antitrypsin secreted by human monocytes treated with tunicamycin formed a complex with elastase. In monocytes of PiZZ individuals the secretion of alpha 1-antitrypsin was decreased. 72% of newly synthesized M-type alpha 1-antitrypsin, but only 35% of newly synthesized Z-type alpha 1-antitrypsin were secreted during a labeling period of 3 h with [35S]methionine. The 51-kDa form of Z-type alpha 1-antitrypsin accumulated intracellularly, whereas the 56-kDa form was secreted. Inhibition of oligosaccharide processing by swainsonine did not alter the decreased secretion of Z-type alpha 1-antitrypsin, whereas inhibition of de novo glycosylation by tunicamycin blocked the secretion of Z-type alpha 1-antitrypsin completely.

Long-term stability of international reference preparations for thromboplastins.

Three certified reference materials for thromboplastins are available from the Community Bureau of Reference (BCR) of the European Commission for calibration of commercial thromboplastins used for control of oral anticoagulant therapy. The long-term stability of these reference materials has been monitored by two independent laboratories, using deep-frozen and lyophilized plasma samples. Prothrombin times and prothrombin time ratios measured on 19 occasions in the period 1981-86 have been analysed for trend with time. Although significant trends of prothrombin time and ratio (P less than 0.05) were observed, a consistent pattern of trends could not be recognized. The significant trends of prothrombin time and prothrombin time ratio are most probably due to changes in local laboratory conditions. There is no indication that the reference materials have deteriorated since the beginning of the study. It is recommended that long-term stability monitoring of thromboplastins be performed by at least two laboratories simultaneously.

Repression of alpha 2-macroglobulin and stimulation of alpha 1-proteinase inhibitor synthesis in human mononuclear phagocytes by endotoxin.

Mononuclear phagocytes are a bone-marrow-derived subgroup of white blood cells which circulate as monocytes and, after differentiation into macrophages, become resident in many tissues. By synthesizing the important proteinase inhibitors alpha 2-macroglobulin and alpha 1-proteinase inhibitor mononuclear phagocytes contribute to the control of proteolysis both in blood and tissues. Applying a culture system which enables human blood monocytes to differentiate into macrophages in vitro, synthesis of alpha 2-macroglobulin and alpha 1-proteinase inhibitor was studied. The normal course of monocyte-macrophage maturation is accompanied by a strong increase of specific alpha 2-macroglobulin synthesis and a concomitant slight decrease of alpha 1-proteinase inhibitor. alpha 2-Macroglobulin can be designated as a marker protein of the monocyte/macrophage differentiation. Endotoxin (Salmonella typhi) in a concentration as low as 100 ng/ml strongly represses alpha 2-macroglobulin synthesis both in monocytes and macrophages. Furthermore, endotoxin completely abolishes the induction of alpha 2-macroglobulin synthesis during the course of normal monocyte in vitro cultivation, indicating that endotoxin is a strong inhibitor of the monocyte-macrophage maturation. In contrast to alpha 2-macroglobulin, alpha 1-proteinase inhibitor synthesis is strongly stimulated by endotoxin in monocytes as well as in macrophages.