Thrombocyte substitution in acute leukemia. Effect of histocompatibility on the clinical efficacy

Thrombocyte substitution is an essential prerequisite for intensive cytoreductive therapy in acute leukemia. Evaluating 228 thrombocyte transfusions in 17 patients shows that the clinical effectiveness of thrombocyte concentrates can be increased by making the coordination of HLA antigens of donor and receiver as good as possible. When measured in the corrected increment (CI) 24 hours after transfusion, the effectiveness of A3/B1 match preparations (CI = 7.0 +/- 1.6) is significantly higher than that of random preparations (CI = 3.0 +/- 0.5). With the presence of HLA antibodies an effective substitution (CI24 greater than or equal to 4.5) can only be achieved by A3/B1 match thrombocytes. This can only be realized by applying the fourfold thrombapheresis of single donors.     

Megakaryocytes and thrombocytopoiesis in idiopathic thrombocytopenic purpura

In idiopathic thrombocytopenic purpura (ITP) patients with histological signs of stronger auto-antibody synthesis (spleen follicle hyperplasia) more frequently revealed megakaryocyte changes due to antibodies and less frequently thrombocyte production compensatory increased than ITP patients with signs of weaker auto-antibody synthesis (spleen follicle normoplasia). The observation suggest that insufficient and frequently lacking compensatory increase of thrombocyte production in ITP is caused by an autoallergic disturbance of megakaryocyte maturation.     

HLA compatibility and survival of 51 chromium-labeled allogeneic thrombocytes

In 37 patients with thrombocytopenia (mostly with ITP) the survival time of 51Cr-labeled allogeneic platelets was investigated. The HLA antigens were typed in donors and recipients and the presence of HLA cytotoxins and specific thrombocyte antibodies in sera of patients were examined. In 7 cases the identity of 2 HLA antigens, in 15 cases that of 1 HLA antigen and in 15 cases the HLA incompatibility between donor and patient were found. The survival of platelets did not depend on the degree of HLA compatibility, unless the HLA cytotoxins in sera of patients appeared. The HLA, as well the specific platelet antibodies brought about the shortened platelet survival to 1 day and less. The importance of these observations for platelet kinetics is discussed.     

Thrombocyte counting with the Laborscale (PSL-1/PSA-1) particle-counting instrument by Medicor (Budapest/Hungary) and thrombocyte volume analysis in the diagnosis of selected thrombocytopathies

A thrombocyte counting technique is presented in comparison with the chambre counting according to the German Book of Medicaments (DAB 7 D.L.) GDR on the particle counting device "Laborscale" (PSL-1/PSA-1) of Medicor by using a thrombofuge (70 g). The composition of reagents accounts for problems of measurement which are caused by the device and influences the separation of particles. The possibility of recording the curve of thrombocyte volume distribution and calculating the average thrombocyte volume (VM) is demonstrated by means of a normal collective of test persons as well as by intravascular disorders of haemostasis and during cytostatic therapy.     

Evaluation of clinical efficacy of platelet substitution in acute leukemia using resonance thrombography

For evaluating the clinical effectiveness of thrombocyte substitution, the resonance thrombogram was registered before and after thrombocyte substitution in 50 patients with acute leukemia and the increase of thrombocytes (corrected increment) determined. Thrombocyte substitution led to a significant diminution of the platelet amplitude (p less than 0.01) and platelet value. The highest value of these alterations of parameters was reached 1 hour after transfusion, it was somewhat lower after 24 hours, yet the initial value was not reached again. Correlations could be found to exist between the platelet amplitude and the corrected thrombocyte increase 1 hour and 24 hours after transfusion. The resonance thrombography enables the successful control of thrombocyte substitution to be made and it is suitable for monitoring hemostasis in severe thrombocytopenia due to production.     

Behavior of thrombocytes in low-dose heparin therapy

In a little prospective study it could be shown that a series of 10-51 low dose heparin injections may be followed by slight or unspecific thrombocyte diminutions without specific heparin antibodies being available. Depending on stress and trauma potential complement activations caused by heparin histon complexes may result in insignificant thrombocyte diminutions by liberating those factors activating platelets from granulocytes. These are not bound to interrupt heparin therapy or heparin prophylaxis.     

Selection of HLA compatible donors for patients with HLA antibodies

A strain of 1,507 typised donors enables 15.97 average donors to be selected for a thrombocyte transfusion in one of 75 patients selected at random with HLA antibodies being previously determined (1 to 66 donors per patient). HLA compatible donors were found for 72 patients (97.33 per cent). More than 5 HLA compatible donors could be found for 58 patients. The high number of compatible donors is based on the fact that among 1,507 donors there were 156 HLA homozygotes and 556 donors with 3 HLA-A or B-antigens respectively. Compatibility in the ABO-system was not taken into account.     

Immunomorphologic characterization of chicken thrombocytes

Summary Chicken thrombocytes were enriched for immunization by utilizing their strong capacity to adhere to plastic surfaces. The produced rabbit anti-thrombocyte serum ATS 3 reacted by means of the unlabeled antibody enzyme method (PAP) specifically with thrombocytes of fixed chicken-blood smears, but not with lymphocytes or other blood cells. When ATS 3 (substrate diaminobencidinetetrahydrochloride = DAB) and a 11 mixture of an anti-bursa serum and anti-thymus serum (ABS/ATS; substrate 4-chloro-1-naphthol = 4-Cl-1-N) were used simultaneously, thrombocytes revealed the brown color typical for DAB, whereas lymphocytes showed the blue stain of 4-Cl-1-N. The finding of a thrombocyte surface antigen not shared by lymphocytes is regarded as a further proof of the diversity of both cell systems, i.e., for the existence of a genuine thrombocyte system in chickens.     

Serological techniques for the detection of antibodies to porcine thrombocytes. 1. The agglutination, fluorescent antibody and immunodiffusion techniques

Attempts were made to detect porcine thrombocyte antibodies by three techniques. No antibodies were detected by immunodiffusion; hetero-antibodies were detected by both the fluorescent antibody and thrombo-agglutination techniques; but iso-antibodies were only detected by thrombo-agglutination. With the latter, however, there were ill-defined end-points which made the interpretation of negative results difficult. It was therefore suitable only for positive reactions of strong avidity.     

Multivariate analysis for the understanding of typical diabetic hemostasis criteria. 2. Prevalent thrombocytic markers]

Various thrombocyte functions and metabolic criteria were investigated in 309 test persons, among them 198 Patients affected with diabetes mellitus, 67 of type I and 131 of type II as well as 111 healthy control persons. From the variety of these factors an optimal amount was determined by means of the statistical method of multivariance analysis separating both types of diabetes and healthy control persons. In addition to haemoglobin A1 concentration, thrombocyte parameters have been found together with the degree of capillary fragility, adrenalin-induced aggregation, platelet aggregation test and clot retraction, which allow individuals to be assigned diagnostically to various groups of test persons. Thus, thrombocyte functions found in both types of diabetes and normal persons exhibit differences which may contribute to a diagnostic classification.     

The effect of cyclooxygenase inhibition on the indices of the thrombocyte-vascular link in hemostasis and on the free-radical processes of lipid oxidation in experimental Salmonella intoxication]

Injection of Salmonella typhimurium endotoxin to the laboratory animals (rabbits) in dose of 1 mg/ml (LD84) induces the particular changes in the thrombocyte vessels system of hemostasis: decrease of aggregatory ability of thrombocytes, increase of thromboxane A2 and prostacyclin activation of lipid peroxidation process. Use of indomethacin--the cyclooxygenase inhibitor--leads to less progressive alterations of the studied parameters of the thrombocyte vessels hemostasis and lipid peroxidation processes.     

Studies of thrombocyte function in CPD blood]

The CPD stabilizer according to Gibson with an addition of 1.25 mMol adeninesulfate and 2.50 mMol guanosine is used in blood storage for better preserving 2.3-bis-phosphoglycerate of erythrocytes. Here platelet-rich CPD plasma was investigated before and during a 3 days storage at 4 degrees C or room temperature with regard to preserving the global thrombocyte function. The latter consists in the ability to seal blood vessels and is tested by means of pressure registration in combined thrombocyte-aggregation-adhesion (DKTA method) as an ability to close the pores of a sieve by adding 10(-5) mM/l of ADP. At room temperature this thrombocyte function is approximately 0 following 3 days of storage in CPD plasma excess without shaking. When stored at 4 degrees C it is preserved to a slight degree. Loss of thrombocyte function will depend on pH, thus being particularly evident at room temperature.     

The question of thrombocyte substitution as a prophylactic measure in splenectomy for idiopathic thrombocytopenic purpura (ITP)]

The problem of intraoperative thrombocyte transfusion in splenectomy of patients with ITP is discussed. The indication for splenectomy cannot be equalized with that for intra-operative platelet substitution. Thrombocyte kinetic examinations with a concurrent determination of the thrombocyte turnover enable those patients to be recognized by their bleeding tendency who are particularly endangered by operations. The intraoperative platelet substitution should be limited to those patients with ITP whose turnover is markedly lowered as a manifestation of reduced thrombocytopoiesis. In these cases it is advisable to shift splenectomy to a later date.     

On the production of anti-human-lymphocyte serum (ALS) by means of combined local and intravenous immunization in swine]

Lymphocytes of donors' peripheral blood were used for the study of pig antihuman ALS preparation by means of local and intravenous immunization. The results were compared with those obtained with exclusively local application of the same antigen. Antisera were tested from the aspect of activity (lymphoagglutination, lymphocytotoxicity, the rosette inhibition test) and for the presence of unwanted antibodies (haemagglutinins, thromboagglutinins, precipitins to serum proteins). Of all alternatives tested the following scheme proved to be optimal: subcutaneous application of 2 X 10(9) lymphocytes with adjuvans on day 0, followed by intravenous application of the same dosage on day 13, and serum harvesting on day 20. Antisera thus obtained displayed titres 1: 32 000 to 1 : 130 000 in the RIT - this being maximum level reached by exclusively local immunization only in some animals. Therefore, favourable immunosuppressive effect in vivo can be expected. Even more profound difference was observed in the level of unwanted antibodies that were found to be on markedly lower level after combined immunization. There is a good reason to suppose that with the help of further techniques, more likely absorption, these can be lowered to acceptable level. It is apparent from the results that with the help of a suitable immunization procedure not only highly active ALS can be obtained but also formation of nondesirable antibodies can be suppressed and, despite their thrombocyte contamination peripheral blood lymphocytes can successfully be used.     

Circadian rhythm of 5-fluorouracil (FU) pharmacokinetics and tolerance

In 28 adult patients with unresectable, disseminated gastrointestinal cancer FU pharmacokinetics as well as hematological and gastrointestinal side-effects of the therapy were evaluated to determine it they were circadian-stage dependent. FU was given as a bolus i.v. injection (15 mg/kg) 4 times at 0100, 0700, 1300, and 1900 at intervals of at least 96h in a changing sequence. Pharmacokinetic parameters were calculated on the basis of drug concentration in the serum, measured by the gas-chromatography method. Hematological side-effects were evaluated on the basis of white blood cell and thrombocyte count. Gastrointestinal toxicity (nausea, vomiting and diarrhoea graded in own 0-2 scale) was evaluated after the therapy. The single-cosinor analysis was applied for both detection and characterization of circadian rhythms. The results of the study were as follows: 1. Pharmacokinetic parameters of FU depend on the time of injection with the highest drug biotransformation at midday and early afternoon. 2. The highest white blood cell count following FU administration was noted after the night injection (0100), though the time of administration did not affect thrombocyte count and gastrointestinal side-effects.     

Effect of continuous irradiation on the thrombopoietic system in mice

In this paper the changes of the megakaryocyte number of the spleen and those of thrombocyte number of the peripheral blood were studied during continuous irradiation with a dose rate of 9.57 to 957 mGy/day. Beginning with a dose rate of 95.7 mGy/day the thrombocyte number of the blood and the megakaryocyte number of the spleen of irradiated animals decreased significantly. Whereas the thrombocyte number remained permanently decreased, the cell number of the megakaryocyte type is increased temporarily to a clearly higher level as compared with the controls on the 100th day of irradiation approximately. This is especially true for the middle dose rate. During this time of irradiation nucleolar hyperchromatosis as well as pycnosis were observed in many megakaryocytes.     

Serological techniques for the detection of antibodies to porcine thrombocytes. 2. The complement fixation and antiglobulin consumption techniques

Attempts were made to detect antibodies to porcine thrombocytes by two indirect techniques. Only hetero-antibodies were detected by complement fixation; iso-antibodies were detected by the antiglobulin consumption technique. The latter gave consistent reliable results, the difference between positive and negative results being clear-cut, but was found to be laborious.
The problems of porcine thrombocyte serology are discussed.     

The prostaglandin synthesis in marine fish thrombocyte

1. Prostaglandin (PG) syntheses from labelled highly unsaturated fatty acids were investigated in washed thrombocyte suspensions of four species of marine fish, flounder (Paralichthys olivaceus), black seabream (Acanthopagrus schlegeli), black rockfish (Sebastes schlegeli), and red seabream (Pagrus major). 2. Synthesized PGs were analyzed by thin-layer radiochromatogram scanner and high-performance liquid chromatography. 3. When [1-(14)C]arachidonic acid (AA) was incubated with washer thrombocyte suspension, AA was mainly converted to PGF(2alpha), PGE2, and PGD2 in all species. 4. It was suggested that [1-(14)C]eicosapentaenoic acid was mainly converted in PGs only in black rockfish thrombocytes, and the metabolites were mainly PGE3 and PGD3. 5 However, [1-(14)C]docosahexaenoic acid was not cyclized into PGs in all the species.     

淡水石斑外周血细胞显微结构观察

淡水石斑(Cichlasoma managuense)外周血细胞可区分出红血细胞、嗜中性粒细胞、嗜酸性粒细胞、单核细胞、淋巴细胞和血栓细胞,未发现嗜碱性粒细胞。外周血液中还存在少量未成熟的和正在分裂的红血细胞。白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;数量上,血栓细胞最多,而嗜酸性粒细胞则最少。     

Trypanosoma congolense: Thrombocyte survival in infected steers

Charolais steers infected with Trypanosoma congolense developed a thrombocytopenia that was first demonstrated shortly before the onset of parasitemia. The thrombocyte count progressively decreased from a level of 6 × 10⁵/mm³ on the 3rd day postinfection to l × 10⁵/mm³, its most depressed level, on the 11th day postinfection. The mean of the thrombocyte level of five infected bovines at the time of thrombocyte survival analysis was 195.6 ± 83.5 × 10³(± 2 SE) as compared to 998 ± 245.9 × 10³ in the control group. In parallel with depressed thrombocyte levels, the mean of the apparent half-lives of ⁵¹Cr-labeled thrombocytes was 1.3 ± 0.5 days as compared to 3.7 ± 0.5 days in control animals. A similar survival was noted in the apparent half-lives of ⁵¹Cr-labeled autologous and heterologous thrombocytes transfused to normal recipients.