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Background: Molecular biology methods based on reverse transcription and polymerase chain reaction (RT-PCR) are able to detect the presence of BCR-ABL transcripts in chronic myeloid leukemia (CML). In this study we present our experience with monitoring of residual disease using real-time PCR with hybridization probes detection in patients treated with imatinib mesylate and in collected peripheral blood progenitor cells (PBPC)., Methods and Results: We measured the level of BCR-ABL transcripts in peripheral blood cells of 27 subjects before and in the course of the imatinib treatment. The median of relative quantity of BCR-ABL in the blood before imatinib therapy was 2.55%. The number of the transcripts in 23 imatinib-sensitive subjects decreased to 0.02% in 6 months. After 12 months of the treatment the BCR-ABL median was 0.005%. Subsequent levels fluctuated between values below the detection limit (DL, 0.001%) and 0.005%. Three patients were primarily resistant to imatinib with the BCR-ABL range of 0.13%-11.7% during the treatment. One subject showed marks of molecular relapse after 18 months of the treatment. Only two of 16 filgrastim-stimulated patients had BCR-ABL levels in the blood and in collected PBPC below DL. In other subjects BCR-ABL transcripts were determined within the measurable range of RT-PCR., Conclusions: Taking into account prognostic importance, the measurement of BCR-ABL transcripts is an effective approach to monitoring of residual CML kinetics. Evaluation of BCR-ABL levels in collected PBPC can complete information on quality of the cells in potential autotransplants, and choose subsequent therapeutic protocols and patient prognosis.

Background: The interest in aspirin resistance has been increasing during the last few years, with researchers earnestly pursuing alternative anti-platelet therapies and devices for measuring platelet aggregation. The recent studies suggest that 5-45% of patients taking aspirin do not experience adequate anti-platelet effects., Methods and Results: There is scant evidence proving that aspirin resistance has some clinical consequences. To assess the prevalence of aspirin resistance in patients with ischemic heart disease (IHD) two independent methods were used: platelet aggregation in platelet rich plasma (PRP) after induction by propylgallate (CPG), and assessment of platelet function by PFA 100 method. The study population consisted of 424 patients treated for IHD on the 2nd Dept. of Medicine, Teaching Hospital, Hradec Kralove. The age, gender, diagnosis and effect of therapy were characterized in this group of the patients. Daily ASA dose was 100 mg. We used two methods to monitor ASA treatment efficacy: a) thrombocyte aggregation after induction by CPG, b) the assessment of platelet function by PFA 100 method. a) Of the patients studied by CPG platelet aggregation, 51 (12.1%) pts were resistant to ASA dose 100 mg/day, and 32 (7.6%) pts remained resistant even after increasig the dose to 200 mg/day. In 80 (20%) pts, the daily ASA dose of less than 100 mg was sufficient to inhibit platelet function. b) Although the PFA-100 system is not able to detect the difference between low and high ASA dose, we found 53 (15.2%) patients aspirin resistant using this method., Conclusions: In the patients with IHD treated with 100 mg of ASA per day, our study has shown that the prevalence of aspirin resistance was 12.1% using CPG method, and 15.2% using PFA-100 method. Aspirin resistance was dose dependent. Prevalence of ASA resistance in patients treated with 200 mg of ASA per day was only 7.6% using the CPG method.

LDL (low density lipoprotein) - apheresis has been established as an alternative management of severe hypercholesterolaemia after failure of conventional diet and drug therapy. General indication criteria for LDL-apheresis have yet been established. Indication guidelines in USA, Europe and japan are based on whether coronary heart disease is present and on the degree of lDL cholesterol elevation after treatment with diet and maximal drug therapy. It is reasonable to consider LDL apheresis therapy for: 1. patient with coronary heart disease and LDL cholesterol 4.9 mmol/l (190 mg/dl); 2. patients without coronary heart disease, but at high risk for disease (due to an LDL cholesterol above 6.4 mmol/l (250 mg/dl), a first-degree relative with premature coronary heart disease, and the presence of one or more additional risk factor. The therapeutical goal with present coronary heart disease is lDL cholesterol less than 3.4 mmol/l (130 mg/dl), with asymptomatic coronary heart less than 5.2 mmol/l (200 mg/dl). In addition, LDL apheresis is recommended for the management of all patients with homozygous familial hypercholesterolaemia due to the very high risk of coronary heart disease and the poor response to usual lipid-lowering treatments. In the end present two typical cases, treated by LDL-apheresis.

The syndrome of disseminated intravascular coagulation (DIC) is an acquired coagulation disorder which is characterized by a significant activation of haemostasis and the formation of intravascular microthrombi, the consumption of coagulation factors and activation of fibrinolysis. DIC has similar clinical stages and a similar laboratory picture, regardless of the causal factor. The diagnosis of DIC should be based on anamnestic, clinical and laboratory data. The laboratory diagnosis of DIC is possible by using relatively simple tests, which have the character of statim examinations. DIC treatment is based on the principle of treatment of the cause, discontinued consumption of haemostatic material, substitution of lacking factors and restoration of the microcirculation. The possible development of DIC must be taken into account in a number of acute conditions, in particular the development of shock syndrome. The presented article summarizes some views on causes, diagnosis and possible treatment of DIC.

Thrombocyte concentrates are vitally important in thrombocytopenic types of haemorrhage. If repeated transfusions are necessary, they are associated with immunological reactions, which cause side-effects in the recipient and the transfusion is ineffective. Donors should be selected according to the HLA system. The authors prepared a programme in the Redap language for selection of donors according to HLA antigens (loci A and B) which respects cross reactions and split antigens. The donor is selected by the grade of agreement with the recipient. The authors differentiate between nine possibilities from complete agreement to complete disagreement between donor and recipient. The initial experience revealed that the programme is feasible for the user and is sufficiently quick.

The authors assessed the test of platelet aggregation for the selection of a compatible donor. The test was compared with the hitherto used microlymphocytotoxic test. Consistent with data published abroad, the authors confirmed that the aggregation test improves the accuracy of selection of a suitable donor. The test can be performed in a shorter time than the microlymphocytotoxic test, this being important in particular in case of acute haemorrhage.

During intensive treatment of haemoblastoses it is important to administer repeatedly high-quality platelet concentrates. The authors describe the first results obtained with the use of an isoradial chamber of Travenol Co. for separation on a CS-3000 separator. Using the chamber, they achieved in the first 20 patients a reduction of the separation time by 30 mins. (23%) and the thrombocyte yield was not reduced. The resulting product contained, however, more leucocytes. The chamber can be used to reduce the separation period also under Czechoslovak conditions where donors have smaller numbers of thrombocytes than reported by western authors (the microcomputer of the separator is set for a regime suited for larger thrombocyte numbers).