Increasing procoagulant activity of circulating microparticles in patients living with HIV.

OBJECTIVES: HIV-infected individuals are at higher risk of non-AIDS diseases associated with procoagulant status. Microparticles are elevated in disorders associated with thrombosis (e.g., cardiovascular diseases). We investigated the association between microparticle levels in untreated and treated HIV-infected subjects, and determined the association with immune status, viral replication, and duration of antiretroviral therapy. PATIENTS AND METHODS: We included 144 HIV-infected subjects, including 123 on antiretroviral therapy (ART) and 21 before treatment initiation. A control group of 40 HIV-negative healthy adults matched for age and sex was used for comparison of microparticle levels. Treated subjects were divided into five groups depending on the period of antiretroviral exposure. Statistically significant differences were determined by Kruskal-Wallis test and Chi2 test. The relation between microparticles and other parameters was assessed using Spearman's coefficient of correlation. RESULTS: Microparticle levels were significantly higher in treated and untreated HIV-infected subjects than in non-HIV-infected controls (P<0.001). The microparticle level was similar between the groups on treatment (P=0.913). No association between the microparticle level and CD4+ count, CD4+/CD8+ ratio, number of HIV-1 RNA copies, or duration of exposure to antiretroviral treatment was observed. CONCLUSION: Increased levels of microparticles may be due to processes independent of viral replication and CD4+ cell count, and microparticle release might persist even during viral suppression by antiretroviral treatment. Elevated microparticle levels might occur in response to other triggers.

[Chronic disseminated intravascular coagulation (DIC) markers in a patient with multiple angiomatosis during treatment with anti-angiogenics: interferon α, thalidomide and lenalidomide]. / Vývoj koagulacních markeru chronické diseminované intravaskulární koagulace (DIK) u pacienta s mnohocetnou angiomatózou v prubehu lécby antiangiogenními léky: interferonem α, thalidomidem a lenalidomidem.

BACKGROUND: Multiple angiomatosis is a rare disease with angiomatous formations in multiple organs and tissues and associated with a risk of fatal bleeding. CASE DESCRIPTION: In this patient, the bones, pleural and peritoneal cavities and digestive tract were involved. The patient had long-term been administered zoledronate that provided relief from bone pain as early as after the second dose. The effect of antiangiogenics was evaluated on CT and MRI. Since angiomatous proliferation is associated with chronic disseminated intravascular coagulation (DIC) and anaemisation, blood count and fibrinogen as well as D-dimer and soluble fibrin monomer concentrations are also used to assess treatment response. RESULTS: Before treatment, D-dimer levels were in excess of 20 µg/mL, fibrinogen 1.4 g/L and soluble fibrin monomers were at measurable levels. During treatment with interferon α at a dose of 6 million units 3 times a week with the dose reduction after 10 month, the median fibrinogen concentration increased to 1.5 (1.2-2.0) g/L, the median D-dimer levels declined to 17.2 (13.4-20.0) µg/mL and fibrin monomers were still detectable. Thalidomide therapy (100 mg/day) provided reduction in the median D-dimer levels to 6.07 (4.71-10.21) µg/ml and increase in median fibrinogen concentration to 1.9 g/L; soluble fibrin monomers were unidentifiable. CT imaging suggested significant reduction of angiomatous mass. Progressing neuropathy required dose reduction of thalidomide to 50 mg/day, leading to D-dimer increase. Lenalidomide 10 mg/day provided an increase in median D-dimer concentration to 10.8 (10.8-17.35) and decline in the level of haemoglobin to a median of 124 (135-117) g/L. Soluble fibrin monomers became detectable again. Therefore, a low dose of lenalidomide 10 mg/day was combined with thalidomide 100 mg and, subsequently, 50 mg/day. Treatment with lenalidomide 10 mg and thalidomide 50 mg provided median D-dimer levels of 9.32 and the disease has remained stable for 9 months. CONCLUSION: Thalidomide 100 mg/day stabilized multiple angiomatosis better than interferon alfa. Thalidomide 50 mg/day was insufficient to maintain disease stability. Lenalidomide at a dose of 10 mg was tolerated really well but this dose was insufficient to maintain low D-dimer levels and normal haemoglobin concentrations. The combination of lenalidomide 10 mg and thalidomide 50 mg daily stabilized the disease for 9 months.

[Six-year follow-up of a patient with multiple angiomatosis involving skeleton, thoracic and abdominal cavities and the gut wall]. / Sestileté sledování pacienta s mnohocetnou angiomatózou postihující skelet, brisní i hrudní dutinu a stenu trávicí trubice.

BACKGROUNDS: Multiple angiomatosis is a rare disease causing angiomatous lesions in multiple organs and tissues with a risk of life-threatening haemorrhage. OBSERVATION: A young man was diagnosed with multiple angiomatosis at the age of 28 after two years of back and abdominal pain. Laparotomy revealed multiple spongy lesions mostly within the retroperitoneal space. Also, an involvement of the gut wall, bones and mediastinum was evident. After 6 years of treatment, the disease has been stabilized. Bone pain ceased with a significant contribution of zoledronate. Using CT and MR imaging, the effectiveness of antiangiogenic drugs was evaluated. Furthermore, treatment response was evaluated using laboratory values for coagulation and blood count, as angiomatous proliferation is known to be associated with disseminated intravascular coagulation and anaemia. RESULTS: Baseline laboratory examination revealed elevated D-dimer (more than 20 µg/mL), low fibrinogen (1.4 g/L), and the presence of fibrin monomers. After treatment with 6 mil. IU of interferon-alpha thrice weekly, there was only partial improvement in D-dimer (17.2 µg/mL) and fibrinogen (1.5 g/L) concentrations but fibrin monomers remained positive. After thalidomide (100 mg daily), D-dimer decreased to 6.1 µg/mL and fibrinogen levels increased to 1.9 g/L with the disappearance of fibrin monomers. CT scanning showed significant regression of angiomatous lesions. Progressive neuropathy was the reason to lower the dose of thalidomide by half and this caused D-dimer to rise again. Switching to lenalidomide 10 mg daily led to an increase in D-dimer to 10.8 µg/mL and decrease in haemoglobin concentration to 124 g/L. Fibrin monomers became positive again. Combined therapy with thalidomide (50 mg/day) and lenalidomide (10 mg days 1-21 in 28-day cycles) has led to stabilisation of the disease. Median concentration of haemoglobin increased to 131 (84-141) g/l. The median of D-dimer decreased to 9.3 (8.0-17) µg/mL. CONCLUSION: Thalidomide in the dose of 100 mg daily led to better stabilisation of the disease than interferon-alpha. However, lowering the dose because of adverse effects failed to be effective sufficiently. Lenalidomide 10 mg daily was well-tolerated but insufficient to improve D-dimer and haemoglobin concentrations. Therefore, for further treatment we have decided to use the combination of lenalidomide and thalidomide in doses of 10 mg and 50 mg, respectively because both drugs have desirable antiangiogenic activities with different adverse effect profiles. On this therapy, the patients disease has been stable for 9 months.

[Blood lipid changes at long-term antiretroviral treatment]. / Zmeny krevních lipidu pri dlouhodobé antiretrovirové lécbe.

UNLABELLED: The aim of study was to find the development trend of blood lipid concentration in a group of HIV positive patients treated by combination antiretroviral therapy (cART). We followed changes during the therapy and evaluated their aterogennic nature. METHODS: The group included 118 patients stepwise allocated to the AIDS Centre of the Faculty Hospital Brno, with the monitoring period being up to 1 month as the minimum and up to 17 years as the maximum. The patients were divided into cART treated patients and not treated patients. The following parameters were analysed: total cholesterol, triglycerides, HDL-cholesterol, apolipoprotein B, the total cholesterol/HDL-cholesterol index and non-HDL-cholesterol. RESULTS: Our group experienced a statistically significant increase of total cholesterol concentration already in the first months after cART initiation and this value continuously increased in the following years. The recommended target value for total cholesterol (5 mmol/l) was exceeded in the group of patients after 3-4 years of cART initiation. The triglyceride concentration showed a sudden increase already a few months after cART initiation, when the recommended optimum value of triglycerides (1.7 mmol/I) was exceeded. These changes had a further no statistic significance. The average triglyceride value was all around (slightly above) 1.7 mmol/l. Our group experienced a statistically significant increase of HDL-cholesterol concentration in the first two years after cART initiation. A statistically significant change of HDL-cholesterol concentration was not found in the following years. The average HDL-cholesterol value was above optimal value HDL-Ch > 1.0 mmol/l for men (except initial category). A statistically significant change of apolipoprotein B concentration was found after 3-4 years of cART treatment. However, the average apolipoprotein B value did not exceed the target value in any of the followed categories. No statistically significant changes of the total cholesterol/HDL-cholesterol index were found. The resulting value was under 5 in all the followed categories. Statistically significant changes of non-HDL-cholesterol were found in patients with cART already a few months after treatment initiation and its concentration continually increased. However, the recommended target value of non-HDL-cholesterol (3.8 mmol/l) was exceeded only in the category of patients treated 4 - 5 years. The development trend of CD4+ lymphocyte count and HIV-1 RNA copies means high active of cART from standpoind of immunoregeneration (CD4+ lymphocyte count) and viral suppression (HIV-1 RNA copies) even in the group of treated patient with the longest monitoring period. CONCLUSION: Monitoring of our group of HIV-positive patients treated by combination antiretroviral therapy revealed a statistically significant increase of blood lipid concentrations (inclusive of HDL-cholesterol) during the treatment. However, these changes do not have an unequivocally aterogennic nature even in the group of treated patient with the longest monitoring period.

[HIV lipodystrophy]. / HIV lipodystrofie.

Combined antiretroviral therapy results in extraordinary decrease of morbidity and mortality of HIV-infected patients and in an essential change of the HIV/AIDS disease prognosis. However, long-term intake of antiretroviral medicaments is related to occurrence of metabolic and morphological abnormalities, of which some have been combined into a new syndrome--the so called HIV lipodystrophy. The HIV lipodystrophy syndrome covers metabolic and morphological changes. Metabolic changes include dyslipidaemia with hypercholesterolaemia and/or hypertriglyceridaemia, insulin resistance with hyperinsulinaemia and hyperlaktataemia. Morphological changes have the nature of lipoatrophia (loss of subcutaneous fat--on the cheeks, on extremities, on buttocks and marked prominence of surface veins) or lipohypertrophia (growth of fat tissue--on the chest, in the dorsocervical area, lipomatosis of visceral tissues and organs, fat accumulation in the abdominal area). Several HIV lipodystrophy features are very similar to the metabolic syndrome of the general population. That is why this new syndrome represents a prospective risk of premature atherosclerosis and increase of the cardiovascular risk in young HIV positive individuals. The article mentions major presented studies dealing with the relation of antiretroviral treatment and the cardiovascular risk. The conclusions of the studies are not unequivocal--this is, among others, given by the reason that their length is short from the viewpoint of atherogenesis. The major risk of subclinical atherosclerosis acceleration seems to be related to the deep immunodeficiency and low number of CD4+ lymphocytes and florid, uncontrolled HIV infection with a high number of HIV-1 RNA copies actually circulating in the plasma. The question, whether metabolic and morphological changes related to HIV and cART carry a similar atherogenic potential as in the general population, remains open for future.

FEIBA in treatment of acute bleeding episodes in patients with haemophilia A and factor VIII inhibitors: a retrospective survey in regional haemophilia centre.

FEIBA (factor eight inhibitor by-passing activity) is used to achieve haemostasis in haemophiliacs with inhibitor. The aim of this study was to evaluate efficacy and consumption of the product in treatment of haemorrhages in haemophiliacs with factor VIII inhibitor, and determine factors that can influence the results of treatment. We used data from our haemophilia centre from years 2000-2008. Six haemophiliacs with factor VIII inhibitor were treated on demand with FEIBA for 61 bleeding episodes (45 haemarthroses, six muscle bleeds, six other sites bleeds and four multiple sites bleeds). The median cumulative dose of FEIBA per bleeding episode was 205 U kg(-1). Bleeding was stopped in 96.7% (59 of 61) of events but re-bleeding occurred in 3 events (4.9%) within 48 h after cessation of bleeding. In home treatment (20 of 61) bleeding stopped in 90% (18 of 20) without recurrence and the median consumption per event was reduced to 153 U kg(-1). Without the use of home treatment the median consumption was 250 U kg(-1) per event and bleeding ceased definitely in 92.7% (38 of 41) of cases. The cumulative dose of FEIBA was lower for three episodes with re-bleeding: median 96 U kg(-1) but not in the two cases of ineffective treatment: 361 U kg(-1). FEIBA in management of bleeding episodes completely resolved the haemorrhage in 91.8% of events and in a further 4.9% if treatment was restarted. Using home treatment saved expenditure due to the lower cumulative dose needed for treatment of haemorrhage.

[Hemophilia]. / Hemofilie.

Hemophilia is the most spread hereditary bleeding disorder with severe bleeding symptoms. Although the number of hemophiliacs is below 0.1 per thousand in population, the care of these patients consumes a lot of financial expenditures, especially for treatment in appearance of inhibitor. Management of hemophilia is best provided by specialist in a hemophilia treatment centre. However, all physicians can meet the patients with hemophilia in urgent situation or with their chronic problems. The aim of this article is to give an overview of this disease from pathophysiology, clinical manifestation and diagnosis (including prenatal) to treatment. We describe most frequent complications of hemophilia treatment, particularly management of inhibitor.

[Treatment of haemorrhage with transfusion preparations and blood derivatives]. / Lécba krvácení transfuzními prípravky a krevními deriváty.

Appropriate use of blood components and blood products improved both patients care and use of health care resources. In bleeding and/or risk bleeding situations use of these products is often indicated. The authors summarise different guidelines, experiences, indications and problems in hemotherapy concerning bleeding complications.

[Preparation of patients with haemostatic disorder for invasive medical interventions]. / Príprava pacientu s poruchou hemostázy k invazivním lékarským zákrokum.

The current trend in medicine is to sustain the possibility for necessary procedures to be performed in patients who suffer from haemostatic disorders which complicate eventual surgery. Among such disorders are congenital blood coagulation disorders, haemostatic disorders concomitant with other diseases and also therapies which affect haemostasis either on purpose or as part of adverse effects. Among coagulation disorders are congenital haemorrhagic or thrombotic conditions, acquired blood coagulation disorders--combined in the vast majority of cases-- and associated with pregnancy, severe internal diseases and surgery related diseases, severe injuries, wounds, burns, malignancies, systemic connective tissue diseases, inflammatory bowel disease, and a number of other diseases. A separate issue is that ofanticoagulation therapy--both antiplatelet, used in the treatment or prevention of venous thrombosis, and anticoagulation, predominantly used to manage venous thromboembolism. Also considered should be any therapy which may have a negative impact on coagulation due to its adverse effects.

[Von Willebrand disease]. / Von Willebrandova choroba.

Von Willebrand disease is the most spread hereditary bleeding disorder with prevalence up to the 1% in the population. Haemorrhagic symptoms are various intensity, mostly not severe. A large part of affected patients do not come to see a doctor. However these patients are potentially endangered by a serious haemorrhage after a trauma, surgery or during another illness. The estimated number of patients with von Willebrand disease is 1,500-3,500 per 1,000,000 of population; the number of persons with symptomatic von Willebrand disease was estimated to be at least 100 per million. This disease deserves attention not only by haematologists but other physicians too. In our communication we present short overview about von Willebrand disease: there are briefly described pathophysiology, classification, diagnosis and treatment of this illness.

Hemocoagulation disorders in extensively burned patients: pilot study for scoring of the DIC.

GOAL OF THE STUDY: To apply results of the hemocoagulation parameters to the DIC (disseminated intravascular coagulation) score system. Compare parameters of the DIC score in patients with extensive burn trauma (hospitalized at the Intensive Care Unit, ICU) and patients with lesser extent of burn injury (hospitalized at the standard unit). To use these data within the evidence based medicine for the prediction of organ damage and multi-organ failure. TYPE OF STUDY: Prospective study. MATERIAL AND METHODS: We have included total of 36 patients in the group within four months. Twelve patients were hospitalized at the ICU, 24 patients were hospitalized at the intermedial care and standard unit. Repeatedly, we have taken blood from patients to evaluate the hemocoagulation parameters. In patients hospitalized at the ICU, the blood was taken from the central vein (prior to the drawn, the access was flushed with 100 ml of F1/1, the blood was taken from different access than from the one administering heparin), in patients hospitalized at the standard unit the blood was taken from a peripheral vein. The results were then put into the tables established according to the ISTH (International Society on Thrombosis and Hemostasis). Next, the DIC score was calculated to predict severity of hemocoagulation balance disorders in burn trauma, or occurrence of complications during the treatment. RESULTS: Part 1: We have evaluated 12 patients (4 females) hospitalized at the ICU. At the day of injury the overt DIC score reached 1.25 (0-3), fifth day after the injury the average value of overt DIC score was 1.83 (0-3), one day after the autotransplantation it was 2.08 (0-3) and at the day of discharge from the hospital 0. In the deceased patient the DIC score reached value of 2. Part 2: We evaluated 24 patients (10 females) hospitalized at the standard unit. The extent of their burn injury was 5.9% TBSA (0.5-12% TBSA), age 49.13 years (17-94 years). At the day of injury the overt DIC score reached 0.25 (0-2). Fifth day after the injury the average value of overt DIC score was 1.66 (0-2). Day after the autotransplantation it was 1.83 (0-3) and at the day of discharge 0.02 (0-2). CONCLUSION: During the treatment of the patients with various extent of burn injury, none of the patients' scores reached values that would mean occurrence of obvious "overt" DIC.

[HIV/AIDS and pathology in blood count]. / HIV/AIDS a patologie v krevním obrazu.

BACKGROUND: HIV/AIDS pandemy has hit the entire world. With the use of retroviral therapy the disease became chronic. The life prolongation often leads to complications in various organs. The aim of our work was to determine the frequency of blood count pathology at the time when the HIV infection was diagnosed, that means before any antiretroviral treatment was administered, and its relation to the disease stage. METHOD AND RESULTS: Out of 70 patients registered in AIDS centre in Brno University Hospital by 1st March 2006, we have complete blood count results including absolute number of CD4+ lymphocytes. Out of these 70 we evaluated a group of 64 HIV-positive individuals (17 women, 47 men), who were examined at the time the disease was diagnosed. Average and medians of all blood count parameters were within the reference range, only CD4+ lymphocytes in 1 mm(3) were out of range. Pathology in red blood cells count was found 26 times (41.9 % of examined patients), in white cell count 22 times (35.5 %). Thrombocyte number was affected 3 times, and only in one of these cases it was a separate finding. CONCLUSIONS: Blood count pathology is relative frequent in the HIV infected. Therefore HIV infection should be considered as one of the possible causes of unexplained blood count pathology.

[The preparation of a patient with long-term anticoagulant cumarin treatment for invasive surgery]. / Príprava nemocných na dlouhodobé antikoagulacní lécbe kumariny k invazivním zákrokum.

Coumarins belong to drugs widely used and the spectrum of their use is going to grow. From this point of view and/or because the coumarins are adminstrated in patients who are treated for the other diseases--medical or surgical--at the same time, it is necessary to modify, interrupt or replace peroral anticoagulant treatment in the dependence on various aspects. It requires to compound different algorithms for given situations solution. It is always to decide, if the situation is imperative from the view of solution planed, what risk brings proposed treatment and what is the risk of anticoagulant treatment modification.

[Complications of heparin administration]. / Komplikace podávání heparinu.

Heparin as an indirect anticoagulant is used in the clinical practice for more than 60 years. It is an effective drug, but it has limitations. To the side effects belong the heparin resistance, risk of bleeding (haemorrhage), hypersensitivity reactions and heparin-induced osteoporosis. The low-molecular-weight heparins exhibit fewer side effects, but it is still necessary to know about them.

[Molecular study of type 2 von Willebrand disease]. / Molekulární studie von Willebrandovy choroby typu 2.

BACKGROUND: von Willebrand disease is an inherited bleeding disorders caused by mutations in the von Willebrand factor gene. We attempted to characterise the phenotype and the genotype in the first five families in Czech Republic affected by this heterogeneous disorder. METHODS AND RESULTS: The level of FVIII was measured by the one stage assay, the vWF:Ag by the immunoelectrophoresis, vWF:RiCo by aggregometry. For the vWF multimer analysis a western blot based technique was used. The vWF binding to FVIII was evaluated by the ELISA method. Two families were classified as the type 2A, one as the type 2B and two as the combined type 1/2N. Based on that knowledge, parts of the vWF gene were selected for genetic analysis. The previously described mutations Arg1374His and Gly1579Arg were identified in two families with the type 2A. In the family with type 2B a substitution Arg1308Cys was detected. In one family with the type 1/2N, two different previously described defects were found on the separate alleles of the vWF gene: a deletion of cytosine 2435 and a polymorphism Arg854Gln. Compound heterzygotes had the type 1/2N phenotype, while a carriers of the deletion had type 1 phenotype. In the second type 1/2N family, only the amino acid substitutions Thr791Me was found explaining the qualitative defect. A mutation underlying the quantitative deficiency needs to be searched for throughout the entire vWF gene. CONCLUSIONS: Based on the characterisation of the phenotype and genotype, five apparently unrelated families with the von Willebrand disease were diagnosed according to the revised classification. Our work represents laboratory basis for further studies into von Willebrand disease in Czech Republic.

[ABO/H blood groups and factor V Leiden]. / Krevní skupiny AB0/H a faktor V Leiden.

BACKGROUND: Jick and his co-workers published in 1969 an observation showing the prevalence of blood group 0 being lower in people with thrombosis than in the healthy population. During the next years this finding was several times confirmed. The aim of our work was to answer the question on the distribution of blood groups in individuals with thrombophilia compared to the control group and whether the non-0 blood group increases the thrombosis risk in factor V Leiden carriers. We were also interested in mean values of the ProC Global test in different blood groups. METHODS AND RESULTS: Factor V Leiden, prothrombin mutation 20210A and ProC Global were examined in individuals with thrombophilia. The blood group distribution was compared in thrombophilia and control groups. The distribution was statistically different (p = 0.000). The mean value of ProC Global in the non-0 carriers was lower than in persons with blood group 0. CONCLUSION: There is a 1.76 times higher thrombosis risk (1/0.5697 = 1.76) in factor V Leiden carriers with blood group non-0 compared to blood group 0. The ProC Global mean values differ in patients with blood group 0 and in non-0 persons.

[The factor V Leiden mutation in users of hormonal contraceptives]. / Leidenská mutace faktoru V u uzivatelek hormonální antikoncepce.

OBJECTIVE: To assess the frequency of factor V Leiden in oral contraceptive users and to define possible anamnestic data that could predict the presence of factor V Leiden. METHODS: Between 1997 and 1998, 583 users of oral estrogen-progestin contraceptives with no history of thrombotic disease were examined. Factor V Leiden was assessed by PCR after isolating DNA from a peripheral venous blood sample. Among other factors, such things as a family history of thromboembolic disease, myocardial infarction and/or stroke in a first-degree relative were monitored. In 448 users cardiovascular complications were evaluated during six months of oral contraceptive use. The data were analyzed using a MS Excel program. P-values were assessed by pair-tests and chi-square tests. SETTING: 1st Department of Obstetrics and Gynecology of Medical Faculty, Masaryk University, Brno. RESULTS: Factor V Leiden frequency was 6.5% in the study group. There were no differences between carriers and others in age, body weight, body mass index and blood pressure. Carriers had significantly more frequently positive family histories of thromboembolic disease or myocardial infarction or stroke. There were no cardiovascular complications observed in a group of 448 users. The positive family histories of any of the above-mentioned conditions have high specificity (97-99%) and a negative predictive value (0.94) with a low sensitivity (2.6-15.8%) in predicting factor V Leiden presence. CONCLUSION: We found a relatively high incidence of Factor V Leiden among oral contraceptive users without a history of thrombotic disease. Through a positive family history of thromboembolic disease or myocardial infarction or stroke, we can predict a Factor V Leiden presence with high specificity but low sensitivity.

[Molecular basis of hereditary antithrombin defects in 10 Czech families]. / Molekulární podstata vrozeného defektu antitrombinu u deseti ceských rodin.

BACKGROUND: Molecular basis of antithrombin deficiency has not yet been studied in Czech Republic. We looked for the causal mutations throughout the antithrombin gene in 26 patients from 10 unrelated families with antithrombin defect. METHODS AND RESULTS: We screened the gene by conformation sensitive gel electrophoresis and sequenced the mismatched regions using fluorescence technology to characterise mutations and polymorphisms. Mutations were detected in all ten families. Four novel mutations were identified in four families with type I antithrombin defect: Trp-6Arg, 5386-5387delCT, Glu163Stop, and 13246-13248del TGA causing deletion of Glu377 with change of Asn376 to Lys. In other three type I families we found following mutations: splicing site mutation G2777C, Arg197Stop and entire gene deletion. In the family carrying Trp-6Arg mutation antithrombin Vienna (Gln118Pro) was also detected. Leu99Phe recurrent in south-eastern Europe was identified in three families with type II defect. Only the homozygous carries of the mutation were symptomatic, although the heterozygous carries had decreased functional levels. CONCLUSIONS: Four novel mutations in families with type I antithrombin deficiency were characterised. In one family two different genetic defects were identified to be responsible for type I and II phenotypes. Altogether our data agree with the expected heterogeneity of the AT genetic defect.

[Kinetics of hematopoiesis after high-dose chemotherapy and autologous peripheral stem cell transplantation]. / Kinetika obnovy krvetvorby po vysokodávkované chemoterapii a autologní transplantaci periferních kmenových bunek.

BACKGROUND: Autologous peripheral blood stem cell transplantation (APBSCT) is gradually replacing autologous bone marrow transplantation in many clinical settings. The key question is how to evaluate the quality of grafts. We analyzed the relationship between hematopoietic reconstitution and characteristics of patients and grafts. METHODS AND RESULTS: Data from 95 APBSCTs were analyzed. Peripheral stem cells were obtained after mobilization using anti-neoplastic chemotherapy followed by Neupogen (G-CSF). After high dose chemotherapy and APBSCT, patients received Leucomax (GM-CSF). Patients were reinfused with a median of 6.1 x 10(6) (range 0.83-29.3) CD34+ cells/kg, and 25.1 x 10(4) (range 1.0-167.0) CFU-GM/kg of body weight. The median time to engraftment was 12 days (both for granulocytes 1 x 10(9)/l and platelets 50 x 10(9)/l). We found a significant correlation between the number of CD34+ cells and CFU-GM reinfused and also between their respective graft sizes and time to leukocytes, platelets, and granulocytes recovery. We did not find a significant correlation between the number of mononuclear cells reinfused and any analyzed parameter (time to engraftment, age, diagnosis, number of previous chemotherapies, type of mobilization or high-dose regimen). However, administration of preparative high-dose chemotherapy consisted of busulphan and cyclophosphamide was associated with the risk of a transient secondary graft failure. CONCLUSIONS: We conclude that the content of progenitors in PBSC grafts and time to booth leukocyte and platelet recovery are best estimated by the number of CD34+ cells (not less than 1 x 10(6)/kg) and CFU-GM (not less than 1 x 10(4)/kg). The number of mononuclear cells in an autologous PBSC graft is not suitable and useful for prediction of engraftment rate. There is probably no additional benefit of reinfusion of more than 8-10 x 10(6) CD34+ cells/kg and/or 50 x 10(4) CFU-GM/kg, because hematopoietic recovery is not more rapid.