Registry of inherited bleeding disorders in Poland--current status and potential role of the HemoRec database.

We present data collected in HemoRec, an Internet-based platform implemented in 2006 in 15 haemophilia treatment centres in Poland and compare them with the national registry of inherited bleeding disorders established since 1991 at the Institute of Haematology and Blood Transfusion in Warsaw. We also analyse the current status of haemophilia treatment in Poland as well as future perspectives. Data on 1102 patients registered in HemoRec were analysed and compared with 4294 patients in the national registry (status as at 17.08.2009). The number of patients with severe haemophilia, mild/moderate haemophilia and von Willebrand in HemoRec is 530, 328 and 54 (respectively), compared with 1199, 1167 and 1128 in the national registry. The mean age of all haemophilic patients registered in HemoRec is 26.2 years, compared with 37.3 years in the general Polish male population in 2008. The number of haemophilic patients with inhibitor registered in HemoRec is 102 compared with 155 in the national registry (resulting in a prevalence of 14.9% of all severe haemophilia A and 1.6% of all severe haemophilia B patients). HemoRec includes data on a representative group of Polish haemophilic patients, mostly with haemophilia and haemophilia with inhibitor. von Willebrand's disease is largely under-registered in Poland. The survival of Polish haemophilic patients is shorter than that in the general population. The number of inhibitor patients in Poland is relatively large and should be decreased by wider availability of immunotolerance induction in 2010.

Epithelial bone marrow cells in patients with advanced esophageal squamous cell carcinoma.

The aim of the current study was to examine epithelial cells in the bone marrow and peripheral blood of patients with various stages of esophageal squamous cell cancer prior to surgical treatment and to analyze the prognostic significance of these carcinoma cells deposits to the stage of the disease and applied surgical therapy. Thirty-two patients (25 men and 7 women), and 5 healthy bone marrow donors serving as controls were studied. Bone marrow samples were evaluated by light microscopy and examined by flow cytofluorometry. Cells were phenotypically analyzed for the antigens CD45- and CD18+ and/or EMA+. Results are presented as the number of cells revealing the investigated phenotype per 10 (5)analyzed cells. CD18 was expressed in the bone marrow cells of 15 of the 32 (47%) patients and EMA in 20/32 (62%), but not in peripheral blood. In 13 of the 32 pts (41%), co-expression of CD18 and EMA was observed. Patients with the proportion of marrow erythroblasts below 15% had higher numbers of CD18+ and EMA+ cells and there was a negative correlation between the number of erythroblasts and EMA+ cells (r=0.54, p=0.01). In patients with esophageal cancer and anemia, the number of EMA+ cells was higher (p=0.05) and the percentage of erythropoietic cells in the bone marrow was lower (p=0.01). In conclusion, flow cytofluorometry using anti-cytokeratin and anti-EMA antibodies may be useful in evaluating microdeposits of esophageal squamous cells in bone marrow. A dysfunctioning erythropoietic system causing anemia can be a first signal for the presence of malignant cell microdeposits in the marrow of patients with esophageal carcinoma.

Immune Thrombocytopenia and JAK2V617F Positive Essential Thrombocythemia: Literature Review and Case Report.

We present the case where immune thrombocytopenia (ITP) and essential thrombocythemia (ET) sequentially appeared in the space of twenty-one years of follow-up. Impaired platelet production is present in both diseases, but clinical presentation and treatment are different. On the basis of this case history a possible role of autoimmunity as a predisposing factor to myeloproliferation has been discussed.

Stimulation of CFU-Mk colony growth by normal plasma and plasma from myelodysplastic patients.

The ability of plasma from myelodysplastic patients to support the clonal growth of normal megakaryocyte progenitors (CFU-Mk) was compared with that of plasma from normal subjects. The resultant megakaryocyte colonies were expressed as a plasma factor index megakaryocyte (PFI-Mk). All cultures included PHA-LCM and medium conditioned by the human bladder carcinoma cell line 5637, and some of them had EPO. PFI-Mk (MDS) was significantly lower than PFI-Mk (normal), both with and without EPO. A positive correlation was found between megakaryocyte and platelet count in normal subjects, but was not present in MDS patients. There was no correlation between platelet count and PFI-Mk in neither group. In MDS there was a negative correlation between megakaryocyte number and PFI-Mk, both with and without EPO. Although, the mean megakaryocyte number in MDS and in normal bone marrow was similar, the proportion of immature megakaryocytes was much higher in MDS. Previous work indicates an abnormal clonal origin of megakaryocytes in MDS. The present study suggests that abnormal plasma factors affects megakaryocytopoiesis in this condition.

Megakaryocyte progenitors in immune thrombocytopenic purpura (ITP).

Megakaryocyte colony formation was increased in 13 patients with ITP. Out of splenectomized ITP patients Colony Forming Unit-Megakaryocyte (CFU-Mk) number normalized in four but was lower than normal in the remaining two. The proportion of immature megakaryocytes was higher in ITP than in normals or in ITP patients after splenectomy. In conclusion, in ITP accelerated platelet destruction leads to stimulation of megakaryocytopoiesis, but some features of its ineffectiveness can be observed.

Natural killer (NK) activity in normal donors undergoing leukapheresis.

The effect of leukapheresis performed with intermittent flow blood separator on normal donors NK activity is presented. There was statistically significant decrease of NK activity immediately after leukapheresis (p less than 0.001) which did not return to predonation values 1 day after leukapheresis (p less than 0.05), however, none of the donors values fell outside the normal range. It seems that NK cells are not as quickly replaced as T lymphocytes.

Serum thrombopoietin levels in patients with reactive thrombocytosis due to lung cancer and in patients with essential thrombocythemia.

In patients with thrombocytosis normal to increased serum thrombopoietin (TPO) levels have been reported. The aim of this study was to investigate the relationship between serum TPO concentration, platelet number and plasma levels of fibrinogen in patients with reactive thrombocytosis (RT) due to lung cancer and in patients with essential (primary) thrombocythemia (ET). A total of 70 newly diagnosed patients with RT or ET (platelet counts >600 G/l) were studied: 45 with RT due to lung cancer (25 with non-small cell lung cancer, NSCLC and 20 with small cell lung cancer, SCLC), and 25 with ET. Twenty normal volunteers were used as controls. TPO was measured by immunoassay technique (ELISA). Mean serum TPO values in patients with RT due to lung cancer were statistically significantly higher than those in patients with ET or in controls. The highest platelet count was seen in patients with ET, and mean plasma fibrinogen levels were the highest in RT patients. In NSCLC patients mean serum TPO concentrations were higher (not statistically significant) than in SCLC, and a statistically significant relationship between TPO serum concentration and fibrinogen level was observed. No correlations between platelet counts and TPO serum concentrations were found. Our results indicate increased serum TPO levels in patients with thrombocytosis in lung cancer which may be related to the activity of neoplasms. In addition, it is postulated that the relatively low TPO values in patients with ET may result from a dysregulation of the feedback loop involved in platelet production.

[Leukemic meningeal involvement in adults and its treatment]. / Zmiany bialaczkowe opon mózgowo-rdzeniowych u doroslych i leczenie.

The authors report two cases of acute leukaemia in adults with development of sings central nervous system involvement at the time of full haematological remission. In the first case the diagnosis was: acute undifferentiated-cell leukaemia, in the second one: acute lymphoblastic leukaemia. In both cases in the first stage of the disease complete remission of neurological manifestations was obtained; in the second case it was also obtained after the following relapse. In the first case death occurred during successive haematological exacerbation. Ametopterin was given intrathecally or X-ray therapy was applied.

[Susceptibility to roxithromycin of gram-positive cocci isolated from patients with lowered immunity]. / Wrazliwosc na roksytromcyne gram-dodatnich ziarenkowców izolowanych od chorych z obnizona odpornoscia.

The study involved 412 strains of Gram-positive cocci isolated from throat, sputum, urine and stool of patients prepared for bone marrow transplantation at Department of Hematology. Determination of drug susceptibility was performed by disc diffusion method applying discs with roxithromycin, erythromycin, lincomycin, clindamycin and augmentin. Among strains of S. aureus, S. epidermidis and Micrococcus susceptible strains comprised respectively 48, 48 and 45%; among group A, B, C and G streptococci respectively 92 and 52%. Among 108 strains of group D Streptococci only 24% were susceptible to roxithromycin. Comparison of susceptibility of tested strains to roxithromycin and other antibiotics revealed similar susceptibility of staphylococci and streptococci to roxithromycin and erythromycin, while among staphylococci higher percentage of strains and among streptococci lower percentage was susceptible to lincomycin than to roxithromycin.

[The origin of fibrinogen in megakaryocytes and blood platelets]. / Pochodzenie fibrynogenu megakariocytów i plytek krwi.

Fibrinogen is involved in platelet aggregation and clot formation. It is mainly synthesized by hepatocytes. The additional, small pool of this glycoprotein is known to exist in the alpha granules of bone marrow megakaryocytes and blood platelets. Up-date of the origin of fibrinogen in these cells has been discussed.

[Blood serum antiplatelet activity in acute leukemia and idiopathic thrombopenia studied with the aid of the platelet factor 3 availability test]. / Aktywnosc przeciwplytkowa w surowicy chorych na ostre bialaczki i samoistna maloplytkowosc badana za pomoca testu dostepnosci czynnika plytkowego 3.

The effect of the activity present in the globulin fraction of the serum of patients with acute leukaemias and thrombocytopenia on the isologous platelets was investigated. This activity caused, due to increased availability of platelet factor 3, an acceleration of the endogenous clotting pathway with resulting shortening of the clotting time in the test of platelet factor 3 availability. This test was done in 44 patients with acute leukaemia and 19 with thrombocytopenia demonstrating a high-grade shortening of the clotting time as compared with that in a group of 30 healthy controls. In some cases of complete remission of acute leukaemia an effect of this activity on the autologous platelets was demonstrated. The antiplatelet activity was inhibited by rabbit serum against human IgG and Fc fragment of the heavy chain in IgG. These results suggest that the activity present in the globulin fraction of the serum may be one of the causes of thrombocytopenia in acute leukaemia.

[Adrenal function in chronic myeloid leukemia]. / Badania funkcji nadnerczy w przewleklej bialaczce szpikowej

Adrenal function was studied in patients with chronic myeloid leukaemia treated in the past or presently with Busulphan. Adrenocortical function was determined by means of 24-hour profile of 11-hydroxysteroids (11-OHCS) in plasma, and urinary 24-hour 17 hydrocorticosteroids (17-OHCS) and 17-ketosteroids (17-KS). The adrenomedullary function was determined measuring VMA level in 24-hour urine. In most patients normal 24-hour 11-OHCS profiles and 24-levels of 17-OHCS, 17-KS and VMA. Only in some cases these levels were raised. This rise was observed more frequently in patients with blastic crisis of myeloid leukaemia which may indicate that the adrenal reserve was maintained in these patients. The investigations failed to show that the disease itself or Busulphan treatment impaired adrenal function. The possibility of a direct effect of Busulphan on melanin metabolism in the organism is discussed.

[Emperipolesis in megakaryocytes in patients with thrombocytosis in the course of myeloproliferative disorders]. / Emperipoleza w megakariocytach u chorych z nadplytkowoscia w przebiegu zespolu mieloproliferacyjnego.

The incorporation of other marrow cells into megakaryocytes, termed emperipolesis, has been studied in paraffin biopsy sections from 17 untreated patients with myeloproliferative disorders (MPDs). The group consisted of 12 females and 5 males, aged from 34 to 72 years (mean 51.3). Patients with essential thrombocythemia (ET)--9, chronic granulocytic leukemia (CGL)--4, polycythemia vera (PV)--3, and myelofibrosis (MF)--1 were included into the study. Clusters of large polyploid megakaryocytes were observed in anatomic relation to the marrow sinusoidal system. Emperipolesis has been scored as being present or absent per 100 megakaryocytes/slide. Cells found within megakaryocytes were mostly erythroblasts and mature granulocytes. The number of incorporated cells varied from 1 to 7 per one megakaryocyte. Considering the 17 patients with MPDs, emperipolesis was observed in a vast majority of those with ET(8/9) and PV(2/3), in some with CGL(1/4), but not in MF. The mechanism of megakaryocytic emperipolesis remains unclear. Adhesion molecules on megakaryocytes and incorporated cells may possible mediate the cell-to-cell interactions important for emperipolesis.

Human megakaryocytopoiesis--normal and abnormal.

Regulation of megakaryocyte and platelet production remains poorly understood. In culture system two separate activities are needed for maximum production of megakaryocyte progenitors: promotor of clonal expansion and promoter of maturation, other growth factors and cells also contribute to regulation of megakaryocytopoiesis. Increased proliferation of megakaryocytes is observed in myeloproliferative disorders and idiopathic thrombocytopenic purpura, and decreased proliferation is found in aplastic anaemia and hypomegakaryocytic thrombocytopenia. Dysmegakaryocytopoiesis is present in myelodysplastic syndromes and acute leukaemia, and a proliferation of immature megakaryocytes in acute megakaryoblastic leukaemia. Increased understanding of human megakaryocytopoiesis is beginning to help in rational clinical management.

Acquired amegakaryocytic thrombocytopenic purpura (AATP): a study of autologous megakaryocyte progenitors and the effect of patients plasma on normal marrow megakaryocyte colony formation.

Megakaryocyte progenitors (Colony Forming Unit-Megakaryocyte, CFU-Mk) and the effect of plasma on megakaryocyte colony formation in normal human marrow (Plasma Factor Index-Megakaryocyte, PFI-Mk) were studied in six patients with acquired amegakaryocytic thrombocytopenic purpura (AATP) and in ten normal subjects. Assay was based on the method of Messner. In one AATP marrow culture four CFU-Mk were found, in the other two a single CFU-Mk were present, and in the remaining three samples no megakaryocyte colonies were observed. PFI-Mk in AATP patients was significantly higher than in normal subjects. No correlation was found between PFI-Mk and platelet count in either group. The results of this study indicate the presence of an intrinsic defect at the level of CFU-Mk in AATP marrow. PFI-Mk in AATP patients relate to changes in marrow megakaryocyte number rather than to peripheral blood platelet count.

Ability of plasma from patients with immune thrombocytopenic purpura (ITP) to promote the growth of megakaryocyte progenitors from normal bone marrow.

The ability of plasma from ITP patients (before and after splenectomy) to support the growth of megakaryocyte progenitors was compared with that from healthy subjects. Plasma Factor Index-Megakaryocyte PFI-Mk (ITP) which expressed resultant colony growth was significantly lower before splenectomy, but it normalized after splenectomy. (PFI-Mk) (ITP) did not relate neither to megakaryocyte nor to platelet counts. A positive correlation has been observed between megakaryocyte and platelet numbers in healthy subjects and in ITP patients after splenectomy, but not before splenectomy. The proportion of immature megakaryocytes was markedly higher in ITP marrow before splenectomy. This study indicates, that in ITP apart from antibodies directed to platelets and megakarocytes a low plasma stimulatory activity affected megakaryocytopoiesis.