bluuebunny/crossref_metadata_2025 huggingface dataset:
Patient education and communication in non‐inpatient intravenous therapy – a review
Authors: Davies J. M. , Newland A. C. , Finch R. G.
Published: July 2010

Abstract:  This paper reviews patient education and communication elements of non‐inpatient intravenous therapy schemes in the United States and the United Kingdom. Common characteristics of these services include: definition of the components of the scheme at all levels; patient selection; patient education; and good communications. A metaplan carried out among UK healthcare professionals highlighted education and communication issues as important factors for ensuring successful therapy. Communication issues are difficult to define or modify. Specific experience from the United States and the United Kingdom is presented and the common elements in these programmes are emphasized. Results of the assessment of patient education are presented. This collective experience may prove useful for other centres setting up similar therapy schemes.
The management of crisis in sickle cell disease
Authors: Okpala Iheanyi
Published: July 2010

Abstract: The symptoms and signs of sickle cell disease are exacerbated in times of crisis, characterized by tissue infarction or worsening anaemia. Prompt medical intervention is required in these distressing situations to provide relief and comfort to the patient. Effective analgesia is crucial in treating the painful crisis of sickle cell disease. The haemoglobinopathy may cause hyposthenuria with reduced ability to excrete the sodium load in normal saline. A 5% dextrose solution or 5% dextrose in 25% normal saline is therefore recommended for intravenous hydration. As the leading cause of morbidity and mortality in sickle cell disease, infections call for vigorous antibiotic therapy. Oxygen administration should be reserved for hypoxic patients, and blood transfusion given only when really indicated. Acute chest syndrome and cerebrovascular accidents are life‐threatening complications of sickle cell disease whereas priapism can cause important long‐term sequelae; all deserve urgent attention. In the long term, comprehensive care is cost‐effective in reducing the frequency and adverse effects of sickle cell crisis.
Variation in cell yield and proliferative activity of positive selected human CD34+ bone marrow cells along the circadian time scale
Authors: Abrahamsen J. Foss , Smaaland Rune , Sothern Robert B. , Laerum Ole Didrik
Published: July 2010

Abstract: Variations in cell yield and proliferative activity of human bone marrow (BM) progenitor cells were determined with flow cytometry along the 24‐h (circadian) time scale. Equal volumes of BM were aspirated every 5 h, altogether 5 times in 5 healthy men. An average 6‐fold higher yield of positive selected CD34+ cells occurred in each subject when BM was aspirated during the daytime and late afternoon, while a lower yield occurred during the night. Using all CD34+ cell yield data normalized to percentage of mean, a significant time–effect was found by ANOVA (p=0.02) and a significant circadian rhythm was detected by the least‐squares fit of a 24 h cosine (p=0.02). The 95% confidence limits of the acrophase (time of highest values) were computed to be at midday between 10:24 and 14:48 h. A highly significant correlation (p=0.001) was found between proliferation of positive selected CD34+ cells and the more mature myeloid precursor cells from the same BM aspirates, suggesting a common temporal pattern along the circadian time scale. However, no correlation was demonstrated between proliferation and cell yield of CD34+ selected cells, suggesting that mechanisms other than variation in proliferation may cause the circadian rhythm in stem cell yield. These circadian variations in stem cell yield and proliferation suggest that proper timing within 24 h may potentially be important regarding outcome from progenitor cell harvesting and treatment with haematopoietic growth factors.
Lectin‐induced increase in clonogenic growth of haematopoietic progenitor cells
Authors: Vehmeyer Katalin , Hajto Tibor , Hostanska Katarina , Könemann Stefan , Löser Holger , Saller Reinhard , Wörmann Bernhard
Published: July 2010

Abstract: The galactoside‐specific plant lectin, Viscum album agglutinin (VAA‐I) increases cellular parameters of natural host defence. It also binds to a variety of haematopoietic cells, including progenitors. We investigated whether VAA‐I has a stimulatory effect on haematopoietic progenitor cells. Peripheral blood progenitor cells from 7 healthy volunteers were cultured in a colony assay with VAA‐I plus erythropoietin (EPO) and stem cell factor (SCF). At 50 pg/ml VAA‐I induced a significant increase in the cytokine‐dependent clonogenic growth (52% in median, p<0.05). In another set of experiments purified CD34+ cells were isolated from the bone marrow aspirate of 4 patients with non‐metastatic breast cancer using fluorescence‐activated cell sorting. Binding to CD34+ cells was demonstrated by using directly fluorescence‐conjugated VAA‐I. Co‐incubation with d‐galactose significantly abrogated this effect. CD34+ cells were cultured in the presence of EPO, SCF, interleukin‐3, granulocyte/monocyte colony‐stimulating factor and granulocyte colony‐stimulating factor. VAA‐I alone had no measurable effect on the clonogenic growth of the isolated cells. However, at concentrations of 100 and 250 pg/ml VAA‐I increased the cytokine‐dependent proliferation and differentiation of CD34+ cells by a median of 75 and 85%, respectively. The results show that VAA‐I binds to haematopoietic progenitor cells and has a co‐stimulatory effect on their proliferation.
Decreased expression of c‐myc oncoprotein by peripheral blood mononuclear cells in thalassaemia patients receiving desferrioxamine
Authors: Kyriakou D. , Eliopoulos A. G. , Papadakis A. , Alexandrakis M. , Eliopoulos G. D.
Published: July 2010

Abstract: Desferrioxamine (DFX) is an iron chelation agent widely used in the treatment of transfusional iron overload in patients with thalassaemia major and other severe refractory anaemias. DFX has been shown to induce inhibition of DNA synthesis and apoptosis in vitro; however, the molecular targets of DFX action are not well known. The c‐myc proto‐oncogene is involved in a number of cellular processes including proliferation, differentiation and apoptotic cell death. We have examined the expression of c‐myc in peripheral blood mononuclear cells from 71 patients with homozygous β‐thalassaemia in regular transfusion and iron chelation therapy with DFX, 5 non‐transfusion, non‐chelation‐dependent thalassaemic patients, and 15 healthy volunteers using an APAAP immunocytochemical method. We have found that mononuclear cells from thalassaemic patients receiving DFX express significantly lower levels of c‐myc protein compared to control healthy volunteers or thalassaemics receiving no DFX. In vitro treatment of HL60 or K562 leukaemic cells with 100 μl DFX also induced a rapid decrease in c‐myc mRNA and protein levels, followed by apoptosis and inhibition of DNA synthesis. These effects were blocked by simultaneous addition of ferric chloride. Our data suggest that deprivation of cellular iron induces downregulation of c‐myc expression in vitro and in vivo and may influence haemopoietic cell growth and survival.
African iron overload and hepatocellular carcinoma (HA‐7–0–080)
Authors: Moyo Victor M. , Makunike Rudo , Gangaidzo Innocent T. , Gordeuk Victor R. , McLaren Christine E. , Khumalo Hlosukwazi , Saungweme Thoko , Rouault Tracey , Kiire Clement F.
Published: July 2010

Abstract: Although HLA‐linked hemochromatosis greatly increases the risk for hepatocellular carcinoma in people of European ancestry, iron overload in Africa is not thought to be etiologically related to this malignancy. To determine if African iron overload may be associated with hepatocellular carcinoma, we reviewed 320 consecutive diagnostic liver biopsies processed at the University of Zimbabwe from 1992 to 1994 and we selected for analysis 215 biopsies from adults that were suitable for the histological assessment of hepatocellular iron. Subjects were stratified according to hepatocellular iron grades of 0–2+ (normal levels to mild siderosis; n=183) and grades of 3+ and 4+ (distinctly elevated levels consistent with iron overload; n=32). Thirty‐six subjects had hepatocellular carcinoma. Logistic regression modeling revealed a significant association between iron overload and hepatocellular carcinoma after adjustment for age, sex and and the presence of portal fibrosis or cirrhosis (p=0.041). The odds of hepatocellular carcinoma in subjects with iron overload was 3.1 (95% confidence interval of 1.05–9.4) times that of subjects without iron overload. While we could not test for exposure to viral hepatitis or to aflatoxins in this study, our findings suggest that iron overload may be a risk factor for hepatocellular carcinoma in Africa.
Clinico‐prognostic relevance of quantitative immunophenotyping in B‐cell chronic lymphocytic leukemia with emphasis on the expression of CD20 antigen and surface immunoglobulins
Authors: Molica S. , Levato D. , Dattilo A. , Mannella A.
Published: July 2010

Abstract:Expression of CD20, evaluated as antibody binding capacity (ABC) (i.e. absolute number of molecules of antibody per cell), was analyzed using flow cytometry on leukemic cells of 93 previously untreated patients, all fulfilling strict criteria of “immunologically typical” (i.e. CD5+, CD23+) B‐cell chronic lymphocytic leukemia (CLL). Although changes of CD20 antigen density did not correlate with clinical parameters representative of either tumor mass (i.e. clinical stage, histological pattern of bone marrow involvement, absolute peripheral blood lymphocytosis) or disease progression (i.e. lymphocyte doubling time), a trend toward a better life‐expectancy was observed in the low CD20 expression group compared with the high CD20 expression group (p=0.05; relative risk of death, 0.51, 95% confidence interval, 0.24–1.04). Given the correlation between CD20 ABC and mean fluorescence intensity (MFI) of light chain (LC) surface immunoglobulins (Sm Ig) (r=0.481,p<0.0001), as well as the impact of MFI of Sm Ig LC on overall survival (p=0.01; relative risk of death 0.44; 95% confidence interval, 0.10 to 0.76), we tried to verify whether a combination of B‐cell markers, evaluated in a quantitative manner, could have additive prognostic properties. To this purpose we gave a value of 1 or 0 to each B‐cell marker according to whether it was expressed at a low (i.e. CD20 ABC<17.9times103molecules/cell, MFI of LC Sm Ig<100) or high (i.e. CD20 ABC≥17.9times103molecules/cell, MFI of LC Sm Ig≥100) level thus allowing patient stratification into two groups with scores of 2 and 0–1, respectively. Survival of patients who scored 2 was significantly longer than that of patients who scored 0–1 (p=0.02; relative risk of death, 0.44; 95% confidence interval, 0.22–0.72). However, when quantitative changes of CD20 antigen and LC Sm Ig expression, either alone or in combination, were simultaneously analyzed in a Cox model which included usual clinico‐hematological features, only absolute peripheral blood lymphocytosis (p=0.0001) and Binet clinical stages (p=0.0001) maintained their prognostic power unmodified. Although variability of CD20 and Sm Ig expression make it possible to appreciate biological heterogeneity of B‐cell CLL better, however, they cannot substitute well‐established clinico‐hematological features in the prognostic assessment of B‐CLL patients.
Delineation of erythropoiesis in normal and malignant bone marrow using monoclonal antibody AS‐E1 directed against transferrin receptors (CD71)
Authors: Olesen Gitte , Carlsen Ingrid , Skovbo Anni , Hokland Marianne , Hokland Peter
Published: July 2010

Abstract: We have delineated the erythropoietic compartment in normal and malignant bone marrow (BM) by using the monoclonal antibody (mAb) AS‐E1 directed against the transferrin receptor by flow cytometric (FCM) analysis. In normal BM we found a bimodal expression in antigen density with a minor subset (˜3%) expressing AS‐E1high and a larger subset (˜15%) expressing AS‐E1low. By fluorescence activated cell sorting, morphological examination of smears stained by immunocytochemistry and by BFU–E assays the AS‐E1high fraction was shown to contain cells of erythroid origin (proerythroblasts, basophilic erythroblasts and polychromatic erythroblasts), whereas the AS‐E1low fraction consisted mainly of promyelocytes and myelocytes. In patients with malignant hematological disorders we found a more pronounced heterogeneity in the density and the degree of AS‐E1low expression compared to normal BM, and to further characterize the AS‐E1low cells in patients and to exclude that this broad reactivity interfered with the identification of the AS‐E1high cells, we employed triple‐color FCM assays with mAbs directed against the myeloid surface markers CD13 and CD66 in addition to AS‐E1. In all patients we found that 80–90% of the AS‐E1low cells co‐expressed CD13 and/or CD66 and thus were of myeloid origin. Finally, we evaluated 2 methods for determination of the AS‐E1high subset and found an assay involving forward light scatter and logAS‐E1 density to be sufficient. We conclude that AS‐E1high is a valid FCM marker for the normal erythropoiesis.
Intravenous immunoglobulin treatment in hematological diseases
Authors: Otten A. , Bossuyt P. M. M. , Vermeulen M. , Brand A.
Published: July 2010

Abstract: In the last decade large amounts of intravenous immunoglobulin (IVIg) have been used worldwide. Doubts exist as to whether this increased use is paralleled by a comparable growth of reliable data on the therapeutic effectiveness of IVIg. We performed a literature search using MEDLINE from January 1981 to January 1997 and analysed articles on the use of IVIg in hematological disorders and searched for published guidelines. For most hematological disorders, evidence to use IVIg as first line therapy is not very strong. For many disorders no controlled trials have been performed. In published guidelines, IVIg is only recommended, with a few exceptions, when other treatments have failed or are contraindicated. Therefore the increase of consumption of IVIg can not be explained by an increase in established indications in hematology.
FLT‐3 ligand mobilizes hematopoietic primitive and committed progenitor cells into blood in mice
Authors: Ashihara Eishi , Shimazaki Chihiro , Sudo Yoshikazu , Kikuta Takehisa , Hirai Hideyo , Sumikuma Toshiya , Yamagata Noboru , Goto Hideo , Inaba Tohru , Fujita Naohisa , Nakagawa Masao
Published: July 2010

Abstract: We investigated the effects of the administration of FLT‐3 ligand (FL) on mobilization of primitive and committed progenitor cells in mice. C57bI/6J mice were injected subcutaneously with FL once a day for 5 d at doses of 20, 100 and 200 μg/kg. After the collection of peripheral blood, we determined the number of white blood cells (WBCs) with the differential counts. The formation of colony‐forming cells (CFCs) in peripheral blood, bone marrow and spleen was evaluated. Although the administration of FL, 20 μg/kg, did not stimulate leukocytosis, its administration at doses of 100 and 200 μg/kg increased the number of WBC up to 1.7‐ and 2.4‐fold, respectively. Committed progenitor cells were mobilized into the peripheral blood dose‐dependently and the number of CFCs was increased up to 5.5‐fold by the administration of FL at 200 μg/kg on d 5. The number of CFCs in the bone marrow increased, but not dose‐dependently. The number of CFCs in the spleen also increased up to 32‐fold at a dose of 200 μg/kg FL. Mobilized peripheral blood mononuclear cells were transplanted into lethally irradiated mice and the number of CFU–S (d 12) was scored. A dose‐dependent mobilization of CFU–S (d 12) into peripheral blood was also observed. These observations suggest that FL can mobilize hematopoietic primitive and committed progenitor cells into the peripheral blood of mice and those cells mobilized by FL may be applicable to peripheral blood stem cell transplantation.
Paradoxical haemolysis in a patient with cold agglutinin disease
Authors: Ulvestad E.
Published: July 2010

Abstract: A patient with classical cold agglutinin disease initially experienced haemolytic episodes during cold exposure. However, with advancing disease cold‐induced haemolysis ceased and was substituted with a haemolytic disposition at elevated body temperatures. To investigate this paradoxical development of disease manifestations, we performed a clinical and immunological study. Our results indicate that the patient's complement system became exhausted during the later phase of his disease, probably due to continual consumption of complement components. Initially, the patient had slightly decreased C4 concentrations and moderately reduced total haemolytic activity (CH50). Later C4 fell to undetectable levels and CH50 declined to zero. The increased haemolytic activity experienced during febrile episodes is probably due to a cold agglutinin with a high thermal amplitude, combined with enhanced synthesis of complement molecules during the acute phase response. Although C4 concentrations never increased to detectable levels during infections or inflammations an acute phase reaction was determined each time, as evidenced by increased concentrations of CRP. By reconstituting the patient's serum with active complement from donor serum or plasma, increased haemolytic activity was observed. These results indicate that some patients with cold agglutinin disease may experience deleterious haemolytic consequences if transfused with plasma‐containing blood products.
Successful treatment of platelet transfusion refractoriness: the use of platelet transfusions matched for both human leucocyte antigens (HLA) and human platelet alloantigens (HPA) in alloimmunized patients with leukaemia
Authors: Kekomäki S. , Volin L. , Koistinen P. , Koivunen E. , Koskimies S. , Ruutu T. , Timonen T. , Kekomäki R.
Published: July 2010

Six patients, 4 with acute myeloid leukaemia and 2 with a myelodysplasia syndrome who were refractory to random donor platelet transfusions and alloimmunized to human leucocyte antigens (HLA) and human platelet alloantigens (HPA), were treated with HLA‐and HPA‐matched platelet transfusions. In all the patients refractoriness and alloantibodies to HLA as well as HPA‐1b or HPA‐5b were detected simultaneously. Sixty‐seven transfusions (445 units) of HLA‐and HPA‐matched platelets were given and responses to them were, in general, satisfactory in all the patients. No major spontaneous bleeding occurred. Four patients underwent bone marrow transplantation despite alloimmunization. The percentages of platelet transfusion days with a platelet nadir below 20×109/l were 88% for the last 3 random donor platelet transfusions and 39% for the first 3 HLA‐and HPA‐matched platelet transfusions, respectively (p=0.009, Fisher's exact test). Four patients received also HLA‐matched platelets, but responses to them were poor. The small number of transfusions with HLA‐matched platelets precluded comparisons to either the random donor or HLA‐and HPA‐matched platelet transfusions. It seems that HLA‐and HPA‐alloimmunized patients can be successfully supported with HLA‐and HPA‐matched platelet concentrates.
A comparison between intravenous iron polymaltose complex (Ferrum Hausmann®) and oral ferrous fumarate in the treatment of iron deficiency anaemia in pregnancy
Authors: Singh K. , Fong Y. F. , Kuperan P.
Published: July 2010

Abstract: Anaemia is the most common medical disorder in pregnancy with iron deficiency anaemia accounting for the majority of cases. Over 90% of the iron deficiency anaemia is due to red cell iron deficiency associated with depleted iron stores and deficient intake. The two main modalities of treating iron deficiency anaemia are oral or parenteral iron. Ferrous Hausmann® (iron dextrin) is the latest iron preparation which can be used for intravenous parenteral administration as a total dose infusion. This study compares the efficacy of Ferrum Hausmann® with oral ferrous fumarate therapy in the treatment of iron deficiency anaemia in pregnancy. Our study shows that treatment with intravenous Ferrum Hausmann® (iron dextrin) resulted in a significantly better level and rate of increase of haemoglobin (p<0.001). Serum ferritin, which is the best indicator of iron stores, was significantly higher (p<0.001) in the intravenous group. Other indices of iron status such as serum iron, serum transferrin and zinc protoporphyrin also showed a significant improvement in the intravenous group compared to those given oral iron. The results suggest that intravenous iron as a total dose infusion is able to replenish iron stores more efficiently, completely and at a faster rate than oral iron therapy, thus providing the fuel for stimulation of full erythopoiesis compared to oral iron. There were also no reports of any adverse reactions with intravenous iron dextrin, whereas there were a considerable proportion of women on oral iron therapy who reported side effects. In conclusion, intravenous iron therapy with Ferrous Hausmann® (iron dextrin) is a suitable, effective and safe alternative to oral iron therapy in the treatment of iron deficiency anaemia in pregnancy.
Expression of activation markers CD23 and CD69 in B‐cell non‐Hodgkin's lymphoma
Authors: Erlanson M. , Grönlund E. , Löfvenberg E. , Roos G. , Lindh J.
Published: July 2010

Abstract: The activation associated proteins CD23 and CD69 are expressed on cells of different lineages upon mitogenic stimulation. CD23 is a well characterized multifunctional protein in lymphocyte development recognized as a diagnostic marker for chronic lymphocytic leukaemia. CD69 is one of the earliest markers expressed after activation of T cells, but its function is unclear. The aim of this study was to investigate the expression of these antigens in B‐cell non‐Hodgkin's lymphomas (NHL) in relation to clinical behaviour. Ninety samples from 84 patients with NHL of B cell type were studied for the expression of CD23 and CD69 in CD20+ B cells by flow cytometric dual parameter analysis. In individual lymphomas the CD23 and CD69 antigens showed an “on or off” pattern with most or very few cells positive for each antigen. The CD23 antigen was expressed in 23 of 53 (43%) indolent lymphomas and in 2 of 37 (5%) aggressive cases. Most indolent lymphomas (81%) and about half the aggressive cases (53%) expressed the CD69 antigen. Thus, both markers were associated with indolent type. CD23 expression correlated with chronic lymphocytic leukaemia subtype and CD69 expression with male gender, advanced stage, newly diagnosed lymphoma and shorter survival.
Expression of dipeptidylaminopeptidase IV/CD26 in peripheral blood lymphocytes of hemophilic subjects
Authors: Invernizzi R. , Montani N. , Giusto M. , Mosconi E. , Lorenzutti F. , Comolli G. , Pecci A. , Gamba G.
Published: July 2010

Abstract: CD26 antigen, a 110 kDa membrane glycoprotein with exopeptidase activity (DAP IV), is an activation marker of T lymphocytes preferentially expressed on CD4+ memory cells and involved in T cell proliferation and IL‐2 production after antigenic stimulation. We employed cytochemical and immunocytochemical techniques to study DAP IV/CD26 expression in circulating lymphocytes from 40 hemophilic patients, chronically treated with coagulation factors, in order to verify the possible involvement of this molecule in the immunological alterations of hemophilia. In all the hemophiliacs DAP IV activity was significantly lower than in the controls, independently of the quantity of blood transfused and previous exposure to viruses. This reduction may be responsible for the impaired proliferative response of lymphocytes to antigens and mitogens, notoriously observed in hemophilia. Whereas in the group of HIV‐ patients CD26 expression was similar to that of normal controls, in the 8 HIV+ hemophilic patients both percentages of positive lymphocytes and intensity of staining were significantly lower. In only 4 of the 8 cases was this deficit associated with CD4+ cell depletion. The significant selective loss of CD26 expression observed in HIV+ patients is probably an early event after HIV infection and seems to occur even before CD4 cell depletion. In conclusion, evaluation of DAP IV/CD26 might be a useful option for monitoring the immunological alterations of all hemophilic patients, HIV positive or not, chronically treated with coagulation factors.
Circulating CD34+ cells in cord blood and mobilized blood have a different profile of adhesion molecules than bone marrow CD34+ cells
Authors: Asosingh K. , Renmans W. , Van der Gucht K. , Foulon W. , Schots R. , Van Riet I. , De Waele M.
Published: July 2010

Abstract: The expression of adhesion molecules was studied on CD34+ hematopoietic precursors in cord blood, bone marrow and mobilized blood. The samples were labeled in a double immunofluorescence procedure with a CD34 monoclonal antibody and with antibodies against maturation and differentiation antigens and adhesion molecules. Myeloid precursors formed the majority of the CD34+ cells in all samples. In bone marrow a separate cluster of B‐cell precursors with low forward scatter was present. Nearly all CD34+ cells in normal bone marrow expressed VLA‐4 and VLA‐5, PECAM‐1, LFA‐3 and HCAM. The majority of the CD34+ cells also had LFA –1 and L‐selectin on the surface membrane. A small subset was VLA‐2, VLA‐3, ICAM‐1 or Mac‐1 positive. CD34+ cells expressing the vitronectin receptor or the CD11c antigen were rare. Cord blood and mobilized blood CD34+ cells had a lower expression of VLA‐2, VLA‐3 and VLA‐5 and a higher expression of LFA‐1, ICAM‐1 and L‐selectin than bone marrow CD34+ cells. Except for LFA‐1, this was not due to the presence of more myeloid precursors in these samples. Low β1 integrin expression may lead to less adhesion to the extracellular matrix. High expression of L‐selectin may facilitate interaction with endothelial cells. Therefore, this phenotype may favour mobilization.
Inv(16) acute myeloid leukemia cells show an increased sensitivity to cytosine arabinoside in vitro
Authors: Tosi Patrizia , Visani Giuseppe , Ottaviani Emanuela , Testoni Nicoletta , Pellacani Annalisa , Tura Sante
Published: July 2010

Abstract: Karyotype represents the major independent prognostic factor for response and remission duration in acute leukemia. In particular, it has been reported that acute myeloid leukemia (AML) patients with inv(16) abnormality show a better prognosis, especially in case of treatment with high‐dose Ara‐C (HD Ara‐C) containing regimens. In this study we aimed at testing whether leukemic cells from patients showing the inv(16) were more sensitive to Ara‐C in vitro, compared to AML blasts from patients with normal karyotype or chromosomal abnormalities other than t(15;17) or t(8;21). We analyzed blast cells from 30 patients who were diagnosed and treated in our institution. The IC50 of Ara‐C, as tested by the XTT colorimetric assay, was significantly lower in cases with inv(16) (18.5±15.88 μmol/l vs. 38±14.6 μmol/l, in cases with other abnormalities, p = 0.01). This result was confirmed by a higher incorporation of [3H]‐Ara‐C into DNA (p = 0.02 and p = 0.001 compared to samples with normal and abnormal karyotype, respectively). All the same, Ara‐C induced apoptosis was significantly increased in cells from patients with inv(16). Our data suggest a possible interaction between the molecular background of inv(16) and a modification of intracellular metabolism of Ara‐C, and could thus provide a rationale for HD‐Ara‐C‐based schedules for patients with inv(16) AML.
Methyl‐GAG, ifosfamide, methotrexate and etoposide (MIME) as salvage therapy for Hodgkin's disease: a prospective study
Authors: Enblad Gunilla , Hagberg Hans , Gustavsson Anita , Glimelius Bengt
Published: July 2010

Abstract: This study presents the results of a prospective study of methylgag, ifosfamide, methotrexate and etoposide (MIME) as salvage regimen for Hodgkin's disease (HD) in Sweden. Sixty‐four patients with recurrent or refractory HD were treated with MIME between July 1988 and December 1993. All patients except one had, earlier, been treated with and failed consecutive or alternating MOPP and ABVD. Median age was 37 yr (range 14–73). Twenty patients (31%) achieved a complete remission (CR) and 17 (27%) a partial remission (PR), giving an overall response rate of 58%. The 5‐yr survival for all patients was 43%. In a multivariate analysis, the most important factors predicting a poor survival were the presence of extranodal disease at relapse, male gender and high age. Twenty‐nine patients were treated with high‐dose chemotherapy with stem‐cell rescue after MIME. Those patients had a similar survival compared to the patients responding to MIME but not treated with high‐dose chemotherapy. We conclude that MIME induces remissions in a high proportion of patients with recurrent and refractory HD with acceptable toxicity. The remissions probably need consolidation, but the nature of this consolidation is still controversial.
Granulocyte colony‐stimulating factor and drugs elevating extracellular adenosine synergize to enhance haematopoietic reconstitution in irradiated mice
Authors: Pospíšil M. , Hofer M. , Znojil V. , Netíková J. , Vácha J. , Holá J. , Vacek A.
Published: July 2010

Abstract: The activation of adenosine receptors has recently been demonstrated to stimulate haematopoiesis. In the present study, we investigated the ability of drugs elevating extracellular adenosine to influence curative effects of granulocyte colony‐stimulating factor (G‐CSF) in mice exposed to a sublethal dose of 4 Gy of 60Co radiation. Elevation of extracellular adenosine in mice was induced by the combined administration of dipyridamole, a drug inhibiting the cellular uptake of adenosine, and adenosine monophosphate (AMP), an adenosine prodrug. The effects of dipyridamole plus AMP, and G‐CSF, administered either alone or in combination, were evaluated. The drugs were injected to mice in a 4‐d treatment regimen starting on d 3 after irradiation and the haematopoietic response was evaluated on d 7, 10, 14, 18 and 24 after irradiation. While the effects of G–CSF on the late maturation stages of blood cells, appearing shortly after the completion of the treatment, were not influenced by dipyridamole plus AMP, positive effects of the combination therapy occurred in the post‐irradiation recovery phase which is dependent on the repopulation of haematopoietic stem cells. This was indicated by the significant elevation of counts of granulocyte‐macrophage progenitor cells (GM‐CFC) and granulocytic cells in the bone marrow (d 14), of GM‐CFC (d 14), granulocytic and erythroid cells (d 14 and 18) in the spleen, and of neutrophils (d 18), monocytes (d 14 and 18) and platelets (d 18) in the peripheral blood. These effects suggest that the repopulation potential of the combination therapy lies in a common multilineage cell population. The results of this study implicate the promising possibility to enhance the curative effects of G–CSF under conditions of myelosuppressive states induced by radiation exposure.
Circadian‐based effects of AcSDKP, with or without rhG–CSF on hematologic toxicity of chemotherapy in mice
Authors: Li Xiao‐Mei , Soulard Chantai , Filipski Elisabeth , De Paillette Evelyne Deschamps , Lévi Francis
Published: July 2010

Abstract: The hematologic toxicity of arabinosylcytosine (Ara‐C) and carboplatin (CBDCA) as well as the stimulating effect of recombinant human granulocyte colony‐stimulating factor (rhG–CSF) on murine bone marrow vary according to their dosing time along the 24‐h time scale. In the present study, we investigated whether the tolerability of Ara‐C or CBDCA, given at their least toxic circadian time, could be improved further with AcSDKP, a negative regulator of hemopoiesis, rhG–CSF or both. A total of 228 B6D2F1 mice received once‐daily injection of either Ara‐C (42 mg/kg/d s.c.) for 7 d (d 0–6) at 8 hours after light onset‐HALO) or CBDCA (40 mg/kg/d i.p.) for 5 d (d 2–6) at 16 HALO. AcSDKP (24 μg/d) was continuously infused for 7 d (d 0–6), using an osmotic minipump. rhG–CSF (400 μg/kg/d s.c.) was injected for 4 d (d 9–12) at 9 HALO. Subgroups of mice were sacrificed at 3 HALO on various days following treatment. AcSDKP significantly increased CFU–GM count on d 7 and leukocyte, neutrophil and monocyte counts on d 13 and d 16 compared to Ara‐C alone. Also, rhG–CSF produced similar protective effects to those of AcSDKP with regard to leukocyte and CFU–GM counts. The combination of AcSDKP with rhG‐CSF induced a further increase in total leukocytes and their subsets as compared to either agent alone, but did not alter the CFU–GM counts. Neither AcSDKP nor rhG–CSF nor their combination reduced CBDCA‐induced hematological toxicity. In conclusion, AcSDKP or rhG–CSF administration further improved the tolerability of Ara‐C beyond that already achieved with optimal circadian timing, while no such effect was observed in mice receiving CBDCA at the dose used. The results warrant further exploration of chronopharmacologic delivery schedules combining Ara‐C with AcSDKP.
Modulatory function on autologous myeloid progenitor cells of clonal T‐lymphocytes following autologous bone marrow transplantation
Authors: Hokland Peter , Carlsen Ingrid , Hokland Marianne , Nielsen Bendt
Published: July 2010

Abstract: We have studied the regulatory capacity of clonal T‐lymphocytes from patients undergoing autologous bone marrow transplantation (ABMT) on the generation of CFU–GM from their harvest marrow preparations. To this end, T‐lymphocytes from peripheral blood from 5 patients undergoing ABMT isolated 10 d before and 7, 14 and 28 d post‐ABMT were placed in limiting dilution conditions (384 wells for each patient at each time point) and polyclonally stimulated. From more than 1600 wells with growth from the 5 patients, preparations from more than 900 wells could be expanded (range between patients 33–452) and identified by immunophenotyping (IP) and flow cytometry (FCM) by their exclusive expression of CD4 or CD8. This was significantly fewer than seen in normal donors, especially so at d 7 and 14 post‐ABMT. The ratio between CD4+ and CD8+ clones varied between 0.6 and 2.8 (median 1.3) and was significantly lower in the patients compared to normal donors (median 3.1; range 3.0–6.5). When the clonal T‐cell preparations were co‐cultured with autologous bone marrow cells obtained at the time of harvest and depleted for T‐lymphocytes, the vast majority of both CD4+ and CD8+ clones exerted a clear enhancement on the CFU–GM growth with no relation to time of blood sampling in its the magnitude. Moreover, a trend seen in the normal donors towards CD4+ clones being more effective in this enhancement was not observed in ABMT patients. We conclude that clonal T‐cells from ABMT patients, irrespective of their phenotype and time of isolation, exert an enhancement on the growth of autologous CFU–GM, which is equal to that seen in normal donors.
Analysis of the granulocyte colony‐stimulating factor receptor gene structure using PCR‐SSCP in myeloid leukemia and myelodysplastic syndrome
Authors: Shibata Shoichiro , Asano Yoshinobu , Yokoyama Toshihiro , Shimoda Kazuya , Nakashima Hitoshi , Okamura Seiichi , Niho Yoshiyuki
Published: July 2010

Abstract: The membrane‐proximal cytoplasmic region of the granulocyte colony‐stimulating factor receptor (G–CSFR) is known to be essential for the proliferation signal, with a more distal region being required for the differentiation signal. Such a separation of functional domains raises the possibility that mutations occurring at these regions may contribute to cell proliferation in the absence of differentiation, this being the most important characteristic in acute leukemia cells. Therefore, we analysed the structural abnormalities at the transmembrane and cytoplasmic region of G–CSFR in a significant number of patients with various myeloid malignancies. When we examined the genomic DNA of G–CSFR obtained from 41 patients with acute myelogenous leukemia (AML), 18 with chronic myelogenous leukemia (CML), 7 with myelodysplastic syndrome (MDS), 2 with chronic myelomonocytic leukemia and 1 with chronic neutrophilic leukemia, we found a polymorphism in 3 patients, but no significant pathogenic mutations in any patients. The screening for this polymorphism in 100 hematologically normal controls revealed that it may be useful as a linkage marker for population and family studies, because the heterozygosity index is at a high level (0.055). While there have been several reports discussing the leukemogenic potential of mutations in the cytokine/hematopoietin receptor superfamily, genetic alterations in the transmembrane and cytoplasmic region of G–CSFR do not seem to play a pathogenic role in leukemia.
Characterization of the ETO and AML1–ETO proteins involved in the 8;21 translocation in acute myelogenous leukemia
Authors: Le Xiao‐Feng , Claxton David , Kornblau Steve , Fan You‐Hong , Mu Zhao‐Mei , Chang Kun‐Sang
Published: July 2010

Abstract: The AML1 and ETO genes are disrupted by the nonrandom chromosomal translocation t(8;21) in acute myelogenous leukemia (AML). While the AML1 gene encodes a transcription factor indispensable for definitive hematopoiesis, the biological function of ETO is unknown. To understand the role of ETO and AML1–ETO in the pathogenesis of AML, the full length cDNAs of ETO and AML1–ETO were cloned and antibodies against AML1 and ETO proteins have been developed in our laboratory. Western blot analysis showed that ETO and AML1–ETO were identified as 70 kDa and 94 kDa proteins, respectively, and that both proteins, like AML1, were associated with the nuclear matrix. To examine whether the t(8;21)‐positive AMLs expressed a 94‐kDa AML1–ETO, protein fractions isolated from leukemia blasts of 10 patients with t(8;21)‐positive AML and the Kasumi‐1 cells were analyzed by Western blotting. The 94 kDa AML1–ETO fusion protein was detected in all samples. However, this fusion protein was not detectable in all 40 patients with t(8;21)‐negative AMLs. The biological significance of AML1–ETO was examined in K562 cells, which stably overexpress AML1–ETO. We found that AML1–ETO blocked the erythroid differentiation of K562 cells induced by low doses of Ara‐C. Thus, t(8;21)‐positive AMLs appear to overexpress the AML1–ETO fusion protein, which may be responsible for differentiation block and leukemogenesis in AML.
Early circulating erythroid progenitor cells and expression of erythropoietin receptors in sickle cell disease
Authors: Perlingeiro R. C. R. , Costa F. F. , Saad S. T. O. , Queiroz M. L. S.
Published: July 2010

Abstract: The ability of circulating progenitor cells from 22 patients with sickle cell disease (SCD) to develop erythroid colonies was studied in vitro in the presence or absence of growth factors (5637‐CM and erythropoietin). In both conditions, SCD patients presented significantly higher numbers of circulating burst‐forming unit‐erythroid (BFU–E/5×105MNC) when compared to control subjects. The study of the expression of erythropoietin receptors revealed an increased level in SCD patients. Moreover, there was a correlation between both stimulated and autocrine (without stimulus) BFU–E and the expression of erythropoietin receptors. These results are of particular interest since they indicate that the phenomenon of spontaneous BFU–E‐derived colonies observed in SCD patients may be due to an increased expression of erythropoietin receptors.
Antibody‐directed superantigen‐mediated T‐cell killing of myeloid leukaemic cell line cells
Authors: Gidlöf Cecilia , Carlson Barbro , Dohlsten Mikael , Tötterman Thomas H.
Published: July 2010

Abstract: Bacterial superantigens (SAgs) bound to MHC class II molecules on target cells are efficient activators of cytotoxic T cells expressing certain T cell receptor (TCR) Vβ regions. We described earlier that the specificity of the SAg Staphylococcus enterotoxin A (SEA) can be changed by introducing a D227A point mutation in the major MHC class II binding site and by genetically fusing the SEA mutant (SEAm) to protein A (PA). This SEAm‐PA fusion protein can then be used to direct cytotoxic T cells to tumour cells coated with monoclonal antibodies (mAbs). In this communication, we tested the PA‐SEAm fusion protein together with mAbs against the myeloid cell surface antigens CD13, CD15 and CD33. A SEA‐reactive T cell line was used as effector cells against 10 different myeloid leukaemic cell lines. Optimal lysis of antigen positive leukaemic cells was obtained at a PA‐SEAm concentration of 1 ng/ml and effector: target cell ratios of 15 : 1. No correlation between target cell sensitivity and the level of surface antigen expression could be seen. The 6 acute myeloid leukaemia (AML) cell lines tested appeared to be more sensitive than the 4 chronic myeloid leukaemia (CML) cell lines. The sensitivity of the AML cell line HL‐60 could be improved further by stimulation with TNFα. This was accompanied by increased surface ICAM‐1 expression whereas specific target molecule expression (CD13, CD33) was unchanged. This suggests that sensitivity to lysis is related to the leukaemic subtype and ICAM‐1 expression but not to the tumour antigen density. Our results show that it is possible to direct cytotoxic T cells to myeloid leukaemia cells by using SAgs linked to mAbs, and encourage the construction and testing of a recombinant direct SAg‐mAb fusion protein as a candidate drug for therapy of myeloid leukaemias.
Comparable TNF‐alpha, IFN‐gamma and GM–CSF production by purified normal marrow CD3 cells in response to horse anti‐lymphocyte and rabbit antithymocyte globulin
Authors: Piaggio G. , Podestá M. , Pitto A. , Pittaluga G. B. , Isaza A. , Benvenuto F. , Bruno B. , Bacigalupo A.
Published: July 2010

Abstract: In vitro priming of T cell with horse antilymphocyte globulin (HALG) results in cytokine release, and this has been associated with its clinical efficacy in patients with severe aplastic anaemia (SAA). Rabbit antithymocyte globulin (RATG) has been studied less extensively. In this study we compare the in vitro priming effect of HALG and RATG on purified normal marrow T cells: end‐points of the study were 1) levels of TNF‐alpha (TNF‐α), IFN‐gamma (IFN‐γ) GM–CSF in T cell supernatants, and 2) effect of T cell supernatants on colony formation with or without exogenous GM–CSF. TNF‐α, IFN‐γ and GM–CSF levels were comparable for HALG, RATG and phytohaemagglutinin (PHA). T cell supernatants showed comparable enhancement of colony formation in the presence of recombinant human GM–CSF (rhGM–CSF) and supported colony forming unit granulomacrophage (CFU–GM) growth in the absence of growth factor. This study shows that horse and rabbit derived ALG/ATG and PHA have a comparable in vitro priming effect on T cells: both agents should probably be tested for their clinical efficacy in SAA patients.
The estimation of efficacy of oral iron supplementation during treatment with epoetin beta (recombinant human erythropoietin) in patients undergoing cardiac surgery
Authors: Sowade Olaf , Messinger Diethelm , Franke Werner , Sowade Birgit , Scigalla Paul , Warnke Harry
Published: July 2010

Abstract: We estimated the efficacy of oral iron therapy during treatment with rhEPO in patients undergoing cardiac surgery who were contraindicated for autologous blood donation. Seventy‐six patients were enrolled in this double‐blind, placebo‐controlled trial and assigned to the 2 treatment groups (5×500 U/kg body weight rhEPO or placebo intravenously over 14 d before surgery). During the treatment period all patients received 300 mg Fe2+ (iron glycine sulfate) orally per day. rhEPO therapy produced significant increases in hemoglobin concentration (Hb), reticulocyte count, hematocrit (Hct) and the hypochromic red blood cells (HRBC), and a decrease in transferrin saturation (41%) compared to the placebo group before surgery. However, the preoperative increase in HRBC was independent of the baseline ferritin and even correlated positively with the preoperative increase in Hct (r = 0.47, p < 0.01). In rhEPO patients there were inverse correlations between baseline serum iron and the preoperative increases in Hb (r = –0.39, p < 0.05), Hct (r = –0.50, p < 0.01) and HRBC (r = –0.53, p < 0.001). With this treatment regimen the HRBC appear to reflect the degree of erythropoietic stimulation rather than functional iron deficiency. The preoperative increases in reticulocytes, HRBC and Hb/Hct in patients with ferritin < 100 mg/l or transferrin saturation < 16% showed no significant difference compared to their complementary groups. The preoperative decrease in storage iron and the inverse correlation between the baseline ferritin and the preoperative change in ferritin (r = –0.94, p < 0.0001) in the rhEPO group indicate that the iron requirement for hemoglobin synthesis is probably covered by the breakdown of stored iron and an increase in the rate of absorption of orally administered Fe2+. Intravenous rhEPO treatment with 5×500 U/kg body weight in combination with 300 mg oral Fe2+/d given over 14 d before surgery is a suitable regimen to increase Hb by about 1.61 g/dl and Hct by 0.06.
Long‐term therapeutic efficacy and toxicity of recombinant interferon‐alpha 2a in polycythaemia vera
Authors: Foa P. , Massaro P. , Caldiera S. , LaTargia M L. , Iurlo A. , Clerici C. , Fornier M. , Bertoni F. , Maiolo A. T.
Published: July 2010

Abstract: We report on long‐term therapeutic efficacy and toxicity of recombinant interferon‐alpha 2a (rIFN‐alpha) in a series of 38 patients with polycythaemia vera (PV). In all patients haematocrit was first brought into the normal range by venesection; rIFN‐alpha was then begun at a starting weekly dose of 9,000,000 IU. Complete response (CR) was defined as persistence of normal haematocrit without venesection and partial response (PR) as >50% reduction of phlebotomy requirement. Eleven patients (28.9%) achieved CR and 8 (21.0%) PR. Median duration of treatment for all responsive patients was 40 months; 12 patients are still responsive and under treatment after 13, 15, 25, 35, 40, 41, 43, 49, 50, 51, 52 and 52 months of therapy with rIFN‐alpha. In responsive patients, rIFN‐alpha also normalized leucocyte counts, platelet counts and spleen enlargement; rIFN‐alpha also relieved generalized pruritus in all 10 patients displaying this symptom. Early toxicity (flu‐like syndrome) was observed in 23.6% and late toxicity (severe weakness) in 13.1% of patients, requiring rIFN‐alpha treatment suspension in all cases. Progression to leukaemia was observed in none of the 10 patients treated only with rIFN‐alpha and in one of the 12 who received alkylating agents before enrolment in this study. According to these data, rIFN‐alpha seems to be an effective and safe treatment option for PV.