bluuebunny/crossref_metadata_2025 huggingface dataset:
An effective age‐unrestricted m‐AMSA‐based second‐line regimen for poor prognosis acute myeloid leukaemia
Authors: Watson A. M. , Seymour J. , Lee N. , Whiteside M. , Firkin F.
Published: July 2010

Abstract:  The efficacy and toxicity of a regimen consisting of amsacrine (m‐AMSA), cytarabine, and thioguanine for remission‐induction therapy in poor prognosis categories of acute myeloid leukaemia (AML) were determined in a single arm study of 46 patients. The study group consisted of 17 patients with disease refractory to daunorubicin plus cytarabine‐based induction regimens, 22 patients with disease which had relapsed during daunorubicin plus cytarabine maintenance therapy, or following completion of this maintenance programme after receiving > 500 mg daunorubicin/m2, and 7 previously untreated patients where cardiac disease contraindicated anthracycline therapy. Complete remission (CR) was attained in 46%, and probability of survival was comparable to published results for first‐line treatment with daunorubicin plus cytarabine regimens. There was no statistically significant difference in CR rate or probability of survival between these three categories of poor prognosis AML, and cardiotoxic complications were uncommon despite extensive anthracycline exposure in the majority. In the 43% of patients who were 60–76 years of age, there was no statistically significant difference in CR rate or probability of survival relative to patients <60 years. This observation fails to support the view that less myelotoxic regimens with lesser efficacy should be the basic approach to treatment of AML in patients > 60 years of age.
Effect of pyrexia in the formation of intraerythrocytic inclusion bodies and vacuoles in haemolytic crisis of haemoglobin H disease
Authors: Chinprasertsuk Sriprapa , Wanachiwanawin Wanchai , Piankijagum Anong
Published: July 2010

Abstract:  Clinical aspects and in vivo formation of inclusion bodies and vacuoles in red cells of patients with haemoglobin H disease were studied and compared during steady state and after pyrexia of acute infections. During period of pyrexia, there were haemolytic crises with significant drops of the steady‐state haemoglobin levels of 8.6 ± 0.9 g/dl to 5.6 ±1.7 g/dl. The phenomenon was observed with splenic enlargement and significant increase of inclusion body formation. The findings suggested that inclusion body formation in red cells during pyrexia of acute infections played an important role in haemolytic crisis of haemoglobin H disease, probably by causing these cells to be sequestered and destroyed in the spleen.
Molecular genetic studies of two families with X‐linked chronic granulomatous disease: Mutation analysis and definitive determination of carrier status in patients' sisters
Authors: Ariga Tadashi , Sakiyama Yukio , Furuta Hirofumi , Matsumoto Shuzo
Published: July 2010

Abstract:  Molecular genetic studies of two families with X‐linked chronic granulomatous disease (X‐CGD) were performed. The patients showed abnormal patterns on Southern blot analysis using cytochrome b heavy chain (CYBB) cDNA as a probe. Both patterns differed and neither has ever been observed in normal individuals. We applied the results to the diagnosis of the carrier state in the patients' sisters. The results clearly demonstrated that each patient's sister possessed the same abnormal allele as the patient's CYBB gene, as detected by Southern analysis. Thus, the results confirm that both of the patients' sisters are carriers of the disease. Further molecular analysis of the patients' mutation revealed that they were a point mutation, and a partial deletion of the CYBB gene, respectively. These mutations have not previously been reported.
Involvement of HIV nef protein in abnormal hematopoiesis in AIDS: in vitro study on bone marrow progenitor cells
Authors: Calenda V. , Graber P. , Delamarter J‐F. , Chermann J‐C.
Published: July 2010

Abstract:  In previous studies (1–4), we described disrupted hematopoiesis in long‐term human bone marrow cultures infected with HIV. Conditioned medium (CM) from HIV‐1 non‐productively infected liquid cultures inhibits the proliferation of granulo‐monocytic progenitor cells (CFU‐GM) in clonogenic assays. A growth‐inhibitory soluble factor was therefore suspected and we report here that this factor has been identified as the nef gene product. These nef‐containing supernatants are specifically neutralized by anti‐nef antibodies and recombinant nef has the same growth inhibitory properties. We used a nef‐deficient virus to confirm that nef protein is responsible for the in vitro growth inhibition of CFU‐GM.
Chronic lymphocytic leukemia: A retrospective study of 122 cases
Authors: Erlanson Martin , Osterman Birgitta , Jonsson Håkan , Lenner Per
Published: July 2010

Abstract:  During the period 1978–1982 in the three northernmost countries of Sweden all 143 patients with a registered diagnosis of chronic lymphocytic leukemia (CLL) were retrospectively analyzed. After re‐evaluation, 122 cases remained in the study. The mean age was 71 years and the male/female ratio was 2.2:1. Sixty‐one patients were Binet stage A, 29 stage B and 32 stage C. The diagnosis CLL was made after routine check‐up for other diseases in most of the patients and they had no symptoms from the CLL. The median survival was 51 months and there were no differences in crude survival according to stage or other prognostic factors such as hemoglobin, lymphocytes or thrombocytes in peripheral blood. Analysis of CLL as a cause of specific mortality showed the stage of CLL to have a slight prognositic significance. This could be due to the fact that many of the patients suffered from other serious diseases, allowing the detection of early stage or advanced CLL with no symptoms.
Four‐chromosomes complex translocations in acute promyelocytic leukemia: Description of two cases
Authors: Zaccaria .Alfonso , Testoni Nicoletta , Martinelli Giovanni , Pelliconi Susanna , Buzzi Marina , Farabegoli Patrizia , Naldi Stefano , Salvucci Marzia , Tura Sante
Published: July 2010

Abstract:  Two cases of acute promyelocytic leukemia with variant translocations involving 4 chromosomes are described. The karyotypes were 47,XX, + 8,t(13;15;17;20)(q22;q22;q12;q13) and 46,XY,t(5;15;16;17)(q22;q22;p13;q12), respectively. Variant translocations in APL apparently do not follow any preferential routes since no recurrent breakpoint additional to those of chromosomes 15 and 17 has been found in any of the cases reported in the literature and in those described here. Moreover, it seems that the translocation of the RARα gene from chromosome 17 to chromosome 15 is directly involved in the pathogenesis of the disease, while the reciprocal one is not, as demonstrated by variant translocations where 15q migrates to chromosomes other than 17.
Acute leukaemia after hydroxyurea therapy in polycythaemia vera and allied disorders: Prospective study of efficacy and leukaemogenicity with therapeutic implications
Authors: Weinfeld Aleksander , Swolin Birgitta , Westin Jan
Published: July 2010

Abstract:  Fifty consecutive patients, 30 of whom had polycythaemia vera (PV), 10 essential thrombocythaemia (ET), and 10 myelofibrosis (MF), entered a long‐term prospective study of hydroxyurea (HU) therapy. The indication for treatment was mainly thrombocytosis or symptomatic splenomegaly. Control of erythrocytosis and thrombocytosis was achieved in 70% of the patients. Continuous maintenance treatment was required. In 15% of responding patients with thrombocytosis, unexpected rises of the platelet count occurred during maintenance therapy. Severe thrombo‐embolic events occurred in 6 patients. The size of the spleen decreased in all patients who did not develop thrombocytopenia and could absorb adequate HU doses. Acute leukaemia (AL) was diagnosed in 9 patients and a myelodysplastic syndrome in one. Seven of them had been treated with HU alone. Among the patients with PV and ET, 6 developed AL and 4 of them were treated with HU alone (3 PV and 1 ET), giving an incidence of 10.5%. In previously untreated patients with initially normal karyotypes (n = 19), chromosome abnormalities developed during HU therapy in 7 (37%). Our results indicate that HU should be regarded as leukaemogenic, at least when used for treatment of PV and allied diseases. Since myelosuppressive treatment of PV does not prolong survival, the use of HU should be restricted to patients in whom the treatment indication outweighs the risk of leukaemia induction.
Uptake and storage of retinol and retinyl esters in bone marrow of children with acute myeloid leukemia treated with high‐dose retinyl palmitate
Authors: Skrede Bjørn , Lie Sverre O. , Blomhoff Rune , Norum Kaare R.
Published: July 2010

Abstract:  Twenty‐one children with AML, who achieved complete remission with cytostatics, were treated with high doses of retinoids after remission was achieved. They were given 52 μmol retinyl palmitate/m2 (50000 International Units/m2) daily for at least 2 years thereafter. Thirteen of the children are still in complete remission with a mean observation time of 103 months. Due to the positive effects of retinoids seen in the treatment of AML, we have studied uptake of lipoprotein‐associated retinyl esters in bone marrow cells and peripheral leukocytes in vivo. An oral load of 104 μmol/m2 (100000 International Units/m2) retinyl palmitate resulted in a doubling of the concentration of total retinol in bone marrow cells and peripheral leukocytes after 5 hours. However, in the fasting state no significant difference was observed between the content of total retinol in bone marrow cells from controls and from patients receiving retinyl palmitate daily for years. Our study suggests that bone marrow cells take up retinyl esters postprandially, but do not store retinoids.
B‐cell chronic lymphocytic leukaemia stage 0. An immunophenotypic study of 66 cases and comparison with B small cell lymphomas*
Authors: Stramignoni Aldo , Valente Guido , Geuna Massimo , Stramignoni Daniela , Novero Domenico , Ragona Riccardo , Resegotti Luigi , Palestro Giorgio
Published: July 2010

Abstract:  The cells of 66 B‐CLL stage 0 patients were analyzed using a large panel of monoclonal antibodies in order to better define the immunophenotype of B‐CLL. The data were compared with the immunophenotypes of lymph node cells from 51 patients with diffuse B small cell lymphoma or leukaemia with lymph node enlargement. The most frequent immunophenotype of B‐CLL stage 0 was SIgM(D)+ EmR+ CD5+ CD9+ CD21+ CD23+ CD35– CD38–. Among the lymphomas, EmR‐positive and EmR‐negative cases were identified. The vast majority of the EmR‐positive cases usually showed the leukaemic pattern, immunophenotype and lymph node histology of B‐CLL. The EmR‐negative cases usually had immunophenotypes quite different from those of B‐CLL and the histology of indented cell lymphoma (centrocytic or intermediate) or features of lymphoplasmacytoid/cytic lymphoma. More than 20% of EmR‐positive cells proved to be the most important marker to distinguish B‐CLL from other lymphocytic lymphomas. Indeed this was a sign of better prognosis. Lymphoplasmacytoid/cytic lymphomas were EmR‐positive with the immunophenotype of B‐CLL or EmR‐negative with a definitely different immunological profile. This suggests two morphologically similar but biologically different subgroups of these diseases.
Repeated PCR in CML during IFN‐α therapy
Authors: Liberati A. M. , Donti E. , Rosso C. , Coco F. Lo , Dore F. , Pardini S. , Cocco S. , Addis M. , Saglio G.
Published: July 2010

Abstract:  Repeated PCR analysis was performed on bone marrow and/or peripheral blood samples from 4 CML patients in complete cytogenetic remission during treatment with IFN‐α. Two patients became PCR‐negative. One was negative for the analyses carried out from the 9th to the 30th months, but reverted to PCR positivity 10 months after IFN was reduced from 1.5 × 106 IU/day to 1 × 106 IU and given on alternate days. Although the dose was again raised to 3 × 106 IU/day, 8 months later her peripheral blood cells were still PCR‐positive, but remained persistently Ph'‐negative. Another patient became PCR‐negative at the 42nd month and remained so at the last analysis performed 3 months later. Two patients were persistently PCR‐positive. Cytogenetic relapse was documented in both, in one while still on full therapy. Ph'‐positive metaphases reappeared in the other patient 7 months after discontinuing IFN‐α therapy.
Differential effects of transferrin receptor antibodies on growth and receptor expression of human lymphocytic and myelocytic cell lines
Authors: Keyna Ulrike , Platzerv Erich , Woith Walter , Nüsslein Ina , Kalden Joachim R. , Manger Bernhard
Published: July 2010

Abstract:  J64, a monoclonal antibody against the human transferrin receptor, has been shown to induce interleukin‐2 production by HUT78 cells. It also causes growth inhibition of several cell lines and stimulated lymphocytes. These effects were also present using transferrin‐free culture conditions. In this paper, we dissect cell membrane and intracellular events after binding of J64 and other transferrin receptor antibodies. Incubation of HUT78 and several other cell lines with J64 resulted in an increased number of receptor molecules expressed on the cell surface in contrast to a downmodulation seen with other monoclonal antibodies to the transferrin receptor. This upregulation after treatment with J64 was not due to an increased concentration of transferrin receptor mRNA in these cells or a higher protein synthesis rate. We therefore suggest that J64 causes a redistribution of transferrin receptor molecules from intracellular pools to the cell surface. Additional experiments investigating signal transduction mechanisms revealed no influence of J64 on intracellular Ca2+ concentrations or translocation of protein kinase C. However, an increase of transferrin receptor phosphorylation was seen in HL60 cells after treatment with phorbolester or J64. This phosphorylation of the transferrin receptor might be a signal transduction pathway involved in activation and growth control.
Differential toxicity of α‐keto hydroxypyridine iron chelators and desferrioxamine to human haemopoietic precursors in vitro
Authors: Cunningham J. M. , Al‐Refaie F. N. , Hunter A. E. , Sheppard L. N. , Hoffbrand A.V.
Published: July 2010

Abstract:  Compliance with iron chelation therapy improves life expectancy in transfusion‐dependent haematological disorders. However, failure of compliance with parenteral desferrioxamine (DF) therapy and the expense incurred makes this drug unavailable for most patients in the developing world. We have been evaluating the orally active iron chelator 1,2‐dimethyl‐3‐hydroxypyrid‐4‐one (L1) in both preclinical and clinical trials. Five patients have developed reversible agranulocytosis during treatment with this agent. We have now studied the effects of L1, other α‐ketohydroxypyridines and DF on bone marrow myeloid progenitors using the CFU‐GM system. The results show that L1 is less toxic than DF to normal bone marrow myeloid progenitors (ID50:130 μmol/l versus 7.9 μmol/l). The L1 ID50 is within the previously reported range of peak plasma values (80–450 μmol/l). When saturating concentrations of iron were added to the cultures, the mean toxicity of all the chelators was significantly decreased over the range of doses tested, e.g. L1 ID50, 567 μmol/l; DF ID50, > 1000 μmol/l. The toxicity of L1 in vitro was similar for marrows from 3 normal donors and for the recovery marrow from a patient with thalassaemia major who had experienced agranulocytosis. Further studies are required to elucidate the mechanisms of L1 ‐induced agranulocytosis.
Clonal T‐lymphocytes from untreated hairy‐cell leukaemia patients enhance the growth of BFU‐E
Authors: Nielsen Bendt , Hokland Peter
Published: July 2010

Abstract:  The anaemia in hairy cell leukaemia (HCL) has been suggested to be mediated in part by T lymphocytes. Consequently, we have investigated the role of T lymphocytes in this disease by cloning pre‐treatment HCL T lymphocytes and testing their influence on the in vitro growth of BFU‐E from autologous post‐treatment and normal donors T‐lymphocyte depleted peripheral blood mononuclear cells (PBMNC‐T). Altogether, 24 CD4+ /CD8‐ and 17 CD4‐/CD8+ T‐lymphocyte clones from 3 different HCL patients were tested. All were found to enhance the growth of BFU‐E, the degree of enhancement being independent of the length of IFN treatment at the time of the post‐treatment sampling. Likewise, no difference in the extent of enhancement was seen between CD4+ /CD8‐ and CD4‐/CD8+ clones, or whether the clones were tested for modulation of BFU‐E from PBMNC‐T of autologous or allogeneic origin. Finally, no differences could be observed between different patients in the extent of clonal enhancement. These findings, which are in line with our previous ones in normal donors, indicate that the T‐lymphocyte function with regard to regulation of the erythropoiesis is normal in HCL, arguing against a T‐lymphocyte mediated suppression of the erythropoiesis in HCL.
Life expectancy in primary myelodysplastic syndromes: A prognostic score based upon histopathology from bone marrow biopsies of 569 patients
Authors: Maschek H. , Gutzmer R. , Choritz H. , Georgii A.
Published: July 2010

Abstract: The retrospective evaluation of bone marrow biopsies of 569 patients with primary myelodysplastic syndrome ‐ pMDS ‐ revealed 256 refractory anemias ‐ RA ‐, 52 refractory anemias with ringed sideroblasts ‐RARS ‐, 133 refractory anemias with excess of blasts ‐ RAEB ‐, 52 refractory anemias with excess of blasts in transformation ‐ RAEB‐t ‐, and 53 chronic myelo‐monocytic leukemias ‐ CMMOL ‐ according to FAB‐criteria, 23 patients were not otherwise specified (myelodysplastic syndrome: not otherwise specified ‐ MDS.NOS ‐). RARS‐patients had the best prognosis (median survival 41.9 months, incidence of leukemia 3.8%), followed by RA‐patients (26.5 months, 16.4%), MDS.NOS‐patients (22.4 months, 21.7%), CMMOL‐patients (12.5 months, 49.1%). RAEB‐and RAEB‐t‐patients had the worst prognosis (median survival time 8.5 and 4.6 months, incidence of leukemia 42.1% and 57.7%, respectively). But the survival times showed a considerable range in each FAB‐subgroup with 0–154 months in RA or 0–52 months in CMMOL. To forecast life expectancy more precisely, a scoring system was developed using nine histopathological parameters, among which the three most important ones were determined: quantity of myeloblasts, myelofibrosis and ALIP's. The scoring system allows a determination of three risk groups with significantly different survival times. It is valid also for patients without increase of myeloblasts (<5% myeloblasts in the bone marrow) and identifies high‐risk MDS patients in this group. By this proposed scoring system, a prognostic approval in primary MDS can be achieved applying histopathology without regarding further methods herewith presenting a system which could be considered independently from hematologic, cytological or laboratory data.
Lineage involvement and karyotype in a patient with myelodysplasia and blood basophilia
Authors: El‐Rifai Wa'el , Pettersson Tom , Larramendy Marcelo L. , Knuutila Sakari
Published: July 2010

Abstract:  We report a 63‐year‐old woman with myelodysplastic syndrome (MDS), refractory anaemia with ring sideroblasts (RARS), and blood basophilia with pathological forms. Karyotype analysis revealed a complex rearrangement: 46,XX,del(3) (p13p25),del(5) (q13q33),der(16)t(1;16) (p 13;q 12)/47,idem,add(20) (?p1l)/49,idem, + add(1) (q32),add(20) (?p11), + mar/46,XX. Karyotype, immunophenotype and in situ hybridization studies by the MAC (morphology antibody chromosomes) combination technique revealed the chromosomal abnormality in granulocytic/monocytic and erythrocytic metaphase cells. Also mature basophils and other granulocytes were involved with the abnormality. We suggest that our patient with the MDS has a stem cell disorder affecting all myeloid cell lineages and that basophilia constitutes a part of the malignant process.
Flow cytometric reticulocyte quantification in the evaluation of hematologic recover
Authors: Remacha Angel F. ,   Spanish multicentric study group for hematopoietic recovery
Published: July 2010

Abstract:  The role of flow cytometric reticulocyte (RET) counting and the immature RET fractions (IRF) in the evaluation of hematopoietic recovery following chemoradiotherapy‐induced aplasia was studied. RET counts and IRF were studied using an automated flow cytometric reticulocyte counter (Sysmex R‐2000) in three groups of patients: 58 patients undergoing an autologous bone marrow transplantation (ABMT group), 28 of whom received granulocyte colony‐stimulating factor (G‐CSF); 28 patients undergoing an allogeneic bone marrow transplantation (BMT group); and 28 patients receiving remission‐induction chemotherapy for acute leukemia (CHEMO group). To evaluate the IRF the percentages of RET fractions with middle and high fluorescence reticulocyte (MFR and HFR, respectively) were used. A rising IRF (expressed as the percentage of MFR ± HFR) was the first sign of hematopoietic recovery (ABMT group, IRF 9 days versus 18 days for the absolute neutrophil count (ANC); BMT group, 15 versus 18 days; CHEMO group, 9 versus 11 days). When recovery of the ANC (>0.5 times 109/1) was compared with that of the IRF (MFR ± HFR > 5%), statistically significant differences were found in all three groups. Additionally, 93.1% of the ABMT, 92% of the BMT and 91.2% of the CHEMO recovered the IRF before the ANC. In conclusion, an elevation in the percentage of IRF is the first sign of hematologic recovery in the majority of patients receiving remission‐induction chemotherapy and the first sign of engraftment in those submitted to ABMT or BMT. Serial automated flow cytometric quantitative reticulocyte counting provides a useful and early measure of erythropoiesis indicative of hematopoietic reconstitution or successful bone marrow engraftment following marrow transplantation.
Deferiprone‐associated myelotoxicity
Authors: Al‐Refaie F. N. , Wonke B. , Hoffbrand A. V.
Published: July 2010

Abstract:  Agranulocytosis developed in a 63‐year‐old patient with myelodysplasia 6 weeks after commencing treatment with the oral iron chelator deferiprone (L1, 1,2‐dimethyl‐3‐hydroxypyrid‐4‐one, CP20) at a daily dose of 79 mg/kg. This was the 3rd case of agranulocytosis (neutrophils 0 times 109/***1) in clinical trials of L1 at the Royal Free Hospital. The neutrophil count recovered 7 days after stopping and commencing G‐CSF at a dose of 300 μg daily. Three other patients with milder degrees of neutropenia (neutrophils <1.5 times 109/1) have also been observed in our trials. The case histories of these 4 patients are described here; other reported cases of neutropenia or agranulocytosis are reviewed. Based on worldwide long‐term clinical trials the incidence of agranulocytosis is about 1.6% and of neutropenia 2%.
Interferon‐gamma modulates cytosolic free calcium in human neutrophilic granulocytes
Authors: Røtnes J. S. , Aas V. , Iversen J. G.
Published: July 2010

Abstract: To investigate the role of cytosolic free calcium ([Ca2+]i in interferon‐γ (IFN‐γ) pre‐activation (priming) of human neutrophilic granulocytes (PMN) we used three different fluorescence methods, i.e. digital imaging of single, adherent, Fura‐2 loaded cells, flow cytometric measurements of single, non‐adherent, Fluo‐3 loaded cells, and spectrofluorometry of Indo‐1 loaded PMN in suspension. IFN‐γ increased the [Ca2+]i level in single, adherent PMN during the second phase of the fMLP response. The bacterial peptide fMLP (N‐formyl‐L‐methionyl‐L‐leucyl‐L‐phenylalanine) is a known stimulant of the calcium/inositol phosphate system. The [Ca2+]i increase was abolished in Ca2+‐free test buffer. Furthermore, the baseline [Ca2+]i level was found to be slightly increased in IFN‐γ primed PMN as analysed with flow cytometry. On the other hand, these [Ca2+]i responses were not detectable with the other methods used. We suggest that IFN‐γ increases the plasma membrane permeability for calcium in PMN, and substantiate this by demonstrating compliance with a capacitative model for intracellular calcium regulation. Mathematical modeling also suggested that IFN‐γ primed human PMN may sequester 13% more Ca2+ than unprimed cells in fMLP‐insensitive intracellular stores. Thus, the Ca2+ responses to IFN‐γ are modest and not easily detectable with some of the methods currently in use. They nevertheless explain why fMLP elicits brisker responses from PMN after IFN‐γ priming.
Immunocytochemical detection of bone marrow‐invasive neuroblastoma cells
Authors: Nagai Junichi , Kigasawa Hisato , Tomioka Kayoko , Koga Noriko , Nishihira Hirokazu , Nagao Takeshi
Published: July 2010

Abstract: We evaluated the utility of an immunocytochemical technique employing the commercially available anti‐CD56 monoclonal antibody, NKH 1. The utility and sensitivity of this technique in the detection of invasive neuroblastoma (NB) cells in the bone marrow were compared with those of Wright‐Giemsa staining. The correlation coefficient for the percent NB cells detected using Wright‐Giemsa staining with the percent NKH 1 immunoreactive cells was 0.78. In the analysis of specificity, this monoclonal antibody showed slight cross‐reactivity with normal bone marrow cells, including macrophages, lymphocytes and osteoblasts. In the evaluation of the sensitivity of the NKH 1 immunocytochemical technique, SK‐N‐DZ and SK‐N‐SH NB cell lines were added to morphologically normal bone marrow mononuclear cells from patients without NB to the final NB cell line at concentrations of 2%, 1% and 0.1%. NB cells at the final concentration of 0.1 % could be detected by the immunocytochemical technique. We conclude that the NKH 1 immunocytochemical staining technique is useful in the detection of metastatic NB cells in bone marrow.
Unassembled (soluble) vimentin in human myeloid leukemia cell line HL60
Authors: Takahashi Noriko , Breitman Theodore R.
Published: July 2010

Abstract: The intermediate filament proteins which include vimentin, desmin, and the keratins are one of three major classes of cytoskeletal proteins in eukaryotic cells. In this study we found that most of the vimentin of undifferentiated HL60 and cells induced to differentiate either along the monocytoid pathway by 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) or along the granulocytic pathway by retinoic acid was soluble in a buffer containing 1% Triton X‐100/0.6 mol/l KCl in which the intermediate filament proteins usually are not soluble. HL60 vimentin separated on Polyacrylamide gel electrophoresis into two proteins of Mr 55000 and 54000 that we detected by immunoblotting. The Mr 55000 species was the major form in undifferentiated HL60 cells and cells induced by retinoic acid. The distribution of both forms of vimentin changed during induction of differentiation by TPA and after 24 h the Mr 54000 species was predominant. After an additional 24 h exposure to TPA the relative levels of the two forms of vimentin approached equivalence and a high level of vimentin degradation products was seen. These results suggest that TPA may increase vimentin degradation along a pathway that has a Mr 54000 intermediate. In addition, the high levels of soluble vimentin in HL60 cells suggests that these cells may be a good model for studying components involved in vimentin assembly.
Differential expression of two ICAM‐1 epitopes and LFA‐1 chains in B‐cell non‐Hodgkin's lymphomas
Authors: Vacca Angelo , Ranieri Girolamo , Ribatti Domenico , Di Stefano Rita , Caloro Daniela , Serio Gabriella , Di Loreto Michela , Silvestris Franco , Dammacco Franco
Published: July 2010

Abstract: B‐cell non‐Hodgkin's lymphomas (B‐NHL) and B‐cell areas of reactive lymphadenopathies were investigated immunohistochemically for expression of two distinct ICAM‐1 epitopes, Me14/D12 and P3‐58, and the LFA‐1 α and β chains. Partial or total loss of expression of one or both epitope(s) and/or chain(s) was evident in all B‐NHL in function of increasing Working Formulation (WF) malignancy grade, with most defects in the high‐grade tumors, namely the lowest detectability of the ICAM‐1 Me14/D12 and LFA‐1 α chain, the lowest co‐expression of ICAM‐1 epitopes and LFA‐1 chains, and the most frequent simultaneous loss. The ICAM‐1 and LFA‐1 profiles overlapped within the low‐ and intermediate‐grades, whereas striking differences between the high‐grade subtypes were detected. Specifically, Burkitt's and lymphoblastic tumors always lost both epitopes and both chains. Large cell, immunoblastic tumors occasionally did so, and also showed either uncoordinated expression or co‐expression of these constituents. It is suggested that expression defects of this type may help differentiate malignant from benign lymphoproliferations, and also be involved in the progression of B‐NHL, since most are observed in high‐grade tumors, whose ICAM‐1 and LFA‐1 profiles indicate that their subtypes are the expression of distinct normal B‐cell differentiation stages.
Leukocyte‐depleted blood components prevent platelet refractoriness in patients with acute myeloid leukemia
Authors: Oksanen K.
Published: July 2010

Abstract: Platelet refractoriness arising from HLA immunization develops in 20–50% of multitransfused patients with hematologic malignancies. We retrospectively analyzed the efficiency of leukocyte‐depleted blood components in preventing refractoriness in 140 adult patients with acute myeloid leukemia (AML), treated according to a standardized cytostatic protocol. Sixty‐eight patients received leukocyte‐depleted (L‐D) platelet concentrates (PCs) and red cells (RBCs), with fewer than 106 leukocytes per unit, and 72 patients received standard (STD) blood components. Two of 67 (3%) evaluable patients in the leukocyte‐depleted group and 14 of 68 (21%) in the standard group became refractory during the median follow‐up time of 229 days. Nine of 24 (37%) previously pregnant women in the STD group but none of 17 in the L‐D group became refractory. Twenty patients were shifted during later treatment from L‐D to STD PCs; none became refractory. Corrected platelet increments (CI) at 18 hours were higher after STD PCs (6.5) than L‐D PCs (5.2), but more PCs and RBCs were transfused per patient in the STD group. It is concluded that effective leukocyte depletion prevents platelet refractoriness in patients with AML, even in those with previous immunization, and reduces the consumption of blood components.
Treatment of hypogammaglobulinemia in chronic lymphocytic leukaemia by low‐dose intravenous gammaglobulin
Authors: Jurlander Jesper , Geisler Christian Hartmann , Hansen Mogens Mørk
Published: July 2010

Abstract: Intravenous immunoglobulin replacement therapy reduces the number of bacterial infections in B‐cell chronic lymphocytic leukaemia (B‐CLL) patients. However, due to the complexity of immunodeficiency in B‐CLL and the cost‐effectiveness of replacement therapy, it is important to identify patients who are likely to benefit from the treatment and to investigate which dose should be used. 15 patients with hypogammaglobulinaemia and a history of recurrent infections received a fixed dose of 10 grams of gammaglobulin intravenously every 3 weeks. Serum IgG levels were significantly higher after three doses (p = 0.0002), and stabilized just above lower reference value after 11 doses. The total number of infection‐related events during 168 months before therapy was compared to the total number of infection‐related events in 169 months during therapy. The number of antibiotic prescriptions was reduced from 78 to 54 (N.S.), the number of admissions to hospital due to infections was reduced from 16 to 5 (p = 0.047) and the number of febrile episodes was reduced from 63 to 31 (p = 0.004). We conclude that a fixed low dose of gammaglobulin intravenously can restore normal serum IgG levels in hypogammaglobulinaemic B‐CLL patients, and leads to a decreased number of febrile episodes and admissions to hospital due infections.
Prognostic relevance of a histologic classification system applied in bone marrow biopsies from patients with multiple myeloma: A histopathological evaluation of biopsies from 153 untreated patients*
Authors: Sailer M. , Vykoupil K.‐F. , Peest D. , Coldewey R. , Deicher H. , Georgii A.
Published: July 2010

Abstract:A total of 153 diagnostic bone marrow biopsies from patients with advanced stages of multiple myeloma corresponding to stages II and III according to the Durie/Salmon classification were evaluated prior to any treatment in a prospective therapy trial of the German Myeloma Treatment Group. Histologic sections were analyzed according to a pre‐defined system of criteria microscopically by 2 observers, determining three criteria: 1) grading by histopathology, regarding the cytologic differentiation of neoplastic cells and quantifying the percentage of plasmacytic, pleomorphic, and plasmablastic myeloma cells distributed within the sections; 2) the volume of infiltration; and 3) the pattern of neoplastic growth. Furthermore, four other criteria, namely hematopoiesis, fiber increase, osteomalacia, and micro‐osteo‐lesions, were evaluated. A cluster analysis using the three histological criteria revealed three groups of patients with significantly different survival times based on histological criteria only; the three criteria were mentioned above. It is concluded from these results that bone marrow biopsies, when evaluated histologically by grading and staging according to the three criteria, provide most valuable prognostic parameters in myeloma patients.
Effects of monocyte‐macrophage colony‐stimulating factor (M‐CSF) on in vitro erythropoiesis of marrow progenitor cells from patients with renal anemia
Authors: Kawano Yoshifumi , Takaue Yoichi , Minakuchi Jun , Kawashima Shuh , Kimura Fumihiko , Motoyoshi Kazuo , Abe Takanori , Sato Junko , Hirao Atsushi , Saito Shin‐ichi , Suzue Takeshi , Kuroda Yasuhiro
Published: July 2010

Abstract: We examined the influence of monocyte‐macrophage colony‐stimulating factor (M‐CSF) on erythropoiesis both in vitro and in vivo in 98 patients with chronic renal failure who were undergoing hemodialysis. Serum levels of M‐CSF and the clinical response to therapy with human recombinant erythropoietin (Epo) were analyzed. The following results were obtained: 1) The serum level of M‐CSF was 6.90 ± 2.41 ng/ml in the patient population (n = 98), but only 2.0 ± 0.3 ng/ml in 10 healthy donors. 2) 41 of the 98 anemic patients were treated with various doses of Epo for 3 months, and the average increase in the blood hemoglobin level during this period was 26.1 ± 12.5 mg/dl/unit of Epo/kg patient's b.w./week. Lower levels of M‐CSF before treatment significantly predicted a better response to subsequent Epo therapy (r = –0.496, p < 0.001). 3) When cultured with a maximally stimulatory amount of Epo (10 IU/ml), the number of marrow early erythroid progenitor cells (burst‐forming unit for erythroid, BFU‐E) in patients was identical to that in normal donors, while the number of late progenitors (colony‐forming unit for erythroid, CFU‐E) was relatively lower in patients. 4) The addition of recombinant M‐CSF to the culture resulted in suppression of erythroid progenitor cell growth in the patient population, but induced enhancement in normal donors. The inhibitory effect of M‐CSF on the patients' cells was not eliminated by the addition of antibodies against interleukin‐1α/β, tumor necrosis factor‐α, or interferon‐α/β/γ. Supernatants from marrow mononuclear cells cultured in the presence of M‐CSF carried this inhibitory effect on marrow CD34+ cells obtained from patients. Together, these results suggest that M‐CSF aggravates a previously existing decreased sensitivity of erythroid progenitor cells to Epo in some patients with renal anemia.
Incidence of myelodysplastic syndromes in a Swedish population
Authors: Rådlund Anders , Thiede Torben , Hansen Sören , Carlsson Magnus , Engquist Leif
Published: July 2010

Abstract: The myelodysplastic syndromes (MDS) have attracted great interest during the last decade. Unfortunately there is not much material published concerning the incidence of these conditions. We present epidemiologic data on MDS based on case registration of patients in a well‐defined population and as a comparison similar data on acute myeloid leukaemia (AML). Between the years 1978–1992 we registered 120 cases of MDS and 146 cases of AML. The median age for all the MDS cases was 74.1 years for men and 78.2 years for women. Among haematologists there is a suspicion that the incidence of MDS is rising. Our study does not support this opinion. We have divided the study period into 5‐year periods and the crude incidence has been 3.2, 4.1 and 3.5/100 000/year for each period. In the age group over 70 years MDS was more frequent than AML and in the last 5‐year period the incidence was 15.0/100 000/year for MDS compared to 10.2/100 000/year for AML. In conclusion MDS is quite common among elderly people and there is no evidence for a rising incidence during the last 15 years.
Pulmonary infiltrates in patients with haematologic malignancies: Clinical usefulness of non‐invasive bronchoscopic procedures
Authors: von Eiff M. , Zühlsdorf M. , Roos N. , Thomas M. , Büchner T. , van de Loo J.
Published: July 2010

Abstract: In a prospective study 90 patients with haematologic malignancies (57 acute leukaemias, 6 Hodgkin's Diseases, 15 Non‐Hodgkin Lymphomas, 12 other diseases), with fever exceeding 38.4°C and newly developed pulmonary infiltrates underwent bronchoscopy obtaining bronchoalveolar lavage, bronchial washings and protected brush specimen (n = 71). Pneumonias due to gram‐negative bacteria (n = 38) and fungi (n = 34) were most frequent. Bronchoscopic specimens yielded 226 isolates (2 different organisms/bronchoscopy on average). 112 organisms were finally regarded as causing pneumonia. Sensitivity of bronchoscopy in diagnosing infectious episodes was 66%, but only 4 out of 13 non‐infectious pulmonary infiltrates could be identified. Bronchoscopy was most effective in the diagnosis of Pneumocystis carinii and herpes virus pneumonia, whereas sensitivity and specificity of detecting fungal and bacterial pneumonia were low. Empirical antimicrobial therapy was verified by evaluation of bronchoscopic samples in 25 out of 90 cases. Empirical therapy was successfully changed according to the results of invasive samplings in 34 out of 90 cases. Early identification of causative pathogens had a significant impact on survival.
Automated measurement of reticulated platelets in estimating thrombopoiesis
Authors: Watanabe Kiyoaki , Takeuchi Kei , Kawai Yohko , Ikeda Yasuo , Kubota Fumio , Nakamoto Hiroyuki
Published: July 2010

Abstract: We described a fully automated measurement of reticulated platelets using a fluorescent dye, auramine O, and a reticulocyte counter, the R‐3000, equipped with special software. Reproducibility and linearity were shown to be good. In the normal subjects studied (n = 60), the mean value for reticulated platelets was 0.98% ± 0.41% and the mean absolute count was 2.12 ± 0.69 × 109/l. The absolute count for reticulated platelets was significantly lower (p < 0.05) in patients with reduced thrombopoiesis as seen in acute myeloblastic leukemia, aplastic anemia or chemotherapy‐induced thrombocytopenia and it was elevated (p < 0.05) in essential thrombocythemia and in chronic myelocytic leukemia with thrombocytosis. All 20 patients with chronic idiopathic thrombocytopenic purpura had a high percentage of reticulated platelets. The percentage of reticulated platelets was significantly increased (p < 0.05) in patients with impaired thrombopoiesis despite the reduction in the absolute count. In 2 leukemic patients, an apparent rise was noticed in the percentage of reticulated platelets which preceded by several days a progressive increase in the platelet count at the recovery phase of thrombocytopenia. The results suggest that an automated measurement of reticulated platelets can be applied to routine laboratories for clinical use.
Synergism of interleukin‐12 and interleukin‐3 on development of hematopoietic progenitors
Authors: Fardoun‐Joalland D. , Teixeira‐Lebrun G. , Lenormand B. , Dzondo‐Gadet M. , Vannier J. P.
Published: July 2010

Abstract: The recently cloned cytotoxic lymphocyte maturation factor [CLMF] also called NK cell stimulatory factor [NKSF] or interleukin‐12 [IL‐12] has been described as a growth factor for mature lymphoid cells. The present study investigated whether purified recombinant human IL‐12 could stimulate CFU colony growth. Source of progenitor cells were peripheral blood cells depleted of adherent, CD2‐ and CD56‐positive cells. RhIL‐12 was investigated either alone or in combination with rhIL‐3, rhIL‐6 and rhGM‐CSF. RhIL‐12 alone did not support colony formation of myeloid or erythroid progenitors. RhIL‐12 in combination with rhIL‐3 increased the numbers of BFU‐E and CFU‐GM. No synergism or additive effect was seen with the combination of rhIL‐12 and rhGM‐CSF or rhIL‐12 and rhIL‐6. An additive increase in the number of granulocytic colonies was observed when rhIL‐3, rhIL‐6 and rhGM‐CSF were used together with rhIL‐12. Our result therefore suggest that, in addition to being a potent lymphopoietic stimulator, IL‐12 acts synergistically with IL‐3 in enhancing the sensitivity of hemopoietic progenitors to IL‐3.
Haemostatic and fibrinolytic parameters in septic patients with leukopenia or leukocytosis
Authors: Saito Masanori , Asakura Hidesaku , Jokaji Hiroshi , Uotani Chika , Kumabashiri Ichiro , Morishita Eriko , Yamazaki Masahide , Yoshida Tomotaka , Aoshima Keiji , Matsuda Tamotsu
Published: July 2010

Abstract: Induction of leukocytopenia by cytotoxic drugs protects against the generalized Shwartzman reaction induced by endotoxin. To elucidate the relationship between leukocyte number and in haemostatic and fibrinolytic disturbances in human sepsis, we studied 32 septic patients with abnormal leukocyte counts. Twenty patients had sepsis in the setting of leukopenia after chemotherapy for haematological malignancies. Twelve patients with leukocytosis developed sepsis associated with benign disorders. Concentrations of thrombin‐antithrombin III complex (TAT), plasminogen activator inhibitor‐1 (PAI‐1) and plasma thrombomodulin (TM) in the leukocytosis group of (12.0 ± 11.0, 40.2 ± 27.0 and 5.5 ± 2.3 ng/ml, respectively) were significantly elevated compared to the leukopenia group of (3.8 ± 2.3, 18.0 ± 15.0 and 3.1 ± 1.0 ng/ml, respectively) and controls (3.3 ± 0.4, 10.5 ± 5.3, 3.0 ± 0.5 ng/ml, respectively). On the other hand, there were no significant differences in these values between leukopenia group and controls. Thus leucocytes may play important roles in thrombin generation, PAI‐1 release and injury to endothelial cells.
HIV Tat protein potentiates in vitro granulomonocytic progenitor cell growth
Authors: Calenda V. , Chermann J. C.
Published: July 2010

Abstract: The direct involvement of hematopoietic progenitor cells in AIDS which results in dysregulated hematopoiesis, probably due to regulatory factors yet to be determined, has been reported by us and others. In this study we demonstrate that the HIV‐2 Tat gene product is released in supernatant of HIV‐2‐infected long‐term bone marrow cultures (LTBMC). These Tat‐containing supernatants specifically enhanced growth of CFU‐GM in agar culture system and this promoting activity was specifically neutralized with anti‐Tat antibodies. The use of different recombinant Tat proteins confirmed that Tat has been responsible for enhanced in vitro growth of CFU‐GM after HIV‐2 infection of LTBMC. Moreover, limiting dilution analysis showed that Tat acts directly on the CD34+ cell population in which it increases the frequency of IL3‐responding cells.
Deep venous thrombosis and pulmonary artery embolism in high‐grade non Hodgkin's lymphoma: incidence, causes and prognostic relevance
Authors: Ottinger H. , Belka C. , Kozole G. , Engelhard M. , Meusers P. , Paar D. , Metz K. A. , Leder L.‐D. , Cyrus C. , Gnoth S. , Gerhartz H. , Huhn D. , Siegert W. , Thiel E. , Aydemir Ü. , Tintrup W. , Lennert K. , Brittinger G.
Published: July 2010

Abstract: To analyse incidence, risk factors, causes and prognostic significance of venous thromboembolism (VTE) in high‐grade non‐Hodgkin's lymphoma (HG‐NHL) a prospective clinical trial (N = 593), also undertaken to analyse other aspects of HG‐NHL, a study of haemostasis (N = 25) and a post‐mortem analysis (N = 70) were performed. Clinical analysis documented a 6.6% incidence of VTE, and 77% of all cases occurred before or within the first 3 months of chemotherapy. Ann Arbor stage IV and B‐mediastinal clear cell histology were risk factors for VTE, while rapid changes in tumour load or application of consolidation chemotherapy were not. Vessel compression by HG‐NHL was the leading cause of VTE, whereas a significant (paraneoplastic or chemotherapy‐induced) thrombophilic state was not disclosed by haemostatic tests. While VTE‐related fatality was found to be low in the clinical trial (1.7%) and at necropsy (8.5%), the occurrence of VTE was associated with an unsatisfactory response of HG‐NHL to chemotherapy and a high incidence of treatment‐related mortality due to diffuse alveolitis. Thus, fatal VTE in HG‐NHL is rare, but VTE is associated with an unfavourable clinical course of HG‐NHL.
Assessment of minimal residual disease in acute promyelocytic leukaemia with t(15;17) by chromosome painting
Authors: Temperani P. , Vaccari P. , Giacobbi F. , Gandini G. , Ferrari M. G. , Emilia G.
Published: July 2010

Abstract: To detect the minimal residual disease (MRD) in acute promyelocytic leukaemia patients treated with all‐trans retinoic acid, we compared the sensitivity of metaphase fluorescence in situ hybridization (FISH) with conventional analysis of G‐banded metaphases. 5 out of 6 patients studied at diagnosis showed the t(15;17) translocation. 4 out of 5 patients carrying t(15;17) achieved complete remission and conventional cytogenetic conversion. In 3 cases the whole chromosome painting (WCP) probe 17 discovered one normal chromosome 17 and two fragments indicative of t(15;17) persistence. The FISH‐WCP technique seems to be highly sensitive and recommendable in monitoring leukaemias with specific chromosome rearrangements.
Bone marrow progenitor cell growth and karyotype changes in healthy 88‐year‐old subjects
Authors: Nilsson‐Ehle Herman , Swolin Birgitta , Westin Jan
Published: July 2010

Abstract: Previous studies have indicated a decline in bone marrow progenitor cell function in subjects aged 75–82 years, possibly causing lower Hb concentrations. We studied the bone marrow with in vitro colony assays and cytogenetic analysis in 24 apparently healthy 88‐year‐olds with Hb concentrations ranging from moderate anaemia to normal levels. Twenty‐two healthy younger subjects, aged 21–57 years, were used as a control group. The 88‐year‐olds showed significantly lower numbers of myeloid bone marrow progenitors than the controls, and the elderly men had lower numbers of both erythroid and myeloid progenitors than the elderly women. There were no in vitro growth differences between elderly subjects with “low” or “normal” Hb concentrations. Ten out of 14 men had bone marrow cells with a missing Y‐chromosome, which did not seem to have any relationship to the erythroid function. No morphological or other cytogenetic indications of a clonal progenitor cell disorder were found. A more rapid decline in Hb concentrations in healthy elderly men as compared to elderly women might be explained by differences in bone marrow progenitor cell function. However, progenitor cell abnormalities do not seem to explain differences in Hb concentrations within groups of apparently healthy men and women of advanced age.