bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effect of macrophage colony‐stimulating factor on haemopoietic recovery after autologous bone marrow transplantation
Authors: Khwaja Asim , Yong Kwee , Jones H. Mark , Chopra Rajesh , McMillan Andrew K. , Goldstone Anthony H. , Patterson Keith G. , Matheson Catherine , Ruthven Karen , Abramson Stephan B. , Linch David C. 
Published: March 2008

Summary.Macrophage colony‐stimulating factor (M‐CSF) is active in the late stages of monocyte maturation, activates mature monocyte‐macrophages and enhances their production of various other cytokines. We have examined the effects of a 21 d course of escalating doses of M‐CSF purified from human urine (hM‐CSF) on recovery following autologous bone marrow transplantation (ABMT) in 20 patients with malignant lymphomas. Four patients were treated at each dose level of 4, 8, 16, 32 and 64.1099 U/m2/d and results compared to 46 concurrent controls. There was no significant difference in recovery to an absolute neutrophil count (ANC) of 0.5×109/1 (median 20 d in hM‐CSF group versus 22 in controls) or in recovery of platelets to 50.109/1 (32 d versus 39 d, 0.05< P <0.1); hM‐CSF patients received a median of 81 platelet units following ABMT (controls 112 units, P = NS). hM‐CSF patients had a median of 5.5 d with fever >37.5°C (control 8, P=NS), received parenteral antibiotics for 14.5 d (control 17, P=NS) and had a 50% incidence of bacteraemia (control 48%). hM‐CSF treated patients were discharged by a median of day 29 following transplantation (control 33, P<0.05). Platelet and neutrophil recovery correlated significantly with the number of marrow mononuclear cells (MNC) reinfused in the hM‐CSF group (P=0.05 and P=0.014 respectively) but not in controls. Subgroup analysis showed that hM‐CSF patients receiving > 2.108 MNC/kg body weight reached an ANC of 0.5×109/1 by a median of day 16.5 (control 18.5, NS), became platelet transfusion independent by day 17 (control 29, P<0.05) and reached a platelet count of 50.109/1 by day 21 (control 40, P<0.05). No significant toxicity attributable to hM‐CSF treatment was seen. These results suggest that hM‐CSF accelerates platelet recovery following ABMT and that relatively large marrow innocula are required to see this effect.
Trisomy 8 detection in granulomonocytic, erythrocytic and megakaryocytic lineages by chromosomal in situ suppression hybridization in a case of refractory anaemia with ringed sideroblasts complicating the course of paroxysmal nocturnal haemoglobinuria
Authors: PArlier V. , Tiainen M. , Beris Ph. , Miescher P. A. , Kimuutila S. , Bellomo M. Jotterand 
Published: March 2008

Summary.Paroxysmal nocturnal haemoglobinuria (PNH) was diagnosed in a 20‐year‐old male patient who suffered from anaemia since the age of 11. Eighteen years after diagnosis, PNH transformed into refractory anaemia with ringed sideroblasts (RARS). Trisomy 8 was observed in 27%, 45% and 53% of the bone marrow metaphase cells analysed in 1987, 1988 and 1990 respectively. In order to determine which bone marrow cell lineages were affected by trisomy 8 and at which stage of stem cell differentiation, MAC (Morphology, Antibody, Chromosomes) and CISS (Chromosomal in Situ Suppression) hybridization techniques were combined. The MAC technique enables karyotypic analysis of morphologically and immunologically classified mitotic cells. CISS hybridization makes it possible to detect individual chromosomes and chromosome aberrations using recombinant DNA libraries from sorted human chromosomes. Trisomy 8 was detected in granulomonocytic (50.6%), erythrocytic (67.2%) and megakaryocytic (one megakaryocyte with trisomy 8, one normal) lineages, providing evidence for the occurrence of trisomy 8 in early haematopoietic cell precursors, at the GEMM or pluripotent level. Cytogenetic and clinical data suggest that the sideroblastic clone originated from a mutation affecting a cell of the PNH clone, progressively replaced by the PNH/RARS clone, due to proliferative advantage.
Identification of CD71 (transferrin receptor) expressing erythrocytes by multiparameter‐flow‐cytometry (MP‐FCM): correlation to the quantitation of reticulocytes as determined by conventional microscopy and by MP‐FCM using a RNA‐staining dye
Authors: Serke Stefan , Huhn Dieter 
Published: March 2008

Reticulocytes express the CD71‐defined antigen, the transferrin receptor. This report describes how by means of dual‐colour immunofluorescence using MP‐FCM (multiparameter‐flow‐cytometry) CD71+ erythrocytes can be detected regularly in blood of healthy adults. Percentages of these CD71+ erythrocytes were compared to the percentages of reticulocytes as determined by conventional microscopy using brilliant cressyl blue, and to the percentages of erythrocytes with high RNA content, as detected by MP‐FCM using a RNA‐staining dye (thiazole‐orange). Only about two‐thirds of the percentages determined by the two latter methods were detected by MP‐FCM using the CD71 expression for definition of reticulocytes. Studying clinical samples, however, including both specimens with very low and very high numbers of reticulocytes. almost identical percentages were determined by all the three methods described.Studying reticulocytes in vitro, a rapid decline of the expression of the transferrin receptor was observed on reticulocytes. Due to the differential expression of the transferrin receptor on reticulocytes, different subsets of reticulocytes could be identified.The dual‐colour MP‐FCM method described allows for the characterization and enumeration of immature erythrocytes, representing the major subset of reticulocytes as determined by conventional methods. Furthermore, it allows for subset dissection of reticulocytes.
Impairment of GM‐CSF production in myelodysplastic syndromes
Authors: Visani G. , Zauli G. , Tosi P. , Fogli M. , Cenacchi A. , Finelli C. , Gamberi B. , Ottaviani E. , Manfoi S. , Tura S. 
Published: March 2008

Summary. In this study we evaluated the production of granulocyte/macrophage‐colony stimulating factor (GM‐CSF), interleukin‐6 (IL‐6) and tumour necrosis factor‐α (TNF‐α) by enriched bone marrow (BM) macrophages in 15 patients affected by myelodysplasia and 20 normal BM donors.The presence of GM‐CSF, IL‐6 and TNF‐α in the culture supernatants of BM macrophages was detectable only after stimulation with lipopolysaccharide (LPS), whereas no differences were present in the amount of IL‐6 and TNF‐α between myelodysplastic patients and normal controls, GM‐CSF production appeared eight‐fold reduced in BM macrophage culture supernatants from myelodysplastic patients with respect to normal controls. After further experiments, we concluded that the impaired release of GM‐CSF by BM macrophages could not be due to a different production kinetic in myelodysplastic patients.Moreover, the number of multipotent (CFU‐GEMM), granulocyte/macrophage (CFU‐GM) and erythroid (BFU‐E) progenitors was significantly impaired in myelodysplastic patients.In conclusion, we demonstrated that the production of GM‐CSF by purified adherent cells from MDS patients is markedly impaired in spite of the peripheral blood cytopenia. This selective defect in GM‐CSF production, along with an intrinsic defect of haematopoietic progenitor cells, might contribute to the impairment of haematopoiesis always observed in myelodysplastic patients.
Quantitation of follicular non‐Hodgkin's lymphoma cells carrying t(14;18) by competitive polymerase chain reaction
Authors: Meijerink J. P. P. , Smetsers T. F. C. M. , Raemaekers J. M. M. , Bogman M. J. J. T. , de Witte T. , Mensink E. J. B. M. 
Published: March 2008

Summary. A competitive polymerase chain reaction (PCR) technique was developed to quantify residual malignant cells in the peripheral blood and bone marrow of patients with low‐grade follicular non‐Hodgkin's lymphoma carrying a translocation between chromosomes 14 and 18. Artificial segments were constructed imitating a translocation between chromosome 14 and 18. These artificial translocation segments were used as competitor molecules in the quantitative PCR. Serial dilutions of a known amount of patient‐derived translocation segments were coamplified with a fixed number of competitor molecules, and a patient specific calibration curve was constructed. Several patient samples were coamplified with an equal number of competitor molecules and the number of t(14;18) translocations within the samples was calculated by comparison with the calibration curve.The method was demonstrated on samples of four follicular non‐Hodgkin's lymphoma (NHL) patients. In a patient transplanted with allogeneic bone marrow declining numbers of residual lymphoma cells were observed.We conclude that the method is accurate, relatively fast and the general principle of this method can be applied to all malignancies with characteristic abnormalities on DNA or RNA level that are detectable by PCR.
The effect of methyldopa and procainamide on suppressor cell activity in relation to red cell autoantibody production
Authors: Garratty George , Arndt Patricia , Prince Harry E. , Shulman Ira A. 
Published: March 2008

Summary. Kirtland et al (1980) suggested that methyldopa caused the production of red cell (RBC) autoantibodies by causing a persistent increase in lymphocyte cyclic AMP, which inhibited suppressor T cell function, leading to unregulated autoantibody production in some patients. They showed that significantly higher lymphocyte cyclic AMP concentrations were generated by lymphocytes from healthy donors after adding methyldopa, and by lymphocytes from patients who were receiving methyldopa compared to lymphocytes from healthy donors without methyldopa present. They also showed that methyldopa affected suppressor cell activity. We measured the effect of methyldopa and procainamide on suppressor cell activity, using a similar approach to Kirtland et al (1980). Suppressor cell activity was measured by measuring the amount of IgG, produced in vitro, by B cells following mitogen stimulation preceded by a 24 h incubation period. We found no significant increase in the amount of IgG generated by normal donor lymphocytes, when methyldopa or procainamide was present during the preincubation period. This is in contrast to the findings of Kirtland et al (1980). We also measured the amount of IgG generated in vitro by mitogen‐stimulated lymphocytes from patients (with and without positive direct antiglobulin tests) taking methyldopa and compared this to the amount of IgG generated by lymphocytes from normal donors and patients (with and without positive direct antiglobulin tests). The results were similar for each group. This does not agree with the findings of Kirtland et al (1980) who found that lymphocytes from patients taking methyldopa produced more IgG in vitro than lymphocytes from normal donors.Our results do not support the hypothesis that methyldopa and procainamide induce autoantibodies by affecting suppressor cell function.
Increased serum levels of transforming growth factor β‐1 in patients affected by thrombotic thrombocytopenic purpura (TTP): its implications on bone marrow haematopoiesis
Authors: Zauli Giorgio , Gugliotta Luigi , Catani Lucia , Vianelli Nicola , Borgatti Paola , Belmonte Monica Mattioli , Tura Sante 
Published: March 2008

Summary. In this study we evaluated the effect of serum collected from seven thrombotic thrombocytopenic purpura (TTP) patients, either in the acute phase of the disease or in clinical remission, on the in vitro growth of bone marrow haematopoietic progenitor cells, obtained from the same TTP patients in clinical remission and from normal donors.The addition to the cultures of autologous sera collected from TTP patients in acute phase of the disease showed a clear‐cut dose‐dependent inhibition of immature haematopoietic progenitor cells (BFU‐E, CFU‐meg and 14th day CFU‐GM). On the other hand, no inhibitory effects were observed on more mature 7th day CFU‐GM. Interestingly, also sera collected from TTP patients in clinical remission still maintained some inhibitory activity on the growth of immature progenitor cells. A similar inhibitory activity was noticed when TTP sera were tested on normal bone marrow haematopoietic progenitor cells.Such inhibitory activity was significantly reduced in blocking experiments by the addition of a polyclonal neutralizing anti‐TGF‐β1 antibody and the presence of increased levels of both bioactive and latent TGF‐β1 in TTP sera was confirmed in a bioassay on CCL64 cells.These data contribute to explain the lack of a clear compensatory haematopoiesis observed in some patients with active TTP and add further evidence to the notion of the existence of a state of latent platelet activation in TTP patients in clinical remission.
Inhomogeneity of the circulating BFU‐E regulation in sickle cell anaemia: accessory cells properties and BFU‐E growth factor response pattern
Authors: Croizat Helena , Nagel Ronald L. 
Published: March 2008

Summary. Sickle cell anaemia (SS) patients with low (<9%) HbF levels (LFSS) are characterized by an increased number of circulating BFU‐E in active DNA synthesis, and release of burst promoting activity (BPA) by unstimulated low density (LD) adherent cells. In contrast, circulating BFU‐E from SS patients with high (>9%) HbF levels (HFSS) are normal in number, largely in resting phase, and their LD cells do not release BPA‐like activity.We report now that in LFSS patients, adherent cell depletion decreases BFU‐E growth in culture and apparent BFU‐E cycling. Furthermore, addition of conditioned media (CM) from LD cells of LFSS patients restored cycling BFU‐E expression in culture. Neutralization analysis with anti‐GM‐CSF antibody demonstrated that GM‐CSF is, at least, one factor responsible for BPA activity present in this CM. Thus, GM‐CSF is constitutively produced by unstimulated monocytes in LFSS patients. In contrast, HFSS patients' adherent cell depletion increases cycling of BFU‐E in culture. CM from HFSS patients inhibits BFU‐E expression in culture. Hence, LD adherent cells from HFSS patients may release a yet unknown inhibitor factor(s). In addition, we report a distinct response pattern in SS patients' BFU‐E to growth factor (GM‐CSF, IL‐3): (a) LFSS patients have a BFU‐E population, equally responsive to GM‐CSF and IL‐3; (b) HFSS patients, have a subset of BFU‐E exclusively dependent on IL‐3 (20–40% of the circulating BFU‐E). This pattern is very similar to that of normal BFU‐E.In conclusion, BFU‐E from LFSS patients represent an actively proliferating population, equally responsive to GM‐CSF and IL‐3, controlled by constitutively produced GM‐CSF, suggesting a unique BFU‐E behaviour in SS patients with low HbF levels and high haemopoietic stress. The heterogeneous regulation of BFU‐E in SS disease seems to be the epiphenomenon of HbF levels, and not vice versa.
Induction of TNF‐α in patients with myelodysplastic syndromes undergoing treatment with interleukin‐3
Authors: Seipelt Gernot , Ganser Arnold , Duranceyk Heidi , Maurer Alexander , Ottmann Oliver G. , Hoelzer Dieter 
Published: March 2008

Summary. The study was undertaken to analyse whether the presence or the induction of TNF‐α, a potent inhibitor of haemopoiesis, might affect the clinical response to treatment with interleukin‐3 in patients with myelodysplastic syndromes. A total of 15 patients were treated with IL‐3. Baseline serum TNF‐α levels were elevated in MDS patients (14·2 ± 2·4 pg/ml) compared to healthy controls (9·1 ± 1·1 pg/ml). During IL‐3 therapy TNF‐α levels remained unchanged in 3/14 patients in whom platelet counts increased, while in non‐responders TNF‐α levels increased 1·9‐fold (P < 0·025).These findings indicate that TNF‐α not only is induced during IL‐3 therapy in MDS patients but that this elevation might be associated with a poor platelet response to therapy.
Reduced responsiveness of bone marrow megakaryocyte progenitors to platelet‐derived transforming growth factor β1, produced in normal amount, in patients with essential thrombocythaemia
Authors: Zauli Giorgio , Visani Giuseppe , Catani Lucia , Vianelli Nicola , Gugliotta Luigi , Capitani Silvano 
Published: March 2008

Summary In this study we evaluated the amount of transforming growth factor‐β1 (TGF‐β1) in platelet lysates obtained from 12 patients affected by essential thrombocythaemia (ET) in comparison with five patients affected by myelofibrosis with myeloid metaplasia (MMM) and 15 healthy donors.The levels of both bioactive and latent TGF‐β1, evaluated in a bioassay on CCL64 cells, before and after transient acidification, were similar in platelet lysates from ET patients and normal donors and significantly (P < 0.01) elevated in platelet lysates from MMM patients. Moreover, platelet lysates from ET patients and normal controls, showed a similar degree of colony suppression when tested on haematopoietic progenitor (CD34+) cells, purified from normal bone marrows, whereas platelet lysates from MMM patients showed a higher (P < 0.01) inhibitory activity on normal CFU‐meg and BFU‐E growth.In parallel, platelet lysates form ET patients and normal controls were tested on CD34+ cells, purified from ET bone marrows. ET bone marrow BFU‐E, similarly to normal bone marrow BFU‐E, were markedly inhibited by platelet lysates, whereas ET bone marrow CFU‐meg were significantly (P < 0.05) less responsive to the inhibitory activity of platelet lysates than normal bone marrow CFU‐meg. The main factor responsible for the inhibitory activity contained in platelet lysates was transforming growth factor‐β1 (TGF‐β1), as demonstrated by the ability of a polyclonal neutralizing anti‐TGF‐β1 antibody to almost completely reverse the suppressive effect of platelet lysates on CFU‐meg and BFU‐E growth.Our data demonstrate that the amount of intraplatelet TGF‐β1 is similar in ET patients and normal controls, whereas it is increased in platelets from MMM patients. Moreover, megakaryocyte progenitors in ET show a reduced sensitivity to platelet‐derived inhibitors and, in particular, to TGF‐β1.
Quantification of leukaemic cells based on heteroduplex formation of tal‐1 gene sequences after PCR coamplification
Authors: Borkhardt A. , Repp R. , Harbott J. , Kreuder J. , Lampert F. 
Published: March 2008

Summary Based on the high sensitivity of the polymerase chain reaction (PCR) several assays have already been described which can be applied to monitor minimal residual disease in patients with leukaemia. However, most of these approaches are only qualitative or semiquantitative at best. Moreover, the semiquantitative assays require rather largescale procedures such as oligonucleotide hybridization or sampling of aliquots during the exponential phase of PCR amplification, which is time consuming and may be error prone.Using the deletion in the tal‐1 gene as a clonal marker of malignant cells, which can be found in about 10–25% of T‐cell acute lymphoblastic leukaemia, we have developed a rapid and simple PCR assay for the quantification of residual leukaemic cells. The assay is based on heteroduplex formation between standard and template DNA sequences after coamplification of both DNA sequences in the same PCR reaction. The sensitivity of this PCR approach allows the detection of neoplastic cells at frequency of at least 10‐4. Application of such an assay needs not to be restricted to patients harbouring a tal‐1 gene deletion. It may easily be adapted to other clonal DNA markers of blast cells and therefore facilitate monitoring of minimal residual disease in many different kinds of leukaemia.
Characterization of an acute micromegakaryocytic leukaemia: evidence for the pathogenesis of myelofibrosis
Authors: Reilly J. T. , Barnett D. , Dolan G. , Forrest P. , Eastham J. , Smith A. 
Published: March 2008

Summary The current hypothesis for the pathogenesis of myelofibrosis involves the intramedullary release of growth factors from defective or abnormal megakaryocytes. We describe a case of an acute micromegakaryocytic leukaemia, in a patient with chronic myelofibrosis, that provides additional evidence for this concept. The micromegakaryocytes, which reached 223 × 109/1, were characterized morphologically by both light and electron microscopy, immunocytochemically and by platelet peroxidase activity. The cells were shown to have a mature cytoplasm, containing alpha granules and the associated proteins: vWF:Ag, fibrinogen, fibronectin and protein S. DNA analysis, by both a Seescan Solitaire Plus image analysis system and flow cytometry, revealed nuclear immaturity, with 92% of cells being diploid.Serum markers of connective tissue synthesis, namely carboxy terminal peptide of procollagen I (PICP), procollagen terminal peptide III (PIIIP) and laminin all increased significantly following transformation and were associated with an increase in platelet‐derived growth factor (PDGF) and transforming growth factor‐β (TGF‐β). These observations support the current hypothesis for bone marrow fibrosis formation and provide, for the first time, a link between in vivo growth factor release, bone marrow stromal turnover and megakaryocyte mass. In addition, the release of biologically active TGF‐β may explain both the increased fibronectin and angiogenesis characteristic of myelofibrotic bone marrow.
Acquired sideroblastic anaemia during treatment of Wilson's disease with triethylene tetramine dihydrochloride
Authors: Condamine Luce , Hermine Oliver , Alvin Patrick , Levine Martine , Rey Caroline , Courtecuisse Victor 
Published: March 2008

Summary We report one case of acquired sideroblastic anaemia in a patient treated for Wilson's disease with triethylene tetramine dihydrochloride (TTH). No other cause of acquired sideroblastic anaemia was found, and neither iron nor pyridoxine therapy could correct this anaemia. In contrast, decreasing the dose of TTH led to disappearance of ringed sideroblasts.Thus TTH should be added as a further cause of secondary acquired sideroblastic anaemia. The pathophysiology of this finding, probably linked to an abnormality of mitochondrial iron metabolism, is briefly discussed.
Long‐term bone marrow culture in Fanconi's anaemia
Authors: Stark R. , Thierry D. , Richard P. , Gluckman E. 
Published: March 2008

Summary. Fanconi's anaemia (FA) is the most common of the constitutional aplastic anaemias; the mechanisms leading to aplasia in this disease are poorly understood. A number of mechanisms have been implicated in the pathogenesis of acquired aplastic anaemia (AA), including a stem cell defect, an immune reaction against haematopoietic cells or defective function of the marrow microenvironment. To investigate the pathophysiology of this disorder we have performed bone marrow colony forming unit‐granulocyte macrophage (CFU‐GM) assays and long‐term bone marrow culture (LTC) in 22 cases of FA compared with 17 cases of acquired AA.Defective in vitro haematopoiesis was observed in all patients with FA, including several cases with normal peripheral blood counts. The mean CFU‐GM value for the FA group was approximately 15 times lower than for the normal group. A correlation was seen between CFU‐GM and the severity of neutropenia in FA. In LTC an adherent layer formed in all cases of FA; despite this fact CFU‐GM were either not generated or rapidly fell to zero in all patients. LTC is a sensitive method for the detection of impaired granulopoiesis in FA and reveals defects in all patients with this disease.