bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polyethylene glycol (PEG) modification of granulocyte‐macrophage colony stimulating factor (GM‐CSF) enhances neutrophil priming activity but not colony stimulating activity
Authors: Knüsli Claudio , Delgado Cristina , Malik Farooq , Dómine Manuel , Tejedor M. Cristina , Irvine Alexandra E. , Fisher Derek , Francis Gillian E. 
Published: March 2008

SummaryPEG‐modified proteins have numerous advantages over their unmodified counterparts (increased half life, reduced antigenicity, improved solubility), but, almost without exception, they show a modest to marked reduction in biological or enzymatic activity. However, while investigating a new protocol for the preparation of PEG‐proteins, we compared PEG‐modified and unmodified GM‐CSF with respect to their polymorphonuclear neutrophil granulocyte (PMN) priming activities. PEG‐GM‐CSF was unexpectedly more active than GM‐CSF in its ability to prime neutrophils to respond to the synthetic peptide n‐formyl‐methionyl‐leucyl‐phenylalanine (FMLP) with an oxidative burst (assessed both by nitroblue tetrazolium reduction and ferricytochrome c reduction). These results were in contrast to the findings for colony stimulating activity and with GM‐CSF induced thymi‐dine uptake, where the biological activity was unchanged or reduced. The enhanced neutrophil priming activity of PEG‐GM‐CSF was confirmed using FPLC fractionated PEG‐modified GM‐CSF. This showed changes in the bioactivity profile consistent with both the shift in protein elution profile and enhanced activity of the PEG‐modified material (reflected in the increased area under the bioactivity curve). We also excluded a neutrophil priming action for PEG‐modified fetal calf serum proteins, carrier proteins and‘irrelevant’cytokine, erythropoietin. The dissociation of the two bioactivities was confirmed using individual FPLC fractions. These results suggest the presence of differences in either binding, receptor/ligand processing or signal transduction for neutrophils versus progenitors, that are differentially affected by PEG‐modification of GM‐CSF. The demonstration that PEG‐modification can partially dissociate two biological activities suggests the feasibility of using PEG‐modification to produce proteins with subtly altered spectra of biological activity and hence new ranges of clinical applications.
Effects of recombinant human granulocyte colony‐stimulating factor on neutrophil functions in aged animals
Authors: Yoshino Takeshi , Tamura Masahiko , Kawabe Mari , Nomura Hitoshi , Imai Nobuo , Ono Masayoshi 
Published: March 2008

SummaryWe have studied the effects of recombinant human granulocyte colony‐stimulating factor (rG‐CSF) on granulopoiesis and neutrophil functions in aged rats and aged mice. We subcutaneously injected rG‐CSF or control vehicle into aged rats (22 months old and 25 months old) for 7 consecutive days. counted the peripheral neutrophils and evaluated the functions of neutrophils isolated from venous blood. The peripheral neutrophil count in aged rats tended to be increased as compared with that in young rats (11 weeks old). However, the neutrophils in aged rats exhibited a decline of superoxide anion (O2‐‐) release and phagocytic activity as compared with young rats. The peripheral neutrophil count in aged rats was significantly increased 5–6‐fold as many as the control value by rG‐CSF treatment, which was accompanied by a significant enhancement of O2‐release and of phagocytic activity being restored to normal levels or better.In another series of experiments, we subcutaneously injected rG‐CSF or control vehicle into aged mice (24–28 months old) or young mice (8 weeks old) for 7 consecutive days, and evaluated the functions of neutrophils isolated from peritoneal cavity. The peritoneal exudate neutrophils from the aged mice exhibited a decline of phagocytic and chemo‐tactic activity as compared with the young mice. These functions in both young and aged mice were significantly enhanced by rG‐CSF‐treatment, and these functions in rG‐CSF‐treated aged mice were restored to a level higher than the level in control young mice.These findings demonstrate that rG‐CSF is capable of enhancing granulopoiesis and restoring the age‐related decline of neutrophil functions.
Long‐term treatment results for acute megakaryoblastic leukaemia patients: a multicentre study
Authors: Ruiz‐Argüelles Guillermo J. , Lobato‐Mendizábal Eduardo , San‐Miguel Jess F. , González Marcos , Caballero María D. , Ruiz‐Argüelles Alejandro , Orfao Alberto , Gómez‐Almaguer David , Vidriales Belén , Ruiz‐Reyes Guillermo , López‐Borrasca Antonio 
Published: March 2008

SummaryThe prognosis and long‐term results of a group of 57 acute megakaryoblastic leukaemia (M7‐AML) patients was analysed from a multicentre perspective. Ages ranged from 4 to 83 years, median 49 years: 30 were males and 27 were females. The median follow‐up time was 7 months, range 1–24 months. Early exits occurred in 12 cases, their median age being 71 years. Forty‐five patients were treated with combined aggressive chemotherapy (CT) (n= 26) or low‐dose cytarabine (LD‐AraC) (n= 19). The following results were obtained with combined CT or AraC, respectively. Complete remission (CR) rates were 73% and 84%. 12‐month survival (SV) were 37% and 26%, 24‐month SV were 12% and 11%. median SV 10 and 4 months, and relapse rates (RR) were 68% and 94%. These differences were not statistically significant. Irrespective of the treatment modality, the results were better for children (n = 10) than for adults (n = 35): RR rates were 90% and 74%, median SV: 7 and 5 months, 12‐month SV: 40% and 22%, 24‐month SV: 30% and 9%, and RR: 78% and 81%. respectively: these differences also were not statistically significant. In addition, a literature review of 42 patients from 18 previous reports is presented, including seven cases treated with allogeneic bone marrow transplantation (BMT). The best results were obtained with BMT: 12 and 24 month SV was 86% and the RR was 0%. On the above‐mentioned basis, we feel that children and young adults with M7‐AML should be offered BMT. In patients over 60 years old or not eligible for aggressive chemotherapy or BMT, an interesting possibility would be the use of LD‐AraC which allows a high CR rate, followed by a classical consolidation regimen in order to prevent early relapses.
Immunosuppressive property of a very high purity antihaemophilic preparation: a low molecular weight component inhibits an early step of PHA induced cell activation
Authors: Vermot‐Desroches Claudine , Rigal Dominique , Blourde Christian , Bernaud Jeanine 
Published: March 2008

Summary.Immune deficiency has been reported in haemophiliac patients receiving antihaemophilic factor VIII preparations, but the mechanisms involved in the immunosup‐pression are not fully understood. By using the proliferative response of peripheral blood mononuclear cells to phyto‐haemagglutinin (PHA) as a test system, we investigated the inhibitory influence of a very high purity antihaemophilic factor (AHF) preparation on T cell proliferation and on T lymphocyte activation molecules. We observed that this preparation reduced significantly the PHA‐induced mono‐nuclear cell proliferation, independently of the monocyte concentration. The AHF preparation did not act through a cytotoxic mechanism or a steric hindrance of PHA. The AHF preparation had no effect on the immediate expression of T lymphocyte activation molecules such as CD54 (ICAM‐1). In contrast, the very high purity AHF reduced the induced expression of two early T cell activation molecules: CD25 (interleukin‐2 receptor) and CD71 (transferrin receptor). The very high purity AHF also had the capacity to inhibit the up‐regulation of two late activation antigens, CD38 and CD11a/CD18, and to inhibit the induced expression of HLA‐DR molecule, defined also as a late T cell activation molecule. The CD45R expression level, used as a control marker, was not changed after AHF exposure. The very high purity AHF therefore influenced an early step of cell proliferation.We have also shown that the immunoregulatory properties of the preparation were not restricted to the factor VIII itself, but resulted from the presence of dialysable and low molecular weight components in the preparation.
Myelodysplastic syndrome (MDS)‐associated inhibitory activity on haematopoietic progenitor cells: contribution of monocyte‐derived lipid containing macrophages (MDLM)
Authors: Ohmori Masami , Ueda Yasunori , Masutani Hiroshi , Hirama Toshiyasu , Anzai Naoyuki , Yoshida Yataro , Okuma Minoru 
Published: March 2008

We studied myelodysplastic syndrome‐associated inhibitory activity (MDS‐IA), which inhibited colony formation in vitro of normal granulocyte‐macrophage progenitors (CFU‐GM). When adherent marrow cells were incubated with fetal calf serum for 21–24 d. monocyte‐derived lipid containing huge macrophages (MDLM) developed. MDLM from MDS marrow (MDS‐MDLMs) and their conditioned medium (MDLM‐CM) consistently suppressed the growth of normal CFU‐GM colony formation. MDS‐IA was active on CFU‐GM during the S‐phase and relatively resistant to heating. Monoclonal antibody against H subunit (acidic) ferritin and polyclonal antibody against placental ferritin neutralized the inhibitory activity of MDS‐MDLMs. In addition, cell lysates of MDS‐MDLMs reacted to both monoclonal anti‐H subunit ferritin and polyclonal anti‐placental ferritin in Western blotting analysis, indicating that the inhibitory activity was predominantly acidic isoferritin.On the other hand, MDLMs obtained from normal bone marrow had a CFU‐GM enhancing activity. These results suggest that MDS‐MDLMs may be responsible for the suppression of granulopoiesis in patients with MDS and that the suppression may be mediated by soluble factors, notably H subunit isoferritin.
Cellular sites of extrarenal and renal erythropoietin production in anaemic rats
Authors: Schuster S. J. , Koury S. T. , Bohrer M. , Salceda S. , Card J. 
Published: March 2008

Summary.The cellular origins of erythropoietin were investigated in the rat using a probe derived from a cloned rat erythropoietin cDNA. In anaemic‐hypoxic rat liver, in situ hybridization detected erythropoietin mRNA primarily in hepatocytes and less frequently in nonparenchymal sinusoidal or perisinusoidal liver cells. An RNase protection assay was used to compare the erythropoietin mRNA contents of separated rat liver cell fractions and also suggested that hepatocytes are the major source of extrarenal erythropoietin with nonparenchymal liver cells contributing less than 1% to total hepatic erythropoietin production. In kidney, in situ hybridization localized erythropoietin mRNA in nonepithelial cells, as yet of undefined lineage, in the cortical and outer medullary interstitium. These results indicate that, in the rat, the primary sources of erythropoietin in liver and kidney are different types of cells.
Mutations of the p53 tumour suppressor gene in haematologic neoplasms
Authors: Mori Naoki , Wada Makio , Yokota Jun , Terada Masaaki , Okada Michiko , Teramura Masanao , Masuda Michihiko , Hoshino Shigeru , Motoji Toshiko , Oshimi Kazuo , Mizoguchi Hideaki 
Published: March 2008

Summary.Mutations of the p53 tumour suppressor gene have frequently been observed in several types of solid tumours and are believed to be implicated in the development of these tumours. To determine the relevance of p53 mutations in haematologic neoplasms, we performed polymerase chain reaction‐single strand conformation polymorphism analysis on the p53 gene in 45 patients with various types of haematologic neoplasms. In exons 5–8 containing highly conserved regions, mobility shifts indicating sequence alterations were detected in four of the 45 patients, and subsequent sequencing was performed. A point mutation resulting in a novel stop codon was detected at codon 213 in one of 23 cases of chronic myelogenous leukaemia (one of five cases of blast crisis). Point mutations causing amino acid substitutions were detected in one of four cases of myelodysplastic syndrome at codon 195, one of three cases of adult T‐cell leukaemia at codon 281, and one of eight cases of acute lymphoblastic leukaemia at codon 281, and these missense mutations were accompanied by loss of the wild type allele. Patients harbouring these nonsense and missense mutations were in advanced disease stages. These findings suggest that mutational inactivation of the p53 gene is infrequent but is involved in the tumorigenesis of several types of haematologic neoplasms at least in some cases.
Electrophoretic and functional characteristics of the von Willebrand factor in human fetal plasma
Authors: Mazurier Claudine , Daffos Fernand , Forestier Francois 
Published: March 2008

Summary.Although the haemostatic role of von Willebrand factor (vWF) in adults is well known, there is little information currently available about its possible contribution to in utero haemostasis. We have investigated the distribution of vWF multimers in 27 pure fetal platelet‐poor plasma (FP) samples aged from 20 to > 36 weeks, by using electrophoresis in both low‐and high‐resolution agarose gels in the presence of sodium dodecyl sulphate. Our data confirm that most FP samples contain higher molecular weight (HMW) vWF multimers than those present in normal adult plasma, the proportion of these HMW forms being lower in FP samples aged >36 weeks. However, we found that the multimeric unit and the protomeric form of vWF were similar in normal fetal and adult plasma. Functional assays of vWF were performed on three pooled FP samples. In all cases, fetal vWF was able to interact with factor VIII and to bind to GP Ib platelet receptor in the presence of ristocetin and to types I and III collagen. These results indicate that plasma vWF has already acquired in fetal life the functional activities required for its role in both coagulation and primary haemostasis.
The resistance of macrophage‐like tumour cell lines to growth inhibition by lipopolysaccharide and pertussis toxin
Authors: Xie Yue , von Gavel Stephanie , Cassady A. Ian , Stacey Katryn J. , Dunn Timothy L. , Hume David A. 
Published: March 2008

Summary. The process of tumorigenesis is frequently associated with resistance to growth inhibition by physiological regulators of normal cells. Murine macrophage‐like cell lines BAC1.2F5, RAW264, J774.1A and PU5/1.8 were resistant to growth inhibition by bacterial lipopolysaccharide (LPS) and pertussis toxin, agents that blocked growth of primary bone marrow‐derived macrophages (BMDM) in the presence of macrophage colony‐stimulating factor (CSF‐1). The resistance of the CSF‐1‐dependent cell line BAC1.2F5 to growth inhibition by pertussis toxin argues against the possibility that pertussis toxin‐sensitive G proteins are essential for the pathway of growth stimulation by CSF‐1. Conversely, these data add further weight to the argument that LPS mediates some of its biological activities by mimicking the action of pertussis toxin and inhibiting G protein function. The resistance of cell lines to LPS and pertussis toxin was not correlated with any alteration in the expression of mRNA encoding any of three pertussis‐toxin sensitive G protein α subunits. The pattern of G protein expression was consistent between primary cells and tumour cells, suggesting that this is a differentiation marker. In particular, Giα2 mRNA was expressed at remarkably high levels in all of the cells. The specificity of LPS resistance was investigated by studying down‐regulation of CSF‐1 binding and induction of protooncogene c‐fos and tumour necrosis factor (TNF) mRNA. BAC1.2F5 cells were LPS‐resistant in each of these assays. In CSF‐1 binding. RAW264 and J774.1A responded in the same way as bone marrow‐derived macrophages but required higher doses of LPS, whereas c‐fos and TNF mRNA were induced in these cells at concentrations that did not inhibit growth.In PU5/1.8 cells, CSF‐1 binding was already very low and was not further down‐regulated, but c‐fos and TNF mRNA was inducible by LPS. By contrast to primary macrophages, the cell lines did not respond to LPS with down‐regulation of c‐fms mRNA, which encodes the CSF‐1 receptor. Hence, the resistance of macrophage‐like tumour cells to LPS and pertussis toxin was specific to the pathways controlling growth, and was correlated with altered regulation of the CSF‐1 receptor.
Peripheral blood progenitor cells mobilized by chemotherapy plus granulocyte‐colony stimulating factor accelerate both neutrophil and platelet recovery after high‐dose VP16, ifosfamide and cisplatin
Authors: Brugger Wolfram , Birken Rainer , Bertz Hartmut , Hecht Thomas , Pressler Klaus , Frisch Jürgen , Schulz Gregor , Mertelsmann Roland , Kanz Lothar 
Published: March 2008

Summary. We report on the chemotherapy plus granulocyte colony‐stimulating factor (G‐CSF) induced mobilization of peripheral blood progenitor cells (PBPCs) and their impact on haematopoietic recovery following high‐dose chemotherapy. Twenty‐four patients with advanced solid tumours or lymphomas received standard‐dose chemotherapy with VP16, ifosfamide and cisplatin (VIP) followed by filgrastim (G‐CSF; 5 μg/kg s.c. daily for 14 d) for the prevention of chemotherapy induced neutropenia and for the simultaneous mobilization of PBPCs. Maximal numbers of progenitors of different lineages were reached at day 11 (range 9–14) after VIP chemotherapy. A median of 0·415 × 109/1 CD34+ cells (range 0·11–1·98), 9000 CFU‐GM/ml (range 2800–17700). 3500 BFU‐E/ml (range 400–10800) and 200 CFU‐GEMM/ml (range 0–4400) were recruited. One single apheresis yielded a median of 1·6 × 108 mononuclear cells/kg (range 0·2–5·4) or 5·4 × 106 CD34+ cells/kg body weight (range 0·2–24·2).Fourteen patients who showed at least a partial remission after two cycles of the standard‐dose chemotherapy regimen were subjected to high‐dose VIP chemotherapy (cumulative doses of 1500 mg/m2 VP16, 12 g/m2 ifosfamide and 150 mg/m2 cisplatin) with or without PBPC support. The first six patients were treated with growth factors only (IL‐3/GM‐CSF) and did not receive PBPCs, whereas the following eight patients were supported with PBPCs in addition to IL‐3 and GM‐CSF. Neutrophil recovery as well as platelet recovery were significantly faster in patients receiving PBPCs with a median of 6·5 d below 0·1 × 109 neutrophils/1 and 3 d below 20 × 109 platelets/1 as compared to 10·5 d and 8 d in control patients receiving growth factors only. The accelerated platelet recovery in patients supported with PBPCs might be explained—in the absence of detectable colony‐forming units megakaryocyte—by the presence of glycoprotein IIb/IIIa+, non‐proliferating endomitotic megakaryocytic precursor cells within G‐CSF mobilized PBPCs. Our data demonstrate that chemotherapy plus G‐CSF mobilized PBPCs accelerate both neutrophil and platelet recovery after high‐dose VIP chemotherapy in patients with solid tumours or lymphomas.
Whole blood platelet aggregation and coagulation factors in patients with systemic sclerosis
Authors: Goodfield M. J. D. , Orchard M. A. , Rowell N. R. 
Published: March 2008

Summary. It is unclear whether the changes in platelet function which are observed in systemic sclerosis are a primary characteristic of this disease or whether they occur secondary to vascular changes. Whole blood platelet aggregation was studied in 26 patients with systemic sclerosis, normal subjects matched for age, sex and secondary characteristics, 19 patients with Raynaud's disease and 19 patients with systemic lupus erythematosus. Plasma levels of fibrinogen, von Willebrand factor antigen and factor VIII:C were also measured.Systemic sclerosis was associated with a significant (P> 0·001) enhancement of the sensitivity of platelets to collagen. In contrast, significant enhancement of the response to either ADP or adrenaline was not observed. Enhanced sensitivity to collagen was not associated with the presence of either Raynaud's disease or systemic lupus erythematosus.Systemic sclerosis was associated with significantly raised levels of von Willebrand factor antigen and fibrinogen. On an individual patient basis, von Willebrand factor antigen was related to the severity of the disease whereas platelet sensitivity to collagen was not. In conclusion, this study suggests that the enhanced sensitivity to collagen which occurs in systemic sclerosis is due to a primary change in the platelet and that this change can combine with elevated levels of adhesive proteins.
Relapse of aplastic anaemia after immunosuppressive treatment: a report from the European Bone Marrow Transplantation Group SAA Working Party
Authors: Schrezenmeier H. , Marin P. , Raghavachar A. , McCann S. , Hows J. , Gluckman E. , Nissen C. , Veer‐Korthof E. T. van't , Ljungman P. , Hinterberger W. , Lint M. T. Van , Frickhofen N. , Bacigalupo A. 
Published: March 2008

Summary This study was designed to determine the incidence of relapse and factors predictive for relapse in 719 patients with severe aplastic anaemia (SAA) after immunosuppressive treatment (IS). Patients developing myelodysplasia or acute leukaemia after IS, and patients receiving a transplant, were excluded from this analysis. Response was defined as reaching complete independence from transfusions, relapse was defined as becoming again transfusion dependent. This criteria was validated by similar figures when using other ‘relapse criteria’ such as drop in neutrophil or platelet counts.Of 358 patients responding to IS, 74 patients relapsed after a mean time of 778 d after treatment. The actuarial incidence of relapse is 35.2% at 14 years after IS. The risk for relapse was higher in patients responding within 120 d from IS (48%) compared to patients responding between 120 and 360 d (40%) and only 20% for slow responders (>360d from IS) (P<0.00001). In multivariate analysis this factor still proved significant (P<0.0001). The mean time between diagnosis and treatment was significantly longer in patients relapsing compared to patients who did not relapse (260 v 134 d, P=0.037). Relapse was not predicted by the severity of the disease, age, and sex. In 39 of the 74 relapsing patients a second response could be achieved. Responses after relapse were associated in univariate analysis with early response to previous IS and early occurrence of relapse. The actuarial survival of patients not relapsing is significantly better than survival of patients relapsing (79.8%v 67.1%, P = 0.0024). However, the actuarial survival of 39 relapsing patients who responded again to IS was similar to patients not relapsing (86%) and significantly better than in 35 patients not reaching a second response after relapse (49.3%, P=0.0015). This study indicates that relapse is a relevant problem in the treatment of aplastic anaemia, and does have an impact on overall survival. Prospective studies of immunosuppressive regimens, looking at responses, should also address this problem in the future.