bluuebunny/crossref_metadata_2025 huggingface dataset:
Malignant transformation and life expectancy in monoclonal gammopathy of undetermined significance
Authors: Blade Joan , Lopez‐Guillermo Armando , Rozman Ciril , Cervantes Francisco , Salgado Camino , Aguilar Josep‐Lluis , Vives‐Corrons Joan‐Lluis , Montserrat Emili
Published: March 2008

The actuarial probability of malignant transformation and the impact on expected survival were analysed in a series of 128 persons diagnosed with monoclonal gammopathy of undetermined significance (MGUS) over a 20‐year period. At a median follow‐up of 56 months the M‐component remains stable in 101 patients (78·9%), 14 patients (10·9%) have died from non‐related disorders and 13 (10·2%) have developed malignant transformation of MGUS (multiple myeloma, 10; primary amyloidosis, two; Waldenström's macroglobulinaemia, one). The actuarial probability of malignant transformation at 5 and 10 years was 8·5% and 19·2%, respectively. When different presenting features were analysed for predictive value of the malignant transformation, the IgA type of MGUS was the only variable associated with a higher probability of such an event (P<0·025). Although no significant difference was observed between the survival probability of persons with MGUS and that of the control population, the development of malignant transformation was associated with a shorter survival (P<0·001).
Unique expression of von Willebrand factor by type IIA von Willebrand's disease endothelial cells
Authors: Wilbourn Barry , Harrison Paul , Lawrie Andrew , LeRoy Frances , Rowley Megan , Bevan David , Inbal Aida , Savidge Geoffrey
Published: March 2008

Endothelial cells (EC) were cultured from the umbilical cord of a male neonate whose mother was previously diagnosed with type IIA von Willebrand's disease (vWd). The diagnosis of type IIA vWd in the proband was confirmed by low ristocetin activity and the absence of the highest molecular weight (MW) forms of von Willebrand factor (vWf) in his platelet poor plasma. The vWf of EC cultured from the neonate's umbilical cord differed from that of control EC and the cell line EA.hy926 in two respects. Firstly, the full range of molecular weight forms was present in the patient EC lysate and, secondly, vWf:Ag expression was approximately seven‐fold greater than that of control cells, Platelet lysates prepared from other affected members of the type IIA vWd family in the presence or absence of proteolytic inhibitors demonstrated a near normal vWf multimeric distribution. Resistance of these high MW forms to heat degradation was conferred by the presence of proteolytic inhibitors. Moreover, the full plasma vWf multimeric distribution could not be restored by the inclusion of EDTA, N‐ethylmaleimide and leupeptin in the anticoagulant during the rapid preparation of platelet poor plasma. These findings lend support to the heterogeneous nature of type IIA vWd and has possible implications in the understanding of the intracellular processes involved in the biosynthesis and storage of the vWf macromolecular complex as well as the pathogenesis of type IIA vWd.
Plasmodium falciparum: diversity of isolates from Malawi in their cytoadherence to melanoma cells and monocytes in vitro
Authors: Goldring J. D. , Molyneux M. E. , Taylor T. , Wirima J. , Hommel M.
Published: March 2008

We observed considerable diversity in the cytoadherence of Plasmodium falciparum isolates from Malawi to melanoma cells, U937 cells and human peripheral monocytes. Each isolate exhibited a unique cytoadherence profile for the three human cell types. These isolates generally adhered well to U937 cells and fresh monocytes, moderately to melanoma cells and poorly to TE 671, MIA‐Pa‐Ca, WI 38, PLC/PRF/5 and HeLa cells. An antimalarial immunoglobulin pool inhibited binding to melanoma cells by 50% or more and to U937 cells by 25% or less. There was no correlation between in vitro cytoadherence to the three cells and clinical disease. These results suggest that malarial adherence ligands exposed on the surface of infected erythrocytes vary from one isolate to another.
Further investigations on the expression of HLA‐DR, CD33 and CD13 surface antigens in purified bone marrow and peripheral blood CD34± haematopoietic progenitor cells
Authors: Pierelli Luca , Teofili Luciana , Menichella Giacomo , Rumi Carlo , Paoloni Alessandra , Iovino Stefania , Puggioni Pier Luigi , Leone Giuseppe , Bizzi Bruno
Published: March 2008

Summary. We evaluated the HLA‐DR, CD33 and CD13 antigen expression on CD34± haematopoietic progenitor cells (HPC) isolated from the bone marrow (BM) and peripheral blood (PB) of normal donors. The majority of both BM and PB CD34± HPC expressed CD13 and HLA‐DR. The coexpression of CD34 and CD33 was found in a minor CD34± subset. After 7 d of culture in the presence of interleukin‐3 and granulocyte‐macrophage colony‐stimulating factor, CD33 expression was detected in about 50% of HPC. At this point CD34 antigen expression was lost and CD13 and HLA‐DR expression was partially lost. After 14 d of culture, the majority of HPC were CD33±. HPC maintained the capacity to generate colony forming unit granulocyte‐macrophage but they lost the capacity to generate burst forming unit‐erythroid. A correlation was found between the percentage of CD34±/HLA‐DR± cells and the total number of colony forming cells in unfractionated samples from BM and PB of patients with malignancies. These studies demonstrate that, in normal conditions, only a minor subset of CD34± cells coexpress CD33 antigen either in BM or in PB and CD33 antigen is a lineage marker which is coexpressed with HLA‐DR and CD13 on a progenitor committed to the granulocytic‐macrophagic lineage.
Expression of T‐cell receptor α and β variable genes in normal and malignant human T cells
Authors: Hu Huaizhong , Queirò Màrio Rui , Tilanus Marcel G. J. , de Weger Roel A. , Schuurman Henk‐Jan
Published: March 2008

Summary. A PCR method was developed to analyse each of 29 families of the T cell receptor Vα gene and 20 families of the Vβ gene at the mRNA level in heterogenous cell populations. All Vα and Vβ families were detectable in blood mononuclear cells from four of six healthy donors. In two donors only Vα22 was missing, and all other Vα and Vβ families were detected. Vβ family expression was observed in T‐leukaemic cell lines Jurkat, HSB, Molt‐3 and Molt‐4. In contrast, Vα family expression was not detectable in any cell line except Jurkat cells. In T‐cell malignancies (non‐Hodgkin's lymphoma and mycosis fungoides), one or two Vα and Vβ families were detectable. Four of 10 cases investigated showed two Vα transcripts and one Vβ transcript. This fits with concepts in literature that allelic exclusion for the genes encoding α chains is not strictly required in the DNA rearrangement, or that this exclusion is a post‐translational event. Using a limited series of antibodies to Vβ gene family products, blood mononuclear cells from healthy donors were analysed by flow cytometry in a follow‐up study. Two of four donors were rather stable in proportions of T cells expressing distinct Vβ families, and two other donors showed variation in one or more families. When analysed on frozen tissue sections of normal lymph node and tonsil, there was no preferential location of lymphocytes expressing a distinct Vβ gene family in different compartments (interfollicular area, follicle, or tonsillar epithelium).
Low molecular weight dermatan sulphate (Desmin 370) does not cross the ovine placenta
Authors: Dawes Joan , Lumbers Eugenie R.
Published: March 2008

Summary. Transplacental passage of the low molecular weight dermatan sulphate Desmin 370 was investigated in pregnant sheep, using 125I‐labelled Desmin 370 to optimize the sensitivity of the study. Chronically catheterized crossbred pregnant ewes at approximately 120 d gestation received 3700 kBq 125I‐labelled Desmin 370 with 1 mg/kg carrier unlabelled Desmin 370 intravenously. Early clearance from the maternal circulation was biexponential, and the volume of distribution corresponded closely with the theoretical value for distribution in total body water. Soon after injection low levels of radioactivity were detected in the fetal circulation and accumulated over the next 2 h, so that as the concentration of 125I‐Desmin 370 in the maternal circulation declined with time the fetal level of radiolabel rose to represent a significant concentration in relation to that in the mother. Radioactivity was also excreted into the fetal urine. However, while 50% of the radiolabeled material present in maternal plasma 150 min post‐injection was intact Desmin 370 and the remaining 50% represented degradation products, fetal urine contained only these fragments. By contrast, intact Desmin 370 was readily excreted into fetal urine after direct introduction to the fetal circulation. Thus molecules of intact Desmin 370 with anticoagulant activity cannot cross the ovine placenta, and low molecular weight dermatan sulphates may be valuable for prophylaxis and treatment of thrombotic disease during pregnancy.
Mechanisms of platelet aggregation by Streptococcus sanguis, a causative organism in infective endocarditis
Authors: Ford I. , Douglas C. W. I. , Preston F. E. , Lawless A. , Hampton K. K.
Published: March 2008

Summary. The ability of certain strains of Streptococcus sanguis to aggregate human platelets in vitro may be related to their virulence in the pathogenesis of infective endocarditis. We have studied the mechanisms of aggregation of human platelets by S. sanguis strain NCTC 7863. Platelet aggregation follows incubation of S. sanguis cells with platelet‐rich plasma from normal, healthy adults, after a lag of 7–19 min. Platelet aggregation was accompanied by 5‐hydroxytryptamine release and thromboxane B2 production. Aggregation was prevented by aspirin and by EDTA. Platelets from two patients with Glanzmann's thrombasthenia did not respond to bacteria. Fixed, washed platelets resuspended in normal plasma were not agglutinated by S. sanguis.

Blocking the glycoprotein Ib receptor with a monoclonal antibody inhibited aggregation of PRP. However, S. sanguis did not induce von Willebrand factor (vWF) binding to platelets; nor did the bacteria prevent ristocetin‐induced platelet agglutination or vWF binding. The aggregation response was not related to plasma vWF activity levels in normal subjects or in patients with von Willebrand's disease.

The platelet response to S. sanguis therefore resembles true aggregation, requiring the cyclo‐oxygenase pathway and the presence of glycoprotein IIb/IIIa. The mechanism also involves glycoprotein Ib, but not apparently through irreversible binding of vWF.
PADGEM/GMP‐140 expression on platelet membranes from homozygous beta thalassaemic patients
Authors: Del Principe Domenico , Menichelli Adriana , Di Giulio Stefano , De Matteis Walter , Cianciulli Paolo , Papa Giuseppe
Published: March 2008

Summary.Thromboembolic events, which are associated with significant morbidity and mortality, occur in betathalassaemia. We studied the expression of the platelet selectin PADGEM/GMP‐140 on intact cells from thalassaemic patients, as a marker ofin vivoplatelet activation. The mean of positive cells (%) was 38 · 143 · 20 · 65 in the patients versus 5 · 048 | 1 · 8 in the controls.n= 21.P< 0<001.

No correlation was found between GMP‐140 expression and splenectomy, platelet counts, plasma ferritin and natural coagulation inhibitors. Instead an indirect correlation was found between GMP‐140 expression and HDL‐cholesterol. Moreover platelet activation was directly correlated with prebeta lipoproteins. Our data indicate that thalassaemic patients present anin vivoplatelet activation, which possibly depends on the dyslipidaemia, which is now regarded as a frequent feature of this disease.
Membrane properties of erythrocytes in subjects undergoing multiple blood donations with or without recombinant erythropoietin
Authors: Brugnara Carlo , Kruskall Margot S. , Johnstone Rose M.
Published: March 2008

Summary. To examine the characteristics of ‘young’human red cells, we studied blood from seven healthy male volunteers who developed systemic reticulocytosis during a 3‐week blood donation period. Each of these subjects donated a total of 6 units (450 ml/unit) of blood (2 units/week for 3 weeks) with subcutaneous recombinant erythropoietin (SC rEPO; 200 U/kg daily for 3 weeks). Two of these subjects were also studied with a similar protocol in the absence of rEPO (4 · 5 · 0 · 5 units donated). SC rEPO administration was associated with an increased K content of the erythrocyte and with the appearance of hypochromic cells, which were initially normocytic and then became normochromic and microcytic. Measurements of cation transport revealed that, with the exception of the Na‐K‐Cl cotransport, all the systems studied increased their activities following blood donations with or without SC rEPO. The increase was highest in the K‐Cl cotransport (2‐ and 5‐fold for control and rEPO parts of the study, respectively), while the Na‐K pump increased slightly in control and 40% with rEPO. The Na‐Li countertransport increased 40% and 100% in the control and rEPO parts of the study, respectively. Concomitant with increased ion transport activity, electron microscopic studies of plasma and red cells of subjects receiving SC rEPO showed the presence of circulating exosomes and cytoplasmic multivesicular bodies. The transferrin receptor was detected in the circulating exosomes, thereby providing evidence that, as do nonhuman red cells, maturing human reticulocytes shed exosome‐associated transferrin receptors.
ICAM‐1 tissue overexpression associated with increased serum levels of its soluble form in Hodgkin's disease
Authors: Pizzolo G. , Vinante F. , Nadali G. , Ricetti M. M. , Morosato L. , Marrocchella R. , Vincenzi C. , Semenzato G. , Chilosi M.
Published: March 2008

Summary. We investigated ICAM‐1/CD54 tissue immunoreactivity and serum levels of its soluble form (sICAM‐1) in patients with Hodgkin's disease (HD) at diagnosis. ICAM‐1 was strongly expressed in involved tissues, and sICAM‐1 serum levels were higher in HD (79 patients) than in controls (P < 0 · 01), and in patients with more advanced or more active disease (stages III ± IV vI ± II: P= 0 · 002; stage ‘B’v‘A’: P <0 · 0001; ‘bulky’disease v non‐‘bulky’: P= 0 · 042). We suggest that tissue ICAM‐1 overexpression leading to increase of circulating sICAM‐1 may interfere with the lymphocyte adhesion machinary thus contributing to the well‐known immune derangement of HD.
Functional analysis of the 4 bp deletion identified in the 5’untranslated region of one of the β‐globin genes from a Chinese β‐thalassaemic heterozygote
Authors: Francès V. , Morlé F. , Godet J.
Published: March 2008

Summary. Two plasmids have been constructed in which a β‐galactosidase/phleomycin‐resistance fusion gene reporter is placed under the control of the human β‐globin gene promoter and 5’untranslated region including or not including nucleotides 40–43 previously found deleted in one Chinese β‐thalassaemic allele. Transient expression assays of these two plasmids failed to reveal any significative effect of this 4 bp deletion either on the level of the β‐galactosidase activity produced in HeLa cells transfected in standard conditions, or on the rate of synthesis of the β‐galactosidase protein in transfected HeLa cells submitted to increasing osmotic shocks. These results suggest that this 4 bp deletion is not responsible for the β‐thalassaemic phenotype in vivo.
Megakaryocyte colony‐stimulating factor (Meg‐CSF) is a unique cytokine specific for the megakaryocyte lineage
Authors: Erickson‐Miller C. L. , Ji H. , Parchment R. E. , Murphy M. J.
Published: March 2008

Summary. The regulation of megakaryocytopoiesis and platelet production has not yet been clearly elucidated. Several cytokines have been shown to be capable of producing megakaryocyte colonies from bone marrow [i.e. Inter‐leukin (IL)‐3, granulocyte‐macrophage (GM)‐ colony‐stimulating factor (CSF), erythropoietin (Epo)]. In addition, other activities have been reported to stimulate megakaryocyte precursors, yet a megakaryocyte‐CSF (Meg‐CSF) has not been purified to homogeneity and IL‐3, GM‐CSF and/or Epo often contaminate purification attempts which could account for the activities. A Meg‐CSF has been isolated from the urine of patients with aplastic anaemia and purified by sequential ultrafiltration, cation exchange, G‐50 chromotography, pre‐parative PAGE, chromatofocusing and cation exchange HPLC. The activity of this material is 2−4 × 104 CFU‐Meg/mg as measured in a murine marrow, serum‐containing assay. This activity also stimulates CFU‐Meg in the absence of adherent accessory cells and in serum‐free cultures, indicative of the direct stimulation on CFU‐Meg. Immunoassays, colony forming assays, and proliferation assays demonstrate that purified Meg‐CSF has no GM‐CSF, IL‐3, M‐CSF, G‐CSF or IL‐1a, −3, −6, −9 and −11. In confirmation of these results, neutralizing antibody to IL‐6 also did not abrogate Meg‐CSF activity. Therefore the previously‐reported megakaryocyte colony‐stimulating activity in purified aplastic anaemia patient urine is due to a unique cytokine: Meg‐CSF.
High doses of intravenous recombinant erythropoietin for the treatment of anaemia in myelodysplastic syndrome
Authors: Goy A. , Belanger C. , Casadevall N. , Picard F. , Guesnu M. , Jaulmes D. , Poisson D. , Varet B.
Published: March 2008

Summary. We studied the efficacy of high doses (100 000 IU intravenously (IV)/twice a week) of human recombinant erythropoietin (rHuEpo) in patients with transfusion dependent myelodysplastic syndromes (MDS). Rationale for such dose of IV Epo was the poor in vitro response of MDS erythroid progenitors (CFU‐E) to physiological concentrations of Epo, and the usual high endogenous serum Epo levels of MDS patients. Seventeen patients (nine males, eight females) were included, five refractory anaemia (RA), six RA with blasts excess (RAEB), five RA with ringed sideroblasts (RARS).

Tolerance was good, except in three patients who experienced severe flu‐like syndrome after Epo injection. None of the patients showed hypertension or developed anti rHuEpo antibodies. Three patients (17.6%) with RAEB had 35–60% reduction of transfusion requirements. No progression of disease occurred. Percentage of erythroblasts, endogenous baseline Epo level and in vitro cultures of erythroid progenitors did not correlate with response to Epo treatment. This study shows that very high IV doses induce only seldom and partial improvement in the status of transfusion dependent MDS. This rate of response, not higher than described with lower dosage, probably represents the maximum expectable response to rHuEpo in this category of patients.
Immunotyping of blasts in refractory anaemia with excess of blasts
Authors: Oertel Joachim , Kleiner Sonja , Huhn Dieter
Published: March 2008

Summary. A study of immunological markers was performed in 16 patients with newly diagnosed refractory anaemia with excess of blasts (RAEB) and RAEB in transformation (RAEB‐T) and in 12 other patients with acute myeloid leukaemia evolving from RAEB or RAEB‐T. Immunocytochemical investigation of bone marrow blasts was done using a modified indirect immunoperoxidase technique. This method permitted accurate morphological identification of blasts and other cells in bone marrow. The monoclonal antibodies used in RAEB and RAEB‐T samples were anti‐CD34, ‐c‐kit, ‐HLA‐DR and ‐CD13.

The range of CD34 expression of blasts in RAEB samples was 1–14% (mean 6·2%) and in RAEB‐T samples 29–48% (mean 35·5%). CD34 positivity was detected in 3·94% (mean 47·4%) of the bone marrow blasts in acute myeloid leukaemia evolving from RAEB and RAEB‐T. Expression of c‐kit was demonstrated only in a low percentage of blast cells in RAEB, RAEB‐T and acute myeloid leukaemia following myelodysplasia. A high percentage (> 30%) of blasts in most patients with RAEB, RAEB‐T and acute myeloid leukaemia was HLA‐DR and CD13 positive. We observed the transformation from RAEB to acute myeloid leukaemia in three patients. The proportion of CD34 positive blasts increased to 25% and 32% in two patients. The third patient showed an unchanged percentage of CD34 positivity of blasts.

These findings indicate that the CD34 positivity of blasts increases with the progression of myelodysplasia to RAEB‐T and acute myeloid leukaemia demonstrating the instability of the clonal defect in myelodysplasia.
Effect of cytotoxic drugs on mature neutrophil function in the presence and absence of granulocyte‐macrophage colony‐stimulating factor
Authors: Humphreys John M. , Stringer Rebecca E. , Hart C. Anthony , Edwards Steven W.
Published: March 2008

Summary. The effects of the cytotoxic drugs, adriamycin, cyclophosphamide, daunomycin (daunorubicin), prednisolone, actinomycin D. azacytidine and vincristine at concentrations of 1 μM on mature neutrophil function were examined. Up to 5 h incubation with adriamycin, azacytidine, cyclophosphamide, daunomycin and prednisolone had no effect on either luminol chemiluminescence or superoxide secretion. However, after 15 min or 1 h (but not 5 h) incubation vincristine enhanced fMet‐Leu‐Phe stimulated chemiluminescence, whilst after 5 h incubation with actinomycin D the ability of neutrophils to generate reactive oxidants in response to all stimuli tested was impaired: after 5 h incubation with adriamycin reactive oxidant production was also impaired, but only when fMet‐Leu‐Phe was used as stimulant. All of the drugs tested except azacytidine inhibited neutrophil oxidant production after 5 h incubation in the presence of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Actinomycin D and cyclophosphamide also inhibited GM‐CSF stimulated protein biosynthesis. These data indicate that cytotoxic drugs may compromise the potentially beneficial effects of CSFs on mature neutrophil function during therapy.
Heparin‐induced thrombocytopenia and thrombosis: clinical and laboratory studies
Authors: Boshkov Lynn K. , Warkentin Theodore E. , Hayward Catherine P. M. , Andrew Maureen , Kelton John G.
Published: March 2008

Summary. Heparin‐induced thrombocytopenia is one of the most common and important immunological complications of drug therapy. Most patients with heparin‐induced thrombocytopenia have isolated thrombocytopenia, which by itself seldom causes serious morbidity. However, a small proportion of patients also develop an acute arterial thrombotic episode which can be fatal. It remains uncertain why some patients have only isolated thrombocytopenia, whereas others have thrombotic complications. In this report we describe 53 patients with heparin‐induced thrombocytopenia in whom the diagnosis was confirmed using the platelet 14C‐serotonin release assay. The intent of the study was to look for laboratory or clinical characteristics that could be used to predict which patients will have the less serious thrombocytopenia and which patients will have thrombocytopenia plus thrombotic complications. The laboratory markers included AT‐III, protein C, protein S and heparin cofactor II. No serological result identified whether a patient was at risk of having isolated thrombocytopenia or an acute thrombotic event. However, during the acute thrombocytopenic episode, there was evidence of global activation of the coagulation cascade as evidenced by reductions in the level of protein C, heparin cofactor II and antithrombin III. Following resolution of the thrombocytopenia, these inhibitory factors returned to normal indicating that the thrombotic complications were not caused by a familial deficiency.

We did observe a highly significant association (P < 0·001) between concomitant cardiovascular complications and the occurrence of an arterial thrombosis in patients with heparin‐induced thrombocytopenia. Recent surgery of any type was strongly associated with venous thrombi (P < 0·001). Our data suggest that heparin‐induced thrombocytopenia is a procoagulant disorder with thrombosis tending to occur at sites of pre‐existing pathology.
The spectrum of β‐thalassaemia in Algeria: possible origins of the molecular heterogeneity and a tentative diagnostic strategy
Authors: Bennani C. , Tamouza R. , Rouabhi F. , Benabadji M. , Malou M. , Elion J. , Labie D. , Beldjord C.
Published: March 2008

Summary. We report here on the final results of an epidemiological survey involving 177 β‐thalassaemic chromosomes in Algeria. Four common mutations account for 86% of the chromosomes, the other ones carrying nine other rare mutations. Combination of these results with those of other smaller regional epidemiological studies indicates the existence of still a wider range of mutations. The nature and frequencies of these mutations, their linkage with RFLP‐haplotypes, agree well with the history of the region. Knowledge of this spectrum of mutations enables the design of a diagnosis strategy that takes into account the local economical constraints.
A pilot study of low‐dose recombinant human granulocyte‐macrophage colony‐stimulating factor in chronic neutropenia
Authors: Kaczmarski R. S. , Pozniak A. , Lakhani A. , Harvey E. , Mufti G. J.
Published: March 2008

Summary. Recombinant human granulocyte macrophage colony stimulating factor (rhGM‐CSF) is under investigation for the treatment of a wide range of haematological disorders. At commonly used doses of > 120 μg/m2/d, extramedullary toxicity is common. We report the effects of low‐dose (LD) rhGM‐CSF in patients with chronic neutropenia related to HIV infection, myelodysplastic syndrome and idiopathic neutropenia. Nine patients with a mean pre‐treatment neutrophil count of 0·6 × 109/1 (range 0·2–1·4 × 109/1) received daily rhGM‐CSF at doses of between 5 and 15 μg/m2, Eight patients responded with a mean post‐treatment ANC of 3·2 × 109/1 (range 1·9–4·6 × 109/1). There was no significant therapy‐related morbidity. We conclude that in chronic neutropenia, LD rhGM‐CSF is an acceptable treatment which has important cost/benefit implications.
Disseminated intravascular coagulation in a patient with haemophilia B during factor IX replacement therapy
Authors: Ohga Shouichi , Saito Mitsumasa , Matsukazi Akinobu , Kai Takeshi , Ueda Kohji
Published: March 2008

Summary. We report a case of haemophilia B, who developed disseminated intravascular coagulation (DIC) with massive bleeding following an administation of factor IX complex concentrates. A 22‐year‐old male with severe haemophilia B had a bone fracture of the fibula and haemorrhage in the ankle joint because of a traffic accident. Factor IX complex concentrates were given to keep the plasma factor IX level more than 80% for surgery. 60 h after the infusion he showed epistaxis, haematuria and swelling of the injured ankle with evidence of DIC. A small dose of monoclonal antibody (mAb)‐purified factor IX concentrates with heparin and gabexate relieved the haemorrhage. A high dose of the mAb‐purified product attained more than 90% levels of plasma factor IX without coagulopathy. This observation emphasizes that currently available factor IX concentrates have thrombogenicity to induce DIC. Highly purified factor IX concentrates are needed for the high‐dose replacement in haemophilia B.
Chronic administration of transforming growth factor‐beta suppresses erythropoietin‐dependent erythropoiesis and induces tumour necrosis factor in vivo
Authors: Chuncharunee Suporn , Carter Candace D. , Studtmann Karl E. , Caro Jaime , Coffey Robert J. , Dessypris Emmanuel N.
Published: March 2008

Summary. Transforming growth factor beta is a known inhibitor of the proliferation and differentiation of early haematopoietic progenitors but has no effect on mature erythroid cells in vitro. Mice injected with rhTGFβ1 exhibited severe and progressive suppression of erythropoiesis manifested by a decline of reticulocyte count, marrow erythroblasts and marrow and spleen CFU‐E, which could be prevented by administration of erythropoietin. This suppression of erythropoiesis was associated with the appearance of tumour necrosis factor in the blood, development of pronounced cachexia and depression of serum erythropoietin levels. TGFβ induces TNF in vivo that leads to cachexia, decrease of serum erythropoietin levels and suppression of erythropoietin dependent erythropoiesis.
The clinical outcome of autoimmune thrombocytopenic purpura patients is related to their T cell immunodeficiency
Authors: Garcia‐Suarez J. , Prieto A. , Reyes E. , Manzano E. , Merino J. L. , Alvarez‐Mon M.
Published: March 2008

Summary. In this work we have furthered the understanding of the alterations of T lymphocytes from 29 patients with active autoimmune thrombocytopenic purpura (ATP) and the clinical significance of their lymphocytes. An increased percentage of in vivo activated (CD25+ and DR+) T lymphocytes was found in ATP patients with respect to that found in 22 healthy controls. The function of these T cells measured as the proliferative response to polyclonal mitogenic signals is heterogeneously impaired in ATP patients. T lymphocytes from 65·5% (19/29) of the ATP patients showed a decreased proliferative response to these mitogenic signals. This functional alteration is associated with a redistribution of the T cell compartment in these patients' peripheral blood since a significant decrease of CD4+ T lymphocytes was found.

We have also found that the impairment of the T cell function is different in the diverse clinical situations of the disease. Those with stable, untreated disease showed a marked decrease in the T cell proliferative response to mitogens. Furthermore, those patients who did not respond either to steroids or to splenectomy showed significantly reduced T lymphocyte blastogenesis after phytohaemagglutinin (PHA) stimulation in comparison to that found in responding patients.
Fixation with formaldehyde induces expression of activation dependent platelet membrane glycoproteins, P selectin (CD62) and GP53 (CD63)
Authors: Cahill M. R. , Macey M. G. , Newland A. C.
Published: March 2008

Summary. Platelet activation in vivo is important in the pathogenesis of thrombosis. Accurate measurement is difficult due to artefactual in vitro preparation related activation. This can be overcome by using whole blood techniques, such as flow cytometry. However, there is little consensus on methods of platelet preparation, procedures for use of fixation, or the optimal types/amounts of fixative. The use of unfixed platelets has received little attention.

We describe a series of experiments comparing platelet activation antigen expression detected by flow cytometry in fixed and unfixed samples. Formaldehyde increased the expression of both CD62 and CD63. We recommend that fixation with formaldehyde should not be used when studying platelet activation.
Instability of repeats of the von Willebrand factor gene variable number tandem repeat sequence in intron 40
Authors: Eikenboom Jeroen C. J. , Reitsma Pieter H. , van der Velden Pieter A. , Briët Ernest
Published: March 2008

Summary. The von Willebrand factor gene contains a highly polymorphic variable number tandem repeat sequence (VNTR) in intron 40. We observed the spontaneous mutation to a new length allele. Sequence analysis revealed that the change in length corresponds to one tetranucleotide repeat unit. This observation emphasizes the risk of errors in family studies and prenatal diagnosis due to VNTR instability.
An unusual variant of congenital dyserythropoietic anaemia with mild maternal and lethal fetal disease
Authors: Roberts D. J. , Nadel A. , Lage J. , Rutherford C. J.
Published: March 2008

Summary. An unusual variant of congenital dyserythropoietic anaemia is described presenting as mild haemolytic anaemia with multinucleated erythroblasts in the marrow of the proband. The outcome of her non‐consanguineous pregnancy was a third trimester, in utero, fetal demise. The hydropic fetus had dyserythropoiesis with circulating multi‐nucleated erythroblasts, and haemosiderosis. Re‐evaluation of the proband revealed a variant of congenital dyserythropoietic anaemia with an inheritance pattern, and with morphological, serological and biochemical features which are not consistent with any of the three described variants of congenital dyserythropoietic anaemias.
ZB51: a monoclonal antibody reactive with human plasma cells
Authors: Webster R. , Pulford K. , Jones M. , Micklem K. , de Waele M. , Mayne K. , Tse A. , Mason D. Y.
Published: March 2008

Summary. Several monoclonal antibodies have been raised in the past which react with human plasma cells but they have all shown additional reactions with other cell types. In this paper we describe a new monoclonal antibody, ZB51, which recognizes an intracellular antigen in normal plasma cells in cryostat tissue sections and cell smears, and which also reacts with neoplastic cells in most cases of myeloma and with plasma cell lines. The antibody shows minimal reactivity with a few eosinophils and myelocytes in bone marrow and stains a myeloid cell line. Whilst normal epithelium is not labelled, antibody ZB51 stains two carcinoma cell lines. Although it was not possible to characterize the target antigen in terms of molecular weight, the reactivity of antibody ZB51 with normal and neoplastic plasma cells makes it a useful new immunocytochemical reagent.
Impact of leucocyte‐depleted blood components on the haematological recovery and prognosis of patients with acute myeloid leukaemia
Authors: Oksanen Kalevi , Elonen Erkki
Published: March 2008

Summary. Use of leucocyte‐depleted blood components reduces refractoriness to platelet transfusions, but the cost‐effectiveness of this policy has been in doubt. We retrospectively analysed data of 115 patients with acute myeloid leukaemia (AML), treated according to a standardized cytostatic protocol. Fifty patients had received standard (STD) platelet concentrates (PCs) and red blood cell concentrates (RBCs). Sixty‐five patients had received leucocyte‐depleted (L‐D) components, with fewer than one million leucocytes per unit. Refractory patients (22% of the STD group and 3% of the L‐D group) were excluded from further observations. There were no differences between the groups during the first cytostatic treatment period. Thereafter, significantly fewer PCs and RBCs were transfused to the L‐D group. In the L‐D group, granulocytopenia (<0·5 × 109/l) and thrombocytopenia (< 50 × 109/l) were shorter, the lowest leucocyte counts were higher, serious infections were less common (44%v 59%), and the patients spent fewer days in hospital. The median relapse‐free survival (RFS) was longer in the L‐D group than in the STD group. We conclude that leucocyte depletion of blood components has favourable effects on the recovery of haematopoiesis, consumption of blood components, occurrence of serious infections, and relapse‐free survival in AML.
Blood platelet heterogeneity: evidence for two classes of platelets in man and rat
Authors: Behnke O. , Forer A.
Published: March 2008

Summary. The platelet population of man and rat can be divided into two classes of about equal size on the basis of presence/absence of an acid phosphatase which acts on paranitrophenylphosphate (a PNPase), at pH 5. The cytochemical reaction product is in the platelet cytoplasmic matrix, without apparent association with organelles or membrane systems.

We could not relate differences in staining to differences in function: all cells responded the same to activation by thrombin, ADP, or collagen, in fibrinogen binding to activated platelets, by endocytosis of fluid‐phase tracers, and in internalization of latex particles.

With respect to possible physiological substrates for the PNP‐ase, there was no reaction product from β‐glycerophosphate, AMP, ADP, ATP, GTP, CMP, IMP, cAMP, creatine phosphate, and inositol phosphates, and the enzyme was not inhibited by 40 mm lithium. There was reaction product from tyrosine phosphate suggesting that the physiological substrate for PNP‐ase is tyrosine phosphate.

In rat bone marrow, megakaryocytes also were of two classes, PNPase positive and PNPase negative, suggesting that different classes of platelets arise from different classes of megakaryocytes.
Platelet activation induced by an antiplatelet autoantibody against CD9 antigen and its inhibition by another autoantibody in immune thrombocytopenic purpura
Authors: Yanabu Mutsumasa , Nomura Shosaku , Fukuroi Tsutomu , Suzuki Masahiko , Kawakatsu Toshihiro , Kido Hirofumi , Yamaguchi Kazuyuki , Kokawa Terutoshi , Yasunaga Kojiro
Published: March 2008

Summary. In a patient with immune thrombocytopenic purpura (ITP), we found a novel platelet‐activating IgG (act‐IgG) and an inhibitory IgG (inhi‐IgG) that prevented activation induced by both CD9 monoclonal antibody (mAb) and the act‐IgG. Purified IgG from the patient plasma caused a rise in [Ca2+]i and the aggregation of normal platelets, and bound to a 24 kD membrane protein. This aggregation was inhibited by aspirin, staurosporine, an inhibitor of protein kinase C, and F(ab′)2 fragments of MALL13, a CD9 mAb. When the platelet count of this patient rose to normal range, the act‐IgG disappeared. About 2 weeks later, the relapse of thrombocytopenia was observed. The purified IgG obtained in this period did not activate platelets but inhibited both the rise in [Ca2+]i and platelet aggregation stimulated by NNKY 1–19, a CD9 mAb, as well as the act‐IgG, and bound to a 40 kD membrane protein. The inhi‐IgG prevented the binding of IV‐3, a mAb against Fcγ receptor II (FcγRII), but did not prevent the binding of NNKY 1–19 to its antigen. We suggest that the activating autoantibody recognized CD9 antigen and activated both the thromboxane‐ and phospholipase C‐dependent pathways, while the inhibitory autoantibody recognized the FcγRII and inhibited CD9 antibody‐induced platelet activation mediated via this receptor.
Soluble P‐selectin is present in normal circulation and its plasma level is elevated in patients with thrombotic thrombocytopenic purpura and haemolytic uraemic syndrome
Authors: Katayama Masahiko , Handa Makoto , Araki Yohko , Ambo Hironobu , Kawai Yohko , Watanabe Kiyoaki , Ikeda Yasuo
Published: March 2008

Summary. P‐selectin is an integral membrane glycoprotein stored in the secretory granules of platelets and endothelial cells. To determine whether soluble P‐selectin may be present in the circulation of healthy humans, we used a sandwich immunoassay to assess citrated plasma from 50 subjects. P‐selectin was present in concentrations ranging from 19 to 521 ng/ml (mean ± SD = 121 ± 84 ng/ml). The apparent molecular weight of P‐selectin immunoisolated from platelet‐poor plasma was similar to that of the detergent‐soluble form isolated from platelet membrane. Plasma levels of P‐selectin were unaffected by the following procedures: (1) drawing of blood in the presence of protease inhibitors; (2) stimulation of platelet‐rich plasma with aggregating agents; (3) ultracentrifugation at 100000 g for 120 min at 4°C or filtration through a 0·22 μm membrane; or (4) preincubation of platelet‐poor plasma with immobilized anti‐platelet glycoprotein Ib monoclonal antibodies. It appeared that plasma P‐selectin did not result from the in vitro activation of platelets, nor was it derived from platelet microparticles. We also found that plasma P‐selectin levels were significantly elevated in patients with thrombotic thrombocytopenic purpura (12 patients, 332 ± 184 ng/ml, P<0.001) and haemolytic uraemic syndrome (17 patients, 297 ± 191 ng/ml, P < 0.0001), as compared to the normal levels. Thus, these data should facilitate the study of the pathophysiological significance of circulating P‐selectin.
A phase I/II study of prostaglandin E1 for the prevention of hepatic venocclusive disease after bone marrow transplantation
Authors: Bearman Scott I. , Shen Danny D. , Hinds Mary S. , Hill Harlan A. , McDonald George B.
Published: March 2008

Summary. We conducted a phase I/II trial of prostaglandin E1 (PGE1) for the prevention of hepatic venocclusive disease (VOD). Twenty‐four patients at high risk for VOD received PGE1 at four dose levels ranging from 1·25 to 10 ng/kg/min. Severe toxicity was experienced at all dose levels and was manifested as cutaneous erythema and desquamation, severe pain in dependent extremities, fluid retention and grade 4 oedema, or hypotension. In 12/24 patients, PGE1 administration was stopped prior to day 15 due to severe toxicity attributed to the drug. Three of 12 patients who received PGE1 for the entire course of treatment and 9/12 patients whose PGE1 was stopped prior to day 15 developed severe toxicity which was attributed to PGE1 (P= 0·039). Severe toxicity attributed to PGE1 was not related to either PGE1 concentration or PGE1, clearance. Six of 12 patients whose PGE1 infusion was stopped prior to day 15 and 2/12 patients who received PGE1 for the full course of treatment developed severe VOD (P= 0·19). We conclude that PGE1 causes significant toxicity in patients undergoing marrow transplantation. The ability of PGE1 to prevent severe VOD in patients at high risk remains unproven.
Avascular necrosis after treatment of aplastic anaemia with antilymphocyte globulin and high‐dose methylprednisolone
Authors: Marsh J. C. W. , Zomas A. , Hows J. M. , Chapple M. , Gordon‐Smith E. C.
Published: March 2008

Summary. Avascular necrosis of bone (AVN) occurring in patients with aplastic anaemia (AA) treated with antilymphocyte globulin (ALG) followed by high‐dose methylprednisolone (HDMP) has been studied retrospectively. Out of 49 patients treated at two centres, seven have developed AVN at a median of 14 months (range 6–30) following treatment. The cumulative incidence of AVN is 21% (95% confidence intervals 7–35%). The hip was involved in six patients, bilaterally in five. Two patients had more than two joints affected. Surgical intervention was necessary in five patients, three of whom required total hip replacement. In contrast, there were no cases of AVN in a historical group of 61 patients with aplastic anaemia treated with an indentical regimen of ALG but using a short course of low‐dose prednisolone. Lack of convincing evidence for benefit, the considerable morbidity from AVN, and increased risk of early life‐threatening infection, suggest that high‐dose methyl prednisolone should be omitted from ALG treatment protocols for aplastic anaemia.
Prognostic factors in elderly patients with acute myeloid leukaemia: development of a model to predict survival
Authors: Johnson P. R. E. , Hunt L. P. , Yin J. A. Liu
Published: March 2008

SummaryWith an increasing number of elderly patients presenting with acute myeloid leukaemia and with recent reports of worthwhile remission rates and survival times following treatment with intensive chemotherapy, there is a pressing need to identify criteria to assist in the selection of appropriate therapy for elderly patients. We have performed multivariate Cox proportional hazard regression analysis of data prospectively collected during the treatment of 104 patients aged 60 and over treated in a multi‐centre study with a standardized regimen of mitozantrone and cytosine arabinoside for induction and consolidation. Four readily available parameters, namely urea, performance status, peripheral blood blast count and presence of hepatomegaly, were identified from which a prognostic model to predict survival has been developed. The model was found to be accurate in predicting survival in the cohort of patients from which it was developed, but needs to be validated in a further test population studied prospectively. Its simplicity suggests that it may be particularly useful in the selection of elderly patients with AML most likely to benefit from intensive chemotherapy.
Anti‐fibrinogen antibody mediates fibrinogen binding to platelet membrane glycoprotein IIb‐IIIa
Authors: Mohri Hiroshi , Tanabe Juichi , Fujita Hiroyuki , Kanamori Heiwa , Ohkubo Takao
Published: March 2008

Summary The binding of fibrinogen to platelets requires the agonist activation of platelet membrane glycoprotein IIb/IIIa. We have now found an anti‐fibrinogen polyclonal antibody (YCU‐R3) that increases the fibrinogen affinity of GPIIb/IIIa‐binding function (activation) and subsequent platelet aggregation. The addition of intact IgG, F(ab)2 fragments or Fab fragments induced platelet aggregation. The antibody‐mediated fibrinogen binding was specific and saturable. This binding was inhibited by native fibrinogen, the RGDS peptide, the peptide of the C‐terminus γ chain of fibrinogen (γ397–411), and the anti‐GPIIb/IIIa monoclonal antibody (LJ‐CP8). The antibody‐dependent fibrinogen binding was similar to that induced by ADP. Moreover, after pretreatment with the anti‐fibrinogen antibody and fibrinogen, formalin‐fixed platelets bound to fibrinogen saturably. These results suggest that this anti‐fibrinogen antibody may function as partial agonist.
Haemopoietic stem/progenitor cell transplant in Fanconi anaemia using HLA‐matched sibling umbilical cord blood cells
Authors: Kohli‐Kumar Mudra , Shahidi Nasorullah T. , Broxmeyer Hal E. , Masterson Margaret , Delaat Cynthia , Sambrano James , Morris Christopher , Auerbach Arleen D. , Harris Richard E.
Published: March 2008

Summary There have only been a few reports documenting the use of umbilical cord blood as a source of stem cells for haemopoietic reconstitution. We report our experience with a child with Fanconi anaemia (FA) who underwent a stem cell transplant using umbilical cord blood cells from his HLA matched sibling. Although the engraftment was somewhat slow, it was complete and comparable to other transplants performed in FA patients using HLA matched sibling marrow. There was no graft‐versus‐host disease. The post‐transplant period was uncomplicated and, at a follow‐up of 36 months, this child is well with normal blood counts and immune function.
Biotinylated platelets: a new approach to the measurement of platelet life span
Authors: Heilmann E. , Friese P. , Anderson S. , George J. N. , Hanson S. K. , Burstein S. A. , Dale G. L.
Published: March 2008

Summary The in vivo life span of dog platelets was determined by derivatizing whole blood with N‐hydroxysuccinimido biotin, reinfusing the biotinylated blood and subsequently monitoring the survival of the biotinylated platelets with flow cytometry. We found that the biotinylated platelets had a mean life span of 6.0 ± 1.1 d as determined by curve‐fitting the platelet disappearance data to gamma functions. These data are in good agreement with literature values of platelet life span for canine platelets labelled with either 111Indium‐oxine or 51Chromium. Biotinylated platelets were analysed after reinfusion and found to aggregate normally in response to the agonists adenosine diphosphate and phorbol myristate acetate. These experiments demonstrate that biotinylated platelets survive normally in vivo and that this labelling method can be used for determining platelet life spans.
Preliminary study to identify cancer patients at high risk of venous thrombosis following major surgery
Authors: Falanga A. , Ofosu F. A. , Cortelazzo S. , Delaini F. , Consonni R. , Caccia R. , Longatti S. , Maran D. , Rodeghiero F. , Pogliani E. , Marassi A. , D'Angelo A. , Barbui T.
Published: March 2008

Summary This study investigated whether the pre‐surgical plasma levels of TAT and F1 + 2 of patients undergoing major surgery for localized tumours could identify patients at higher risk of thrombosis, and how heparin prophylaxis affected in vivo coagulation after cancer surgery. We measured the pre‐and post‐operative levels of TAT, F1+2, total factor VII (FVIIt) and zymogen FVII (FVIIz) in 117 cancer patients, with and without heparin prophylaxis. The end points of this study were DVT. ihitially detected by 125I‐fibrinogen uptake test and confirmed by ascending venography. Pre‐operative [TAT] and [Fl+2] of the cancer patients were significantly higher than those of age‐matched control subjects (n=50) (P<0.005 and P<0.05, respectively): pre‐operative [FVII] was not significantly different. One of the 83 patients receiving prophylaxis, and 8/34 not receiving prophylaxis developed post‐operative DVT. Of the parameters evaluated, only the pre‐operative [TAT] > 3. 5 ng/ml identified patients at higher risk for post‐operative DVT. Heparin reduced plasma TAT levels and FVII consumption following surgery, suggesting that heparin modulates coagulation associated with cancer surgery. The results of this study also suggest that the pre‐operative [TAT] may identify patients with higher risk for post‐operative DVT.