bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum erythropoietin in patients with myelofibrosis with myeloid metaplasia
Authors: Barosi G. , Liberato L. N. , Guarnone R.
Published: March 2008

Summary. Serum erythropoietin levels (s‐Epo) were measured by radioimmunoassay in 61 consecutive anaemic patients (Hb<12 g/dl) with myelofibrosis with myeloid metaplasia (MMM). S‐Epo was inversely correlated with Hb (r= ‐0.48, P<0.0001). When observed s‐Epo values were compared with predicted levels based on the relationship between s‐Epo and Hb in control subjects, all but eight patients (87%) had s‐Epo levels appropriate for the degree of anaemia. The observed/predicted (O/P) s‐Epo ratio was significantly lower in patients with signs of active disease, and a significant inverse correlation was found between the O/P ratio and erythrokinetic measurement of the extent of erythropoiesis (r=0.31; P=0.02). Circulating Epo levels were appropriate for the variations in Hb during the postsplenectomy period in three patients. In conclusion, this study does not support the idea that therapy with erythropoietin should be extensively used in anaemic patients with MMM, but rather that it should be considered only in selected cases.
Expression of CD18 (integrin β2 chain) correlates with prognosis in malignant B cell lymphomas
Authors: Erikstein Bjørn K. , Holte Harald , Kvaløy Stein , Andersson Kristin B. , Steen Harald B. , Hannisdal Einar , Smeland Erlend B.
Published: March 2008

Summary. Several studies have shown that cell cycle related parameters including DNA synthesis and activation antigen expression can predict patient survival in lymphoma patients. In this study of 69 malignant B cell lymphomas we have examined the cell surface expression of several cell interaction and activation molecules by flow cytometry. Expression of CD18 (integrin β2 chain) was found to correlate strongly with patient survival (median follow up 50 months) even when adjusting for other important prognostic factors (P= 0.0001). The percentage of cells positive for CDw75 proved important both as a single parameter and in the multivariate analysis. Histology, classified as low versus high grade malignancy, bulky versus not bulky disease and high versus low thymidine incorporation, was also found to correlate with prognosis in this study.
The significance of detection of minimal residual disease in childhood acute lymphoblastic leukaemia
Authors: Potter Michael N. , Steward Colin G. , Oakhill Anthony
Published: March 2008

Summary. In acute lymphoblastic leukaemia (ALL), minimal residual disease (MRD) can be defined as disease occurring at a subclinical level and beyond detection by conventional methods of assessment. Application of the polymerase chain reaction (PCR) to the hypervariable segment of the immunoglobulin heavy chain (IgH) gene, allows detection of MRD at a level of one leukaemic cell in 104‐105 normal marrow cells. We have performed a retrospective study using this technique in the assessment of children with precursor B‐cell ALL in whom the clinical outcome is known. In the early treatment period MRD is commonly detected in children who remain in complete remission on subsequent follow up. Thus, the detection of MRD at this time may have little value in the prediction of future relapse. However, at the end of treatment, children who remain in complete remission have no evidence of MRD. Conversely, detectable MRD at this time would seem to predict for future relapse, though this can be a delayed event. Remarkably, in two children who suffered a bone marrow relapse 8.5 and 9 years after completing therapy for their initial disease, MRD was detected, in their end of initial treatment marrow samples. Clearly PCR technology is changing the definition of the remission state in childhood ALL. and may have predictive value in the assessment of children who are at a high risk of future relapse. Large prospective studies of molecular monitoring are now required to confirm these preliminary results.
Mutations in haemophilia A
Authors: Schwaab R. , Oldenburg J. , Tuddenham E. G. D. , Brackmann H. H. , Olek K.
Published: March 2008

In the present study DNA from 281 unrelated haemophilia A patients including 15 inhibitor patients has been analysed by Southern blotting technique. Using various restriction enzymes, cloned factor VIII cDNA probes and genomic fragments we have identified 14 mutations. Six of the mutations are novel partial factor VIII gene deletions. One deletion affects exon 1, two deletions concern exon 6, another deletion, of which breakpoints are sequenced, takes part of exon 16 and two deletions affect exon 26. Besides the deletions, eight point mutations have been found at the TaqI restriction sites of exons 18, 24 and 26. Five C → T mutations resulted in nonsense mutations, one in exon 18, one in exon 26 and three in exon 24. Two G → A mutations caused a missense mutation in exon 24 leading to an arginine/glutamine exchange. Although two patients showed this mutation, their clinical phenotypes were different, possibly due to an additional unidentified sequence polymorphism. A G → T mutation in exon 26 substituted the arginine with leucine. All deletions and seven of the point mutations are associated with severe disease with a detectable inhibitor in the patient with the TaqI‐point mutation in exon 18. One of the G → A mutations is associated with mild haemophilia but the patient also has developed an inhibitor. Amongst these mutations the origin of the mutation could be determined in four kindred, one of which showed maternal mosaicism.
Alpha interferon in the treatment of chronic hepatitis C infection in thalassaemia major
Authors: Donohue S. M. , Wonke B. , Hoffbrand A. V. , Reittie J. , Ganeshaguru K. , Scheuer P. J. , Brown D. , Dusheiko G.
Published: March 2008

Summary. Hepatitis C virus (HCV) is responsible for the majority of cases of post transfusion non‐A non‐B (NANB) hepatitis in thalassaemia major (TM). Twelve multi‐transfused TM patients with serological, biochemical, histological and molecular biological evidence of HCV infection have been treated for 6 months with recombinant α‐interferon (IFN). Ten (83%) responded as assessed by a fall of at least 50% of pre‐treatment serum transaminase levels. Histological improvement was observed in 6/7 responders tested. Natural killer (NK) cell activity 24 h after the first dose of IFN was significantly increased in responders as compared to non‐responders (P<0.05). HCV RNA disappeared from serum in 5/12 and from liver tissue in 2/5 of the responders. The degree of induction of peripheral blood mononuclear cell 2′5’oligoadenylate synthetase messenger RNA (2‐5 OAS mRNA), an enzyme induced by IFN, after the first dose of IFN did not correlate with response. IFN was generally well tolerated. We conclude that the response rate in multi‐transfused TM patients infected with HCV and treated with IFN is similar to that in non‐multi‐transfused patients.
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.
Clonality in myelodysplastic syndromes: demonstration of pluripotent stem cell origin using X‐linked restriction fragment length polymorphisms
Authors: Tsukamoto Norifumi , Morita Kimio , Maehara Tadashi , Okamoto Kiyoshi , Karasawa Masamitsu , Omine Mitsuhiro , Naruse Takuji
Published: March 2008

Restriction fragment length polymorphisms (RFLP) of the X‐chromosome genes phosphoglycerate kinase (PGK) and hypoxanthine phorphoribosyltransferase (HPRT) were used to determine the clonal nature of myelodysplastic syndromes (MDS) in 22 patients. These included eight with refractory anaemia (RA), four with RA with ring sideroblasts (RARS), six with RA with an excess of blasts (RAEB), three with RAEB in transformation (RAEB‐T), and one with chronic myelomonocytic leukaemia (CMML). Monoclonal X‐inactivation patterns were observed in 19/22 patients. The remaining three cases, one each with RA, RARS and RAEB, were of polyclonal composition. Separated T‐lymphocyte and granulocyte fraction analyses in six patients of the former cases revealed that T‐lymphocyte as well as granulocyte fractions showed a monoclonal pattern of X‐inactivation. These results support the view that the majority of MDS arise from a pluripotent stem cell capable of myeloid and lymphoid differentiation.
Porcine platelets contain an increased quantity of ultra‐high molecular weight von Willebrand factor and numerous alpha‐granular tubular structures
Authors: Wilbourn Barry , Harrison Paul , Lawrie Andrew , Savariau Elisabeth , Savidge Geoffrey , Cramer Elisabeth Martin
Published: March 2008

Summary. Immunoelectronmicroscopy of human platelet α‐granules reveals that von Willebrand factor (vWf:Ag) colocalizes with a small number of discrete tubular structures which appear identical to those observed within the WeibelPalade bodies of endothelial cells. Although it is likely that tubules are composed of vWf:Ag as they are absent in severe vWD porcine platelets, their exact structural and functional nature is still unclear. In this study quantitative/qualitative analysis of vWf:Ag was undertaken in a series of platelet preparations obtained from normal pigs, normal humans and various vWD patients. Electron microscopy confirmed that normal pig platelet α‐granules contain numerous, regularly spaced tubular structures eccentrically located and coincident with immunogold staining of vWf:Ag. In contrast, normal human platelet α‐granules contain significantly fewer tubules (usually four to six) which are absent or reduced in number within various vWD platelet sections. Furthermore, the pig platelet lysates not only contained a full complement of multimers but also demonstrated significant intense staining of ultra‐high MW material, irrespective of the presence or absence of proteolytic inhibitors. This ultrahigh MW vWf appears similar to that observed within lysates prepared from endothelial cells and is susceptible to degradation to lower MW multimers. This study suggests that the tubular structures within α‐granules and Weibel‐Palade bodies may be composed of, or structurally related to, the ultra‐high MW intracellular form of vWf:Ag.
Unreliability in immunoassays of erythropoietin: anomalous estimates with an assay kit
Authors: Bechensteen A. G. , Lappin T. R. J. , Marsden J. , Muggleston D. , Cotes P. M.
Published: March 2008

Summary. We report the finding of inappropriately high estimates of erythropoietin (Epo) in 11% of serum samples tested by one commercial assay kit. These estimates were not confirmed when the same samples were tested independently in four other systems for the immunoassay of Epo. As yet no explanation has been found to account for the anomalously high estimates.
Reversible adult‐onset cyclic haematopoiesis with a cycle length of 100 days
Authors: Birgens Henrik S. , Karle Hans
Published: March 2008

Cyclic neutropenia is the most frequent of the cyclic haematopoietic disorders characterized by its regular 21 d cyclic fluctuations in the number of blood neutrophils, and in many cases simulataneous fluctuations in the other blood cell lines.

In this paper we describe a 77‐year‐old woman with a cyclic pancytopenia including all the myeloid cell lines and to some extent the lymphocytes with a constant and predictable oscillation period of about 100d. Serial bone marrow biopsies and plasma lactoferrin measurements indicated a similar fluctuating pattern in the bone marrow production of neutrophils. Serial measurements of plasma GM‐CSF concentration pointed at a simple feed‐back inhibitory system. The condition was present for at least 4 years, after which it gradually improved, although the thrombocyte count still showed a fluctuating tendency after a further 4 years of observation.

The clinical consequences were mild symptoms of anaemia and a few episodes of respiratory infections occurring during pancytopenic periods.

We think this is the first case described in the literature with this variant of a cyclic haematopoietic disorder. The precise pathophysiological mechanism behind this condition is obscure, but probably it is due to a regulatory disturbance at a very early step in the haematopoietic stem cell hierarchy.
Vascular complications of essential thrombocythaemia: a link to cardiovascular risk factors
Authors: Watson Kathleen V. , Key Nigel
Published: March 2008

Essential thrombocythaemia (ET) is a myeloproliferative disorder characterized by absolute thrombocytosis and increased incidence of thrombosis and haemorrhage. We report higher morbidity in patients with ET due to arterial ischaemic complications when cardiovascular risk factors are present. In this retrospective analysis of 46 patients, arterial complications occurred in 20/46 patients (43.4%); patients with cardiovascular risk factors, especially cigarette smoking, had more than twice as many arterial complications than patients without risk factors (62.5%v 22.7%, P<0.05). Neither age, gender nor degree of thrombocytosis were linked to the number of complications. In contrast, we observed no association between cardiovascular risk factors and venous thrombosis or haemorrhagic complications of ET.
Childhood acute lymphoblastic leukaemia presenting with relapsing hypoplastic anaemia: progression of the same abnormal clone
Authors: Liang Raymond , Cheng Gregory , Wat M. S. , Ha S. Y. , Chan L. C.
Published: March 2008

A 13‐year‐old girl presented with two spontaneously reversible episodes of marrow hypoplasia. She subsequently developed acute lymphoblastic leukaemia (ALL) 8 months later. Southern analysis showed identical clonal immunoglobulin heavy chain gene rearrangement bands in the leukaemic cells as well as the marrow cells obtained at the two hypoplastic episodes. Hypoxanthine phosphoribosyl transferase polymorphism studies showed that the ALL blast cells, bone marrow and peripheral blood cells during the two hypoplastic episodes all exhibited clonal haematopoiesis with the same X‐chromosome inactivated. This case provides strong evidence that aplastic anaemia and ALL may represent evolution of the same abnormal clone.
Differential sensitivity of human myeloma cell lines and normal bone marrow colony forming cells to a recombinant diphtheria toxin‐interleukin 6 fusion protein
Authors: Chadwick D. E. , Jean L. F. , Jamal N. , Messner H. A. , Murphy J. R. , Minden M. D.
Published: March 2008

Summary. The cytotoxicity of a recombinant interleukin 6 (IL‐6)‐diphtheria toxin (DT) fusion protein towards human myeloma cell lines was investigated. DAB389‐IL‐6 inhibited protein synthesis and methylcellulose colony formation by U266 myeloma cells. In the clonogenic assay, the fusion protein approached the level of cytotoxicity achieved by native DT. The specificity of killing by DAB389‐IL‐6 was demonstrated by inhibition of cytotoxicity by a molar excess of free rhIL‐6.

The effect of DAB389‐IL‐6 on colony formation by six OCI‐My cell lines was assessed. Similar to U266 cells, colony growth by the OCI‐My 5 and ‐My 2 cell lines was inhibited in a simple dose dependent manner. However, a biphasic effect was observed for the IL‐6 dependent OCI‐My 4 cells: DAB389‐IL‐6 stimulated colony formation at low ( 10‐11 m) concentrations, yet was inhibitory at higher doses. Three other cell lines whose growth was not altered by IL‐6 were relatively unaffected by DAB389‐IL‐6, despite their sensitivity to native DT. Flow cytometric analysis for IL‐6 receptor expression using phycoerythrin‐conjugated IL‐6 demonstrated specific binding sites on both DAB389‐IL‐6 sensitive and certain insensitive cell lines, suggesting that other factors in addition to the expression of IL‐6 receptors are involved in killing by the fusion toxin.

Despite evidence for a role of IL‐6 in myeloid cell development, normal bone marrow was insensitive to the IL‐6 fusion toxin. In cultures containing both normal bone marrow and U266 cells DAB389‐IL‐6 effectively inhibited the growth of U266 myeloma colonies but had little effect on normal bone marrow erythroid, granulocyte and mixed erythroid/granulocyte colony growth. From these experiments we conclude that DAB389‐IL‐6 is specifically cytotoxic towards a subset of IL‐6‐responsive human myeloma cell lines and may be useful, in some cases, in the selective elimination of tumour cells from mixed populations of normal and malignant cells.
Dynamic modulation of the cell surface expression of the granulocyte‐macrophage colony‐stimulating factor receptor
Authors: Khwaja Asim , Carver Julia , Jones H. Mark , Linch David C.
Published: March 2008

Summary. The GM‐CSF receptor (GM‐CSFR) is composed of α and β subunits. Surface expression of the α chain alone leads to low affinity GM‐CSF binding and of both subunits to high affinity binding: the β chain is required for transducing a proliferative signal. Studies of GM‐CSFR expression have concentrated largely on static events occurring under conditions of binding equilibrium. We have examined the dynamic regulation of high and low affinity GM‐CSFR expression in neutrophils (1100 ± 200 R/cell, KD 50 ± 15 pm) and a GM‐CSF dependent human leukaemic cell line, TF‐l (2000 ± 450 R/cell KD 15 ± 5 PM) and 8600 ± 150 R/cell KD 1.8 ± 0.3 nm). The addition of GM‐CSF to TF‐1 cells (350 PM, 4 h at 37°C) caused a reduction in subsequent binding of 125 I‐GM‐CSF at low ligand concentration (100 pm) (following a low pH wash to remove surface bound ligand) to 16 ± 4% and a reduction in binding at high ligand concentration (2 nm 125I‐GM‐CSF) to 36 ± 9% of control. Scatchard analysis showed complete down‐regulation of high affinity GM‐CSFR and a significant reduction in low affinity GM‐CSFR. In neutrophils, concentration‐response curves of ligand induced receptor down‐regulation at 37°C showed that observed down‐modulation was more than 10‐fold greater than predicted by static equilibrium binding data and correlated closely with GM‐CSF priming of the neutrophil respiratory burst. The addition of IL‐3 to TF‐1 cells at 37°C reduced 100 PM 125I‐GM‐CSF binding to 18 ± 4% and 2 nm 125I‐GM‐CSF binding to 46 ± 5% of control. TF‐1 cells, but not neutrophils, were able to re‐express GM‐CSFR following removal of GM‐CSF from medium. TF‐1 proliferation assays showed that pulsed GM‐CSF (0.35–3.5 nm) for up to 4 h did not cause a significant increase in 3H‐thymidine incorporation which required the continued presence of GM‐CSF (control 2875 ± 208 cpm, pulsed GM‐CSF 5 ng/ml 49 72 ± 1344. continuous GM‐CSF 5 ng/ml 17249 ± 2982). Therefore, proliferation of TF‐1 cells required the continued presence of GM‐CSF at a time when there was no detectable surface high affinity GM‐CSFR. This shows that signal transduction can take place via the GM‐CSFR at a time when there is no detectable high affinity GM‐CSF binding and introduces a further layer of complexity in the analysis of GM‐CSF receptor function.
Different membrane expression of CD11b and CD 14 on blood neutrophils following in vivo administration of myeloid growth factors
Authors: Hansen Per Boye , Kjærsgaard Erik , Johnsen Hans E. , Gram Jesper , Pedersen Michael , Nikolajsen Kirsten , Hansen Niels Ebbe
Published: March 2008

Summary. During the administration of recombinant human granulocyte colony‐stimulating factor (rhG‐CSF) or granulocyte‐macrophage CSF (rhGM‐CSF) we studied the early and late changes of membrane antigen density on neutrophils. RhG‐CSF and rhGM‐CSF both caused an early transient reduction in blood neutrophilic granulocyte‐concentration within the first 30 min after treatment followed by a marked later increase during the subsequent 24 h. During the early neutropenia quantitative flow cytometry showed an associated marked increase in the density of membrane CD11b from 169 × 103 before to 568 × 103 A.U. per cell induced by rhGM‐CSF but a non‐significant change by rhG‐CSF, suggesting that different mechanisms may be responsible for the transient neutropenia. The subsequent neutrophil granulocytosis was followed by a significantly (P<0.05) increased density of the CD14 antigen from 6.1 × 103 before to 15.9 × 103 A.U. per cell during treatment with rhG‐CSF. but not by rhGM‐CSF administration.

These results demonstrate that the two cytokines may affect the function of neutrophilic granulocytes in different ways. The increased expression of CD1 1b could explain some of the side‐effects during treatment with rhGM‐CSF. The upregulation of CD14 induced by rhG‐CSF may be clinically relevant, as CD14 is an opsonic receptor for lipopolysaccharide binding proteins, acting in the defence against Gramnegative bacterial infections.
Results of Medical Research Council trial UKALL IX in acute lymphoblastic leukaemia in adults: report from the Medical Research Council Working Party on Adult Leukaemia
Authors: Durrant I. J. , Richards S. M.
Published: March 2008

Summary. The MRC UKALL IX trial for patients with untreated ALL aged 14 years and over was open to new patients from July 1980 to April 1985. 266 patients were randomized between two induction schedules, M (involving intermediate dose rnethotrexate with folinic acid rescue) and D (involving daunorubicin). Schedule M resembled that used in the previous MRC adult ALL, trial (UKALL VI). while schedule D was somewhat more intensive. No difference in disease‐free survival was found between the treatment arms, but patients on the daunorubicin arm went into remission earlier.

The overall remission rate was 8 7%, which is at least as good as in contemporary studies elsewhere: factors predictive of a lower remission rate were older age and higher WBC. For those who entered remission, WBC, age and sex were the most important prognostic factors. Time to achieve remission was not a significant factor after allowance was made for these.

An historical comparison does not show any improvement over the preceding MRC adult trial, although the subsequent trial does show a modest improvement at present. Because the improved outlook seen in children is not apparent in adults, and no other randomized trial has demonstrated substantial benefit for any particular regimen, the next trial, UKALL XII, will be investigating the benefit or otherwise of bone marrow transplantation.
Reversibility of thrombin‐induced decrease in platelet glycoprotein Ib function
Authors: Lu He , Menashi Suzanne , Garcia Isabel , Cramer Elisabeth Martin , Li Hong , Romeuf Christophe De , Soria Jeannette , Soria Claudine , Tenza Danièle
Published: March 2008

Summary. Thrombin induces a redistribution of glycoprotein (GP) Ib/GP IX complex from the platelet surface into the surface connected canalicular system (SCCS). This redistribution results in a reduced interaction of platelet GP Ib with von Willebrand factor (vWF) bound to subendothelium leading to impaired platelet adhesion. In this study we show that the platelet aggregation and degranulation require concentrations of thrombin above 0.05 U/ml, while the decrease in GP Ib function (about 50% of control value), as determined by ristocetin induced platelet agglutination, can be induced by lower concentrations (0.01–0.04 U/ml). Moreover, we show that when adding thrombin inhibitors to the platelets preincubated with <0.04 U/ml thrombin for 5 min, their agglutinability by ristocetin was gradually recovered within 30 min. indicating that in these conditions the decrease in platelet adhesiveness is reversible. Immuno‐electromicroscopic study showed that this restoration of platelet GP Ib function was associated with a reversed translocation of GP Ib from the SCCS to the plasma membrane. The data obtained from counting gold particles showed that the ratio of GP Ib immunolabelling on the external membrane versus that on the SCCS was 3.31±0.90 for resting platelets, down‐regulated to 0.84±0.13 (P>0.05 versus resting platelets) for the platelets treated with 0.04 U/ml thrombin and returned to 2.63±2.21 (P>0.05 versus resting platelets) after incubation for 30 min with hirudin. However, the translocation of GP Ib was poorly reversed by thrombin inhibitors when higher concentrations of thrombin were used which induced platelet aggregation and large extent of degranulation. We conclude that thrombin affects platelets in a dose dependent manner, and that at low concentrations the decrease in platelet GP Ib related function is a reversible phenomenon.
Study of a protein S gene polymorphism at DNA and mRNA level in a family with symptomatic protein S deficiency
Authors: Marchetti G. , Legnani C. , Patracchini P. , Gemmati D. , Ferrati M. , Palareti G. , Coccheri S. , Bernardi F.
Published: March 2008

Summary. A protein S gene polymorphism, detectable by restriction analysis of amplified exonic sequences, was investigated in a family with members affected by protein S deficiency, deep vein thrombosis and ictus. The clinical laboratory findings as well as RFLP analysis were consistent with the presence of a type WP III protein S deficiency clearly marked by a polymorphic allele, thus enabling us to determine the carrier status in several subjects. The RFLP analysis, extended to platelet mRNA after reverse transcription and amplification, demonstrated that the mRNA produced by the putative defective gene was present in a subject affected by thrombosis.
Treatment of acute monoblastic leukaemia by combination of recombinant human macrophage colony‐stimulating factor and low dose of ara‐C
Authors: Kitano Kiyoshi , Kobayashi Hikaru , Maeyama Hironobu , Miyabayashi Hidegaru , Furuta Seiichi
Published: March 2008

Summary. We present a patient with acute monoblastic leukaemia (AMoL) who achieved a complete remission on combination therapy with macrophage colony‐stimulating factor (M‐CSF) and low dose of cytosine arabinoside (ara‐C). This 26‐year‐old man was admitted with a relapse of AMoL which proved refractory to several chemotherapeutic regimens. To kill dormant leukaemic cells, we administered 20 mg/m ara‐C by continuous intravenous infusion and 800 × 104 unit M‐CSF by 30 min drip intravenous infusion together for 14 d. The blasts disappeared, followed by a recovery of normal blood cells. The patient continued the complete remission for 5 months. This observation suggests that a combination M‐CSF and a low dose of ara‐C may be useful in treating some patients with AMoL.
Haemophilia B Liverpool: a new British family with mild haemophilia B associated with a — 6 G to A mutation in the factor IX promoter
Authors: Stowell K. M. , Figueiredo M. S. , Brownlee G. G. , Jones P. , Bolton‐Maggs P. H. B.
Published: March 2008

Summary. We report a Lancashire family with mild haemophilia B associated with a — 6 G to A mutation in the factor IX promoter. This mutation has been reported previously in Britain in one other affected family. The factor IX haplotype of these two unrelated patients was determined by PCR analysis of seven polymorphic sites within or close to the factor IX gene. This analysis involved the intragenic Msp I polymorphism which we have now located precisely to allow the design of suitable PCR primers. The haplotype of both of these patients was identical suggesting but not proving a possible common origin for the mutation.
Hereditary ovalocytosis with compensated haemolysis
Authors: Reardon David M. , Seymour Carol A. , Cox Timothy M. , Pinder Jennifer C. , Schofield Ann E. , Tanner Michael J. A.
Published: March 2008

Summary. The clinical and laboratory phenotype of compensated haemolysis in a patient with hereditary ovalocytosis is reported. Clinical presentation was intermittent jaundice and abdominal pain due to pigment gall stones. Haematological analysis revealed an absolute reticulocytosis with an otherwise normal full blood count and biochemical evidence of haemolysis. Variable results were observed with blood grouping reagents. The patient's red cells were stomatocytic ovalocytic, rigid, resistant to malarial parasite invasion, defective in anion transport, and had the characteristic two linked mutations in the red cell band 3 gene.
Peripheral blood stem cell mobilization using high‐dose cyclophosphamide and G‐CSF in pretreated patients with lymphoma
Authors: Craig Jenny I. O. , Anthony Robert S. , Stewart Anne , Thomson Eric B. , Gillon Jack , Parker Alistair C.
Published: March 2008

Summary. Peripheral blood stem cell (PBSC) mobilization for autologous transplantation is more difficult in treated patients and those with bone marrow involvement. In 17 pretreated lymphoma patients, cyclophosphamide (4 g/m2) and G‐CSF mobilized a median circulating peak of 1959 CFU‐GM/ml on day 12.5. PBSC harvesting commenced when WBC was 1×109/l at day 10.5 collected a median of 21.2 × 104/CFU‐GM/kg. 13/17 (76%) patients exceeded the 10 × 104 CFU‐GM/kg threshold for engraftment. The CFU‐GM yield was significantly higher in patients whose WBC recovered from 1–5 × 109/1 in less than 3 days and correlated with the maximum WBC pre and post the cyclophospharnide induced nadir. This regime safely mobilized adequate PBSC in the majority of pretreated lymphoma patients.
Exposure to lipophilic industrial solvents leads to increased P‐glycoprotein expression in peripheral blood cells
Authors: Hegewisch‐Becker Susanna , Szudra Annemarie , Hossfeld Dieter K.
Published: March 2008

Summary. The MDRl gene product P‐glycoprotein (P‐gp), a transmembrane efflux pump, is expressed in many normal tissues including subpopulations of normal peripheral blood. To test the potential impact of cigarette smoke and/or a variety of lipophilic industrial solvents present in paint and varnish, P‐gp expression in unsorted peripheral blood cells of 51 healthy volunteers was measured by means of immuno‐cytochemistry and daunorubicin uptake studies. 10 of these volunteers were heavy smokers and 10 were working as painters or sprayers, and consequently in regular and intense contact with industrial solvents. Cigarette smoke had no impact with regard to increased P‐gp expression and function. However, extensive contact with industrial solvents of lipophilic origin was shown to cause a significant increase in the level of P‐gp expression (P<0.001), thus providing new evidence for the broad substrate specificity of the P‐gp pump.
Autografting for patients with CML in chronic phase: an update
Authors: Hoyle Christine , Gray Richard , Goldman John
Published: March 2008

Between 1984 and 1992, 21 patients with chronic myeloid leukaemia (CML) in chronic phase (CP) were treated with high‐dose chemotherapy (or chemoradiotherapy) followed by autografting with unmanipulated peripheral blood stem cells (PBSC). 12 of these patients survive at a median of 82 months from the time of autografting (range 9–105 months). Nine patients died, six of leukaemia in transformation and three from other causes. Survival of these 21 autograft patients was compared to that of 636 age‐matched controls on the Medical Research Council's (MRC) data base, who had been treated with conventional chemotherapy over the same period. Autografted patients had a significantly longer survival at 5 years post autograft than chemotherapy patients (56%v 28%) even after appropriate allowance for time at risk before autograft (Mantel‐Byar, 2P= 0·003). There was no difference in survival whether autografting was performed early in the disease or later or whether the PBSC had been harvested soon after diagnosis or later. If the benefits of autografting in chronic phase seen in this non‐randomized series can be confirmed in a randomized study, autografting might be the treatment of choice for younger CML patients who do not have suitable donors for allogeneic transplant.
No evidence for protein modifications in fresh frozen plasma after photochemical treatment: an analysis by high‐resolution two‐dimensional electrophoresis
Authors: Tissot Jean‐Daniel , Hochstrasser Denis F. , Schneider Bruno , Morgenthaler Jean‐Jacques , Schneider Philippe
Published: March 2008

To study if photochemical treatment of human plasma by methylene blue in combination with visible light induces protein alterations, we analysed by high‐resolution two‐dimensional gel electrophoresis (2‐D PAGE) untreated and photochemically treated fresh frozen plasma collected by apheresis from the same donor (n= 8). Polypeptides were separated according to their charges by isoelectric focusing and to their sizes in polyacrylamide gels in presence of sodium dodecyl sulphate, and visualized by silver staining. Despite their apparent complexity, protein maps of untreated plasma and photochemically treated fresh frozen plasma revealed identical spot patterns. Thus, photochemical treatment did not induce apparent protein pattern modifications.
The variation with gestational age of the rheological properties of the blood of the new‐born
Authors: Anwar M. A. , Rampling M. W. , Bignall S. , Rivers R. P. A.
Published: March 2008

A study has been made of the variation of blood viscosity and related factors with the gestational age of neonates from 24 weeks to normal term. Viscosity increases significantly over this period by 36% at high shear rate (128·5 s‐1) and 250% at low shear (0·277 s‐1). The high shear rate changes can be explained largely by the effects of variations in haematocrit and plasma viscosity. At low shear rate the same factors are involved together with changes in the plasma protein composition, in particular the age‐related increase in the concentration of the proteins known to induce rouleaux formation. The variation in the degree of sialination of fibrinogen with gestational age may also play a part.
Randomized double‐blind, placebo‐controlled trial of twice‐daily zidovudine in asymptomatic haemophiliacs infected with the human immunodeficiency virus type 1
Authors: Mannucci P. M. , Gringeri A. , Savidge G. , Gatenby P. , Laurian Y. , Pabinger‐Fasching I. , Martinez‐Vazquez J. M. , Hessey E. W. , Steel H. M.
Published: March 2008

In this double‐blind, placebo‐controlled trial of HIV‐infected asymptomatic haemophiliacs, the efficacy of 2‐year zidovudine therapy (1000 mg daily in two divided doses) in preventing progress of HIV infection was prospectively evaluated. Drug tolerance was also studied. 143 haemophiliacs from five European countries and Australia with p24 antigenaemia and/or CD4 cell counts of 0·1–0·4 × 109/l were enrolled. The main measures of outcome were progression to AIDS, CDC group IV disease, symptomatic HIV‐related disease, and a decrease in CD4+ T‐lymphocyte count to fewer than 0·2 × 109/l. There were no significant treatment differences in the proportion of patients progressing to AIDS, CDC group IV or symptomatic disease. Analysis of time to CD4+ counts less than 0·2 × 109/l showed a non‐significant trend in favour of zidovudine. Haemoglobin concentrations were less than 8 g/dl in 4% of zidovudine recipients; neutropenia was less than 0·75 × 109 cells/l in 5% of zidovudine recipients; alanine aminotransferase levels were greater than 10 times the upper normal limit in 3% of zidovudine recipients, but also in 4% of placebo recipients. Hence there was a very low prevalence of side‐effects in haemophiliacs, despite the use of a higher zidovudine dosage than is currently widely used.
Localized, late‐onset, high‐grade lymphoma following bone marrow transplantation: response to combination chemotherapy
Authors: O'Riordan J. M. , Molloy K. , O'Briain D. S. , Corbally N. , Devaney D. , McShane D. , Considine N. , McCann S. R.
Published: March 2008

Non‐Hodgkin's lymphoma is the commonest secondary cancer following bone marrow transplantation (BMT). We report the case of a 42‐year‐old man who developed a laryngeal high‐grade B‐cell lymphoma 5 years following a matched T depleted BMT for CML. Polymerase chain reaction (PCR) analysis using the microsatellite marker Cyp 19 demonstrated the donor origin of involved tissue. Epstein‐Barr virus (EBV) genomic sequences were identified by PCR. Although EBV related B‐cell lymphoproliferative disorders (BLPD) post BMT are difficult to treat, there was a complete remission in this patient following three courses of chemotherapy (CHOP) administered with G‐CSF. This case of late‐onset BLPD appears clinically distinct from the well‐defined, aggressive, early post‐transplant BLPD.
Pancytopenia and vacuolation of marrow precursors associated with necrotizing encephalopathy
Authors: Blatt Julie , Katerji Ammar , Barmada Mamdouha , Wenger Sharon L. , Penchansky Lila
Published: March 2008

Subacute necrotizing encephalopathy (SNE) or Leigh disease is an autosomal recessive disorder associated with various defects of oxidative phosphorylation. Two reports have described the concurrence of SNE with pancytopenia and vacuolation of bone marrow precursors, and have raised the possibility that this symptom complex may be part of a spectrum of diseases which includes Pearson's syndrome (vacuolation of bone marrow precursors, sideroblastic anaemia, exocrine pancreatic dysfunction). We describe a case of Pearson's syndrome in which haematological manifestations antedated progressive neurological deterioration by several years. Cytogenetic studies showed an inverted duplication of chromosome 9 (qh) [inv dup (9) (qh)]. We suggest that cytopenia associated with vacuolation of bone marrow precursors even without clinically apparent central nervous system pathology should prompt consideration of SNE, or related diseases. Conversely, a diagnosis of SNE should prompt evaluation of other organ system functions including bone marrow. Cytogenetic evaluation of other patients with SNE may determine whether the 9 (qh) findings are pathogenetic.
Acute myeloid leukaemia with t(9;11)(p22;q23) in a patient treated for adult T cell leukaemia
Authors: Nakamura Hideo , Ishizaki Takeshi , Itoyama Takahiro , Soda Hisashi , Yoshida Yoshiharu , Yamada Yasuaki , Kuriyama Kazutaka , Sadamori Naoki , Tomonaga Masao
Published: March 2008

A 37‐year‐old male patient with adult T cell leukaemia (ATL) began receiving chemotherapy in March 1992. He achieved complete remission in May 1992, but developed acute myeloid leukaemia (AML, FAB subtype M2) with t(9;11)(p22;q23) in May 1993. The presence of chromosome 11 abnormality at band 11q23 in this patient suggests that the AML was related to the chemotherapy with etoposide for ATL. Furthermore, the combination of etoposide with two cytostatic drugs, cyclophosphamide and carboplatin, possibly induced the leukaemia early (14 months) after the start of chemotherapy. To our knowledge, this is the first report of therapy‐related AML after chemotherapy for ATL.
Clonality in chronic myeloproliferative disorders defined by X‐chromosome linked probes: demonstration of heterogeneity in lineage involvement
Authors: Tsukamoto Norifumi , Morita Kimio , Maehara Tadashi , Okamoto Kiyoshi , Sakai Hirotaka , Karasawa Masamitsu , Naruse Takuji , Omine Mitsuhiro
Published: March 2008

Summary The restriction fragment length polymorphisms (RFLP) of the X‐chromosome phosphoglycerate kinase (PGK) and hypoxanthine phosphoribosyltransferase (HPRT) genes were used to study the clonal basis of the chronic myeloproliferative disorders (CMPD). Analyses were performed on granulocyte and T‐lymphocyte fractions obtained from 24 females; 13 had essential thrombocythaemia (ET), eight polycythaemia vera (PV) and three myelofibrosis with myeloid metaplasia (MMM). All 24 of these patients had monoclonal patterns of X‐inactivation in the granulocyte fraction. For the T‐lymphocyte fraction, non‐clonal patterns of Xinactivation were observed in 8/13 patients with ET, 7/8 with PV and 1/3 with MMM, while the remaining eight subjects were found to have monoclonal patterns of N inactivation. Our findings suggest that the majority of the CMPD in these patients originated from a relatively committed progenitor cell without the capacity to differentiate into T cells, and convincingly demonstrated heterogeneity of lineage involvement.
T‐cell receptor β gene rearrangements in leukaemic B‐cells from patients with chronic lymphocytic leukaemia: association with chromosome 6 deletions
Authors: Merup Mats , Juliusson Gunnar , Hammarström Lennart , Smith C. I. Edvard , Gahrton Gösta
Published: March 2008

Summary Rearrangement of immunoglobulin genes is a characteristic finding in normal B‐cells and in leukaemic cells of B‐cell origin. In some leukaemic cells simultaneous crosslineage rearrangement of immunoglobulin‐ and T‐cell receptor (TcR) genes occur. We have analysed TcR β gene rearrangement in 100 patients with B‐cell chronic lymphocytic leukaemia. All samples expressed CD5, CD19 and CD20 and six patients had rearrangements of both immunoglobulin and TcR β genes. Analysis of gene expression in cells with TcR β gene rearrangement indicated production of truncated TcR β transcripts but no expression of the T‐cell markers CD3, CD4, CD8, TcR αβ or δ on the cell surface. Three of the patients with both Ig and TcR β rearrangement (50%) were 44 years or younger at diagnosis and cells from three such patients expressed IgG. Three of the six patients had a terminal deletion of the long arm of chromosome 6 with different breakpoints, with or without other chromosomal abnormalities, whereas 6q deletions were found in 4/94 patients without TcR β gene rearrangement (4·3%) (P=0·001). This study indicates a correlation between TcR β gene rearrangement and deletion of chromosome 6q.
Detection of clonal T‐cell populations in gastrointestinal lymphomas by analysis of cRNA conformational polymorphisms of rearranged T‐cell‐receptor‐gamma genes
Authors: Koch O. M. , Probst M. , Tiemann M. , Jakob I. , Volkenandt M. , Wienecke R. , Buer J. , Atzpodien J. , Lenz H. J. , Danenberg P. V. , Bertino J. R. , Parwaresch R.
Published: March 2008

Summary Analysis of complementary RNA molecules of junctional regions of rearranged T‐cell‐receptor‐gamma genes show a pattern of conformational polymorphisms which is specific for an individual lymphocytic clone. In a blinded study we analysed formalin‐fixed, paraffin‐embedded histological specimens from gastrointestinal lymphomas and control tissues (lymphomas: pleomorphic T‐cell 10, anaplastic large cell [Ki1+] 9, centroblastic 5, immunocytoma 1, B‐CLL 2, Hodgkin's 2, centroblastic‐centrocytic 1, MALT [mucosa associated lymphoid tissue] 1, T‐cell acute lymphoblastic leukaemia 1, non‐lymphoid or polyclonal lymphoid tissues 5). Junctional regions of rearranged TCR‐gamma genes were amplified by the polymerase chain reaction and the products were transcribed into cRNA. Conformational patterns of cRNA molecules were analysed by polyacrylamide gel electrophoresis. 13/20 T‐lineage lymphomas and the T‐cell acute lymphocytic leukaemia displayed a distinct cRNA band pattern, all B‐lineage lymphomas and the nonlymphoid control tissues were negative. Only one case of nasopharyngeal (lymphoepithelial. Schmincke‐Regaud) carcinoma showed a faint cRNA banding pattern. This novel and non‐radioactive assay allows for the rapid detection and molecular characterization of clonal lymphoid populations in minute histological biopsy specimens.
Characterization of haemolyser‐resistant cells increased in the blood of erythropoietin‐treated mice
Authors: Said Ahmed A. , Yamaguchi Teruhide , Uchida Eriko , Hayakawa Takao
Published: March 2008

Summary Recently, we reported a new in vivo assay method for erythropoietin (Epo) by means of counts of the number of haemolyser‐resistant cells (HRCs) increased in Epo‐injected mice. Here, we attempted to characterize the HRCs. Flow‐cytometric studies revealed that HRCs obtained from Epo‐injected mice expressed the transferrin receptor on their surface membranes. Furthermore, a fluorophotometric study suggested that the number of transferrin receptor‐positive cells increased in a dose‐dependent manner in response to treatment with Epo. On the other hand, flowcytometric and fluorophotometric studies of glycophorin A on HRCs using a rabbit antiglycophorin A antibody also showed a high expression of glycophorin A on them as compared with on HRCs from vehicle‐treated animals (control). The results indicated that HRCs could be defined by their expression of both transferrin receptors and glycophorin A. We concluded that HRCs might be immature reticulocytes.
Incidence of autoantibodies to GPIIb/IIIa in chronic autoimmune thrombocytopenic purpura may be overestimated by the MAIPA
Authors: Hewitt John , Burton Ian E.
Published: March 2008

Summary Twenty‐one serum samples from patients with thrombocytopenia and five from normal individuals were analysed by the MAIPA test to determine specificity of autoantibody reactivity against platelet glycoprotein IIb/IIIa using murine monoclonal antibodies. The sera were also analysed by dot blot to determine their anti‐murine IgG activity. Two of the three patient samples positive in the MAIPA test were positive in the dot blot test for anti‐murine IgG activity, and on repeating the MAIPA following preabsorption of the serum with mouse ascites proteins they gave negative results. Reactivity of the sample which did not correct by this procedure was shown to be due to an anti‐HPA1a alloantibody. The potential problem of human heterophile antibodies reacting with mouse antibodies in the MAIPA has been identified and a modification which corrects this has been demonstrated.
Increased in vitro neutrophil adherence in a case of chronic idiopathic neutropenia
Authors: Biasi Domenico , Bambara Lisa Maria , Carletto Antonio , Caramaschi Paola , Ortolani Riccardo , Perona Giuseppe , Bellavite Paolo
Published: March 2008

Summary In this report we describe a patient with persistent neutropenia whose neutrophils showed increased adhesion in a microplate assay. In three separate assays, from 12·5% to 13·7% of the patient's blood neutrophils exhibited spontaneous (unstimulated) adhesion to fetal bovine serum‐coated microplate wells, much higher than adhesion of cells from healthy controls (1·9% 2·5 SD, n= 20). The difference of spontaneous adhesion between the patient's and control neutrophils was even higher when cells from a skin‐window exudate were examined (patient: 42·1–100% adhesion: control: 3·6% 3·5 SD, n= 20). Over 80% inhibition of the increased adhesion was produced by the 60.3 anti‐CD 18 monoclonal antibody, suggesting an involvement of β2‐integrins.
Autoimmune haemolytic anaemia presenting 9 years prior to Castleman's disease
Authors: Rojas Rafael , Martin Carmen , Roman Jose , Garcia Jose M. , Marchal Trinidad , Torres Antonio
Published: March 2008

Summary Castleman's disease (CD) is a rare disease of unknown aetiology and pathogenesis. We present an unusual case of abdominal CD in whom the first manifestation was an autoimmune haemolytic anaemia presenting 9 years before the diagnosis. The Coombs test became negative 2 months after surgical resection of the mass, suggesting that the mass was the source of the autoantibody. CD may be present in autoimmune haemolytic anaemia in patients with no evidence of any other disease.