bluuebunny/crossref_metadata_2025 huggingface dataset:
The treatment of thrombotic thrombocytopenic purpura: plasma infusion or exchange?
Authors: Novitzky Nicholas , Jacobs Peter , Rosenstrauch Wilhelm
Published: March 2008

SUMMARY. Classic thrombotic thrombocytopenic purpura has substantial mortality and, because the pathogenesis is uncertain, multiple therapies are often used. These include corticosteroids and antiplatelet drugs, with plasma infusion or exchange most dramatically influencing outcome. To compare the relative efficacy of these latter two options, the records of 20 patients were retrospectively analysed. The groups were well matched for size and disease severity and received equivalent volumes of plasma. No significant difference in response rate or survival was demonstrated, although plasmapheresis may be preferable in the presence of impaired renal function with fluid overload.
Age‐related alterations in erythroid and granulopoietic progenitors in Diamond‐Blackfan anaemia
Authors: Casadevall Nicole , Croisille Laure , Auffray Isabelle , Tchernia Gil , Coulombel Laure
Published: March 2008

SUMMARY. Mechanisms involved in the erythroid failure characterizing Diamond‐Blackfan anaemia (DBA) remain unidentified. The general consensus is that the defect is intrinsic to the marrow erythroid progenitor, but the target progenitor cell has not been precisely identified, and in vitro studies have revealed considerable heterogeneity between patients. In order to understand better the meaning of such a biological heterogeneity, we examined the in vitro response of erythroid progenitors CFU‐E (colony‐forming unit‐erythroid) and BFU‐E (burst‐forming unit‐erythroid) to erythropoietin (Epo), interleukin‐3 (IL‐3) and stem cell factor (SCF) in a large series of 24 patients from 1 month to over 20 years of age. Results of colony assays revealed a striking correlation between the age of the patient and the extent of the abnormalities detected in vitro. Therefore, despite profound anaemia, 80% (7/10) of the patients studied within 1 year of diagnosis had normal numbers of both CFU‐E and BFU‐E which exhibited a normal response to cytokines. In contrast, 12/14 patients followed up for more than 3 years had decreased numbers of erythroid progenitors, in seven cases associated with decreased colony‐forming unit granulocyte‐macrophage (CFU‐GM). The number of CFU‐E and BFU‐E was not normalized even by the addition of high concentrations of combined Epo, IL‐3 and SCF. These data strongly support the idea that the haemopoietic defect in DBA involves a pluripotent progenitor and worsens with time: it is masked by the culture conditions at the onset of the disease, whereas overt expression of intrinsic alterations occurs only at later stages of the disease and these are not restricted to the erythroid lineage.
Congenital haemolytic anaemia associated with adenylate kinase deficiency
Authors: Toren Amos , Brok‐Simoni Frida , Ben‐Bassat Isaac , Holtzman Fanny , Mandel Mathilda , Neumann Yoram , Ramot Bracha , Rechavi Gideon , Kende George
Published: March 2008

SUMMARY. Chronic haemolytic anaemia associated with adenylate kinase (AK) deficiency is very rare and only seven cases in five families have been described. We present six children of one family who are deficient of this enzyme and in three of them a combined G6PD deficiency was found. AK deficiency was transmitted by an autosomal recessive gene and heterozygous state was not accompanied by disease, whereas homozygously affected individuals present a congenital chronic non‐spherocytic haemolytic anaemia with haemoglobin levels of 8‐9 g/dl. Patients also deficient in G6PD suffer from a more severe haemolytic anaemia with haemoglobin levels around 6 g/dl. The AK‐deficient children are also mentally retarded. Splenectomy performed in five of the six patients resulted in complete remission of the haemolytic process.
Refractory anaemia and mitochondrial cytopathy in childhood
Authors: Bader‐Meunier Brigitte , ROutig Agnes , Mielot Francoise , Lavergne Jean Maurice , Croisille Laure , Rustin Pierre , Landrieu Pierre , Dommergues Jean Paul , Munnich Arnold , Tchernia Gil
Published: March 2008

SUMMARY. We report two cases of childhood myelodysplasia (MDS) related to a mitochondrial (mt) cytopathy that illustrate the difficulty in recognizing such disorders in patients with solely haematological signs. Both patients have refractory anaemia with ring sideroblasts and vacuolization of haemopoietic precursors. These cytological features are similar to those observed in Pearson's disease, recently identified as a mitochondrial disease, and are strongly suggestive of a mitochondrial enzyme defect. The diagnosis of mitochondrial cytopathy was established on Southern blotting of mt DNA, showing a mt DNA deletion, or on the impairment of the respiratory chain enzyme activities. The absence of cytogenic abnormalities, and the polyclonal pattern of peripheral neutrophil and lymphocyte fractions, suggest that, in mt cytopathies, MDS cannot be considered as a truly malignant disorder.
Late listeriosis after fludarabine plus prednisone treatment
Authors: Girmenia Corrado , Mauro Francesca Romana , Rahimi Siavash
Published: March 2008

SUMMARY. Fludarabine plus prednisome treatment in patients with chronic lymphocytic leukaemia has recently been correlated with an increased incidence of Listeria monocytogenes infections. We observed a patient with chronic lymphocytic leukaemia who received fludarabine plus prednisone and developed a fatal listeriosis 7 months later. A further case of listeriosis has recently been described in a man who developed the infection 2 years after successful fludarabine treatment while he had a normal CD4 lymphocyte count. These cases suggest that the risk period for the development of such an unusual infection could be unexpectedly and unpredictably long. This data could be important in the evaluation of a possible antibacterial prophylaxis schedule and in the wariness about the potential danger of food known to contain L. monocytogenes.
Possible role of granulocyte‐macrophage colony stimulating factor (GM‐CSF) on the rapid progression of AIDS‐related Kaposi's sarcoma lesions in vivo
Authors: Hermans Philippe , Gori Andrea , Lemone Marc , Franchioly Patricia , Clumeck Nathan
Published: March 2008

SUMMARY. Drug‐related neutropenia is a common observation in AIDS patients. Haematological growth factors are therefore increasingly used in combination with myelotoxic agents to reduce the risk of infection and to improve the haematological tolerance of these regimens.

We report a case of an AIDS patient with Kaposi's sarcoma who received GM‐CSF for severe neutropenia due to antitumour chemotherapy combining α‐interferon 2b and zidovudine. During GM‐CSF administration there was a marked increase in the size of the Kaposi's sarcoma lesions as confirmed by ultrasonographic examination. As GM‐CSF in vitro has been shown to promote Kaposi's sarcoma‐like cell cultures, we discuss the potential role of this growth factor in increasing Kaposi's sarcoma lesions in vivo.
PCR‐based determination of Rhc and RhE status of fetuses at risk of Rhc and RhE haemolytic disease
Authors: Le Van Kim Caroline , Mouro Isabelle , Brossard Yves , Chavinie Jacques , Cartron Jean Pierre , Colin Yves
Published: March 2008

Summary. After anti‐RhD, anti‐Rhc is the most important red cell alloantibody which can cause haemolytic disease of the newborn (HDN) when the mother is Rhc‐negative and the fetus Rhc‐positive. We report here the development of polymerase chain reaction (PCR) assays which detect the presence of the Rhc alleles in amniotic cells by the use of allele‐specific primers (ASP). It is expected that such determination will help in the management of pregnancies at risk of Rhc haemolytic disease. In the course of this study we have similarly performed PCR‐ASP experiments to detect fetal RHE alleles since, in rare cases, anti‐RhE can also cause HDN.
Acute myeloid leukaemia with +i(12p) shortly after treatment of mediastinal germ cell tumour
Authors: Vlasveld L.T. , Splinter T. A. W. , Hagemeijer A. , Van Lom K. , Löwenberg B.
Published: March 2008

Summary. We report a patient who developed acute myeloid leukaemia (M2) shortly after successful treatment of a mediastinal germ cell tumour. The leukaemia was preceded by a documented myelodysplastic phase. Complex cytogenetic abnormalities were found in bone marrow and peripheral blood cells including +i(12p), typical of germ cell malignancy. Fluorescence in situ hybridization revealed the presence of +i(12p) in myeloblasts, erythroblasts and neutrophils but not in lymphocytes. This case provides further evidence for a common clonal origin of haematological malignancies and mediastinal germ cell tumours.
Further evidence that activated protein C resistance can be misdiagnosed as inherited functional protein S deficiency
Authors: Cooper P. C. , Hampton K. K. , Makris M. , Abuzenadah A. , Paul B. , Preston F. E.
Published: March 2008

Summary. A recent report that activated protein C (APC) resistance interferes with functional protein S (PS) assays prompted us to re‐investigate two pedigrees previously diagnosed as having functional PS deficiency. APC resistance was demonstrated in all individuals with apparent functional PS deficiency. The latter diagnosis was shown to be due to the assay being non‐linear, functional protein S becoming normal at higher dilutions. This observation, taken in conjunction with results of in vitro recovery studies with purified PS, leads us to conclude that APC resistance was the primary disorder in both pedigrees. The misdiagnosis of APC resistance as functional PS deficiency can be prevented by performing the PS assay at several dilutions, including concentrations lower than those recommended by PS assay manufacturers. Subjects previously diagnosed as having functional PS deficiency should be re‐investigated for APC resistance.
Fluctuating symptomatic cryofibrinogenaemia in a patient with left atrial myxoma
Authors: Cobcroft Ralph G. , Shearer Andrew , Gill Devinder S. , Forgan‐Smith Jeffery
Published: March 2008

Summary. We report a case of left atrial myxoma associated with a fluctuating level of cryofibrinogen. Her initial symptoms and signs were consistent with primary cryofibrinogenaemia, but a repeat episode occurred without the cryofibrinogen being detectable. A more typically embolic cerebro‐vascular accident (CVA) occurred, with the subsequent discovery of the patient's myxoma. The cryofibrinogen was again present but it disappeared with resection of the myxoma. We propose that the degree of activation of the coagulation system around the myxoma fluctuated, causing variable conversion of fibrinogen to fibrin, so that at times partially cross‐linked fibrin(ogen) resulted in cryofibrinogen, but at other times frank emboli from more complete clotting occurred.
The effects of IL‐1 and IL‐4 on the Epo‐independent erythroid progenitor in polycythaemia vera
Authors: de Wolf J. Th. M. , Hendriks D. W. , Esselink M. T. , Halie M. R. , Vellenga E.
Published: March 2008

Summary. Human recombinant interleukin‐1 (IL‐1) was studied for its effects on the erythroid progenitors from normal subjects and from patients with polycythaemia vera (PV). No supportive effect of IL‐1 was noticed on the normal erythropoietin (Epo) dependent, erythroid burst‐forming unit (BFU‐E) using peripheral blood or bone marrow. In contrast, the Epo‐independent BFU‐E from peripheral blood of PV patients could be stimulated significantly. This enhancing effect of IL‐1 was not only observed with unsorted but also with sorted CD34+ cells. In addition, it was shown that IL‐1 indirectly stimulated the Epo‐independent BFU‐E because anti‐GM‐CSF could abrogate the supportive effects of IL‐1. In contrast to the Epo‐independent BFU‐E, the Epo‐dependent erythroid colony formation from PV patients could not be augmented by IL‐1. Finally, we studied the effects of IL‐4 on the Epo‐independent BFU‐E, because IL‐4 can affect the erythroid colony formation and modulate the effects of IL‐1. IL‐4 suppressed the Epo‐independent BFU‐E. This effect could be counteracted by the addition of IL‐1 to the culture medium. However, the suppressive effect of IL‐4 was not related to a decline in spontaneous release of IL‐1, because an anti‐IL‐1 antibody did not modify the spontaneous erythroid colony formation. These data indicate that IL‐1 and IL‐4 exert separate influences on the Epo‐independent erythroid colony formation in PV.
All‐trans retinoic acid promotes a differential regulation of adhesion molecules on acute myeloid leukaemia blast cells
Authors: Noto R. Di , Schiavone E. M. , Ferrara F. , Manzo C. , Pardo C. Lo , Vecchio L. Del
Published: March 2008

Summary. In the present study we investigated the membrance expression of selectin ligands (CD15/Lex, CDw65/VIM2, CD15s/sLex), β2 integrins (CD11a/LFA‐1, CD11b/Mac‐1) and CD45 phosphatase isoforms (CD45RA, CD45RO on leukaemic cells from 28 patients with acute myeloid malignancies cultured with and without all‐trans retinioc acid (ATRA). Within each adhesion system, ATRA was bale to differentially regulate distinct molecules. Furthermore, it was able to exert effects specific for acute promyelocytic leukaemia (APL) blast cells, as well as to induce a series of non‐cytotype‐restricted phenotypic changes. An impressive feature of ATRA induction was a simultaneous increase in the expression of CD15, CDw65 and CD11b on leukaemic promyelocytes. The sialylated antigen CD15s, however, Showed results independent from the other two carbohydrates (CD15 and CDw65), suggesting a differential enzymatic regulation within the selectin ligands system.

In spite of the well‐recognized expression of CD11a throughout all stages of normal myeloid differentiation, APL blast cells were found to virtually lack LFA‐1 expression. Moreover, ATRA was unable to promote and up‐regulation of this antigen in APL, while inducing a frequent down‐modulation in non‐APL cases constitutively expressing this antigen. In APL cases ATRA generated an asynchronous phenotype (CD15+, CDw65+, CD11b+, CD11a‐), undetectable on normally maturing myeloid cells, but consistent with the concept that incomplete differentation, in terms of surface molecule expression, can be sufficient to obtain therapeutic results.
Recombinant human immunodeficiency virus type‐1 (HIV‐1) Tat protein sequentially up‐regulates IL‐6 and TFG‐β1 mRNA expression and protein synthesis in peripheral blood monocytes
Authors: Gibellini Davide , Zauli Giorgio , Re Maria Carla , Milani Daniela , Furlini Giuliano , Caramelli Elisabetta , Capitani Silvano , Placa Michele La
Published: March 2008

Summary. In this study we evaluated the effect of human immunodeficiency virus type 1 (HIV‐1) recombinant Tat protein on mRNA expression and protein synthesis of two inflammatory cytokines ‐ interleukin‐6 (IL‐6) and transforming growth factor‐β1 (TGF‐β1) ‐ by peripheral blood (PB) monocytes. Whereas maximal levels of IL‐6 protein were recovered in PB monocyte culture supernatants after 24–48 h from the addition of 1 μg/ml of recombinant Tat, TGF‐β1 showed a slower and progressive increase, reaching maximal levels only after 72–96h of culture. Consistently, the analysis of the steady‐state levels of mRNA showed a sharp increse of IL‐6 mRNA expression after 24h of culture, with a slow decline thereafter. On the other hand, TGF‐β1 mRNA expression showed a slow increase only after 72–96h of culture. Moreover, IL‐6 appeared involved in the up‐regulation of TGF‐β1, because the addition of a neutralizing anti‐IL‐6 antibody to Tat‐treated PB monocyte cultures significantly reduced the amounts of TGF‐β1 recovered in the culture supernatants after 96 h.

The present demonstration that HIV‐1 Tat protein directly up‐regulates IL‐6 expression and stimulates TGF‐β production both directly and indirectly, through early IL‐6 production, could have important implications in the pathogenesis of HIV‐1 disease.
Interferon‐alpha up‐regulates bcl‐2 expression and protects B‐CLL cells from apoptosis in vitro and in vivo
Authors: Jewell A. P. , Worman C. P. , Lydyard P. M. , Yong K. L. , Giles F. J. , Goldstone A. H.
Published: March 2008

Summary. The bcl‐2 oncoprotein, which is involved in the t(14.18) translocation, protects cells against apoptosis. We examined the effects of interferon‐alpha (IFN‐α) on bcl‐2 protein expression and apoptosis in B‐chronic lymphocytic leukaemia (B‐CLL) cells. None of 12 patients with B‐CLL examined expressed the t(14,18) translocation; however, all these, and seven other patients, expressed significant levels of bcl‐2 protein. In vitro, IFN‐α (500U/ml over 18 h) increased bcl‐2 expression on CLL cells (to 200±23% of control MCF, as determined by indirect immunofluorescence and flow cytometry. n = 10, P < 0.001). All of eight patients who received IFN‐α (3 megaunits subcutaneously three times a week) demonstrated an increase in bcl‐2 expression on circulating malignant cells. CLL cells undergo apoptotic cell death when cultured in vitro (35.6 ± 10.3% DNA fragmentation after 18 h, n = 10). In the presence of IFN‐α, however, DNA fragmentation was reduced to 6.6 ± 5.8% (n = 10, P < 0.001). IFN‐α also protected CLL cells against apoptosis induced by hydrocortisone and gamma irradiation (reducing DNA fragmentation from 63.9 ± 12.6% to 10.8 ± 4.5 % and from 80 ± 2.9% to 5.4 ± 1.6%, respectively, P < 0,001 for both). The protective effect of IFN‐α was dose dependent, and maintained for up to 24 h. Our data demonstrate that bcl‐2 expression and apoptosis of CLL cells can be influenced by cytokines. In addition, it seems unlikely that the observed clinical responses to IFN‐a in patients with CLL are due to a direct effect on the malignant cells.
Aplastic anaemia following exposure to 3,4‐methylenedioxymethamphetamine (‘Ecstasy’)
Authors: Marsh J. C. W. , Abboudi Z. H. , Gibson F. M. , Scopes J. , Daly S. , O'Shaunnessy D. F. , Baughan A. S. J. , Gordon‐Smith E. C.
Published: March 2008

Summary. We report two cases of aplastic anaemia following exposure to ‘Ecstasy’ (MDMA, 3,4‐methylenedioxymethamphetamine). In both cases the aplastic anaemia resolved spontaneously 7–9 weeks after presentation. Long‐term bone marrow culture study of one patient demonstrated complete normalization of haemopoiesis at time of haematological recovery, suggesting either that damage to the haemopoietic stem cell had been only transient, or that a more mature, committed progenitor cell was the target. Because MDMA may have been a factor in the aetiology of the bone marrow suppression in these two cases, we recommend close haematological monitoring of young adults presenting with toxicity from MDMA, and a detailed history of exposure to recreational drugs in all new patients presenting with aplastic anaemia.
The differences in quantities of α2‐and α1‐globin gene variants in heterozygotes
Authors: Molchanova T. P. , Pobedimskaya D. D. , Huisman T. H. J.
Published: March 2008

Summary. We have identified through sequencing of amplified DNA the mutations in the α2‐ and α1‐globin genes in 63 individuals with a heterozygosity for an α chain abnormal haemoglobin (Hb). Moreover, we developed a reverse transcription/polymerase chain reaction (RT/PCR) based procedure for the determination of the α2‐ and α1‐ mRNA ratio in normal individuals. The numbers of α2 and α1 variants were nearly the same. The average precentage of the abnormal Hb in heterozygotes with α2 mutations (23.5%) was slightly higher than that in heterozygotes with α mutations (19.7%) (stable Hbs only). These percentages correspond to a ratio of α2 to α1 of 1.19 to 1 at the protein level. Variations in the number of active α‐globin genes and in the stability of the variants (greatly) affected the percentages of the abnormal protein. The average ratio between the α2‐ and α1‐mRNAs in 12 normal individuals was 2.6–2.75 to 1, about as expected from published data. and 2.0 to 1 for two persons with an α‐thalassaemia‐2 (α‐thal‐2) (‐3.7 kb) heterozygosity. The high relative mRNA (α2) level which is about twice the relative level of the α2 protein suggests a less efficient translation of the α2‐mRNA.
Sequential acquisition of trisomy 8 and N‐ras mutation in acute myeloid leukaemia demonstrated by analysis of isolated leukaemic colonies
Authors: Price Cathy M. , Marshall Christopher J. , Bashey Asad
Published: March 2008

Summary. Specific chromosomal aberrations and point mutations of the N‐ras proto‐oncogene are characteristic genetic alterations in acute leukaemias. However, the relationships between these two different genetic changes are unclear. Here we have determined the order of genetic events in a patient with acute myeloid leukaemia characterized by trisomy 8 and a point mutation of N‐ras at codon 12 (N12‐cys) and codon 61 (N61‐his). 30 colonies obtained by in vitro clonogenic assay of leukaemic cells from a patient with AML were individually analysed for the presence of trisomy 8 and each of two different N‐ras mutations by fluorescence in situ hybridization (FISH) and the polymerase chain reaction (PCR). Trisomy 8 was detected in 25/26 evaluable colonies. 19/26 colonies contained the N12‐cys mutation. The N61‐his mutation was not detected in any of the colonies obtained. All the colonies with the N12 cys mutation were also trisomic from chromosome 8, whereas 6/25 colonies with trisomy 8 had no N‐ras mutation. These data suggest that trisomy 8 was acquired before N12 cys mutation in the pathogenesis of this leukaemia and that two genetic events can co‐operate within a single subclone.
Distinct lysozyme content in different subtypes of acute myeloid leukaemic cells: an ultrastructural immunogold study
Authors: Resnitzky Peretz , Shaft Dina
Published: March 2008

Summary. Using an ultrastructural immunogold method, we performed a quantitative study on cellular lysozyme (LZ) content in young normal bone marrow cells and in 14 cases of acute myeloid leukaemia (AML) of the M2, M3, M4 and M5 types. In five cases of M2 we found significantly lower LZ content than in normal promyelocytes and than in nine cases of M3, M4 and M5. In M3, M4 and M5 cells a very high LZ content was observed whereas the serum LZ activity was high in M4 and M5 and normal in M3. The intragranular LZ content was especially high in M5 and in most granules of M4 cells. The immunogold reaction (IGR) for LZ was also performed in cells previously reacted for myeloperoxidase (MPO). In M2 the granules showed definite positive MPO reactivity and low LZ density (granulocytic pattern), whereas in M5 we found high granular LZ content and weak or almost negative MPO activity (monocytic pattern). In M4 we found ‘granulocytic’ and ‘monocytic’ type of granules in the same cell. The IGR for LZ performed in post‐embedded M5 cells which were previously subjected to phagocytosis of latex particles, showed granules that had moved toward the phagosome, releasing LZ without degranulation. The above findings and those showing normal serum LZ in M3 despite their high cellular LZ content, definitely indicate that only leukaemic M4 and M5 cells secrete LZ into their environment, explaining the high serum LZ observed in those leukaemias.
Platelet procoagulant complex assembly in a tissue factor‐initiated system
Authors: Monroe Dougald M. , Roberts Harold R. , Hoffman Maureane
Published: March 2008

Summary. The aim of this study was to examine the assembly of the factor IXa/VIIIa (Xase) and factor Xa/Va (IIase) complexes on the platelet surface in a system designed to mimic tissue factor‐initiated coagulation. The experimental system contained tissue factor‐bearing monocytes, unactivated platelets, and plasma concentrations of factors V, VIII, IX, X, prothrombin, tissue factor pathway inhibitor (TFPI), antithrombin III (ATIII), and small amounts of factor VIIa. The time courses of platelet activation, coagulation factor binding and thrombin generation were compared. In this system, thrombin generation by the combination of monocytes and platelets was synergistic compared to each cell type alone. Platelet activation and thrombin generation were minimal in the absence of prothrombin or factor X. After a lag period, platelet activation began, followed by progressive binding of factors Va and VIIIa. This was followed by factor IXa and Xa binding and the onset of thrombin generation. Unexpectedly, a transient early increase in platelet‐associated factor IX and X was also seen, that was due to release from platelets. The amount of factor IX bound to isolated activated platelets was increased by addition of factor VIIIa, or by activation of factor IX to IXa. In contrast, factor VIIIa binding was not altered by the presence of factor IX or IXa. We conclude that in a tissue factor‐initiated system, assembly of the procoagulant complexes on the platelet surface begins after platelet activation occurs. Platelet activation requires thrombin generation in the vicinity of the tissue factor bearing cells. The cofactors Va and VIIIa bind to the platelets and facilitate subsequent binding of factors IXa and Xa to form functional procoagulant complexes.
IgD monoclonal gammopathy with long‐term follow‐up
Authors: Bladé Joan , Kyle Robert A.
Published: March 2008

Summary. The presence of a serum IgD monoclonal protein (M‐protein) is usually indicative of a malignant plasma cell disorder. However, one case of well‐documented benign monoclonal gammopathy (BMG) of IgD type has been reported. We describe another patient with IgD monoclonal gammopathy of undetermined significance (MGUS) with long‐term follow‐up. In this patients an IgD λ M‐protein was found on routine serum electrophoresis. She is alive and has no evidence of multiple myeloma or amyloidosis after 8 years of follow‐up. Although IgD MGUS is exceedingly rare, this case demonstrates that the presence of a serum IgD M‐protein is not necessarily synonymous with a malignant plasma cell process.
HCV RNA levels and HIV infection: evidence for a viral interaction in haemophilic patients
Authors: Telfer Paul T. , Brown David , Devereux Helen , Lee Christine A. , Sheiko Geoffrey M. Du
Published: March 2008

Summary. In order to investigate a possible interaction between HIV and HCV infections, we compared HCV RNA levels in 29 matched pairs of haemophilic patients seropositive for HCV and serodiscordant for HIV. Levels were assayed using the new Chiron Quantiplex bDNA assay and were found to be significantly higher in HIV seropositive patients. There was no association between HCV RNA and age, duration of HCV infection, concentrate usage, markers of HIV progression, or use of zidovudine. Our study supports the hypothesis that HIV infection facilitates HCV replication and leads to more severe liver damage.
Infection of donor lymphocytes with human T lymphotrophic virus type 1 (HTLV‐I) following allogeneic bone marrow transplantation for HTLV‐I positive adult T‐cell leukaemia
Authors: Ljungman Per , Lawler Mark , ÅSjo Birgitta , Bogdanovic Gordana , Karlsson Karin , Malm Claes , McCann Shaun R. , Ringdén Olle , Gahrton Gösta
Published: March 2008

Summary. Human T lymphotrophic virus type 1 (HTLV‐I) associated leukaemia has a poor prognosis even with chemotherapy. We describe a patient with adult T‐cell leukaemia treated with allogeneic bone marrow transplantation from an HTLV‐I negative identical sibling donor. During follow‐up after bone marrow transplantation, HTLV‐I could be repeatedly isolated inspite of anti‐viral prophylaxis. The patient died of an acute encephalitis and HTLV‐I could be detected in autopsy material from the brain. By a PCR‐based technique using short tandem repeats (STRs) it was shown that the patient's haemopoiesis was of donor origin. This shows the infection of donor cells in vivo by an aetiological agent which has been implicated in the leukaemogenic process for adult T‐cell leukaemia.
Complement receptor type 1 (CR1) deficiency on neutrophils in myelodysplastic syndrome
Authors: Ohsaka Akimichi , Saionji Katsu , Watanabe Norimichi , Yokomichi Hironao , Sugahara Yuichi , Nagayama Reizo , Igari Jun
Published: March 2008

Summary. We present a patient with myelodysplastic syndromes (MDS) whose neutrophils exhibited defective expression of complement receptor type 1 (CR1). A 73‐year‐old man was admitted with an evolution of MDS from RA into RAEBT according to the FAB classification of MDS. The neutrophil alkaline phosphatase (NAP) score was zero. The surface expression of membrance effector melecules on neutrophils was determined by indirect immunofluorescence using flow cytometry and monoclonal antibodies. The expression of CR1 on neutrophils as identified by staining with CD35 was defective in the patient, and the expression of other complement receptors (CR3 and CR4), Fc receptors and adhesion molecules was normal. CR1 deficiency and defective NAP score on neutrophils in the patient might account for impairment of common storage pool, presumably novel intracellular secretory vesicles.
The kinetics of colony formation by CFU‐GM in vitro
Authors: Lewis J. L. , Blackett N. M. , Gordon M. Y.
Published: March 2008

Summary. Video‐recordings of whole normal bone marrow granulocyte‐macrophage colony (CFU‐GM) cultures were made after 7, 14 and 21 d. Retrospective viewing of the tapes allowed the relationships to each other of the colonies scored on the three different occasions to be documented. The results show that, according to our scoring criteria, there is very little overlap between the numbers of colonies scored on days 7. 14 and 21. Moreover. only about 10% of the progenitors in a sample form day 21 colonies. The remaining progenitors form colonies earlier in the culture period and either disappear before day 21 or remain small.
Interleukin‐3 is an autocrine growth factor of human megakaryoblasts, the DAMI and MEG‐01 cells
Authors: Chen Yuan Zhong , Gu Xue Fan , Caen Jacques P. , Han Zhong Chao
Published: March 2008

Summary. Interleukin‐3 (IL‐3), a cytokine konw to be produced by activated T lymphocytes, mast cells, eosinophils and neutrophils, is a potent stimulator of normal haemopoiesis, particularly magakaryocytopoiesis. However, it remains unknown whether leukaemic magakaryoblasts can produce IL‐3 and whether IL‐3 is involved in the pathological process of megakaryoblastic leukaemia. In this study, several human leukaemia cell lines with or without megakaryocytic features the DAMI, MEG‐01, HEL, K562, HL‐60 and U937, were chosen as the models. It was first demonstrated by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and indirect immunofluorescence assay that IL‐3 was expressed in DAMI and MEG‐01 cells, but not in other cell lines, although two erythroleukaemic cells, the HEL and K562, also possess some megakaryocytic features. Interestingly, the mRNA for IL‐3 receptor was detected in nearly all the cell lines except K562 cells, suggesting that expression of IL‐3 and its receptor may be dissociated in most of the cell lines and that co‐expression of IL‐3 and its receptor exists in megakaryoblastic cell lines, the DAMI and MEG‐01. Of the cell lines which did not express IL‐3 under unstimulated condition, only HEL cells were able to express IL‐3 mRNA after treatment with PMA for 72 h. Furthermore, the proliferation of DAMI and MEG‐01 cells could be enhanced in the presence of IL‐3 and suppressed by the anti‐IL‐3 antibody and the IL‐3 antisense oligodexyonucleotides (ODNs). These findings indicate that IL‐3, as an autocrine growth factor, is involved in the growth of some megakaryocytic leukaemia cell lines.
Haemopoiesis of transplanted patients with autologous marrows assessed by long‐term marrow culture
Authors: Domenech J. , Gihana E. , Dayan A. , Truglio D. , Linassier C. , Desbois I. , Lamagnere J. P. , Colombat P. , Binet C.
Published: March 2008

Summary. We assessed the effect of antitumoural therapy at intensive doses on the haemopoietic system using long‐term marrow cultures (LTMC) established from 33 patients (25 with haematological diseases and eight with solid tumours) after autologous bone marrow transplantation (ABMT). When compared to 42 pre‐graft patients, a decreased CFU‐GM Production and a defect in stromal layer (SL) confluence were found after ABMT on day 90 but also on day 365. However, these abnormalities were observed only in patients with haematological diseases and no differences between pre‐graft and post‐graft results were found in patients with solid tumours. Among the patients with haematological diseases, on day 90 those with acute lymphoid leukaemias showed lower CFU‐GM production whereas patients with non‐Hodgkin's lymphomas developed more frequently subconfluent or confluent SL. Other factors studied such as sex, patient age, disease status, marrow purging and post‐graft administration of growth factors did not appear to influence post‐graft LTMC results. Multivariate analysis including all the patients has shown (a) that solid tumours were associated with higher CFU‐GM production, and (b) that conditioning regimens with total body irradiation (TBI) or busulfan led more frequently to non‐confluent SL. In conclusion, high‐dose therapy followed by ABMT can induce a persistent impairment of the stem cell and stromal cell compartments, particularly in patients with haematological diseases conditioned with TBI, despite the absence of any alloimmune reaction and post‐graft immunosuppressive therapy.
In vivo metabolic studies of glucose, ATP and 2,3‐DPG in β‐thalassaemia intermedia, heterozygous β‐thalassaemic and normal erythrocytes: 13C and 31P MRS studies
Authors: Ting Y. L. T. , Naccarato S. , Qualtieri A. , Chidichimo G. , Brancati C.
Published: March 2008

Summary. 13C and 31P magnetic resonance spectroscopy was used to characterize the in vivo kinetics of glucose metabolism and intracellular ATP and 2,3‐DPG concentrations in erythrocytes obtained from β‐thalassaemia intermedia, heterozygous β‐thalassaemic and normal individuals and maintained in suspension. Except for an upfield chemical shift in the 2P and 3P resonance of 2,3‐DPG in the thalassaemia intermedia erythrocytes, the 31P spectra were comaparable between all three blood types, showing similar concentrations of ATP (from 4.5 to 5.2 μmol/g Hb) and 2,3‐DPG (from 17.2 to 19.7 μmol/g Hb). However, the profile of glucose metabolism was quite different in β‐thalassaemia intermedia erythrocytes, where glucose was consumed at a rate of 0.089 ± 0.035 fmol/cell/h, significantly higher than that of normal (0.032 ± 0.018 fmol/cell/h; P= 0.01) and heterozygous (0.025 ± 0.004 fmol/cell/h; P= 0.01) erythrocytes. This near 3‐fold faster rate of glucose metabolism in the thalassaemia intermedia erythrocytes could not be accounted for by any increase in glucose flux via the Embden‐Meyerhof pathway, since no significant difference in 3‐13C‐lactate synthesis was observed among the three blood types (in units of fmol/cell/h, normal, 0.021 ± 0.013; heterozygous, 0.021 ± 0.006; β‐thalassaemia intermedia 0.045 ± 0.025). These results reflect an accelerated rate of glucose metablolism in thalassaemia intermedia erythrocytes because the contribution of reticulocytes to this altered pattern of metabolism could be excluded. As the only other route of glucose metabolism in erythrocytes is the pentose phosphate pathway (PPP), these results indicate that the PPP is more active in β‐thalassaemia intermedia erythrocytes, perhaps as a consequence of their elevated intracellular oxidative state.
Homozygous β‐thalassaemia resulting in the β‐thalassaemia carrier state phenotype
Authors: Rosatelli Maria Cristina , Pischedda Alessandra , Meloni Alessandra , Saba Luisella , Pomo Anna , Travi Maurizio , Fattore Silvia , Cao Antonio
Published: March 2008

Summary. This paper describes the phenotypic manifestations of a very mild β‐thalassaemia mutation detected in several members of two families of Italian descent. The molecular defect, defined by denaturing gradient gel electrophoresis analysis and direct sequencing. consists of a C G substitution at position 844 of IVSII of the β‐globin gene within the consensus sequence of IVSII acceptor splice site. Heterozygotes for this mutation show a haematological phenotype ranging in severity from silent β‐thalassaemia to that of a mild β‐thalassaemia carrier silent β‐thalassaemia to that of a mild β‐thalassaemia carrier state, whereas homozygotes have the typical manifestations commonly resulting from heterozygosity for a β‐thalassaemia mutation. Compound heterozygotes for the IVSII nt844 (C G) mutation and a severe β‐thalassaemia mutation have the phenotype of thalassaemia intermedia.

This paper indicates that the presence of borderline red blood cell indices or HbA2 values should make one suspect the presence of a very mild or silent β‐thalassaemia.
Factor VIII, ABO blood group and the incidence of ischaemic heart disease
Authors: Meade T. W. , Cooper J. A. , Stirling Y. , Howarth D. J. , Ruddock V. , Miller G. J.
Published: March 2008

Summary. Relations of factor VIII activity, FVIIIC, and von Willebrand factor antigen (vWFAg), with ischaemic heart disease (IHD) were examined in 1393 men aged between 40 and 64 years at entry to the Northwick Park Heart Study (NPHS) who experienced 178 first major episodes of IHD during an average follow‐up period of 16.1 years. After allowing for the large factor VIII differences between the main ABO blood groups, FVIIIC was probably associated with IHD incidence, possibly more strongly with fatal than non‐fatal episodes. Thus, an increase of 1 standard deviation in FVIIIC raised the risk of fatal IHD by about 28%. vWFAg was also significantly associated with fatal events. The observed relation of FVIIIC with IHD incidence probably underestimates the true strength of the association because of the considerable within‐person and laboratory variability in factor VIII measurements. FVIIIC and vWFAg were strongly correlated (r= 0.57) and in statistical terms there may be little to choose between them in long‐term studies of IHD. Taking account of evidence that haemophiliacs seem to experience less IHD than expected, high factor VIII levels may contribute to the incidence of IHD by increasing thrombogenic potential. The incidence of IHD was significantly higher in those of blood group AB than in those of groups O, A or B, particularly for fatal events. There was no evidence that the FVIIIC and vWFAg associations with IHD are determined by ABO group. The factor VIII and ABO blood group effects therefore appeared to be independent. Group AB may be a genetic marker of characteristics influencing other indices of IHD risk such as short stature, NPHS men (though not women) of group AB being about 2 cm shorter than those of other groups.
Epstein‐Barr virus in familial Hodgkin's disease
Authors: SCHLAIPER DANIEL , RIGAL‐HUGUET FRANCOISE , ROBERT ALAIN , ATTAL MICHEL , ABBAL MICHEL , FONCK YVETTE , LAURENT GUY , PRIS JACQUES , DELSOL GEORGES , BROUSSET PIERRE
Published: March 2008

Summary Hodgkin's disease (HD) has been found to be linked to Epstein‐Barr virus (EBV). Familial HD (FHD) may be related to a possible unknown agent. We have determined whether EBV small RNAs (EBERs) were found in Reed‐Sternberg cells from FHD. Five families were studied for histological subtype and EBER presence. There was a striking similarity in FHD subtypes of each family and 3/11 (27%) of the cases were EBER positive. In conclusion, EBV EBERs are only infrequently found in FHD and other factors including viruses different from EBV should be further investigated in FHD.
Successful treatment of essential thrombocythaemia and recurrent abortion with alpha interferon
Authors: Williams Jonathan M. , Schlesinger Peter E. , Gray Atherton G.
Published: March 2008

Summary. A 27‐year‐old woman with essential thrombocythaemia (ET) had three miscarriages. She was treated with alpha interferon which was continued after she became pregnannt again. The platelet count remained well controlled and at term she delivered a healthy baby with normal blood counts. The case demonstrates the effectiveness of IFN in preventing recurrent miscarriage associated with ET and the safety of IFN with respect to fertility and fetal development.
First report of fludarabine in gamma‐heavy chain disease
Authors: Agrawal Samir , Abboudi Zaid , Matutes Estella , Catovsky Daniel
Published: March 2008

Summary. We describe a case of gamma‐heavy chain disease presenting with lymphadenopathy, splenomegaly, lytic lesions and bone marrow infiltration. Chemotherapy with epirubicin, chlorambucil and prednisolone produced a partial remission which lasted 5 months. The patient was then treated with fludarabine. After six courses of fludar‐abine a good partial remission was achieved which converted to complete remission with disappearance of the monoclonal IgG3 in the ensuing 12 months without further therapy. This response has, so far, been sustained for 2½ years after stopping treatment.
A flow cytometric assay of CD34‐positive cell populations in the bone marrow
Authors: Syrjälä M. , Ruutu T. , Jansson S.‐E.
Published: March 2008

Summary CD34+ BM cells form a heterogenous population of primitive stem cells and more mature progenitors committed to different lineages of differentiation. By combining CD45 expression with SSC, it is possible to separate immature cells from more diferentiated BM cells, and, by three‐colour flow cytometry, analyse the antigens expressed in various subsets of cells. In this paper we show that in the normal BM at least four distinct CD34+ cell populations can be identified by their different patterns of CD45 expression and SSC. The most immature CD34+ cell population (0.1% of the BM cellularity) lacked all signs of lineage commitment and was CD45RA negative and only weakly CD45 positive. With increasing expression of the CD45 antigens, a second CD34+ population (0.2% of the BM cellularity) was formed expressing mainly primitive lymphatic antigens. However. 30% of the cells co‐expressed B‐cell line antigens and myeloid antigens. Cells committed to the myeloid cell line lost B‐cell line antigens, gained CD45 antigen expression and SSC and formed two CD34+ cell population (0.2% and 0.1% of the BM cellularity. respectively) differing only with respect to the pattern of myeloid antigen expression and SSC characteristics. Similarly, differentiation along the lymphatic pathway implicated down‐regulation of myeloid antigens, loss of the stem cell antigen and immature lymphatic antigens and gain of CD45 expression and mature lymphatic antigens.
Long‐term safety of treatment with recombinant human granulocyte colony‐stimulating factor (r‐metHuG‐CSF) in patients with severe congenital neutropenias
Authors: Bonilla Mary Ann , Dale David , Zeidler Cornelia , Last Linda , Reiter Alfred , Ruggeiro Mary , Davis Mark , Koci Barbara , Hammond William , Gillio Alfred , Welte Karl
Published: March 2008

Summary . Congenital neutropenias include a heterogenous group of diseases characterized by a decrease in circulating neutrophils. In phase I/II/III studies in patients with severe congenital and cyclic neutropenia, treatment with recombinant human granulocyte colony‐stimulating factor (r‐metHuG‐CSF) resulted in a rise in the absolute neutrophil counts (ANC) and a reduction in infections. We report the effects of long‐term safety of subcutaneous r‐metHuG‐CSF administration in 54 patients (congenital n= 44, cyclic n= 10) treated for 4–6 years. A sustained ANC response was seen in 40/44 severe congenital neutropenia patients and 10/10 cyclic neutropenia patients. Two patients required an increase of > 25% in dose to maintain a clinical response; one patient became refractory to therapy. A significant decrease in the incidence of severe infections and the need for intravenous antibiotics was noted. Significant adverse events noted which may or may not be related to therapy included: osteopenia (n= 15), splenomegaly (n= 12), hypersplenism (n= 1), vasculitis (n= 2), glomerulonephritis (n= 1), BM fibrosis (n= 2), MDS/leukaemia (n= 3), and transient inverted chromosome 5q with excess blasts (n= 1). R‐metHuG‐CSF has been well tolerated in the majority of patients and resulted in a longterm improvement in their clinical status.
Haemophagocytic lymphohistiocytosis: experience at two U.K. centres
Authors: Hirst W. J. R. , Layton D. M. , Singh S. , Mieli‐Vergani G. , Chessells J. M. , Strobel S. , Pritchard J.
Published: March 2008

Summary Haemophagocytic lymphohistiocytosis (HLH) is a rare disorder of inappropriate macrophage activation. Both familial and sporadic forms, which may be infection‐associated, are recognized. Between 1985 and 1991 we treated 23 cases of HLH (12 male, 11 female). There were eight familial cases, defined by a previously affected sibling and/or history of consanguinity, age 3 d to 15 months at presentation. The age of the remaining 15 cases varied from 1 month to 9.5 years. A potential viral trigger was identified in four cases (EBV, two; parvovirus B19, one; echovirus II, one) including one familial case. Six of eight (75%) patients who received supportive care alone, including all four familial cases, died within 6 months of presentation. Both long‐term survivors in this group presented at an older age (7.5 and 8 years) and had proven or suspected virus‐associated HLH. 15 patients were treated with etoposide (150–250 mg/m2 days 1–3 every 21 d) and methylprednisolone; 10 patients received intrathecal methotrexate in addition. In nine (60%) of these cases a complete (six) or partial (three) response was achieved, though one child suffered a fatal ‘tumour lysis’ syndrome. Overall mortality in the treated group was 66.6%, being highest (75%) in patients under 2 years at presentation compared to 33% in those over 2 years. Two of three familial and one of five sporadic cases relapsed and died 3 d to 20 months from diagnosis. Only one familial case survives at follow‐up of 11 months. Of the five remaining survivors, two received allogeneic bone marrow transplantation (one matched related, one haploidentical) and are alive at 11 and 29 months. Three cases aged 2.5, 7.5 and 9.5 years remain in remission at 11, 20 and 25 months respectively. The high mortality of HLH supports a role for allogeneic BMT in selected cases, particularly those with a familial basis or under 2 years at presentation.
Expression and functional role of c‐kit ligand (SCF) in human multiple myeloma cells
Authors: Lemoli Roberto M. , Fortuna Alessandra , Grande Alexis , Gamberi Barbara , Bonsi Laura , Fogli Miriam , Amabile Marilina , Cavo Michele , Ferrari Sergio , Tura Sante
Published: March 2008

Summary. In this study we investigated the proliferation of three well‐documented MM lines and 10 bone marrow samples from myeloma patients in response to rh‐SCF alone and combined with Interleukin‐6 (IL‐6), IL‐3 and IL‐3/GMCSF fusion protein PIXY 321. Neoplastic plasma cells were highly purified (>90%) by immunomagnetic depletion of T, myeloid. monocytoid and NK cells. The number of S‐phase cells was evaluated after 3 and 7d of liquid culture by the bromodeoxyuridine (BRDU) incorporation assay. The proliferation of RPMI 8226 and U266 cell lines was also assessed by a clonogenic assay. All the experiments were performed in serum‐free conditions. RPMI 8226 cell line was not stimulated by SCF which also did not augment the proliferative activity of IL‐6, IL‐3 and PIXY‐321. Conversely, SCF addition resulted in 2.4‐fold increase of the number of U266 colonies and in a higher number of U266 and MT3 cells in S‐phase (24.5 ± 2% SEM v 14.5 ± 1% SEM and 32 ± 3% SEM v 21 ± 4% SEM, respectively; P < 0.05). The c‐kit ligand also enhanced the proliferation of MT3 and U266 cells mediated by the other cytokines. Anti‐SCF polyclonal antibodies completely abrogated the proliferative response of MT3 cells to exogenous SCF and markedly reduced the spontaneous growth of the same cell line. Reverse transcriptase‐polymerase chain reaction amplification (RT‐PCR) did detect SCF mRNA in MT3 and RPMI 8226 cells. Moreover, secreted SCF was found, in a biologically active form, in the supernatant of the two cell lines by the M07e proliferation assay.

When tested on fresh myeloma samples, SCF increased the number of S‐phase plasma cells (4.7 ± 1.6% v 3.4 ± 1.3% in control cultures: P= 0.02). Significant proliferation was also induced by IL‐6 (7 ± 2.3% of BRDU+ cells: P = 0.006), IL‐3 (5.3 ± 1.3%; P=0.01) and PIXY‐321 (5.4 ± 1.6%; P = 0.02). The addition of SCF significantly enhanced the proliferation of myeloma cells responsive to IL‐6. In summary, our results indicate that SCF is expressed in MM cells and stimulates the proliferation of neoplastic plasma cells.