bluuebunny/crossref_metadata_2025 huggingface dataset:
An oxaliplatin‐based chemotherapy in patients with relapsed or refractory intermediate and high‐grade non‐Hodgkin's lymphoma
Authors: Chau Ian , Webb Andrew , Cunningham David , Hill Mark , Rao Sheela , Ageli Suad , Norman Andrew , Gill Kairen , Howard Anne , Catovsky Daniel
Published: March 2003

This study was designed to assess the efficacy and safety of substituting cisplatin with oxaliplatin in the DHAP (dexamethasone, cytarabine and cisplatin) regimen for patients with relapsed or refractory non‐Hodgkin's lymphoma. Twenty‐four evaluable patients with intermediate or high‐grade non‐Hodgkin's lymphoma were treated at 3‐weekly intervals with oxaliplatin (130 mg/m2, d 1), cytarabine (2 g/m2 for two doses, d 2) and dexamethasone (40 mg, d 1–4). The median age of the patients was 58 (range 18–70). Histological subtypes were diffuse large B cell, 20; mantle cell, two; anaplastic large cell, one; and peripheral T cell, one. The overall objective response rate (RR) was 50% [95% confidence interval (CI) = 29–71%] including four complete responses and eight partial responses. RR for those patients treated at first relapse was higher than those treated at second and subsequent relapse (77% versus 29%). Grade 3 and 4 toxicity was mainly haematological: anaemia 17%, neutropenia 75% and thrombocytopenia 75%. No grade 4 non‐haematological toxicity was reported. No significant renal and neurotoxicity was demonstrated. Median survival was 10·6 months. Probabilities of 1‐year progression‐free survival and overall survival were 47% (95% CI = 26–66%) and 50% (95% CI = 23–72%) respectively. In conclusion, dexamethasone, cytarabine and oxaliplatin (DHAX) is a novel combination in salvage therapy for relapsed or refractory non‐Hodgkin's lymphoma. It has clinically significant activity with an acceptable toxicity profile. Lack of renal toxicity makes DHAX an attractive cytoreductive regimen before high‐dose chemotherapy.
Real‐time reverse transcription polymerase chain reaction detection and quantification of t(1;19) (E2A–PBX1) fusion genes associated with leukaemia
Authors: Curry John D. , Glaser Mathew C. , Smith Martyn T.
Published: March 2003

A real‐time reverse transcription polymerase chain reaction (RT‐PCR) method is described that enabled the detection and quantification of E2A–PBX1 fusion gene transcripts associated with t(1;19). The method was highly reproducible and offered exceptional sensitivity at 5 fg of fusion transcript per reaction, without the need for a nested PCR primer design. To illustrate the usefulness of this new technology the E2A–PBX1 fusion gene transcript expression level for several human leukaemia cell lines that are positive and negative for cytogenetically detectable t(1;19) was determined. The RCH–ACV had a threefold higher expression of E2A–PBX1 transcripts (600 transcripts per cell) than the other t(1;19) positive 697 (150 transcripts per cell). The only other cell line with detectable E2A–PBX1 was CEM, but the level of expression was < 1 transcript per cell.
Genotype at the Secretor blood group locus is a determinant of plasma von Willebrand factor level
Authors: O'Donnell James , Boulton Frank E. , Manning Richard A. , Laffan Michael A.
Published: March 2003

Summary. Previous reports on the effect of Secretor and Lewis blood groups on plasma factor VIII–von Willebrand factor (FVIII–VWF) levels have produced conflicting findings. To determine whether either or both loci can influence plasma FVIII–VWF complex levels, we studied the relationship between Secretor and Lewis genotypes, determined definitively using polymerase chain reaction–restriction fragment length polymorphism analysis, and plasma FVIII coagulant activity (FVIII:C) and VWF antigen (VWF:Ag) levels in 136 healthy volunteers. Overall, significantly higher VWF:Ag levels were found in those individuals homozygous for the Se allele (genotype SeSe) than in those heterozygous for the Se allele (P < 0·001). To minimize any confounding influence of ABO genotype/phenotype, we investigated the relationship between Secretor genotype and plasma FVIII–VWF levels within individuals of the same ABO blood group genotype. In the subgroup analysis of group O1O1 individuals alone, VWF:Ag levels were again significantly higher in those individuals with Secretor genotype SeSe than in those either heterozygous or homozygous for the se null allele. Among A1O1 subjects, homozygous Secretors also had significantly higher VWF:Ag levels. In contrast, we found no relationship between Lewis genotype and either VWF:Ag or FVIII:C levels. This study is the first based on genotypic rather than serological analysis, and resolves the previously confounding effects of the Lewis and Secretor loci on plasma FVIII–VWF complex levels.
Fetal haemopoietic cells display enhanced migration across endothelium
Authors: Yong Kwee L. , Fahey Anne , Pahal Gurmit , Linch David C. , Pizzey Arnold , Thomas N. Shaun B. , Jauniaux Eric , Kinnon Christine , Thrasher Adrian J.
Published: March 2003

Summary. Fetal haemopoietic cells continually circulate and migrate into tissues, and thus may have specialized homing capabilities. In this study we investigated the in vitro features of haemopoietic cells in fetal blood and liver which are relevant to homing and engraftment. Fetal cells were examined for long‐term culture‐initiating cell (LTC‐IC) and progenitor content, adhesion molecule expression, cell cycle behaviour and transendothelial migratory activity. The LTC‐IC content of fetal CD34+ cells is similar to that of CD34+ cells from cord and adult mobilized blood. In contrast to adult and cord blood CD34+ cells, fetal CD34+ cells were actively cycling (11·0 ± 1·7% and 28 ± 1·1% of fetal blood and liver CD34+ cells, respectively, in S+G2M, P < 0·001, compared with cord and adult cells). The striking finding was that fetal haemopoietic cells (both LTC‐ICs and committed progenitors) displayed significantly higher levels of migration across endothelium (P < 0·05 compared with cord, P < 0·01 compared with adult blood and bone marrow CD34+ cells), which were further increased by chemokines and growth factors. The superior migratory activity of fetal haemopoietic cells may underlie a more efficient homing ability, in keeping with their physiological role.
Nitric oxide donor properties of hydroxyurea in patients with sickle cell disease
Authors: Gladwin Mark T. , Shelhamer James H. , Ognibene Frederick P. , Pease‐Fye Margaret E. , Nichols James S. , Link Beth , Patel Daksesh B. , Jankowski Marcin A. , Pannell Lewis K. , Schechter Alan N. , Rodgers Griffin P.
Published: March 2003

Summary. Hydroxyurea therapy reduces the rates of vaso‐occlusive crisis in patients with sickle cell anaemia and recent data suggest that hydroxyurea treatment can generate nitric oxide (NO). Nitric oxide has been proposed as a novel therapy for sickle cell disease via a number of pathways. We therefore sought to determine whether hydroxyurea has NO donor properties in patients with sickle cell anaemia and explore potential mechanisms by which NO production could be therapeutic. Venous blood was collected from 19 fasting sickle cell anaemia patients, on chronic hydroxyurea therapy, at baseline and 2 and 4 h after a single morning dose of hydroxyurea, as well as 10 patients not taking hydroxyurea. The plasma and red cell NO reaction products nitrate, nitrite and nitrosylated‐ haemoglobin were measured using ozone‐based chemiluminescent assays (using vanadium, KI and I3– reductants respectively). Consistent with NO release from hydroxyurea, baseline levels of total nitrosylated haemoglobin increased from 300 nmol/l to 500 nmol/l (P = 0·01). Plasma nitrate and nitrite levels also significantly increased with peak levels observed at 2 h. Glutathionyl–haemoglobin levels were unchanged, while plasma secretory vascular cellular adhesion molecule‐1 levels were reduced in patients taking hydroxyurea (419 ± 40 ng/ml) compared with control patients with sickle cell anaemia (653 ± 55 ng/ml; P = 0·003), and were inversely correlated with fetal haemoglobin levels (r = −0·72; P = 0·002). These results demonstrate that hydroxyurea therapy is associated with the intravascular and intraerythrocytic generation of NO. The role of NO in the induction of fetal haemoglobin and possible synergy between NO donor therapy and classic cytostatic and differentiating medications should be explored.
Familial pseudohyperkalaemia Cardiff: a mild version of cryohydrocytosis
Authors: Gore Daniel M. , Chetty Margaret C. , Fisher Julie , Nicolaou Anna , Stewart Gordon W.
Published: March 2003

Summary.  We have investigated a Welsh pedigree showing the ‘familial pseudohyperkalaemia’ phenotype of dominantly inherited, red‐cell‐based, temperature‐dependent pseudohyperkalaemia associated with normal haematology. The ‘passive leak’ to K across the membrane of these abnormal red cells showed a ‘U‐shaped’ temperature dependence, with a minimum at about 23°C, qualitatively similar to that seen in the frankly haemolytic ‘cryohydrocytosis’ variant of the hereditary stomatocytosis group. Like three previous pedigrees with cryohydrocytosis, these patients show an excess of ether lipids in the membrane. However, these patients differ from other ‘familial pseudohyperkalaemia’ pedigrees, in which the leak showed different temperature profiles.
Gastroduodenal lesions in polycythaemia vera: frequency and role ofHelicobacter pylori
Authors: Torgano Giuseppe , Mandelli Clara , Massaro Paolo , Abbiati Carla , Ponzetto Antonio , Bertinieri Giovanni , Bogetto Stefano Ferrero , Terruzzi Elisabetta , De Franchis Roberto
Published: March 2003

Summary. The prevalence of gastroduodenal lesions is higher in polycythaemia vera (PV) than in the general population. However, the role ofHelicobacter pylori(H. pylori) in the pathogenesis of such lesions is unknown. The aim of our study was to evaluate the prevalence of gastroduodenal lesions in PV patients and dyspeptic controls, and to assess the role of PV andH. pyloriinfection in inducing them. Thirty‐five PV patients fulfilling selection criteria and 73 age‐ and sex‐matched controls underwent upper gastrointestinal endoscopy. Six gastric mucosal biopsies were taken in all patients and controls, and analysed for presence ofH. pylori; serum anti‐CagA was assayed by Western blot. Data were analysed with descriptive statistics and multivariate regression analysis. Compared with controls, PV patients showed a significantly higher frequency of erosions (46% versus 12%), ulcers (29% versus 7%),H. pyloripositivity (83% versus 57%), and anti‐CagA positivity (66% versus 37%). Fourteen out of 20 (70%) asymptomatic PV patients had gastroduodenal lesions. At multivariate analysis,H. pylori, presence of PV alone, and both PV and anti‐CagA were significantly and strongly associated with a higher frequency of gastroduodenal lesions (P < 0·05,P < 0·01 andP< 0·05 respectively). Both PV andH. pyloriinfection were independent risk factors for gastroduodenal lesions; the underlying pathogenetic mechanism responsible for gastroduodenal lesions in PV possibly involves blood mucosal flow and trophism. The higher susceptibility ofH. pyloriinfection and the high frequency of asymptomatic gastroduodenal lesions in PV patients suggest a surveillance of these patients.
Regulation of plasminogen activator inhibitor‐1 secretion by urokinase and tissue plasminogen activator in rat epithelioid‐type smooth muscle cells
Authors: Lau Herbert K. F. , Ho Joanne
Published: March 2003

Summary. Tissue plasminogen activator (tPA) and urokinase (uPA) are targets of plasminogen activator inhibitor‐1 (PAI‐1) inhibition. We have previously shown that both proteases can also induce PAI‐1 secretion in rat smooth muscle cells (SMCs). We now report that both proteases appear to use very similar cellular mechanisms for signal transduction. They induced PAI‐1 secretion using a pathway(s) involving protein kinase C (PKC). They also activated the Raf/Mek/mitogen‐activated protein kinase (MAPK) pathway, which lies downstream of PKC activation. Activation of protein kinase A (PKA), however, lowered PAI‐1 secretion induced by uPA and tPA, as a result of an inhibition of the PKC pathway and inhibition of Raf, Mek and MAPK phosphorylations. Src and syk family non‐receptor tyrosine kinases (TK) were also involved in PAI‐1 induction. The mechanisms of interaction of these tyrosine kinases with other pathways appeared to be quite different: src appeared to act within the PKC and PKA pathways, while syk operated independently of these pathways. Furthermore, whereas src inhibition resulted in inhibition of Raf/Mek/Erk phosphorylations, syk inhibition could only inhibit Mek and Erk phosphorylations but not the phosphorylation of Raf. These multiple pathways utilized by uPA and tPA to modulate PAI‐1 secretion might be involved in determining the proteolytic or antiproteolytic potential of the SMCs under different pathophysiological conditions.
Interleukin 6 and tumour necrosis factor alpha serum levels in monoclonal gammopathy of undetermined significance
Authors: Bladé, Joan , Filella, Xabier , Montoto, Silvia , Bosch, Francesc , Rosiñol, Laura , Coca, Francesca , Giné, Eva , Nadal, Elisabeth , Aymerich, Marta , Rozman María , Montserrat Emili
Published: March 2003

Summary.The objectives of the present study were to compare the interleukin 6 (IL‐6) and tumour necrosis factor‐alpha (TNF‐alpha) serum levels of individuals with monoclonal gammopathy of undetermined significance (MGUS) with those of healthy controls, and to ascertain the predictor value of these cytokines in the evolution from MGUS to multiple myeloma. After a median follow‐up of 7 years from the initial cytokine measurements, nine patients with MGUS have evolved to a malignant condition. The actuarial probability of malignant transformation in patients with increased IL‐6 and TNF‐alpha was not significantly higher than in those with normal values.
The endogenous granulocyte colony‐stimulating factor response following autologous peripheral blood stem cell transplantation is impaired in patients with myeloma
Authors: Morris Treen C. M. , Magill Margaret K. , Drake Mary , Price Susan , Ranaghan Lisa , Bridget Stephen , Jordan Anne E. , Irvine Alexandra E.
Published: March 2003

Summary. Granulocyte colony‐stimulating factor (G‐CSF) levels were studied in 23 patients (10 myeloma, 13 relapsed Hodgkin's disease, non‐Hodgkin's lymphoma or germ cell tumours), post autologous peripheral blood stem cell transplantation (PBSCT). The two groups had similar previous chemotherapy and numbers of CD34+ cells transplanted. All patients received G‐CSF by injection starting 8 d post transplantation. Twenty out of 23 patients showed raised endogenous levels of G‐CSF before cytokine administration. Myeloma patients showed significantly lower levels of endogenous G‐CSF than the other patients (0·767 versus 3·262 ng/ml, P < 0·05). Further rises in G‐CSF levels were seen following the administration of exogenous G‐CSF which then fell, despite ongoing administration of G‐CSF, as neutrophil recovery occurred.
The absence of the p15INK4B gene alterations in adult patients with precursor B‐cell acute lymphoblastic leukaemia is a favourable prognostic factor
Authors: Hoshino Koyu , Asou Norio , Okubo Toshiya , Suzushima Hitoshi , Kiyokawa Tetsuyuki , Kawano Fumio , Mitsuya Hiroaki
Published: March 2003

Summary. We examined deletion and methylation of the p15INK4B(p15) and p16INK4A(p16) genes, using Southern blotting and methylation‐specific polymerase chain reaction (PCR), in 70 untreated adult patients with precursor B‐cell acute lymphoblastic leukaemia (PBC‐ALL) and analysed the relationship between their genetic changes and clinical outcome. Methylation and homozygous deletion of the p15 gene were detected in 30 (43%) and 18 (26%) patients, while those of the p16 gene were found in 16 (23%) and 11 (16%) patients respectively. Thirteen out of 17 patients with wild‐type p15 gene showed expression of p15 mRNA, whereas 31 out of 39 patients with alteration (deletion and methylation) of the p15 gene showed no p15 mRNA expression by reverse transcription–PCR, suggesting that alterations of the p15 gene are highly associated with loss of␣p15 mRNA expression. Disease‐free survival (DFS) at 4 years in patients with wild‐type p15 gene is 33%, compared with 4% of those with p15 gene alterations (P = 0·049). Multivariate analysis showed that the absence of p15 gene alterations was an independent significant favourable prognostic factor for longer DFS (P = 0·0001). These results suggest that alterations in the p15 but not p16␣gene can be used as a genetic prognostic indicator in PBC‐ALL.
High level of microsatellite instability but not hypermethylation of mismatch repair genes in therapy‐related and secondary acute myeloid leukaemia and myelodysplastic syndrome
Authors: Mohammad Sheikhha H. , Khalid Tobal  , John Yin A. Liu
Published: March 2003

Summary. Microsatellite instability (MSI) is associated with defects in the DNA mismatch repair (MMR) system, such as mutation or epigenetic silencing of the genes by promoter hypermethylation. We investigated the presence of MSI and promoter hypermethylation of hMLH1 and hMSH2 genes in 82 patients (68 acute myeloid leukaemia, AML; 14 myelodysplastic syndromes, MDS). Twelve separate microsatellite loci, including three mononucleotide repeat markers, were used. Mutator phenotype (RER+) was detected in 20 AML (29·4%) and 3 MDS (21·4%) patients. RER+ rate was much higher in the therapy‐related and secondary cases compared with the de novo cases. Three out of 7 (42·9%) secondary (s‐AML) and 8 out of 17 (47·1%) therapy‐related (t‐AML) showed RER+ in comparison with 9 out of 44 (20·5%) de novo cases. Similar rates were detected in MDS patients (2/2 therapy‐related and 1/12 de novo). The promoter hypermethylation was found in three hMLH1 (3·7%) and two hMSH2 (2·4%) genes. All these five patients had AML and were older than 60 years of age. Two of them had s‐AML and one had t‐AML. RER+ was detected in three of these five patients. Our data suggest that genetic instability is associated with AML and MDS, especially t‐AML and s‐AML. In addition, our results indicate that the hMSH2 and hMLH1 promoter hypermethylation is not a common event in these malignancies, but may play a role in the development of AML in elderly patients.
Ex vivo expansion marginally amplifies repopulating cells from baboon peripheral blood mobilized CD34+ cells
Authors: Norol Françoise , Drouet Michel , Pflumio Françoise , Léonardi Marjorie , Mourcin Frédéric , Debili Najet , Job Agnès , Vainchenker William , Kuentz Mathieu , Hérodin Francis
Published: March 2003

Summary.  The ability of ex vivo expansion to increase the long‐term repopulating capacity of a graft is still unknown. One problem is the most reliable way to quantify transplantable cells. We addressed this point in a baboon model based on autologous transplantation of serial limiting doses of non‐manipulated or ex vivo‐expanded mobilized CD34+ cells and determined the threshold doses of non‐manipulated and expanded cells which supported long‐term multilineage engraftment. In the expansion group, CD34+ cells were cultured for 6 d with a combination of early acting cytokines (Flt3‐ligand, stem cell factor, thrombopoietin and interleukin 3). Grafted cells were characterized by their surface antigens and biological properties [semisolid assays, long‐term culture‐initiating cells (LTC‐IC) and non‐obese diabetic severe combined immunodeficient reconstituting cells (SRC)]. Animals were followed for at least 12 months post transplantation. The expansion protocol yielded 12·3‐fold, 16·9‐fold, 3·7‐fold, 3·5‐fold and 2·2‐fold increases in CD34+ cells, granulocyte‐‐macrophage colony‐forming units (CFU‐GM), megakaryocyte CFU (CFU‐MK), LTC‐IC and SRC respectively. It induced a modest increase in the long term reconstitutive ability of the graft; the threshold value for long‐term engraftment was 0·5 × 106/kg CD34+ cells in the control group and 0·3 × 106/kg CD34+ cells in the expansion group, although one animal in this latter group remained hypoplastic. Frequencies of SRC had a high predictive value of long‐term engraftment (r > 0·80). The main advantage of the protocol was the acceleration of granulocyte recovery, achieved at the different doses tested. In conclusion, these experiments suggest that this ex vivo expansion protocol marginally amplifies long‐term reconstituting cells.
Acquired activated protein C resistance is associated with the co‐existence of anti‐prothrombin antibodies and lupus anticoagulant activity in patients with systemic lupus erythematosus
Authors: Nojima Junzo , Kuratsune Hirohiko , Suehisa Etsuji , Kawasaki Tomio , Machii Takashi , Kitani Teruo ,   Yoshinori Iwatani, Kanakura Yuzuru
Published: March 2003

Summary. Venous thromboembolism (VTE) is one of the common manifestations in the anti‐phospholipid (aPL) syndrome. We examined the levels of IgG antibodies (Abs) to β2‐glycoprotein I (β2‐GP I) and prothrombin, lupus anticoagulant (LA) activity, activated protein C resistance (APC‐R), and factor V Leiden in 96 patients with systemic lupus erythematosus (SLE); 19 with VTE and 77 without VTE. Acquired APC‐R, which was not found in any patient with the factor V Leiden mutation, was present in 33 (34·4%) out of the 96 patients with SLE. The presence of acquired APC‐R was a strong risk factor for VTE. The SLE patients were divided into four groups according to the results of enzyme‐linked immunosorbent assay (ELISA) and LA activity for each aPL Abs: ELISA+, LA+; ELISA+, LA–; ELISA–, LA+; and ELISA–, LA–. A significant association was observed between APC‐R and the co‐existence of anti‐β2‐GP I Abs and LA activity or of anti‐prothrombin Abs and LA activity. There was no association between APC‐R and the presence of anti‐β2‐GP I Abs, anti‐prothrombin Abs, or LA activity alone. However, when multivariate logistical regression analysis was performed, it was clear that only the co‐existence of anti‐prothrombin and LA activity was a significant risk factor for APC‐R. These findings indicate that the co‐existence of anti‐prothrombin Abs and LA activity may be an important factor in the pathogenesis of acquired APC‐R in patients with SLE.
Improvement of interleukin 2 production, clonogenic capability and restoration of stromal cell function in human immunodeficiency virus‐type‐1 patients after highly active antiretroviral therapy
Authors: Isgrò Antonella , Aiuti Alessandro , Mezzaroma Ivano , Addesso Maria , Riva Elisabetta , Giovannetti Antonello , Mazzetta Francesca , Alario Cecilia , Mazzone Annamaria , Ruco Luigi , Aiuti Fernando
Published: March 2003

Summary. Haematological abnormalities frequently occur in patients infected by human immunodeficiency virus‐type 1 (HIV‐1). Increasing evidence indicates that bone marrow suppression (BM) results from viral infection of accessory cells, with impaired stromal function and alteration of haematopoietic growth factor network. We have investigated the effects of antiretroviral therapy on cytokine and chemokine production by BM cells and stromal cells in a group of HIV‐1‐infected subjects before and during treatment. Compared with uninfected controls, an altered cytokine and chemokine production by BM cells was observed before treatment, characterized by decreased interleukin 2 (IL‐2) and elevated tumour necrosis factor‐α, macrophage inflammatory protein (MIP)‐1α, MIP‐1β, and RANTES (regulated on activation, normal T cell‐expressed and secreted) levels, along with a defective BM clonogenic activity. Antiretroviral therapy showed increased BM clonogenic capability, associated with normalization of IL‐2 production and chemokine receptors expression on CD34+ cells. Pre‐therapy, BM accessory cells were represented by macrophage‐like cells, in some cases positive for HIV‐1 DNA, suggesting that these cells are the main target of HIV‐1 infection. During therapy, the stromal cells became predominantly fibroblastoid‐like, as observed in normal controls, and were negative for HIV‐1 DNA. Controlling HIV‐1 replication may produce amelioration of stem cell activity, and restoration of stromal cell pattern and functions, with increased IL‐2 production at BM level.
Diabetes insipidus as first manifestation of acute myeloid leukaemia with EVI‐1‐positive, 3q21q26 syndrome and T cell‐line antigen expression: what is the EVI‐1 gene role?
Authors: Breccia Massimo , Petti Maria C. , Ottaviani Emanuela , Mancini Marco , D'Elia Gianna Maria , Mecarocci Sergio , Alimena Giuliana
Published: March 2003

Summary. Two cases of acute myeloid leukaemia (AML) with CD2 and CD7 expression associated with diabetes insipidus (DI) as the initial symptom are presented. Both patients had t(3;3)(q21;q26) associated with monosomy 7 and EVI‐1 overexpression. No neurohypophysis infiltration was evident. One patient died during induction chemotherapy, the other did not respond to therapy and died with persistent DI. Our findings further support the existence of a distinct AML entity characterized by the presence of DI, abnormalities of chromosome 3q, dysmegakaryopoiesis and poor outcome, and provide evidence of EVI‐1 gene involvement. The possible role of chromosome 3q26 abnormalities in determining this peculiar clinical–biological association is emphasized.
Life‐threatening human parvovirus B19 infection transmitted by intravenous immune globulin
Authors: Hayakawa Fumihiko , Imada Kazumi , Towatari Masayuki , Saito Hidehiko
Published: March 2003

Summary. Infection of human parvovirus B19 (B19) is usually a self‐limiting febrile illness, but can sometimes be life‐threatening under certain circumstances, such as aplastic crisis in patients with haemolytic anaemia, hydrops fetalis in pregnant women and fulminant hepatitis. B19 canbe transmitted through respiratory secretions, transplacentally and by transfusion of blood or blood products. In the present case, administration of intravenous immune globulin (i.v.Ig) transmitted B19 infection and consequently caused pure red cell aplasia and aggravation of hepatitis to fulminant hepatitis. Our case may raise important questions as to the safety of i.v.Ig and possible contamination by B19.
Defective assembly of the B‐cell receptor chains accounts for its low expression in B‐chronic lymphocytic leukaemia
Authors: Payelle‐Brogard Béatrice , Magnac Christian , Alcover Andres , Roux Pascal , Dighiero Guillaume
Published: March 2003

Summary. B‐cell chronic lymphocytic leukaemia (B‐CLL) characteristically displays low amounts of B‐cell receptor (BCR), which mainly consists of the heterodimer CD79a/CD79b bound non‐covalently with the surface immunoglobulin (SIg). This heterodimer is required for SIg expression and BCR signalling. To better define the mechanisms related to low BCR expression, we have investigated transcription, protein synthesis, assembly and transport of the BCR in B‐CLL cells. Our results demonstrated that: (1) there was no major defect in transcriptional expression of the B29 (CD79b) gene; (2) the BCR components were intracellularly detected, thus adequately synthesized, in almost all patients; (3) neither a genetic defect in the transmembrane region of SIg, which associated with CD79a/CD79b, nor a genetic abnormality in the chaperone protein calnexin that is involved in folding and assembly of the BCR were found; (4) a constant defect in the assembly of IgM and CD79b chains occurred leading to abnormal accumulation of both chains in different intracellular compartments; (5) in a majority of CLL patients all of the nascent IgM failed to be processed into mature chains and remained unsuitable for transport. These findings demonstrated that a post‐transcriptional defect located at the BCR intracellular assembly and/or trafficking levels could be involved in its low surface expression in B‐CLL.
Familial transient erythroblastopenia of childhood is associated with the chromosome 19q13.2 region but not caused by mutations in coding sequences of the ribosomal protein S19 (RPS19) gene
Authors: Gustavsson Peter , Klar Joakim , Matsson Hans , Forestier Erik , Henter Jan‐Inge , Rao Sreedhar , Seip Martin , Skeppner Gunnar , Dahl Niklas
Published: March 2003

Summary.  Transient erythroblastopenia of childhood (TEC) is a rare condition, which at onset may be difficult to distinguish from Diamond–Blackfan anaemia (DBA). We have previously shown that mutations in the ribosomal protein S19 gene (RPS19) cause DBA. In order to clarify whether TEC and DBA are allelic, we investigated the segregation of markers spanning the RPS19 gene region on chromosome 19q13.2 and performed sequence analysis of all exons in the RPS19 gene in seven TEC sibling pairs. Linkage analysis supported allelism for TEC and DBA at the RPS19 gene locus and implies molecular mechanisms other than structural mutations in the RPS19 gene.
Factor XIII Val34Leu polymorphism, factor XIII antigen levels and activity and the risk of deep venous thrombosis
Authors: Van Hylckama Vlieg Astrid , Komanasin Nantarat , Ariëns Robert A. S. , Poort Swibertus R. , Grant Peter J. , Bertina Rogier M. , Rosendaal Frits R
Published: March 2003

Summary. Varying results on the effect of factor XIII (FXIII) Val34Leu on venous thrombotic risk have been reported. The probability of a true association between this polymorphism and venous thrombotic risk would be enhanced by a laboratory phenotype associated with this polymorphism and with the thrombotic risk. The aim of this study was to assess the effect of FXIII Val34Leu, FXIII activity and subunit levels on venous thrombotic risk in a large case–control study, The Leiden Thrombophilia study (LETS). We found higher FXIII activity for 34Leu carriers (Leu/Leu: 158·0, Val/Val: 95·0). FXIII subunit levels were not associated with genotype. Higher FXIII activity was associated with a slightly decreased thrombotic risk [Odds ratio (OR): 0·8, 95% confidence intervals (CI): 0·5–1·3]. This effect was not present for elevated FXIII subunit levels. Higher FXIII activity was also associated with a higher dissociation index (percentage A2B2 complex dissociated after activation by thrombin for a fixed time interval). This index was higher for FXIII 34Leu carriers. The risk of deep venous thrombosis was slightly decreased for carriers of the 34Leu allele [OR: 0·9 (95%CI: 0·7–1·1)]. For homozygous 34Leu carriers the OR was 0·7 (95%CI: 0·4–1·3). This finding, suggesting a weak protective effect, was completely restricted to men. An overall estimate of thrombotic risk was calculated by using earlier reports on the risk of FXIII Val34Leu. The overall risk estimate for homozygous 34Leu carriers was 0·8 (95%CI: 0·6–1·0). In this study, a weak protective effect against venous thrombosis was found, of FXIII 34Leu as well as of increased FXIII activity.
Rapid detection of the major Mediterranean β‐thalassaemia mutations by real‐time polymerase chain reaction using fluorophore‐labelled hybridization probes
Authors: Moreno Isabel , Bolufer Pascual , Perez M. Luz , Barragán Eva , Sanz Miguel A.
Published: March 2003

Summary. We present a new method to detect the major β‐thalassaemia mutations of the Mediterranean countries (IVS I‐1, IVS I‐6, IVS I‐110, CD‐37 and CD‐39). The procedure is based upon real‐time polymerase chain reaction (PCR), using specific fluorescently labelled hybridization probes. The melting curves for each of the specific probes obtained after PCR enabled the identification of different alleles. Genotyping of 71 patients with thalassaemia and 20 controls without thalassaemia by the established method and conventional allele‐specific restriction enzyme analysis produced identical results. The established method is a robust, fast and straightforward assay that allows detection of the major Mediterranean β‐thalassaemia mutations simultaneously in less than 60 min.
A family of textilinin genes, two of which encode proteins with antihaemorrhagic properties
Authors: Filippovich Igor , Sorokina Natasha , Masci Paul P. , De Jersey John , Whitaker Alan N. , Winzor Donald J. , Gaffney Patrick J. , Lavin Martin F.
Published: March 2003

Summary. Two peptides, textilinins 1 and 2, isolated from the venom of the Australian common brown snake, Pseudonaja textilis textilis, are effective in preventing blood loss. To further investigate the potential of textilinins as antihaemorrhagic agents, we cloned cDNAs encoding these proteins. The isolated full‐length cDNA (430 bp in size) was shown to code for a 59 amino acid protein, corresponding in size to the native peptide, plus an additional 24 amino acid propeptide. Six such cDNAs were identified, differing in nucleotide sequence in the coding region but with an identical propeptide. All six sequences predicted peptides containing six conserved cysteines common to Kunitz‐type serine protease inhibitors. When expressed as glutathione S‐transferase (GST) fusion proteins and released by cleavage with thrombin, only those peptides corresponding to textilinin 1 and 2 were active in inhibiting plasmin with Ki values similar to those of their native counterparts and in binding to plasmin less tightly than aprotinin by two orders of magnitude. Similarly, in the mouse tail vein blood loss model only recombinant textilinin 1 and 2 were effective in reducing blood loss. These recombinant textilinins have potential as therapeutic agents for reducing blood loss in humans, obviating the need for reliance on aprotinin, a bovine product with possible risk of transmissible disease, and compromising the fibrinolytic system in a less irreversible manner.
Leukaemic relapse of donor origin after allogeneic bone marrow transplantation from a donor who later developed bronchogenic carcinoma
Authors: Au W. Y. , Chan E. C. , Siu L. L. P. , Lau T. C. M. , Lie A. K. W. , Ma S. K. , Kwong Y. L.
Published: March 2003

Summary. Donor‐derived leukaemia is exceptional after allogeneic bone marrow transplantation (BMT). A woman with chronic myeloid leukaemia received an allogeneic BMT from a human leucocyte antigen‐identical brother. The donor, a 50‐year‐old non‐smoker, died of squamous cell bronchogenic carcinoma 1 year later. At 4 years post BMT, the patient became BCR/ABL positive and relapsed with acute myeloid leukaemia, which was shown to be donor‐derived cytogenetically and molecularly. Retrospective analysis showed that the donor‐leukaemic clone had started to evolve as early as 6 months post BMT. Sequencing of p53 ruled out Li–Fraumeni syndrome. Predisposition to malignancy might be an underlying mechanism of donor‐cell leukaemia.
Expression and function of pituitary tumour transforming gene for T‐lymphocyte activation
Authors: Stoika Rostyslav , Yu Run , Melmed Shlomo
Published: March 2003

Summary. Pituitary tumour transforming gene (PTTG) isolated from pituitary tumour cells transforms cells in vitro and causes in vivo tumour formation. PTTG is expressed in several human tumours and cell lines. In normal adult tissues, the testis expresses abundant levels of PTTG mRNA comparable to that found in tumour cells. Although PTTG is not expressed in resting T cells, we showed here that activation of normal adult human T cells using either immobilized anti‐CD3 antibodies or phytohaemagglutinin was accompanied by marked PTTG induction, reaching levels observed in human tumour cells. Inhibitors of T‐cell functions, such as cyclosporin A and hydrocortisone, decreased induction of PTTG mRNA expression. During T‐cell activation, PTTG mRNA abundance corresponded with the increase in S‐phase cells, suggesting PTTG involvement in cell cycle‐dependent processes. These results showed that PTTG‐1 expression follows cell cycling patterns in T lymphocytes, providing a convenient model for studying PTTG functions in normal cells.
Anticardiolipin antibodies as a risk factor for venous thromboembolism in a population‐based prospective study
Authors: Runchey Shauna S. , Folsom Aaron R. , Tsai Michael Y. , Cushman Mary , McGovern Paul D.
Published: March 2003

Summary. Anticardiolipin antibodies, one of the family of ‘antiphospholipid’ antibodies, increase the risk of venous thromboembolism in the presence of autoimmune disease. Our objective was to determine prospectively whether there is a positive association between anticardiolipin antibodies and venous thromboembolism in ostensibly healthy adults. We conducted a nested case–control study (n = 317 patients and n = 655 control subjects) in a longitudinal study of over 20 000 participants. Baseline (prediagnosis) anticardiolipin IgG and IgM antibodies were assessed by enzyme‐linked immunoassays. Venous thromboembolism was validated using standardized criteria for venous thrombosis and pulmonary embolism. There was no association between anticardiolipin antibodies and subsequent venous thromboembolism occurrence, overall or in any subgroup. For example, the multivariate‐adjusted relative risk was 0·88 (95% confidence interval, 0·43, 1·78) for greater than versus less than the 95th percentile of anticardiolipin IgG. In conclusion, in this general population sample, an elevated anticardiolipin antibody level was not a risk factor for venous thromboembolism.
Dissociation between fibrinogen and fibrin interaction with platelets in patients with different subtypes of Glanzmann's thrombasthenia: studies in an ex vivo perfusion chamber model
Authors: Hainaud Patricia , Brouland Jean‐Philippe , André Patrick , Simoneau Guy , Bal dit Sollier Claire , Drouet Ludovic , Caen Jacques , Bellucci Sylvia
Published: March 2003

Summary. To explore the possible role of a residual or variant αIIbβ3 integrin (αIIbβ3) in thrombogenesis, we used a new ex vivo perfusion chamber model to examine blood from patients with different subtypes of Glanzmann's thrombasthenia (GT). Non‐anticoagulated blood was perfused through capillaries coated with type III collagen for 4·5 min (shear rate: 1600/s). Platelet deposition was quantified as platelet adhesion and mean thrombus size volume; fibrin and von Willebrand Factor (VWF) were specifically revealed by immunohistochemistry. In two patients with variant and in one patient with type II GT, platelet adhesion was maximal and we observed an unexpected formation of thrombi that were smaller than normal in size. These thrombi were surrounded by a thick meshwork that displayed a strong staining for fibrin and VWF. In two patients with heterozygous GT, platelet adhesion and thrombogenesis were normal. In two patients with type I GT, there was no thrombus formation, although platelet adhesion was also maximal. These data suggest the existence of a substitute pathway for thrombogenesis mediated by fibrin and possibly αIIbβ3 (αIIbβ3 at a reduced level, as in type II, and/or abnormal) as this fibrin network was not observed in type I GT with no αIIbβ3. These interactions might facilitate haemostasis and even lead to thrombosis under certain favourable conditions. Furthermore, these data might have pharmacological relevance to the development of anti‐αIIbβ3 antithrombotic agents.
Respiratory virus infections in transplant recipients after reduced‐intensity conditioning with Campath‐1H: high incidence but low mortality
Authors: Chakrabarti Suparno , Avivi Irit , Mackinnon Stephen , Ward Kate , Kottaridis Panagiotis D. , Osman Husam , Waldmann H. , Hale Geoff , Fegan Christopher D. , Yong Kwee , Goldstone Anthony H. , Linch David C. , Milligan Donald W.
Published: March 2003

Summary. Respiratory virus infections can cause serious morbidity and mortality after conventional allogeneic stem cell transplantation. However, the incidence and outcome of these infections after reduced intensity conditioning has not been reported. Between 1997 and 2001, 35 episodes of respiratory virus infections were noted in 25 of 83 transplant recipients conditioned with fludarabine, melphalan and Campath‐1H, and 80% of them received early antiviral therapy. Parainfluenza virus (PIV) 3 was the commonest isolate (45·7%) followed by respiratory syncytial virus (37%). Patients with myeloma were more susceptible to these infections [odds ratio (OR) 4·1, P = 0·01] which were often recurrent in patients with severe acute or chronic graft‐versus‐host disease (GVHD) (OR 10·6, P = 0·03). Infection within the first 100 d (OR 5·0, P = 0·05) and PIV 3 (OR 9·2, P = 0·01) isolation were risk factors for developing lower respiratory infection. Although more than half of the episodes progressed to lower respiratory infection, the mortality was only 8%. This could have been due to early initiation of antiviral therapy, but the attenuation of pulmonary damage due to the reduced‐intensity conditioning, low incidence of GVHD and, paradoxically, the low CD4+ T‐cell subset in this setting might also have been contributory factors.
Size variants of beta‐2‐microglobulin‐free human leucocyte antigen class I heavy chain make different contributions to its serum increase in multiple myeloma
Authors: Perosa Federico , Prete Marcella , Luccarelli Grazia , Dammacco Franco
Published: March 2003

Summary. We previously showed that serum beta‐2‐microglobulin (β2m)‐free human leucocyte antigen (HLA) class I heavy chain (FHC) levels were increased in MM and correlate with disease activity. The present investigation, carried out in 124 multiple myeloma patients, studied the expression of the three size variants of FHC, namely the 42 kDa intact heavy chain (A variant, AV), released through a shedding process, and the truncated FHC (tFHC) 39 kDa (BV) and 36–35 kDa (CV) released by means of membrane‐type metalloprotease activity. The increase in FHC correlated with a high expression percentage of BV (r = 0·32, P = 0·0002) and tFHC (r = 0·42, P < 0·0001), which could help to discriminate multiple myeloma from monoclonal gammopathy of undetermined significance (tFHC mean ratio = 3·2; Mann–Whitney U‐test, P < 0·0001). tFHC levels highly correlated with other disease activity markers, namely haemoglobin (r = −0·35; Spearman's rank, P = 0·0001), percentage of bone marrow plasma cells (r = 0·4, P < 0·0001) and β2m levels (r = 0·36, P < 0·0001), while only the last barely correlated (r = 0·2, P = 0·03) with AV. Finally, the 0·4, 0·57 and 0·71 mg/l BV, tFHC and (to a lesser extent) FHC cut‐off values divided patients into two groups with different survival curves (P = 0·0005, P = 0·0025 and P = 0·04 respectively). These data are in favour of a correlation between disease aggressiveness and cleavage of these variants by membrane‐type metalloprotease enzymes.
Prognostic impact of cytogenetic and interphase fluorescence in situ hybridization‐defined chromosome 13 deletion in multiple myeloma: early results of total therapy II
Authors: Shaughnessy John , Tian Erming , Sawyer Jeffrey , McCoy Jason , Tricot Guido , Jacobson Joth , Anaissie Elias , Zangari Maurizio , Fassas Athanasios , Muwalla Firas , Morris Christopher , Barlogie Bart
Published: March 2003

Summary. Cytogenetic abnormalities of chromosome 13 (CA 13) and those detected by fluorescence in situ hybridization (FISH 13) have both been associated with poor prognosis in multiple myeloma (MM) patients. The prognostic implications of CA, FISH 13 and other standard laboratory parameters were examined in the first 231 patients enrolled in Total Therapy II, an intensive cytotoxic chemotherapy programme with tandem autotransplants. Three‐year projections of event‐free survival (EFS) and overall survival (OS) were 71% and 77% respectively. CA 13 was detected in 14% and significantly correlated with FISH 13 (present in 51%), tumour burden, proliferative activity and lactic dehydrogenase (LDH). Both EFS and OS were significantly shorter in patients with CA 13, FISH 13, LDH ≥ 190 U/l, β2 microglobulin ≥ 4 mg/l and C reactive protein ≥ 4·0 mg/l; other CA was an additional risk factor for OS. Two‐thirds of CA 13 patients were identified by FISH 13 and plasma‐cell‐labelling index (PCLI) ≥ 0·4%; however, PCLI failed to identify additional risk groups in FISH subsets. Although present in considerably fewer patients, CA 13 imparted more rapid relapse (61% at 3 years) and death (43% at 3 years) than FISH 13 (38% and 35%; P = 0·02 and 0·1 respectively) and should be part of the initial work‐up of patients with MM.
Successful treatment of acute myelomonocytic leukaemia with NUP98–HOXD11 fusion transcripts and monitoring of minimal residual disease
Authors: Terui Kiminori , Kitazawa Junichi , Takahashi Yoshihiro , Tohno Chikako , Hayashi Yasuhide , Taketani Takeshi , Taki Tomohiko , Ito Etsuro
Published: March 2003

Summary. Patients with haematological malignancies involving the NUP98 gene have been reported to have an aggressive clinical course and a poor outcome. We report successful treatment of a 15‐year‐old Japanese boy with acute myelomonocytic leukaemia having t(2;11)(q31;p15) and a novel fusion transcript, NUP98–HOXD11. He achieved complete remission by combined chemotherapy, and underwent unrelated cord blood transplantation 4 months after diagnosis. He is in complete remission 24 months after diagnosis. Monitoring of minimal residual disease (MRD) showed the absence of fusion transcript 12 months after transplantation. This is the first report of monitoring MRD in a patient with haematological malignancy involving NUP98 fusion transcripts.
Overexpression of complement receptors and related antigens on the surface of bone marrow mast cells in patients with systemic mastocytosis
Authors: Núñez‐López Rosa , Escribano Luis , Schernthaner Gerit‐Holger , Prados Aránzazu , Rodríguez‐González Ramón , Díaz‐Agustín Beatriz , López Antonio , Hauswirth Alexander , Valent Peter , Almeida Julia , Bravo Pilar , Orfao Alberto
Published: March 2003

Summary. Depending on their stage of maturation and other factors, mast cell (MC) subsets differ from each other in terms of the expression of complement‐associated antigens. This study analysed the expression of various complement‐related cell surface antigens (CD11b/CR3, CD11c/CR4, CD35/CR1, CD55/DAF, CD59/MIRL, CD88/C5aR) on bone marrow mast cells (BMMC) in patients suffering from systemic mastocytosis (SM), other haematological diseases and non‐haematological disorders (control groups). Expression of complement‐associated cell surface antigens was analysed by flow cytometry. There were clear immunophenotypic differences between BMMC obtained from patients with SM and those from the control subjects: the percentage of patients expressing surface CD11c, CD35 and CD88 was significantly higher in patients with SM (76%, 100%, 54%) than in the control subjects (58%, 11%, 18%) (P < 0·05). In addition, the levels of CD11c, CD35 and CD88 expressed per MC (sites per cell) were significantly higher (P < 0·05) in SM than in the control group. Expression of the complement regulatory molecules CD55 and CD59 was detected in BMMC in all patients analysed. However, the levels of CD59 per BMMC were higher in patients with SM as compared with the control subjects, which could help to explain the formation of BMMC aggregates in the former group of individuals. Together, our results showed that BMMC in systemic mastocytosis overexpressed the cell surface membrane receptors involved in binding of complement components and complement‐mediated cell activation. Whether this pathological expression of complement receptors is of pathophysiological significance remains to be determined.