bluuebunny/crossref_metadata_2025 huggingface dataset:
Feasibility and recent improvement of autologous stem cell transplantation for acute myelocytic leukaemia in patients over 60 years of age: importance of the source of stem cells
Authors: Gorin, N. C. , Labopin M. , Pichard P. , Sierra J. , Fiere D. , Rio B. , Meloni G. , Sanz, M. A. , Iriondo, A. , Fouillard L. , Frassoni F. ,   for the Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation
Published: March 2003

A total of 193 patients with acute myelocytic leukaemia (AML) [147 in first complete remission (CR1)], ranging from 60 years to 75 years of age (median 63 years), were autografted between January 1984 and December 1998. The source of stem cells was peripheral blood (PB) in 128 patients, bone marrow in 51 patients and a combination of both in 14 patients. Total body irradiation (TBI) was used in 34 cases. Ninety‐seven per cent of patients had successful engraftment of neutrophils on day 15 (range days 7–71) and of platelets on day 30 (range days 9–894). In patients autografted in CR1, the transplant‐related mortality (TRM) was 15 ± 4%, the relapse incidence (RI) was 58 ± 5%, the leukaemia‐free survival (LFS) was 36 ± 5% and the overall survival was 47 ± 5% at 3 years. The source and dose of stem cells were studied in particular; in patients transplanted in CR1, the RI was 44 ± 11% in those receiving marrow compared with 63 ± 6% in those receiving PB (P = 0·04). Patients autografted in CR1 who received higher granulocyte–macrophage colony‐forming units (CFU‐GM) doses (above the median) had a lower RI (47 ± 11% vs. 79 ± 9%,P = 0·009). There was a significant improvement in patients transplanted after March 1996; for those in CR1, the RI was 41 ± 8% vs. 65 ± 6% (P = 0·01), the LFS was 53 ± 8% vs. 28 ± 5% (P = 0·01) and the overall survival was 72 ± 7% vs. 36 ± 6% (P = 0·02). By multivariate analyses, significant factors for the outcome were the date of transplant with recent improvement and the source of stem cells, with a lower RI for marrow. Autologous stem cell transplantation (ASCT) is a potential therapeutic approach in patients with AML over 60 years of age; results have improved recently.
Allogeneic haematopoietic transplantation for Richter's syndrome
Authors: Rodriguez Jose , Keating Michael J. , O'brien Susan , Champlin Richard E. , Khouri Issa F.
Published: March 2003

This study evaluated the efficacy of allogeneic bone marrow transplantation (AlloBMT) in eight patients who had chronic lymphocytic leukaemia (CLL) in Richter's transformation. Five patients were in resistant relapse and three others were in sensitive or untreated relapse. Three out of eight patients (38%) were alive and in remission at 14 months, 47 months and 67 months respectively. Two of these three patients, including one with an unrelated‐donor transplant and a prior, unsuccessful autologous transplantation, received a non‐myeloablative preparative regimen. These data suggest that, compared with conventional chemotherapy, AlloBMT improves the prognosis for patients with Richter's syndrome. Further study of a larger number of patients is needed to confirm these results .
Glycoprotein Ib/IX complex is the target in rifampicin‐induced immune thrombocytopenia
Authors: Pereira Jaime , Hidalgo Patricia , Ocqueteau Mauricio , Blacutt Marcelo , Marchesse Miguel , Nien Yih , Letelier Luz , Mezzano Diego
Published: March 2003

Thrombocytopenia is a major adverse effect of several drug treatments. Rifampicin has been recognized as a cause of immune thrombocytopenia during intermittent high‐dose therapy. We characterized the antibody of a patient who presented with purpura and thrombocytopenia during treatment of tuberculosis with rifampicin. Drug‐dependent binding of the antibody to platelets was demonstrated by flow cytometry. In a glycoprotein‐specific immunoassay, the binding epitope of the IgG antibody was found in the glycoprotein Ib/IX complex, using four different monoclonal antibodies (mAbs) against various epitopes on the GPIb/IX complex, as well as mAbs against GPIIb/IIIa, GPIa/IIa and GPIV. By immunoprecipitation of biotin‐labelled platelets, reactivity of the antibody with GPIb/IX was found only in the presence of the drug. These findings clearly demonstrate that rifampicin induces the formation of drug‐dependent antibodies capable of causing thrombocytopenia. The binding site of the rifampicin‐dependent antibody, located in the GPIb/IX complex, seems to be a favoured target for antibodies induced by different drugs.
Long‐term remission after intensive chemotherapy in advanced myelodysplastic syndromes is generally associated with restoration of polyclonal haemopoiesis
Authors: Aivado Manuel , Rong Astrid , Germing Ulrich , Gattermann Norbert , Kobbe Guido , Rieth Claudia , Haas Rainer , Aul Carlo
Published: March 2003

The clonality of peripheral blood cells was assessed in eight female patients with myelodysplastic syndrome (MDS) by means of the human androgen receptor gene‐based assay (HUMARA). The patients were in complete remission for a median follow‐up time of 83 months after intensive chemotherapy. X‐chromosome inactivation patterns (XCIPs) indicated polyclonal haemopoiesis in five patients. Two patients had skewed lyonization (i.e. unbalanced XCIPs in both granulocytes and T cells) and one patient presented monoclonal granulocytes together with polyclonal T cells. We conclude that long‐term remission in MDS following intensive chemotherapy is usually associated with polyclonal haemopoiesis.
Automated genotyping for accurate assignment of the (AT)xNz(AT)y motif within the β‐globin locus control region‐hypersensitive site 2
Authors: Tatu Thanusak , Thein Swee Lay
Published: March 2003

Within the core region of hypersensitive site 2 of the β‐globin locus control region is the (AT)xNz(AT)y motif, which comprises two short tandem repeats of dinucleotide (AT) interrupted by a short variable DNA sequence (Nz). The motif is usually investigated using direct sequence analysis of specifically amplified DNA, but without family studies, phase alignment of the two tandem repeats, occasionally confounded by a variable N sequence in between, can be erroneous. We combined the technique of automated genotyping with direct sequence analysis to demonstrate that it is possible to overcome this problem; the approach should be applicable for the investigation of other sequence repeats of this nature.
Long‐term outcome of individualized prophylactic treatment of children with severe haemophilia
Authors: Van Den Berg H. M. , Fischer K. , Mauser‐Bunschoten E. P. , Beek F. J. A. , Roosendaal G. , Van Der Bom J. G. , Nieuwenhuis H. K.
Published: March 2003

The development of arthropathy is a serious complication of severe haemophilia. With the use of prophylaxis, bleeds can be prevented and arthropathy delayed. We investigated whether an individually tailored prophylactic regimen can prevent arthropathy and whether it had a similar effect on orthopaedic outcome compared with that of a high‐dose regimen. Efficacy was determined clinically and by radiographs of six major joints. Prophylaxis was started in 70 patients at a mean age of 4·1 years. Mean follow‐up was 15·6 years (range 8–24·5 years). The mean factor VIII consumption was 2319 IU/kg/year. The mean number of joint bleeds was 3·5/year and the mean clinical score (maximum score 90) was 1·0, with a mean Pettersson joint score (maximum score 78) of 3·0 at a mean age of 13·5 years. In conclusion, long‐term, early‐onset, individualized prophylaxis in haemophilia is feasible and prevents arthropathy.
Use of a simplified clinical scoring system and d‐dimer testing can reduce the requirement for radiology in the exclusion of deep vein thrombosis by over 20%
Authors: Janes Sarah , Ashford Nicholas
Published: March 2003

A simplified, robust, clinical assessment, used in conjunction with a d‐dimer assay, has been developed to identify patients with a low risk of deep vein thrombosis. These patients did not undergo radiological examination. The protocol has been used by junior doctors in a district general hospital with a predominantly elderly population. Four hundred and thirty‐one patients with potential deep vein thrombosis of the lower limb were assessed using this method and 98 (22·7%) did not require further investigation. Of these, one was admitted 5 months later with a pulmonary embolus. None of the other patients subsequently required admission for a thromboembolic event or died during a minimum of 3 months follow‐up.
Injecting drug use is a risk factor for deep vein thrombosis in women in Glasgow
Authors: McColl M. D. , Tait R. C. , Greer I. A. , Walker I. D.
Published: March 2003

Three hundred and twenty‐two consecutive women aged 16–70 years who presented with objectively confirmed symptomatic venous thromboembolism (VTE) were studied to determine precipitating factors for thrombosis. One hundred and eighty‐seven presented with deep vein thrombosis (DVT), 116 with either definite or possible pulmonary embolism (PE) and 19 with both DVT and PE. Injecting drug use (IDU) via femoral vein puncture was a common risk factor for DVT, associated with 21·4% of all cases of DVT and 52·4% of cases of DVT in women under 40 years. All women with drug‐related thrombosis presented with DVT. None presented with symptomatic PE. A number of clinically diagnosed DVT associated with IDU were also documented, suggesting that IDU may be the most common risk factor for DVT in our region. DVT associated with IDU presents significant management challenges.
Sensitivity to the abl inhibitor STI571 in fresh leukaemic cells obtained from chronic myelogenous leukaemia patients in different stages of disease
Authors: Gambacorti‐Passerini Carlo , Barni Rossella , Marchesi Edoardo , Verga Magda , Rossi Francesca , Rossi Fabio , Pioltelli Pietro , Pogliani Enrico , Marco Corneo Gian
Published: March 2003

STI571 (CGP57148B) is an inhibitor of BCR/ABL, the cause of chronic myeloid leukaemia (CML). A difference exists between CML patients in chronic phase, in which responses to STI571are durable, and patients in blast crisis, who generally experience only transient responses. Leukaemic cells from six CML patients from whom samples could be obtained during chronic phase and at the time of blast crisis (BC) were compared for sensitivity to STI571, using an in vitro assay. BC samples showed a sensitivity similar to that obtained during chronic phase, suggesting that no substantial intrinsic resistance to STI571 was present in BC.
Efficacy of epoetin alfa in the treatment of anaemia of multiple myeloma
Authors: Dammacco Franco , Castoldi Gianluigi , Rödjer Stig
Published: March 2003

Effects of epoetin alfa on transfusions, haemoglobin (Hb) and quality of life (QOL) were evaluated in a placebo‐controlled study of 145 patients with multiple myeloma and anaemia (Hb < 11 g/dl). During the 12‐week, double‐blind phase, patients received 150 IU/kg epoetin alfa or a matching volume of placebo subcutaneously three times weekly; the dose (or volume) was doubled at week 4 if Hb response was inadequate. Patients completing this phase could enter the subsequent optional 12‐week phase of open‐label epoetin alfa treatment. During double‐blind treatment, epoetin alfa significantly decreased the incidence of transfusion compared with placebo (28% vs. 47%, P = 0·017), regardless of patients' transfusion history, and increased mean Hb (1·8 g/dl vs. 0·0 g/dl, P < 0·001). Univariate analysis showed significant (P ≤ 0·05) improvement in more QOL measures with epoetin alfa than with placebo; multivariate analysis discerned no between‐treatment differences. Significantly (P = 0·038) more epoetin alfa vs. placebo patients had improved performance scores. At the end of the open‐label treatment phase, patients who had continued epoetin alfa maintained Hb status, and placebo patients who were switched to epoetin alfa had mean Hb increases of 2·4 g/dl. Adverse events were similar between treatment groups. Epoetin alfa proved effective and well tolerated for treating anaemia in patients with multiple myeloma.
Graft‐versus‐host disease and survival after ABO‐incompatible allogeneic bone marrow transplantation: a single‐centre experience
Authors: Stussi Georg , Seebach Lutz , Muntwyler Jörg , Schanz Urs , Gmür Jürg , Seebach Jörg D.
Published: March 2003

The effect of ABO‐incompatibility on graft versus host disease (GVHD) and survival was evaluated in 173 consecutive patients receiving allogeneic bone marrow transplantation (BMT). Thirty‐four percent of the patients developed GVHD and univariate analysis suggested a higher incidence of GVHD in minor ABO‐incompatibility than in ABO‐identity (14/30, 47% versus 37/112, 33%; P = 0·02). However, using logistic regression adjusted for potential confounders, the GVHD risk did not differ significantly. During a mean follow‐up time of 59 months, the mortality was 37% and survival was significantly dependent on ABO‐compatibility (P = 0·004). In particular, patients with bidirectional ABO‐incompatibility had an excess mortality rate (RR, 7·6; 95% CI, 2·5–23·2; P = 0·0004). Taken together, these results suggest that ABO‐incompatibility may represent a risk factor in allogeneic BMT.
Polymerase chain reaction protocols for alpha globin haplotype polymorphisms
Authors: Miles K. L. , Norwich J. T. , Martinson J. J. , Clegg J. B.
Published: March 2003

Polymerase chain reaction protocols were designed specifically to amplify regions of the alpha globin complex that contain the nine common polymorphic haplotyping sites. These reactions provided a quicker and more sensitive approach to determining alpha globin haplotypes than Southern blotting methods.
Congenital thrombocytopenia and radio‐ulnar synostosis: a new familial syndrome
Authors: Thompson Alexis A. , Woodruff Kelly , Feig Stephen A. , Nguyen Long T. , Schanen N. Carolyn
Published: March 2003

The association of bone marrow failure and skeletal defects has been frequently noted, however, the genetic basis for most of these syndromes remains unclear. We describe a previously uncharacterized autosomal dominant syndrome of amegakaryocytic thrombocytopenia associated with radial‐ulnar synostosis. The clinical features of this syndrome appear to be distinct from other similar conditions, including Fanconi's anaemia and thrombocytopenia‐absent radii (TAR). The physical findings at diagnosis and clinical management of each case are detailed, as well as a discussion of this disorder in the context of other syndromes in which marrow failure and skeletal defects are prominent features. We also review recent developments in molecular genetics that may provide important clues to the underlying aetiology of this condition.
Epitope mapping of factor VIII inhibitor antibodies of Chinese origin
Authors: Huang Chin‐Chin , Shen Ming‐Ching , Chen Jen‐Yang , Hung Mei‐Hua , Hsu Ting‐Chang , Lin Shu‐Wha
Published: March 2003

Epitopes recognized by factor VIII (FVIII) inhibitors of Chinese origin were analysed by immunoblotting with full‐length recombinant FVIII (rFVIII), thrombin‐activated FVIII (FVIIIa) and 16 FVIII fusion proteins synthesized by bacteria. Twenty‐eight patients, 12 with haemophilia A and 16 with autoimmune diseases, were recruited. Antibodies from 22 patients showed reactivity with rFVIII, 20 with FVIIIa, and one reacted only with FVIII fusion proteins. Of these 22 cases, most were reactive with A2‐a2 and A3‐C1‐C2 of FVIII(a). Of the nine cases that depicted binding to the fusion proteins, three were reactive with the A domains, three with only the B domain, and the other three with both the A and B (or C) domains. An epitope for a neutralizing antibody of a haemophilia A patient, designated TWN‐112, was localized to residues 323–390, specified by FVIII fusion proteins. The same epitope also appeared on an FVIII‐expression phage library screening. Immunoabsorption of antibodies from TWN‐112 with the epitope reduced the neutralizing activity of the inhibitor by 33%. The incidence of a1 of FVIII is higher, and that of a3 is lower, than previously reported. Two novel epitopes, reported for the first time in this paper, were localized on the 8B2 (amino acid residues 1022–1204) and 8A2(V) (residues 673–740) fusion proteins. These two epitopes were able to reduce inhibitory antibody activity by 24% and 25% respectively. Changes of FVIII fragment specificity were also observed in one of six patients for whom multiple samples, collected at different times, were available. Our initial finding showed that the FVIII inhibitors in these Chinese patients shared epitopes with those of patients from very different genetic backgrounds, suggesting a common mechanism for the development of FVIII inhibitors.
True histiocytic lymphoma following B‐acute lymphoblastic leukaemia: case report with evidence for a common clonal origin in both neoplasms
Authors: Bouabdallah Réda , Abéna Pascale , Chetaille Bruno , Aurran‐Schleinitz Thérèse , Sainty Danielle , Dubus Pierre , Arnoulet Christine , Coso Diane , Xerri Luc , Gastaut Jean‐Albert
Published: March 2003

True histiocytic lymphoma (THL) is a very rare type of non‐Hodgkin's lymphoma (NHL) in which neoplastic cells exhibit markers of histiocytic differentiation. Some cases of THL have been reported in patients with previous acute lymphoblastic leukaemia (ALL), especially in children and young adults, in whom the acute leukaemia was of T‐cell origin. The relationship between the initial lymphoid tumour and the secondary THL remains unclear, as a common monoclonal origin shared by both neoplasms has never been definitively demonstrated. We report a patient with B‐ALL who developed a nodal and extranodal tumour with histological and immunohistochemical features of THL 4 years after the initial diagnosis. Genotypic study showed that both neoplasms contained the same immunoglobulin heavy gene rearrangement, which has not been reported previously.
Increased activation of protein C, but lower plasma levels of free, activated protein C in uraemic patients: relationship with systemic inflammation and haemostatic activation
Authors: Mezzano Diego , España Francisco , Panes Olga , Medina Piedad , Pais Edgar , Marshall Guillermo , Tagle Rodrigo , Downey Patricio , Caceres Soledad , González Fernando , Quiroga Teresa , Pereira Jaime
Published: March 2003

Chronic renal failure (CRF) courses with both systemic inflammatory reaction and haemostatic activation. We explored the relationship of these processes with plasma levels of free, activated protein C (APC) and complexes of APC with its inhibitors in patients with CRF under conservative treatment. Plasma concentrations of inflammatory cytokines [tumour necrosis factor alpha (TNFα) and interleukin 8], acute‐phase proteins (C‐reactive protein, fibrinogen, α1‐anti‐trypsin and von Willebrand factor), and markers of haemostatic activation (thrombin–anti‐thrombin complexes, plasmin–anti‐plasmin complexes, and fibrin and fibrinogen degradation products) were higher in patients than in controls. Inflammatory and haemostatic markers were significantly and positively correlated. Total plasma APC and APC:α1‐anti‐trypsin (α1AT) complexes were 44% and 75% higher in patients than in controls (P = 0·0001), whereas free APC was 20% lower (P < 0·015). No significant difference was observed in APC:protein C inhibitor (PCI) complexes between both groups. The free/total APC ratio was significantly lower in patients than in controls (P < 0·0001). Total plasma APC and APC:α1AT were positively correlated with activation markers of haemostasis and acute‐phase proteins, whereas free APC was inversely correlated with plasma levels of creatinine, acute‐phase proteins and fibrin degradation products (FnDP). Systemic inflammation and activation of haemostasis are interrelated processes in CRF. APC generation was increased in response to elevated thrombin production, but the inflammatory reaction, associated with increased synthesis of α1AT, reduced its anticoagulant effect. Lower free plasma APC in CRF may be pathogenically associated with atherothrombosis, a major cause of death in this disease.
Long‐term follow‐up of a prospective, double‐blind, placebo‐controlled randomized trial of clodronate in multiple myeloma
Authors: McCloskey Eugene V. , Dunn Janet A. , Kanis John A. , MacLennan Ian C. M. , Drayson Mark T. ,   the Medical Research Council Working Party on Leukaemia in Adults
Published: March 2003

Oral clodronate (1600 mg/d) has been shown to significantly reduce the incidence of skeletal complications in multiple myeloma. Preliminary analysis of a double‐blind placebo‐controlled trial of this treatment indicated that clodronate might prolong survival in patients without vertebral fractures at presentation. This issue was re‐examined after further follow‐up of the patients recruited into the Medical Research Council (MRC) VIth Myeloma Study. The trial examined the effects of clodronate on the natural history of skeletal disease in multiple myeloma; 619 patients were randomized between June 1986 and May 1992 commencing 15 d after the start of ABCM [adriamycin, BCNU (carmustine), cyclophosphamide, melphalan] chemotherapy or 43 d after ABCMP (ABCM + prenisolone); 535 patients who received clodronate or placebo were included in the analysis. The presence or absence of spinal fractures was assessed centrally from spinal X‐rays; long‐bone fractures were assessed locally. With a median follow‐up of 8·6 years, there was no overall significant difference in survival between the two treatment groups (O/E, χ2 = 0·78,P = 0·38). Among the subgroup of 153 patients with no skeletal fractures at presentation there was a significant survival advantage (O/E, χ2 = 7·52,P = 0·006) in favour of the 73 patients receiving clodronate, with median survivals being, respectively, 59 months (95% CI 43–71 months) and 37 months (95% CI 31–52 months), and 5‐year survivals being 46% and 35%. The original analysis of this study shows that there is a benefit in taking 1600 mg clodronate daily for patients with myelomatosis to prevent the development of new skeletal disease. Bearing in mind the limitations of subgroup analysis, the present study indicates that treatment may prolong survival in patients without overt skeletal disease at diagnosis. These observations, however, require confirmation in prospective clinical trials.
Irradiation enhances the support of haemopoietic cell transmigration, proliferation and differentiation by endothelial cells
Authors: Gaugler Marie‐Hélène , Squiban Claire , Mouthon Marc‐André , Gourmelon Patrick , Van der Meeren Anne
Published: March 2003

Endothelial cells (ECs) are a critical component of the bone marrow stroma in the regulation of haemopoiesis. Recovery of bone marrow aplasia after radiation exposure depends, in part, on the repair of radiation‐induced endothelial damage. Therefore, we assessed the ability of an irradiated human bone marrow EC line (TrHBMEC) to support transmigration, proliferation and differentiation of CD34+ bone marrow cells either irradiated or not in transendothelial migration or co‐culture models. Radiation‐induced EC damage was reflected by an increased release of soluble intercellular adhesion molecule (sICAM)‐1 and platelet endothelial cell adhesion molecule (PECAM)‐1. Irradiation of TrHBMECs with a 10 Gy dose strongly enhanced the transmigration of CD34+ cells, granulo‐monocytic progenitors (CFU‐GM) and erythroid progenitors (BFU‐E). While ICAM‐1 and PECAM‐1 expression on irradiated TrHBMECs was increased, only antibodies against PECAM‐1 inhibited the radiation‐induced enhanced transmigration of haemopoietic cells. Irradiation of TrHBMECs (5–15 Gy) also increased proliferation and differentiation towards the granulo‐monocytic lineage of co‐cultured CD34+ cells, as well as colony formation by those cells and the production of interleukin 6 (IL‐6), IL‐8, granulocyte colony‐stimulating factor (CSF) and granulocyte‐macrophage CSF. Irradiated TrHBMECs were more capable of stimulating irradiated (1,2 Gy) CD34+ cells and haemopoietic progenitors than non‐irradiated TrHBMECs. Together, these results suggest that, despite the radiation‐induced damage, irradiated ECs may favour haemopoietic reconstitution after radiation exposure.
Determination of heparin–platelet factor 4–IgG antibodies improves diagnosis of heparin‐induced thrombocytopenia
Authors: Lindhoff‐Last E. , Gerdsen F. , Ackermann H. , Bauersachs R.
Published: March 2003

Only a few patients with heparin‐induced antibodies develop heparin‐induced thrombocytopenia (HIT). In this study, we investigated whether different immunglobulin classes can be used to differentiate between antibody‐positive patients with and without HIT. Four different patient populations were investigated: 32 patients with the immune type of HIT with thromboembolic complications, 13 patients with HIT without thromboembolism, 24 patients with heparin–platelet factor 4 (PF4) antibodies without clinical symptoms of HIT, and 20 heparin‐treated patients with thrombocytopenia caused by other reasons. In all patients the immunglobulin mixture of IgG, IgM and IgA, and the single immunglobulin classes of heparin–PF4 antibodies, were investigated. No significant differences between HIT patients with thromboembolic complications and patients with isolated HIT were found concerning the different immunglobulin classes. Antibody‐positive patients with HIT had significantly higher levels of IgG antibodies than those without HIT (P < 0·05), while they did not differ concerning IgM and IgA antibodies. By determining IgG antibodies, the specificity of the enzyme‐linked immunosorbent assay (ELISA) system was increased without loss of sensitivity. Heparin–PF4–IgG antibodies can identify patients at risk of developing life‐threatening HIT. Future ELISAs should only include this immunglobulin class, as the determination of the antibody mixture may lead to overestimation of HIT.
RAFTK/Pyk2 involvement in platelet activation is mediated by phosphoinositide 3‐kinase
Authors: Koziak K. , Kaczmarek E. , Park S. Y. , Fu Y. , Avraham S. , Avraham H.
Published: March 2003

Platelet activation by different agonists initiates a signalling cascade involving the phosphorylation of several protein kinases, which control key regulatory events. Previously, we demonstrated that the related adhesion focal tyrosine kinase (RAFTK, Pyk2) was involved in an early phase of platelet activation, independent of integrin and glycoprotein IIb–IIIa activation. In this study, we demonstrate that RAFTK is co‐immunoprecipitated with phosphoinositide 3‐kinase (PI3K) upon platelet activation, and that thrombin, ADP and collagen induced the phosphorylation of both PI3K and RAFTK. A low dose of thrombin (0·015 U/ml) induced RAFTK phosphorylation and platelet aggregation in a PI3K activity‐dependent manner, whereas a high dose of thrombin (0·1 U/ml) induced these events in a PI3K activity‐independent manner. ADP and collagen also induced RAFTK phosphorylation and platelet aggregation in a PI3K activity‐dependent manner, similar to that of the low‐dose thrombin. Furthermore, protein tyrosine phosphatase activity was associated with RAFTK in response to platelet activation, and was found to be that of protein tyrosine phosphatase‐2 (SHP‐2). The association of SHP‐2 with RAFTK was PI3K‐dependent and was increased upon RAFTK phosphorylation. Taken together, our results strongly suggest that the involvement of RAFTK in platelet activation is mediated via the PI3K pathway.
Decreased fibrinolytic potential in South Asian women with ischaemic cerebrovascular disease
Authors: Kain Kirti , Catto Andrew J. , Carter Angela M. , Young John , Bamford John , Bavington John , Grant Peter J.
Published: March 2003

To investigate gender differences in conventional, coagulation and fibrinolytic factors in South Asian ischaemic stroke patients, we compared these variables in 50 South Asian females (SAFP) with 90 South Asian males (SAMP) with ischaemic stroke and in 52 females (SAFC) and 38 males (SAMC) without stroke. Plasminogen activator inhibitor‐1 (PAI‐1) antigen levels were significantly higher in SAFP compared with SAMP (18·2 vs. 13·3 U/ml, P = 0·04) even after adjustment for known covariates, but there was no difference in PAI‐1 antigen levels between males and females in the control group. South Asian females exhibited higher levels of factor VII antigen and FVII:C activity in both stroke patients (114 vs. 99% in males, P = 0·01; 116 versus 104% in males, P = 0·04) and controls (116 vs. 97% in males, P = 0·004; 115 vs. 93% in males, P = 0·01). There were no significant differences in the levels of fibrinogen (3·8 vs. 3·7 g/l), FXIIa (2·2 vs. 2·4 ng/ml), von Willebrand factor (1·8 vs. 1·9 IU/ml) and tissue plasminogen activator (11·4 vs. 12·0 ng/ml) in SAMP and SAFP respectively. These results suggest that South Asian females have increased FVII levels and that females with a history of ischaemic stroke have a decreased fibrinolytic potential in comparison with males.
Philadelphia (Ph) chromosome‐positive chronic myeloid leukaemia relapsing as Ph‐negative leukaemia after allogeneic bone marrow transplantation
Authors: Au W. Y. , Lie A. K. W. , Ma S. K. , Wan T. S. K. , Liang R. , Leung Y. H. , Kwong Y. L.
Published: March 2003

Two patients with Philadelphia chromosome‐positive (Ph+) chronic myeloid leukaemia (CML) relapsed 1·5 and 5 years after allogeneic bone marrow transplantation (BMT). Although the leukaemias were of recipient origin, t(9;22) could no longer be detected using conventional cytogenetics/fluorescence in situ hybridization or molecularly. Both patients responded to immunotherapy with donor lymphocytes/peripheral blood stem cells, although one patient ultimately relapsed again. These patients were similar to the occurrence of Ph− leukaemias previously described in Ph+ CML after treatment with interferon or autologous BMT, and might be relevant in the pathogenesis and monitoring of treatment after BMT in CML.
E1A3 as a unique, naturally occurring BCR–ABL transcript in an indolent case of chronic myeloid leukaemia
Authors: Roman Jose , Jimenez Antonio , Barrios Manuel , Castillejo Juan Antonio , Maldonado Juan , Torres Antonio
Published: March 2003

A woman with Ph‐positive chronic myeloid leukaemia (CML) with an atypical e1a3 BCR–ABL hybrid gene is described. To our knowledge, this is the first report of this transcript type as a unique naturally occurring BCR–ABL fusion in a CML patient. This case was characterized by a low leucocyte count and a very indolent course without treatment. Because the deletion of ABL exon 2 sequences results in deletion of an essential part of the ABL SH3 domain, our case suggests that this ABL SH3 domain is not absolutely necessary for efficient induction of a myeloproliferative disease in the context of BCR–ABL/p190.
Efficient ex vivo generation of dendritic cells from CD14+ blood monocytes in the presence of human serum albumin for use in clinical vaccine trials
Authors: Araki Hiroto , Katayama Naoyuki , Mitani Hidetsugu , Suzuki Hirohito , Nishikawa Hiroyoshi , Masuya Masahiro , Ikuta Yasushi , Hoshino Natsuki , Miyashita Hiroyuki , Nishii Kazuhiro , Minami Nobuyuki , Shiku Hiroshi
Published: March 2003

Dendritic cells (DC) with the potential to induce anti‐tumour immunity represent one of the promising candidates for cancer vaccines. Efficiency of ex vivo DC generation depends on culture conditions, especially protein components in the plasma or serum used. Using human serum albumin (HSA), we devised a constant and reproducible culture method for DC generation from peripheral blood CD14+ cells. The number of DC obtained with 2% HSA‐supplemented cultures containing granulocyte‐macrophage colony‐stimulating factor and interleukin 4 were consistently higher than in cultures with various concentrations of autologous plasma or serum. The concentrations and time points tested for plasma or serum considerably affected the number of DC recovered. DC prepared with HSA acquired the ability to uptake dextran, and expressed high levels of major histocompatibility (MHC) and co‐stimulatory molecules similar to DC cultured with autologous plasma or serum. Although DC cultured with autologous plasma or serum consisted of CD1a+ and CD1a− populations, DC differentiated in the presence of HSA expressed CD1a. DC obtained with HSA primed and induced immunogenic peptide‐specific cytotoxic T lymphocytes against a tumour rejection antigen, HER2. These findings suggest that our method for preparation of DC with HSA should prove valuable in DC generation for immunotherapy.
Interphase fluorescence in situ hybridization and spectral karyotyping reveals hidden genetic aberrations in children with acute lymphoblastic leukaemia and a normal banded karyotype
Authors: Nordgren Ann , Schoumans Jacqueline , Söderhäll Stefan , Nordenskjöld Magnus , Blennow Elisabeth
Published: March 2003

Twenty‐two cases of childhood acute lymphoblastic leukaemia (ALL) with normal G‐ or Q‐banded karyotypes were studied by interphase fluorescence in situ hybridization (FISH) and spectral karyotyping. Probes detecting MLL, BCR/ABL and TEL/AML1 rearrangements were used for the interphase studies, along with centromere‐specific probes from chromosomes 17 and X. In 10 patients (45%), previously undetected aberrations were demonstrable. Specific gene rearrangements and structural changes were found in six cases and numerical changes in five. Five of these aberrations have previously been reported to have an impact on prognosis. Three cases were massively hyperdiploid and, in one, the prognostically important BCR/ABL fusion was detected. In addition, a near‐haploid karyotype with 27 chromosomes was found in one patient and TEL/AML1 rearrangements were detected in two cases. This study indicates that about half of childhood ALL cases with apparently normal karyotypes harbour genetic aberrations that may be detected using interphase FISH and spectral karyotyping.
Severe factor XI deficiency caused by compound heterozygosity for the type III mutation and a novel insertion in exon 9 (codons 324/325 +G)
Authors: Dossenbach‐Glaninger Astrid , Krugluger Walter , Schrattbauer Karl , Eder Sabine , Hopmeier Pierre
Published: March 2003

We investigated a 42‐year‐old Caucasian woman with severe factor XI deficiency and her family members. Restriction enzyme analysis and DNA sequencing revealed compound heterozygosity in the patient for the known type III mutation, which is a Phe283Leu amino acid substitution in the fourth apple domain causing impaired dimerization and secretion, and for a novel frameshift mutation in exon 9 (codons 324/325 +G), leading to premature termination with lack of parts of the fourth apple domain and the downstream serine protease domain.
Vitamin E correlates inversely with non‐transferrin‐bound iron in sickle cell disease
Authors: Marwah S. S. , Wheelwright D. , Blann A. D. , Rea C. , Beresford R. , Phillips J. D. , Wright J. , Bareford D.
Published: March 2003

Decreased serum vitamin E levels are found in homozygous sickle cell disease (SCD). Excessive transfusions may lead high non‐transferrin‐bound iron (NTBI). Hypothesizing a relationship between the two, vitamin E (measured using high performance liquid chromatography) was significantly lower in 30 SCD patients than in 30 age‐/sex‐matched controls (P < 0·001), but NTBI (bleomycin assay) was higher (P < 0·001). Vitamin E was lower in 10 transfused patients than in 20 non‐transfused patients (P < 0·001) with a significant inverse correlation between the NTBI and vitamin E (r = −0·58, P < 0·001). NTBI associated with iron overload in SCD may increase the potential for oxidative damage and low vitamin E activity may compound this effect.
Differentiation of human acute myeloid leukaemia cells in primary culture in response to cotylenin A, a plant growth regulator
Authors: Yamada Kazunari , Honma Yoshio , Asahi Ken‐ichi , Sassa Takashi , Hino Ken‐ichiro , Tomoyasu Shigeru
Published: March 2003

Cotylenin A, which has a diterpenoid tricarbocyclic skeleton, has been isolated as a plant growth regulator, has been shown to affect several physiological processes of higher plants and have differentiation‐inducing activity in several myeloid leukaemia cell lines. We examined the effect of cotylenin A on the differentiation of leukaemic cells that were freshly isolated from acute myeloid leukaemia (AML) patients in primary culture. Cotylenin A significantly stimulated both functional and morphological differentiation of leukaemia cells in 9 out of 12 cases. This differentiation‐inducing activity was more potent than those of all‐trans retinoic acid and 1α,25‐dihydroxyvitamin D3 (VD3). Treatment with a combination of cotylenin A and VD3 was more effective than cotylenin A or VD3 alone at inducing the monocytic differentiation of AML cells.
Intravenous busulphan for conditioning before autologous or allogeneic human blood stem cell transplantation
Authors: Schuler Ulrich S. , Renner Ulf D. , Kroschinsky Frank , Johne Christine , Jenke Andreas , Naumann Ralph , Bornhäuser Martin , Deeg H. Joachim , Ehninger Gerhard
Published: March 2003

This study was undertaken to evaluate the toxicity and pharmacokinetics of a dimethyl sulphoxide (DMSO)‐based intravenous formulation of busulphan in the conditioning of 45 patients undergoing allogeneic or autologous stem cell transplantation (SCT). Busulphan was given as a single daily dose. In 15 patients a single dose of intravenous busulphan, given over 3 h in 1 d, was combined with additional oral (single daily) doses. Thirty patients received all four daily doses intravenously. Busulphan plasma levels were analysed using high performance liquid chromatography. There was no major acute toxicity with daily intravenous doses of 2·8–3·1 mg/kg infused over 3 h. No veno‐occlusive disease (VOD) was seen in 30 patients receiving busulphan as an intravenous formulation over 4 d. In the group of 15 patients receiving three oral doses and one intravenous single daily dose, one patient experienced mild VOD. Pharmacokinetic samples were taken over at least 2 d of treatment in 44 patients. The area under the concentration time curve (AUC) values normalized for a dose of 1 mg/kg were 7000 ng/ml × h on d 1 and 5890 ng/ml × h on d 4, thus showing a moderate decrease over time. This was accompanied by a moderate increase of the clearance from 2·6 to 3·0 ml/min/kg. Administration of busulphan as a DMSO‐based intravenous formulation was well tolerated. The total dose of busulphan can be given in four (rather than the typical 16) doses. With such a regimen, the intravenous administration becomes feasible on an outpatient basis.
Primary breast lymphoma: a report of 20 cases
Authors: Ribrag V. , Bibeau F. , El Weshi A. , Frayfer J. , Fadel C. , Cebotaru C. , Laribi K. , Fenaux P.
Published: March 2003

Limited data are available concerning treatment and outcome of primary lymphoma of the breast (PLB), especially after CHOP (cyclophosphamide, hydroxydoxorubicin, vincristine, prednisone) chemotherapy. We retrospectively reviewed 20 consecutive cases of localized PLB seen at our institution over a 20 year period. All PLB were of B‐cell origin: treatment was CHOP or a CHOP‐like regimen in all patients. Sixteen of the 20 patients achieved complete remission (CR) and two achieved partial remission (> 75% tumour regression). Two patients had progressive disease on therapy. With a median follow‐up of 54 months, six patients relapsed after 8–66 months. Two of the relapses involved the central nervous system (CNS) (isolated in one case, associated with other sites of relapse in the other). The two patients who achieved partial remission also had progression in the CNS, 4 and 8 months after the end of CHOP chemotherapy. All four patients have died as a result of their disease 3, 6, 10 and 13 months after CNS relapse. Of the 16 centroblastic diffuse large B‐cell lymphoma (DLCL), three had CNS disease at relapse. We also observed three (15%) controlateral breast relapses. Thirteen of the initial 20 patients are alive in CR, six patients have died as a result of their lymphoma and one of unrelated disease. In conclusion, we observed a high incidence of CNS relapse in this group of localized extranodal lymphoma, strongly suggesting that CNS prophylaxis should be associated with systemic chemotherapy in localized PLB.
Biological significance of MAPK, AKT and JAK‐STAT protein activation by various erythropoietic factors in normal human early erythroid cells
Authors: Ratajczak Janina , Majka Marcin , Kijowski Jacek , Baj Monika , Pan Zhixing K. , Marquez Leah A. , Janowska‐Wieczorek Anna , Ratajczak Mariusz Z.
Published: March 2003

The aim of this study was to identify signal transduction pathways activated by erythropoietin (EpO) and erythropoietin co‐stimulatory factors (kit ligand), insulin‐like growth factor, thrombopoietin, interleukin 3 and granulocyte‐macrophage colony‐stimulating factor) in normal human bone marrow CD34+ cells and d 11 erythroid burst forming unit derived glycophorin+ cells. The activation of these signal transduction pathways was further correlated with various biological effects such as (i) cell proliferation, (ii) inhibition of apoptosis, (iii) activation of adhesion and (iv) secretion of the matrix metalloproteinases (MMPs) MMP‐9 and MMP‐2, and vascular endothelial growth factor (VEGF). We found that in human CD34+ cells and erythroblasts erythropoietic factors may activate similar but different signalling pathways, and that activation of each of the JAK‐STAT, MAPK p42/44 or PI‐3K‐AKT axes alone is not sufficient either to stimulate cell proliferation or inhibit apoptosis, suggesting that these processes are regulated by orchestrated activation of multiple signalling cascades. Accordingly, we found that although cell proliferation was more related to simultaneous activation of JAK‐STAT and MAPK p42/44, the effect on cell survival correlated with activation of PI‐3K‐AKT, MAPK p42/44 and JAK‐STAT proteins. We also demonstrated that differentiating normal human erythroid cells lose their adhesive properties and secrete angiopoietic factors such as MMP‐9, MMP‐2 and VEGF, and we postulate that this secretion by early erythroid cells may play a role in their maturation and egress from the haematopoietic niches of the bone marrow.
Early recognition of hereditary motor and sensory neuropathy type 1 can avoid life‐threatening vincristine neurotoxicity
Authors: Naumann Ralph , Mohm Johannes , Reuner Ulrike , Kroschinsky Frank , Rautenstrauss Bernd , Ehninger Gerhard
Published: March 2003

Hereditary motor and sensory neuropathy type 1 (HMSN‐1) is an autosomal dominant disorder, which is usually not associated with neoplastic diseases. The disease predisposes to severe vincristine neurotoxicity. We report a 31‐year‐old women with recurrent Hodgkin's lymphoma and unrecognized HMSN‐1 who developed severe motor neuropathy 3 weeks after the first cycle of treatment including 2 mg of vincristine. HMSN is diagnosed in most cases retrospectively, usually suggested by the observation of foot abnormalities or family history. Recognizing early signs of HMSN, such as areflexia and pes cavus deformity, can prevent severe neurotoxicity of polychemotherapy by avoiding vincristine.
Continuous oral cytarabine ocfosfate with interferon‐α‐2b for patients with newly diagnosed chronic myeloid leukaemia: a pilot study
Authors: Del Cañizo M. C. , Fisac M. P. , Galende J. , González Hurtado J. A. , Moro M. J. , Rodriguez J. A. , Hernandez Rivas J. M. , Tabernero M. D.
Published: March 2003

Recombinant® interferon α (r‐IFN‐α) has been shown to be an effective drug for chronic myeloid leukaemia (CML). However, higher response rates can be achieved using cytarabine along with r‐IFN‐α. YNK01 is a derivative of cytosine arabinoside for oral administration. So far, the only published experience with continuous YNK01 was in advanced CML (10 cases). We have performed a pilot study to evaluate the efficacy and toxicity of the combined therapy r‐IFN‐α and daily oral YNK01 in patients with newly diagnosed Ph+ CML. Ten previously untreated patients were included in the study. Among those patients evaluable for cytogenetic response, 87% (seven out of eight) reached a major cytogenetic response with four reaching complete cytogenetic response (50%). The most significant side‐effects were gastrointestinal. Macrocytic anaemia was observed in three patients. In conclusion, continuous oral administration of YNK01 in combination with IFN‐α is safe and can result in high‐cytogenetic response rates.
Response to thalidomide in progressive multiple myeloma is not mediated by inhibition of angiogenic cytokine secretion
Authors: Neben Kai , Moehler Thomas , Kraemer Alwin , Benner Axel , Egerer Gerlinde , Ho Anthony D. , Goldschmidt Hartmut
Published: March 2003

Thalidomide (Thal) is a drug with anti‐angiogenic properties. To explore whether the effect of Thal on angiogenesis is associated with a reduction of angiogenic cytokine levels in progressive multiple myeloma (MM), plasma levels of basic fibroblast growth factor, vascular endothelial growth factor, interleukin 6, tumour necrosis factor‐α and hepatocyte growth factor (HGF) were measured in 51 patients at 0, 3 and 6 months of Thal therapy. After 6 months of treatment, 26 patients were considered to be responsive to Thal therapy, including 17 minimal responses, eight partial responses and one complete response. Only HGF (decreasing, P = 0·02) in the group of responsive patients showed a statistically significant change over a period of 6 months. Because HGF levels are known to correlate to MM tumour burden, we conclude that the mechanism of action of Thal in MM is not caused by a specific inhibition of angiogenic cytokine secretion.
High prevalence of low serum vitamin B12 in a multi‐ethnic Israeli population
Authors: Gielchinsky Y. , Elstein D. , Green R. , Miller J. W. , Elstein Y. , Algur N. , Lahad A. , Shinar E. , Abrahamov A. , Zimran A.
Published: March 2003

This study ascertained serum vitamin B12 levels among patients with Gaucher disease and among healthy Israelis. Serum B12 and metabolites' levels were studied in consecutive adult patients with Gaucher disease not treated with enzyme plus Ashkenazi Jewish neighbour‐controls, together with healthy blood‐donor volunteers of various ethnicities. Each group showed a high incidence of low serum B12 concentrations, with a 22·3% incidence among Ashkenazi Jews and 40% among patients with Gaucher disease. These findings raise questions on the individual and community levels of serum B12. We recommend evaluation of B12 levels among geographically contingent peoples.
Detection of AP12‐MALT1 chimaeric gene in extranodal and nodal marginal zone B‐cell lymphoma by reverse transcription polymerase chain reaction (PCR) and genomic long and accurate PCR analyses
Authors: Yonezumi Masakatsu , Suzuki Ritsuro , Suzuki Hiroko , Yoshino Tadashi , Oshima Kouichi , Hosokawa Yoshitaka , Asaka Masahiro , Morishima Yasuo , Nakamura Shigeo , Seto Masao
Published: March 2003

t(11;18)(q21;q21) has been recognized as a characteristic chromosomal translocation in mucosa‐associated lymphoid tissue (MALT)‐type lymphoma, and recent studies have demonstrated that this translocation results in the chimaeric transcript of API2 (apoptosis inhibitor 2)‐MALT1 (mucosa‐associated lymphoid tissue lymphoma translocation gene 1). In this study, we used reverse transcription polymerase chain reaction (RT–PCR) to analyse the incidence of this fusion product in a large series of MALT lymphoma, nodal marginal zone B‐cell lymphoma (nMZBCL) and extranodal diffuse large B‐cell lymphoma (DLBL) cases. RT–PCR analysis revealed that 17 of the 95 (17·9%) MALT lymphomas but none of the nine nMZBCLs or 16 DLBLs had API2‐MALT1 fusion transcripts. The incidence of API2‐MALT1 varied among MALT lymphomas arising from different sites and was highest for pulmonary MALT lymphomas (10 out of 16 cases, 62·5%). The presence of the API2‐MALT1 fusion gene was also confirmed by long and accurate (LA)–PCR with genomic DNA, and the result correlated well with that obtained with the RT–PCR assay, thus demonstrating the usefulness of LA–PCR for the detection of the API2‐MALT1 fusion gene.