bluuebunny/crossref_metadata_2025 huggingface dataset:
Amplification of cyclin D1 gene in multiple myeloma: clinical and prognostic relevance
Authors: Hoechtlen‐Vollmar Wilma , Menzel Gabriele , Bartl Reiner , Lamerz Rolf , Wick Manfred , Seidel Dietrich
Published: March 2003

Approximately 30% of myeloma patients express cyclin D1 RNA and protein. The low incidence of translocation t(11; 14) detected by conventional cytogenetics suggests that the up‐regulation of cyclin D1 protein might result from other mechanisms as well as from gene amplification. Therefore, the frequency and the clinical and prognostic implications of cyclin D1 amplification were examined. We highly purified myeloma cells from bone marrow by magnetic cell sorting and analysed 50 myelomas by fluorescencein situhybridization (FISH) using probes specific for cyclin D1 and 20 samples by immunoblotting to detect cyclin D1 expression. The amplification of cyclin D1 gene was found in 19 of 50 analysed patients and was associated with expression of cyclin D1 protein. The amplification correlated significantly with the bone marrow infiltration, plasma cell morphology and labelling index as well as serum β2‐microglobulin, C‐reactive protein (CRP) and creatinine levels. In univariate analysis, the amplification of the cyclin D1 gene was a significantly unfavourable parameter with regard to overall survival (P = 0·0064) and progression‐free survival (P = 0·0005). In multivariate analysis, cyclin D1 amplification and serum β2‐microglobulin were independent and well‐suited parameters for predicting survival. The detection of cyclin D1 amplification seems to be of promising prognostic value in multiple myeloma.
Association between plasma viscosity and all‐cause mortality: results from the MONICA–Augsburg Cohort Study 1984–92
Authors: Koenig W. , Sund M. , Löwel H. , Döring A. , Ernst E.
Published: March 2003

Several studies have reported a strong association between various markers of the acute‐phase response and death from cardiovascular diseases and all‐cause mortality. Inflammation at a low level of intensity may be a common phenomenon associated with the majority of causes of death owing to chronic diseases. We sought to investigate the association of plasma viscosity with all‐cause mortality in a cohort of apparently healthy men. The study population consisted of 964 men aged 45–64 years at entry, randomly selected from the general population and taking part in the first MONICA–Augsburg survey 1984–85. The main outcome measure was all‐cause mortality. During 8 years of follow‐up, there were 81 deaths (37 cardiovascular deaths, 23 deaths from cancer and 21 deaths from other causes). There was a strong positive and statistically significant age‐adjusted relationship between plasma viscosity and all‐cause mortality. The relative risk of death for a one standard deviation increase in plasma viscosity (0·070 mPa/s) was 1·45 [95% confidence interval (CI) 1·19–1·76]. After further adjusting for smoking, total cholesterol, body mass index, blood pressure and education, a relative risk of 1·41 (95% CI 1·14–1·74) resulted. Other risk variables had only negligible confounding effects. The relative risk of the median of the top quintile of the plasma viscosity distribution compared with the median of the bottom quintile, computed from the adjusted model, was 2·68 (95% CI 1·63–4·42). These findings suggest that plasma viscosity may have considerable potential to predict death from all causes in middle‐aged men.
Porcine von Willebrand factor and thrombin induce the activation of c‐Jun amino‐terminal kinase (JNK/SAPK) whereas only thrombin induces activation of extracellular signal‐related kinase 2 (ERK2) in human platelets
Authors: Song Seng , Freedman John , Mody Meera , Lazarus Alan H.
Published: March 2003

The interaction of platelets with subendothelial von Willebrand factor (VWF), especially under high shear stress, is considered to be the first activation step which primes platelets for subsequent haemostatic events. The signalling cascade which results from the interaction of VWF and its receptor GPIbIX has only been partially defined. Mitogen‐activated protein kinases (MAPKs) are a family of downstream transmembrane signalling serine–threonine kinases and have been demonstrated to be present and functional in platelets; these include the extracellular signal‐related kinases (ERKs), c‐Jun amino‐terminal kinases (JNKs) and p38 MAPK. Previously, we showed that p38 MAPK was not required in VWF‐induced human platelet activation. It is not known whether VWF‐dependent platelet activation involves the activation of the JNK and ERK family of signalling molecules. This report demonstrates that porcine von Willebrand factor (pVWF) induced a sustained and stable JNK activation measurable by 1 min after activation. Thrombin also induced JNK activation assessed at 1 min after activation. In contrast to thrombin, pVWF did not induce ERK2 activation at any time point tested. To ensure that ERK activation was unnecessary for pVWF‐dependent platelet activation, we functionally inhibited ERK‐dependent signalling with PD98059, a potent and selective inhibitor of the MAP kinase kinase (MEK‐1), which is the upstream kinase of ERK1 and ERK2. Although PD98059 inhibited ERK2 activation in platelets, it had no effect on pVWF‐ or thrombin‐induced platelet alpha or lysozomal granule release, modulation of membrane glycoprotein CD41, microparticle formation, platelet shape change or platelet agglutination. It is concluded that pVWF and thrombin induced JNK activation, but whereas thrombin induced ERK2 activation VWF did not; functional ERK2 activity was also not required for pVWF‐ or thrombin‐dependent platelet activation.
Detection of Gov system antibodies by MAIPA reveals an immunogenicity similar to the HPA‐5 alloantigens
Authors: Berry J. E. , Murphy C. M. , Smith G. A. , Ranasinghe E. , Finberg R. , Walton J. , Brown J. , Navarrete C. , Metcalfe P. , Ouwehand W. H.
Published: March 2003

The glycosylphosphatidylinositol‐linked platelet protein CD109 carries the biallelic alloantigen system Gov. There is limited information on the incidence of Gov alloantibodies in neonatal alloimmune thrombocytopenia (NAITP), post‐transfusion purpura (PTP) and platelet refractoriness. We adapted the monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) assay to the detection of Gov antibodies and determined their incidence in 605 archived samples (112 with HPA antibodies) referred for the aforementioned conditions. Here, we show that CD109 expression was reduced upon platelet storage in saline or by cryopreservation, but was stable when stored as whole blood or therapeutic platelet concentrate. Fourteen of the 605 samples contained Gov alloantibodies (anti‐Gova, n = 10; anti‐Govb, n = 4), with the majority in platelet refractoriness (n = 9) and, of the remaining five, four in NAITP and one in PTP. In seven cases, no other HPA antibodies were detected, three being NAITP cases. The incidence of Gov antibodies was significantly lower than HPA‐1 system antibodies (n = 87), but equalled the number of HPA‐5 system antibodies (n = 14) and outnumbered HPA‐2 and ‐3 system antibodies (10 altogether).
Hereditary cyclic neutropenia in the male members of a Chinese family with inverted Y chromosome
Authors: Peng Hong‐Wen , Chou Chun‐Fen , Liang Der‐Cherng
Published: March 2003

We describe four male members of a Chinese family, including the father and three sons, with hereditary cyclic neutropenia. These patients had all developed cyclic neutropenia in childhood with a cycle of around 21 d. Recurrent mucosa and skin infections with fever had occurred frequently, but gradually decreased in severity on reaching adulthood. Monocytosis was found during the neutrophil nadirs in all four patients. Mildly increased serum immunoglobulin (Ig)A and IgG levels, low levels of serum stem cell factor, as well as decreased sperm count and motility were demonstrated in the two elder sons. Chromosomal analysis showed a pericentric inversion of Y chromosome [46,X, inv(Y)(p11.2;q11.23)] in all of the men. These findings may suggest an association between cyclic neutropenia with oligospermia and inv(Y)(p11.2;q11.23) in this particular family.
Second malignancies in patients with essential thrombocythaemia treated with busulphan and hydroxyurea: long‐term follow‐up of a randomized clinical trial
Authors: Finazzi Guido , Ruggeri Marco , Rodeghiero Francesco , Barbui Tiziano
Published: March 2003

We have previously demonstrated that hydroxyurea (HU) reduces the rate of vascular complications in patients with essential thrombocythaemia (ET) at high risk of thrombosis. However, the relatively short follow‐up (median 27 months) did not enable the evaluation of the risk of developing secondary malignancies. In this study, we report the long‐term outcome of the 114 patients included in the trial: 56 patients randomized to receive HU and 58 patients to receive no cytoreductive therapy. Before randomization, 15 patients had been treated with busulphan. During the observation period, 29 patients (50%) shifted from the control to the HU group mainly because of thrombosis. Median follow‐up was 73 months (range 3–94). Analysis was by intention to treat and, when indicated, by treatment.

When analysed by intention to treat, 46 out of 54 patients (85%) originally randomized in the HU group are alive, compared with 49 of 58 patients (84%) in the control group [not significant (n.s.)]. Five patients (9%) in the HU group and 26 patients (45%) in the control group had thrombosis (P < 0·0001). Seven patients (13%) in the HU group developed secondary acute leukaemia, myelodysplastic syndromes or solid tumours, compared with only one of the control group patients (1·7%) (P = 0·032). The occurrence of secondary malignancies was also analysed by treatment: none of the 20 patients who had never been treated with chemotherapy developed neoplasia vs. three of the 77 patients given HU only (3·9% n.s.) and five of the 15 patients given busulphan plus HU (33% P < 0·0001).

This study showed that: (a) HU reduced the risk of thrombosis in ET patients; (b) the sequential use of busulphan and HU significantly increased the risk of second malignancies; and (c) overall survival was not affected by HU therapy.
Relationship between TNF‐α and iron metabolism in differentiating human monocytic THP‐1 cells
Authors: Scaccabarozzi Anna , Arosio Paolo , Weiss Guenter , Valenti Luca , Dongiovanni Paola , Fracanzani Anna Ludovica , Mattioli Michela , Levi Sonia , Fiorelli Gemino , Fargion Silvia
Published: March 2003

The human monocytic cell line THP‐1 differentiates along the macrophage line after phorbol‐12‐myristate‐13‐acetate (PMA) supplementation and can be stimulated to secrete tumour necrosis factor α (TNF‐α) by interferon γ (IFN‐γ) addition. We found that, in the early stage of differentiation (1–48 h), PMA induction elicited an upregulation of intracellular H ferritin and H ferritin binding sites and a downregulation of transferrin receptor. In addition, we found that iron administration to PMA‐differentiating cells induced the expression of TNF‐α mRNA and TNF‐α secretion to levels even higher than those induced by IFN‐γ alone. The iron chelator desferrioxamine showed the opposite effect and reduced TNF‐α release. In contrast, preincubation of the cells with iron before PMA induction resulted in a decrease of the TNF‐α secretion induced by IFN‐γ, whereas the opposite was true after preincubation with desferrioxamine. The data support a co‐ordinate interaction between iron and TNF‐α in monocyte macrophages, with an iron‐mediated upregulation of TNF‐α in the early phase of differentiation and an iron‐mediated inhibition at later stages. This complex relationship has to be considered in evaluating the effects of iron on inflammation.
Mortality and causes of death in patients with monoclonal gammopathy of undetermined significance
Authors: Gregersen Henrik , Ibsen Jenna Salling , Mellemkjœr Lene , Dahlerup Jens Frederik , Olsen Jørgen Helge , Sørensen Henrik Toft
Published: March 2003

To evaluate the mortality and causes of death in monoclonal gammopathy of undetermined significance (MGUS), we identified 1324 cases of MGUS in the period 1978–93 in North Jutland County, Denmark. Data on mortality were obtained by record linkage to the Danish Death Registry. There were 868 deaths in the MGUS cohort during 7785 years of follow‐up vs. 409·6 expected, giving a standardized mortality ratio (SMR) of 2·1 (95% confidence interval 2·0–2·3). Malignant transformation was the cause of death in 97 patients vs. 4·9 expected, yielding a SMR of 20·0 (16·2–24·4), which explained about 20% of the excess mortality in the cohort. The mortality was increased for several other malignant and non‐malignant causes of death during the first 4 years of follow‐up. For late follow‐up, 5–18 years after detection of the M‐component, the overall SMR was 1·7 (1·5–1·9), and malignant transformation was the only cause of death with a substantial increase of SMR. Although malignant transformation is an important cause of death in MGUS patients, it did not entirely explain the increased mortality. MGUS patients often suffer from coexisting clinical conditions that increase the mortality, especially during the first years after detection of the M‐component.
Novel mutations in the Factor VII gene of Taiwanese Factor VII‐deficient patients
Authors: Shen Ming‐Ching , Lin Jen‐Shiou , Lin Shu‐Wha , Yang Wei‐Shin , Lin Bodo
Published: March 2003

The genetic defects of four Taiwanese patients with factor VII (FVII) deficiency were studied. FVII activity and antigen levels were < 1 u/dl and 125·7 u/dl (patient I), < 1 u/dl and < 1 u/dl (patient II), 3·4 u/dl and 5·9 u/dl (patient III), and 1·2 u/dl and 30·4 u/dl (patient IV) respectively. The 5′ flanking region, and all exons and junctions were amplified using polymerase chain reaction and sequenced. Patient I was homozygous for a 10824C→A transversion with Pro303→Thr mutation in exon 8. In patient II, a heterozygous transversion, 9007+1G→T at the IVS6, a heterozygous decanucleotide insertion polymorphism at −323 (both mutations present in his father) and a heterozygous deletion, del TC (26–27) in exon 1A (originating from his mother) were identified. Patient III had a homozygous 10961T→G transversion with His348→Gln mutation in exon 8. Patient IV had a heterozygous 10902T→G transversion with Cys329→Gly mutation in exon 8 (transmitted to her second son) and a heterozygous decanucleotide insertion polymorphism at −323 (transmitted to her third son). All but one of the FVII gene mutations detected in the four patients have not been previously reported. In conclusion, four novel mutations of the FVII gene in Taiwanese, including two missense mutations in exon 8, one point mutation at the exon 6 splice site and one deletion in exon 1A, were identified.
Three cases of typical aplastic anaemia associated with a Philadelphia chromosome
Authors: Suzan F. , Terré C. , Garcia I. , Bastie J. N. , Baumelou E. , Gluckman E. , Castaigne S.
Published: March 2003

We report three cases of typical aplastic anaemia (AA) associated with a Philadelphia chromosome. This translocation was detected at the time of diagnosis of AA (one patient) and when overt leukaemia was diagnosed (two patients: one chronic myeloid leukaemia and one acute lymphoblastic leukaemia) after AA therapy and recovery of blood counts. We discuss the literature arguments about considering some cases of AA as preleukaemic disorders and suggest that our cases illustrate the association of AA with a clonal malignant disorder. We conclude that cytogenetic analysis is necessary at diagnosis of AA or after recovery of blood counts.
Continuous complete remission in adult patients with acute lymphocytic leukaemia at a median observation of 12 years after autologous bone marrow transplantation
Authors: Abdallah Alhossain , Egerer Gerlinde , Goldschmidt Hartmut , Wannenmacher Michael , Körbling Martin , Ho Anthony D.
Published: March 2003

We report our long‐term experience with autologous bone marrow transplantation (ABMT) for 32 adult patients with acute lymphocytic leukaemia (ALL) in second or later remission (CR), or in first CR but with high‐risk. Bone marrow was purged with mafosfamide (n = 25) or with immunomagnetic beads and monoclonal antibodies (n = 7). Retrospective analysis showed that 12 out of 32 patients were in continuous complete remission (CCR) at a median of 143 months (range 66–181 months). A plateau was reached at 50 months and the disease‐free and overall survival rates were both 37·5%. It was notable that durable CCR could be achieved for patients in second (three out of nine) or third (one out of six) CR. ABMT could produce durable CCR and the long‐term outcome compared favourably with those reported for allogeneic transplantation.
Drug monitoring of low‐dose PEG‐asparaginase (Oncaspartm) in children with relapsed acute lymphoblastic leukaemia
Authors: Vieira Pinheiro J. P , Müller H. J. , Schwabe D. , Gunkel M. , Casimiro da Palma J. , Henze G. , Von Schütz V. , Winkelhorst M. , Würthwein G. , Boos J.
Published: March 2003

Use of asparaginase (ASNase) in the treatment of relapsed childhood acute lymphoblastic leukaemia (ALL) is associated with a high rate of hypersensitive reactions. ‘Silent’ inactivation may additionally reduce treatment intensity. Therefore, PEG‐ASNase (Oncaspartm), a polyethylene glycol conjugate of the native Escherichia coli‐ASNase, was introduced into the Berlin‐Frankfurt‐Münster (BFM) 96 treatment protocol for relaped ALL under drug monitoring conditions. A single i.v. dose of 500 IU/m2 PEG‐ASNase, substituted for the native ASNases, was administered to supply a plasma activity of 100 IU/l for 1 week. From November 1997 to March 2000, 35 patients from 23 BFM‐associated hospitals, with or without a previous allergic reaction to one or both native preparations, underwent monitoring. After 82 applications, a total of 270 samples were submitted to be tested for ASNase activity. The ASNase activity on the day of the administration and the following day ranged between < 20 and 693 IU/l, with a median of 413 IU/l (53 samples). The median on d 7 ± 1 was 199 IU/l (range <20–421 IU/l; 41 samples) and on d 14 ± 1, 105 IU/l (range <20–188 IU/l; 19 samples). An ASNase activity of > 100 IU/l was seen on d 7 in 36 activity time courses of 52 interpretable applications (69%). Intraindividual variability of activity time courses was low. However, a rapid decrease in ASNase activity after repeated applications was observed in 4 out of 20 children. Previously experienced allergic reactions to native ASNases did not influence PEG‐ASNase pharmacokinetics. PEG‐ASNase is a useful alternative to the native ASNases in children with relapsed ALL. Whenever possible, drug monitoring should be performed to identify patients with ‘silent’ inactivation.
A DNA‐based immunization protocol to produce monoclonal antibodies to blood group antigens
Authors: Chu Te‐Hua Tearina , Halverson Gregory R. , Yazdanbakhsh Karina , Øyen Ragnhild , Reid Marion E.
Published: March 2003

A major challenge facing transfusion medicine is the establishment of immunological methods to produce specific and avid blood group typing reagents to the many polymorphic blood group antigens. This is especially true when sources of human antibody are limited. Based on the knowledge that inoculation with plasmid DNA can induce a humoral response in the host animal, we inoculated mice with plasmid DNA followed by a single boost injection with plasmid‐transfected cells that have a high level of expression of the same target protein. Using this method, several hybridoma clones that produced strongly reactive antibodies specific for the Kell polymorphic antigens (anti‐K, anti‐k, anti‐Kpa) were isolated. The monoclonal antibodies that were produced with this method have potential clinical utility for identifying a patient's blood type and for screening for antigen‐negative donor blood.
Symptomatic type 1 protein C deficiency caused by a de novo Ser270Leu mutation in the catalytic domain
Authors: Lind Bent , Koefoed Pernille , Thorsen Sixtus
Published: March 2003

Heterozygosity for a C8524T transition in the protein C gene converting Ser270(TCG) to Leu(TTG) in the protease domain was identified in a family with venous thrombosis. The mutation was associated with parallel reduction in plasma levels of protein C anticoagulant activity and protein C antigen, which is consistent with a type 1 deficiency. Transient expression of mutant protein C cDNA in human kidney 293 cells and analysis of protein C antigen in culture media and cell lysates showed that the secretion of mutant protein compared with wild‐type protein was reduced by at least 97% while the intracellular content of mutant and wild‐type protein was similar. Northern blot analysis of total mRNA from transfected cells showed no reduction of the mutant protein C mRNA compared with wild‐type protein C mRNA. Collectively, these results indicate that the Ser270Leu mutation in the affected family caused the plasma protein C deficiency and are consistent with a disease mechanism that involves synthesis of mutant protein followed by intracellular degradation before its secretion into the extracellular space. The mutation was not present in the parents of the proband, suggesting a de novo mutation. Non‐paternity was excluded after the analysis of three intragenic protein C polymorphisms and six dinucleotide repeat allele sets located in five different chromosomes.
Patient cytomegalovirus seropositivity with or without reactivation is the most important prognostic factor for survival and treatment‐related mortality in stem cell transplantation from unrelated donors using pretransplantin vivoT‐cell depletion with anti‐thymocyte globulin
Authors: Kröger Nicolaus , Zabelina Tatjana , Krüger William , Renges Helmut , Stute Norbert , Schrum Johanna , Kabisch Hartmut , Schafhausen Philippe , Jaburg Nicole , Löliger Cornelius , Schäfer Peter , Hinke Axel , Zander Axel R.
Published: March 2003

We evaluated the cytomegalovirus (CMV) serostatus as a risk factor for survival and treatment‐related mortality (TRM) in 125 patients allografted from an unrelated donor between 1994 and 1999. All patients received pretransplantin vivoT‐cell depletion using rabbit anti‐thymocyte globulin (ATG). Only one patient had primary graft failure and severe grade III/IV graft‐versus‐host disease occurred in 14% of the patients. The overall survival (OS) at 3 years was 70% for CMV‐negative patients (n = 76) and 29% in the seropositive cohort (n = 49) (P > 0·001). In multivariate analyses, CMV seropositivity remained an independent negative prognostic factor for OS (RR: 2·1; CI: 1·2–3·8;P = 0·014), apart from age > 20 years (RR: 2·74; CI: 1·2–3·8;P = 0·004) and late leucocyte engraftment (RR: 2·4; CI: 1·2–4·9;P = 0·015). The TRM for all patients was 27%. Despite monitoring for CMV antigenaemia and preemptive therapy with ganciclovir when reactivation occurred, seropositive patients had a three times higher risk of fatal treatment‐related complications than seronegative patients. In multivariate analyses, CMV seropositivity remained the strongest independent negative factor for TRM (RR: 5·3; CI: 1·9–14·6;P = 0·002), followed by age > 20 years (RR: 4·8; CI: 1·3–18·1;P = 0·02) and delayed leucocyte engraftment (RR: 3·6; CI: 1·2–11;P = 0·02). The TRM was identical in seropositive patients with (n = 27) or without (n = 22) CMV reactivation (44% versus 50%). We conclude that CMV seropositivity, despite preemptive ganciclovir therapy and even without reactivation, is a major negative prognostic factor for survival as well as for TRM in unrelated stem cell transplantation using pretransplantin vivoT‐cell depletion with ATG.
Factors affecting lymphocyte subset reconstitution after either related or unrelated cord blood transplantation in children – a Eurocord analysis
Authors: Niehues Tim , Rocha Vanderson , Filipovich Alexandra H. , Chan Ka Wah , Porcher Raphael , Michel Gerard , Ortega Juan J. , Wernet Peter , Göbel Ulrich , Gluckman Eliane , Locatelli Franco
Published: March 2003

Immune recovery after cord blood transplantation (CBT) is of concern owing to the low number of lymphocytes transferred with the graft and their immaturity. Risk factors influencing lymphocyte subset reconstitution related to disease, patient, donor and transplant were studied in 63 children (< 16 years), given either related (n = 14) or unrelated (n = 49) CBT for malignant (n = 33) or non‐malignant diseases (n = 30). Only children with sustained myeloid engraftment were analysed. Absolute numbers of T (CD3+, CD4+, CD8+), B and natural killer (NK) cells were reported 2–3, 6, 9, 12 and 12–24 months after CBT. Median patient age was 4·0 years (0–15) and median follow‐up was 23 months (1·7–61·0). Twenty‐six patients received human leucocyte antigen (HLA)‐matched CBT and 37 received HLA‐mismatched CBT. The median number of nucleated cells (NCs) collected/recipient weight was 6·1 × 107/kg. In this selected population, the estimate 2 year survival was 85%. Lymphocyte reconstitution (defined as the median time to reach the normal value of age‐matched healthy children) was 3, 6 and 8 months for NK, B and CD8+cells, while it was 11·7 months for both CD3+and CD4+lymphocytes. In the multivariate analysis, factors favouring T‐cell recovery were: related donor (P = 0·002); higher NCs/kg (P = 0·005) and recipient cytomegalovirus (CMV)‐positive serology (P = 0·04). Presence of acute graft‐versus‐host disease (GVHD) delayed T‐cell recovery (P = 0·04). To summarize, in children with sustained myeloid engraftment the concern that lymphocyte recovery after CBT could be delayed does not appear to be substantiated by our results.
Clinical significance of Aspergillus fungaemia in patients with haematological malignancies and invasive aspergillosis
Authors: Girmenia Corrado , Nucci Marcio , Martino Pietro
Published: March 2003

The clinical significance of Aspergillus fungaemia in the setting of a deep‐seated aspergillosis has not been clearly established. Among 107 microbiologically documented Aspergillus infections in patients with haematological diseases observed over a 17‐year period, blood cultures grew Aspergillus species from 10 cases. Aspergillus fungaemia was documented in 9 out of 89 (10·1%) patients with pulmonary aspergillosis at a median of 5 d from the onset of clinical signs of infection, and in one patient with central venous catheter focal infection. Five (50%) patients died as a result of fungal infection a median of 12 d (range 4–48) from the documentation of Aspergillus fungaemia. A comparison between cases of invasive aspergillosis with or without fungaemia showed that fungaemic patients were similar to those without positive blood cultures regarding clinical presentation, risk factors, clinical course and outcome. The diagnostic role of Aspergillus fungaemia in the setting of a deep‐seated infection is limited because blood cultures become positive when a microbiological or clinical diagnosis of aspergillosis has already been performed. Aspergillus fungaemia does not necessarily seem to be correlated with a disseminated infection or a poorer prognosis.
Autologous peripheral blood stem cell transplantation in patients with relapsed lymphoma results in accelerated haematopoietic reconstitution, improved quality of life and cost reduction compared with bone marrow transplantation: the Hovon 22 study
Authors: Vellenga E. , Van Agthoven M. , Croockewit A. J. , Verdonck L. F. , Wijermans P. J. , Van Oers M. H. J. , Volkers C. P. , Van Imhoff G. W. , Kingma T. , Uyl‐de Groot C. A. , Fibbe W. E.
Published: March 2003

The present study analysed whether autologous peripheral blood stem cell transplantation (PSCT) improves engraftment, quality of life and cost‐effectiveness when compared with autologous bone marrow transplantation (ABMT). Relapsing progressive lymphoma patients (n = 204; non‐Hodgkin's lymphoma n = 166; Hodgkin's disease n = 38) were, after induction treatment with the DHAP–VIM (cisplatin, cytarabine, dexamethasone, etoposide, ifosfamide, methotrexate) regimen, randomly (2:1) assigned to the harvest of granulocyte–macrophage colony‐stimulating factor‐mobilized stem cells after the second DHAP course or autologous bone marrow cells before the second DHAP course. These stem cells were reinfused following high‐dose myeloblative chemotherapy. After induction, 118 patients obtained a partial or complete response and were eligible for randomization. In the PSCT arm (n = 76) significantly faster engraftment of neutrophils [≥ 0·1 and ≥ 0·5 × 109/l: 10·7 d (7–36, median, range), 15 (9–45) versus 13 (8–25) and 26 (14–80), P < 0·01] and thrombocytes [≥ 20 × 109/l: 13 d (7–51) versus 18 (11–65), P < 0·01] were observed. In addition, significantly fewer transfusions of red blood cells [6 (0–23) versus 8 (2–24), P < 0·01] and platelets [4 (0–60) versus 8 (2–55), P = 0·01] were required in the PSCT arm. These findings were associated with a significant reduction in the median days of intravenous antibiotics in patients with fever [8·5 (0–30) versus 14 (0–34), P = 0·04] and hospital stay [27 (8–51) versus 34 (24–78), P < 0·05]. Quality of life demonstrated a significant difference in favour of the PSCT arm. Total transplantation costs were significantly lower in the PSCT arm [$13 954 ($4913– 29 532) versus $17 668 ($10 170–44 083) P < 0·05], as a result of the reduced hospital stay and lower antibiotic costs. In summary, these results indicate that PSCT is superior to ABMT with regard to engraftment, supportive care, quality of life and cost.
Mutation screening of the c‐MYB negative regulatory domain in acute and chronic myeloid leukaemia
Authors: Lutwyche Jodi K. , Keough Rebecca A. , Hughes Timothy P. , Gonda Thomas J.
Published: March 2003

Over‐expression of the c‐myb gene and expression of activated forms of myb are known to transform haemopoietic cells, particularly cells of the myeloid lineage. Truncations or mutations that disrupt the negative regulatory domain (NRD) of the Myb protein confer an increased ability to transform cells. Although it has proved difficult to link mutations in c‐MYB to human leukaemia, no studies investigating the presence of mutations within the c‐MYB NRD have been reported. Therefore, we have performed mutational analysis of this region, using polymerase chain reaction–single‐stranded conformation polymorphism and sequence analysis, in 26 patients with acute or chronic myeloid leukaemia. No mutations were detected, indicating that mutation of this region of the Myb protein is not common in the pathogenesis or progression of these diseases.
Therapy‐related acute lymphoblastic leukaemia with MLL rearrangements following DNA topoisomerase II inhibitors, an increasing problem: report on two new cases and review of the literature since 1992
Authors: Andersen Mette Klarskov , Christiansen Debes H. , Jensen Bjarne Anker , Ernst Peter , Hauge Gunnar , Pedersen‐Bjergaard Jens
Published: March 2003

A highly increased risk of myelodysplasia (MDS) and acute myeloid leukaemia (AML) is well established in patients previously treated for other malignancies with alkylating agents or topoisomerase II inhibitors. More recently, single cases of acute lymphoblastic leukaemia (ALL), often presenting balanced translocations involving chromosome band 11q23, have been observed. We present two such cases with t(4;11)(q21;q23), one of whom had previously received only single‐agent chemotherapy with 4‐epi‐doxorubicin. A review of the literature since 1992 including these two patients reveals a total of 23 cases of ALL or lymphoblastic lymphoma after chemotherapy presenting balanced translocations to 11q23. All 23 patients had previously received at least one topoisomerase II inhibitor, and in two patients 4‐epi‐doxorubicin had been administered as single‐agent chemotherapy for breast cancer. The latency period to development of t‐ALL was 24 months or less in 20 out of 22 cases. The MLL gene was found to be rearranged in 14 out of 14 cases, and in three out of six cases the breakpoint was at the telomeric part of the gene, as observed in most cases of AML following therapy with topoisomerase II inhibitors. These results indicate that patients with ALL and balanced translocations to chromosome band 11q23 following chemotherapy with topoisomerase II inhibitors in the future should be included with cases of MDS or AML in calculations of risk of leukaemia.
Protein A sepharose immunoadsorption: immunological and haemostatic effects in two cases of acquired haemophilia
Authors: Guillet B. , Kriaa F. , Huysse M. G. , Proulle V. , George C. , Tchernia G. , D'Oiron R. , Laurian Y. , Charpentier B. , Lambert T. , Dreyfus M.
Published: March 2003

Acquired haemophilia is a life‐threatening disorder caused by circulating auto‐antibodies that inhibit factor VIII coagulant activity (FBIII:C). Immunoadsorption on protein A sepharose (IA‐PA) was performed in two bleeding patients with acquired haemophilia: we observed a dramatic and quick decrease in the anti‐FVIII:C inhibitor titre leading to a normal, albeit transient, haemostatic status. In one case, IA‐PA was the only procedure which succeeded in stopping massive haemorrhage. In the second case, IA‐PA reinforced the haemostatic effect of recombinant activated factor VII by increasing the endogenous plasma factor VIII level. The efficacy of IA‐PA was sustained with immunosuppressive treatment introduced, respectively, 10 and 15 d before the IA‐PA procedures. Our experience with IA‐PA suggests that this extracorporeal anti‐FVIII:C removal procedure is a valuable therapeutic tool for acquired haemophilia and can alleviate life‐threatening haemorrhages.
Oral ciprofloxacin plus colistin: prophylaxis against bacterial infection in neutropenic patients. A strategy for the prevention of emergence of antimicrobial resistance
Authors: Prentice H. Grant , Hann Ian M. , Nazareth Bernadette , Paterson Pamela , Bhamra Amrat , Kibbler Christopher C.
Published: March 2003

Following a 2‐year study, the combination of oral ciprofloxacin and colistin has been used continuously for 10 years without the emergence of resistance. During a 2‐year period (1987–1989), we compared ciprofloxacin + colistin (CIP + COL) with neomycin + colistin (NEO + COL) in a randomized trial – combinations chosen because of the potential for prophylaxis of Gram‐negative infection by ciprofloxacin, with colistin given to reduce the risk of emergence of resistance. Sixty‐four patients with similar demographics in each arm were evaluable for efficacy analysis. Patients on CIP + COL had a significantly lower proportion of neutropenic days with fever (P < 0·001) and neutropenic days on intravenous antibiotics (P < 0·001) than patients on NEO + COL. A total of 54 (15 bacteriologically documented) pyrexial episodes occurred in patients on CIP + COL and 77 (41 bacteriologically documented) in patients on NEO + COL. Only two Gram‐negative bacterial infections occurred in the CIP + COL arm compared with 16 in the NEO + COL arm. No Staphylococcus aureus infections occurred in the CIP + COL group compared with 10 in the other patients. Two CIP‐resistant Gram‐negative bacilli were isolated from patients on CIP + COL compared with 13 NEO‐resistant Gram‐negative bacilli from patients on NEO + COL. Following a subsequent decade of unchanged use of this prophylactic strategy in neutropenic patients, a 2‐year follow‐up study between 1 January 1998 and 31 December 1999 showed 66 significant infections during 350–400 neutropenic episodes. Eight of the 111 (7·2%) isolates were with ciprofloxacin‐resistant organisms, involving 2% of the neutropenic episodes, indicating that the strategy of combining colistin with ciprofloxacin has been effective in the prevention of Gram‐negative sepsis in neutropenic patients without the emergence of significant resistance despite widespread concurrent hospital and community use of the quinolones.
Thalidomide in agnogenic and secondary myelofibrosis
Authors: Canepa Letizia , Ballerini Filippo , Varaldo Riccardo , Quintino Sabrina , Reni Lizia , Clavio Marino , Miglino Maurizio , Pierri Ivana , Gobbi Marco
Published: March 2003

Myelofibrosis with myeloid metaplasia (MMM) is a clonal disorder involving disregulation of angiogenesis and immunomodulatory mechanisms. Thalidomide (Thal) retains antiangiogenic, immunomodulatory and cytokine regulatory properties and recently it has been used successfully in multiple myeloma. Here, we report our experience in 10 MMM patients treated with Thal. Patients with agnogenic MMM treated in an early phase of the disease obtained significant benefits from the therapy and remain transfusion‐free. In contrast, all secondary MMM failed to respond. These preliminary findings confirm that Thal plays a role in MMM therapy, although the efficacy in the different phases of the disease must be further evaluated.
International prognostic scoring system and TP53 mutations are independent prognostic indicators for patients with myelodysplastic syndrome
Authors: Kita‐Sasai Yuri , Horiike Shigeo , Misawa Shinichi , Kaneko Hiroto , Kobayashi Miyako , Nakao Mitsushige , Nakagawa Hitoshi , Fujii Hiroshi , Taniwaki Masafumi
Published: March 2003

We applied the International Prognostic Scoring System (IPSS) to our series of 118 patients with myelodysplastic syndrome (MDS) to determine its validity, and also used univariate and multivariate analyses to evaluate the prognostic significance of TP53 configurations. Sixteen patients with the mutation had a strikingly worse prognosis and the multivariate analysis demonstrated that this alteration was the most significant factor. The prognostic comparison between patients with and without the mutation within each IPSS subgroup showed a significant difference in the intermediate subgroups. A combination of clinical manifestations and genetic configurations provided us with more accurate prognostic information in MDS patients.
Breast cancer resistance protein expression and resistance to daunorubicin in blast cells from patients with acute myeloid leukaemia
Authors: Sargent Jean M. , Williamson Christine J. , Maliepaard Marc , Elgie Alena W. , Scheper Rik J. , Taylor Colin G.
Published: March 2003

Breast cancer resistance protein (BCRP) is a recently described member of the ATP binding cassette transporter superfamily. It has been shown to confer resistance to mitoxantrone, topotecan, doxorubicin and daunorubicin in human tumour cell lines. We describe a study of BCRP expression in blast cells derived from 20 patients with acute myeloid leukaemia (AML). Twelve samples were from patients who had received previous cytotoxic therapy. BCRP expression was measured by immunocytochemistry using the BXP‐34 monoclonal antibody. In vitro drug sensitivity was assessed using the methyl thiazol tetrazoliumbromide assay. BCRP expression varied between patients, and six out of 22 (27%) samples had > 10% cells staining positively (median 37%, range 13–95%). BCRP positivity was seen in both de novo samples and those from previously treated patients. There was a marked variation in the effect of all drugs tested between patients. Although there was no correlation between BCRP positivity and the effect of mitoxantrone, topotecan or doxorubicin, the median daunorubicin LC50 value of BCRP+ cells was fourfold higher than that of BCRP− cells (0·89 µmol/l compared with 0·21 µmol/l, P < 0·05). These results suggest that BCRP may be involved in resistance to the agents commonly used in AML and may explain some of the anomalous results found when studying other membrane transporters, such as P‐gp or MRP.
Comparison of peripheral blood interphase cytogenetics with bone marrow karyotype analysis in myelofibrosis with myeloid metaplasia
Authors: Tefferi Ayalew , Meyer Reid G. , Wyatt William A. , Dewald Gordon W.
Published: March 2003

In a prospective study of 42 patients with myelofibrosis with myeloid metaplasia (MMM), peripheral blood (PB) and bone marrow (BM) interphase cytogenetics and PB CD34 enumeration were performed concomitantly with BM karyotype analysis. Interphase cytogenetics was performed with a panel of fluorescence in situ hybridization (FISH) probes that were capable of detecting most of the known recurrent cytogenetic lesions in MMM. There was a close concordance in the results of interphase cytogenetics between PB and BM, regardless of the PB CD34 count. In general, FISH‐detectable abnormalities were also detected by BM karyotype. Although complementary, interphase cytogenetics may not always provide the necessary karyotypic information in MMM.
Significant linkage and non‐linkage of type 1 von Willebrand disease to the von Willebrand factor gene
Authors: Casaña Pilar , Martínez Francisco , Haya Saturnino , Espinós Carmen , Aznar José A.
Published: March 2003

Significant linkage of types 2A and 2B von Willebrand disease (VWD) to the von Willebrand factor (VWF) gene have been reported, as well as mutations in the VWF gene. However, data for the partial quantitative variant are less consistent. An inconsistency of association between the type 1 VWD phenotype and genotype has been reported recently. We undertook linkage analysis of 12 families with definite or possible type 1 VWD patients. One family with classic type 1 VWD had a high lod score (Z = 5·28, θ = 0·00). A total lod score of 10·68 was obtained for the four families with fully penetrant disease. In two families linkage was rejected, while three families did not show conclusive evidence of linkage. This study corroborates ABO blood group influence, especially in patients with mild deficiencies and/or incomplete penetrance. Indirect genetic analysis may be an option for diagnosing asymptomatic or presymptomatic type 1 VWD carriers, particularly in families showing higher penetrance. The study indicates defects of the VWF locus are to be expected in more than half of the families studied. However, as defects at different loci may be the cause of this phenotype, the results of the segregation analyses should be interpreted with caution, especially in studies involving small families, or mild expressions of the disorder or incomplete penetrance.
Reappraisal of the clinical significance of CD7 expression in association with cytogenetics in de novo acute myeloid leukaemia
Authors: Ogata K. , Yokose N. , Shioi Y. , Ishida Y. , Tomiyama J. , Hamaguchi H. , Yagasaki F. , Bessyo M. , Sakamaki H. , Dan K. , Kuriya S.
Published: March 2003

Debate exists over whether CD7 expression indicates an unfavourable prognosis in de novo acute myeloid leukaemia (AML). Meanwhile, the type of cytogenetics is a strong prognostic factor in AML. We analysed 256 de novo adult AML cases and found that the proportion of CD7+ cases increased stepwise from the cases with favourable cytogenetics to the cases with intermediate and unfavourable cytogenetics (3 out of 69 cases, 51 out of 140 cases and 25 out of 47 cases respectively, P < 0·0001). CD7‐positivity adversely affected the survival only in the cases with unfavourable cytogenetics (P < 0·03). We recommend that CD7 expression in AML be interpreted in association with the cytogenetics.
Mixed chimaerism is common at the time of acute graft‐versus‐host disease and disease response in patients receiving non‐myeloablative conditioning and allogeneic stem cell transplantation
Authors: Mattsson Jonas , Uzunel Mehmet , Brune Mats , Hentschke Patrik , Barkholt Lisbeth , Stierner Ulrika , Aschan Johan , Ringdén Olle
Published: March 2003

We report the clinical outcome and results of chimaerism analysis in various cell lineages of 30 patients given non‐myeloablative conditioning, followed by allogeneic stem cell transplantation (SCT). The commonest diagnoses were chronic myelogenous leukaemia (n = 11) and solid tumours (n = 11). Twenty‐one patients received SCT from human leucocyte antigen (HLA)‐identical siblings and nine from matched unrelated donors. Median patient age was 53 (28–77) years. Four non‐myeloablative protocols were used, including fludarabine (30 mg/m2 × 3–6), busulphan (4 mg/kg × 2), cyclophosphamide (Cy) (30 mg/kg/day × 2) or total body irradiation (2 Gy), and anti‐thymocyte globulin. The patients were analysed by polymerase chain reaction (PCR) analysis of minisatellites on days 14, 21 and 28, then every other week up to 3 months and monthly thereafter. All samples were cell separated for T, B and myeloid cells using immunomagnetic beads. Eighteen patients were alive at a median follow‐up of 11 (6–20) months. Acute graft‐versus‐host disease (GVHD) occurred in 22 patients. Eighteen of the 22 patients with acute GVHD showed mixed chimaerism (MC) in the T‐cell fraction at the time of acute GVHD. However, all patients with acute GVHD showed donor chimaerism (DC) in the T‐cell fraction median 76 (7–414) days after onset versus three out of eight patients without acute GVHD, P < 0·001]. Disease response was diagnosed in 15 patients, median 100 (37–531) days after SCT. At the time of disease response, six out of15 patients showed MC in the T‐cell fraction. In conclusion, mixed chimaerism in the T‐cell fraction is common at the time of acute GVHD and disease response in patients conditioned with non‐myeloablative therapy.
Chronic myeloid leukaemia with marked thrombocytosis in a patient with thalassaemia major: complete haematological remission under the combination of hydroxyurea and anagrelide
Authors: Voskaridou Ersi , Terpos Evangelos , Komninaka Veroniki , Eftyhiadis Eftyhios , Mantzourani Marina , Loukopoulos Dimitris
Published: March 2003

Summary. The co‐existence of thalassaemia major and chronic myeloid leukaemia (CML) is a very rare event. We report a 32‐year‐old man with thalassaemia major whose progressively increasing leukocytosis and thrombocytosis led to the diagnosis of CML confirmed by the characteristic t(9;22)(q34;q11) chromosomal translocation and the bcr‐abl (b3a2) DNA fusion. The patient was treated with hydroxyurea and anagrelide. This combination resulted in the satisfactory control of both the white blood cell and platelet counts, which has continued over the past 14 months with no major side‐effects, albeit with no molecular response. The administration of hydroxyurea was also associated with a significant HbF increase.
Enhanced haemolysis with β‐thalassaemia trait due to the unstable β chain variant, Hb Gunma, accompanied by hereditary elliptocytosis due to protein 4·1 deficiency in a Japanese family
Authors: Maehara Tadashi , Tsukamoto Norifumi , Nojima Yoshihisa , Karasawa Masamitsu , Murakami Hirokazu , Hattori Yukio , Ideguchi Hiroshi
Published: March 2003

Summary. We identified a Japanese family with a β‐thalassaemia trait and hereditary elliptocytosis (HE). We studied five members of this family. One was normal, one had only the β‐thalassaemia trait, one had heterozygous HE, and two had compound heterozygous β‐thalassaemia trait and HE. The last two had already undergone splenectomy. The molecular profile of β‐thalassaemia was consistent with that of Hb Gunma: codon 127/128CAGGCT(Gln–Ala)→ CCT(Pro). Analysis of erythrocyte membrane proteins revealed a partial deficiency of protein 4·1 in all those with HE, whereas the spectrin content was within the normal range. Each heterozygous family member with either the β‐thalassaemia trait or HE was asymptomatic, whereas the two with both β‐thalassaemia and HE had marked red blood cell deformities and haemolysis. The abnormalities of the red blood cells in patients with the β‐thalassaemia trait might be enhanced by association with HE owing to a protein 4·1 deficiency.