bluuebunny/crossref_metadata_2025 huggingface dataset:
Mitoxantrone, a topoisomerase II inhibitor, induces apoptosis of B‐chronic lymphocytic leukaemia cells
Authors: Bellosillo Beatriz , Colomer Dolors , Pons Gabriel , Gil Joan
Published: March 2003

B‐chronic lymphocytic leukaemia (B‐CLL) is characterized by the accumulation of long‐lived CD5+ B lymphocytes. The effect of mitoxantrone, a topoisomerase II inhibitor, on B‐CLL cells was studied. Treatment of B‐CLL cells for 48 h with mitoxantrone (0.5 μg/ml) induced a decrease in cell viability as determined by MTT assay. The IC50 calculated for the cells of three patients was 0.7 μg/ml for two of them and 1.4 μg/ml for the third. In all three patients the maximum effect was observed with 2 μg/ml. An additive cytotoxic effect was observed when mitoxantrone (0.5 μg/ml) was combined with fludarabine (5 μg/ml). Mitoxantrone induced DNA fragmentation and the proteolytic cleavage of poly(ADP‐ribose) polymerase (PARP), a marker of the activation of caspases, in all the patients studied, demonstrating that the cytotoxic effect of mitoxantrone was due to induction of apoptosis. These results suggest that mitoxantrone, and possibly other topoisomerase II inhibitors, may be used in the chemotherapy of B‐CLL, and that combination of mitoxantrone with fludarabine or other drugs could improve the effectiveness of the treatment.
Circulating interleukin‐6 levels are elevated in adult T‐cell leukaemia/lymphoma patients and correlate with adverse clinical features and survival
Authors: Yamamura Masaomi , Yamada Yasuaki , Momita Saburou , Kamihira Shimeru , Tomonaga Masao
Published: March 2003

We measured the circulating levels of interleukin (IL)‐6 in adult T‐cell leukaemia/lymphoma (ATL) patients using an enzyme‐linked immunosorbent assay. The IL‐6 levels in 59 ATL patients (median 8.2 pg/ml; range < 1.0 to 185.7 pg/ml) were significantly higher than in 30 healthy controls (median < 1.0 pg/ml; range < 1.0 to 3.5 pg/ml) (P < 0.0001) or 32 human T‐lymphotropic virus type‐I (HTLV‐I) carriers (median 4.2 pg/ml; range  < 1.0 to 13.3 pg/ml) (P = 0.002). Among the ATL patients, the IL‐6 levels in the acute‐ or lymphoma‐type patients were significantly higher than those in the chronic‐type patients (P < 0.0001). The IL‐6 levels were also higher in the patients with B symptoms than in those without B symptoms (P = 0.039), and were significantly correlated with increased serum lactate dehydrogenase (LDH) (P = 0.0004) and C‐reactive protein (CRP) (P < 0.0001) and decreased serum albumin (P = 0.0003) values. The patients with elevated IL‐6 levels had inferior overall survival periods compared to those with normal IL‐6 levels (P = 0.025). ATL is a single disease entity, although its clinical features are quite diverse; the increased production of cytokines may cause the diversity of clinical features. The results of our study indicate that IL‐6 is one such cytokine.
‘Lymphoid’ blast crisis of chronic myeloid leukaemia is associated with distinct clinicohaematological features
Authors: Cervantes Francisco , Villamor Neus , Esteve Jordi , Montoto Silvia , Rives Susana , Rozman Ciril , Montserrat Emilio
Published: March 2003

It has been suggested that in blast crisis (BC) of chronic myeloid leukaemia (CML) the clinical and laboratory features of patients with ‘lymphoid’ phenotype differ from those of patients with non‐lymphoid BC. In order to assess any differences, 97 patients consecutively diagnosed with BC that followed a known chronic phase of CML were analysed. 19 patients had ‘lymphoid’ BC: in 17 the blasts expressed a B‐lineage phenotype; in the remaining two they corresponded to T lymphoblasts. Four cases of B‐lineage phenotype BC were considered as biphenotypic, due to the co‐expression of myeloperoxidase and one or two other myeloid markers (CD33, CD13 and CD68) on the blast cells; in the other six cases of B‐lineage BC the blasts expressed one or both of the myeloid markers CD33 (n = 4) and CD13 (n = 3). Patients with ‘lymphoid’ BC seldom had an acceler‐ated phase prior to BC (1/19 v 36/78 with non‐lymphoid BC, P = 0.002), had less frequent splenomegaly (9/19 v 59/78, P = 0.03) and hepatomegaly (5/19 v 45/78, P = 0.02) and showed a higher degree of marrow blast infiltration (mean value 74 ± 24% v 38 ± 23%, P < 0.0001), lesser blood basophilia (2.2 ± 2.5% v 8.2 ± 7.8%, P < 0.0001), and higher serum albumin levels (P = 0.001) than those with non‐lymphoid BC. 13 patients with ‘lymphoid’ BC (68.4%) showed a favourable response to chemotherapy regimens including vincristine and prednisone and, overall, ‘lymphoid’ BC patients survived significantly longer than the remainder (median survival 12 months v 4.7 months, P = 0.006). These results indicate that ‘lymphoid’ BC of CML has a distinct clinicohaematological profile and confirm the better prognosis of such patients.
The myeloma cell antigen syndecan‐1 is lost by apoptotic myeloma cells
Authors: Jourdan Michel , Ferlin Martine , Legouffe Eric , Horvathova Mira , Liautard Janny , Rossi Jean FranÇois , Wijdenes John , Brochier Jean , Klein Bernard
Published: March 2003

Syndecan‐1 is a cell membrane proteoglycan that binds extracellular matrix components and various growth factors. It is expressed only on malignant plasma cells in bone marrow samples from patients with multiple myeloma (MM). Several reports have suggested that syndecan‐1 was present only on a part of the myeloma cells. By using either IL‐6‐dependent myeloma cell lines or primary myeloma cells stained by annexin V, we report here that syndecan‐1 was rapidly lost by myeloma cells undergoing apoptosis. In the same experimental conditions, expression of other cell membrane antigens such as CD38, HLA class‐I or CD49d on apoptotic myeloma cells was not affected. In addition, we show that syndecan‐1 loss was independent of activation of the gp130 IL‐6 transducer. Dexamethasone induced a strong apoptosis of myeloma cells associated with the loss of syndecan‐1. Finally, by using freshly‐explanted tumoural samples, we show that syndecan‐1 rapidly disappeared from myeloma cells in association with induction of apoptosis. In conclusion we showed that syndecan‐1 is a marker for viable myeloma cells which is rapidly lost by apoptotic cells. These results emphasize the usefulness of anti‐syndecan‐1 antibodies to purge tumoural cells from haemopoietic grafts or to purify these cells for further manipulations for immuno or gene therapies.
Risk of recurrent venous thromboembolism in patients with the factor V Leiden (FVR506Q) mutation: effect of warfarin and prediction by precipitating factors
Authors: Baglin Caroline , Brown Karen , Luddington Roger , Baglin Trevor ,   the East Anglian Thrombophilia Study Group
Published: March 2003

Three cohorts of patients with the factor V Leiden mutation were recruited independently (heterozygotes, homozygotes and combined thrombophilia). The antithrombotic efficacy of oral anticoagulation and the predictive value for recurrence of an idiopathic as opposed to a precipitated first event were determined. Idiopathic first events occurred at an older age than precipitated events (43 v 26 years, LR = 23.31, P < 0.001). None of the patients had a recurrent event while on warfarin but the median time to recurrence after stopping warfarin was 9 years (95%CI 0.7–17.3 years). The time to recurrence was shorter when the first event was idiopathic as opposed to precipitated (3.5 v 13 years, LR = 4.76, P = 0.029). A calculation of benefit to risk of oral anticoagulation with a target INR of 2.5 does not support the use of long‐term therapy in all patients with the factor V Leiden mutation following a first thrombotic event.
Promising activity of gemcitabine in refractory high‐grade non‐Hodgkin's lymphoma
Authors: Bernell Per , Ohm Lotta
Published: March 2003

Three patients (aged 68–75 years) with histologically confirmed relapsed or refractory high‐grade non‐Hodgkin's lymphoma were entered in this pilot study in which gemcitabine 800 mg/m2 was given as a 30 min i.v. infusion once a week for 3 weeks. One patient responded with complete remission and the other two with partial remission and stable disease for 2 and 3 months, respectively. Haematological toxicity was modest with grade 4 leucopenia (one cycle) and grade 4 thrombocytopenia (two cycles). The activity and mild toxicity seen with gemcitabine suggest that this agent should be further evaluated in the treatment of high‐grade non‐Hodgkin's lymphoma.
Primary central nervous system lymphoma in a child with acute B‐cell lymphoblastic leukaemia: consecutive Epstein‐Barr virus‐related malignancies
Authors: Buckingham Steven C. , Benaim Ely , Sandlund John T. , Gan Yan‐Jun , Freiden Pamela J. , Behm Fred G. , Ribeiro Raul C. , Sixbey John W. , Slobod Karen S.
Published: March 2003

Primary central nervous system lymphoma (PCNSL), observed among immunocompromised AIDS patients, has not been reported during chemotherapy for acute lymphoblastic leukaemia (ALL). We report a case of PCNSL occurring in a child receiving intensive multiagent chemotherapy for B‐cell ALL. In situ hybridization studies demonstrated Epstein‐Barr virus genome in both tumours, suggesting a possible link between the two diseases. The clinical response of the PCNSL to conservative therapy highlights the importance of accurately diagnosing such EBV‐related disorders, especially in patients where immune compromise can be reversed.
High incidence of anti‐heparin/platelet factor 4 antibodies after cardiopulmonary bypass surgery
Authors: Trossaërt Marc , Gaillard Anne , Commin Pierre Louis , Amiral Jean , Vissac Anne‐Marie , Fressinaud Edith
Published: March 2003

Fifty‐one patients undergoing cardiopulmonary bypass (CPB) were studied on day 0 and day 8 for heparin‐induced thrombocytopenia (HIT). The platelet aggregation test (PAT) and tests for anti‐heparin‐platelet factor 4 (anti‐H.PF4), anti‐IL8 and anti‐neutrophil activating peptide 2 (anti‐NAP2) antibodies (Ab) were performed by ELISA. On day 8, 27% of patients were positive for anti‐H.PF4Ab. None of these results were found to influence thrombotic complications or platelet counts after CPB. Our results suggest that IgG to H.PF4 may be considered a risk factor, but that additional factors must be required for HIT to develop. We conclude that assays based on platelet activation would be more appropriate for the diagnosis of HIT after CPB.
Changing bone marrow micro‐environment during development of acute myeloid leukaemia in rats
Authors: Thing Mortensen BØrge , Østrup Jensen Peter , Helledie Niels , Ole Iversen Per , RalfkiÆR Elisabeth , Knud Larsen JØrgen , Thorup Madsen Mads
Published: March 2003

The Brown Norwegian rat transplanted with promyelocytic leukaemic cells (BNML) has been used as a model for human acute myeloid leukaemia. We have previously shown that both the blood supply to the bone marrow and the metabolic rate decrease in relation to the leukaemic development in these rats. Here we have investigated how the development and progression of this leukaemia affect oxygenation, pH and proliferation of normal and leukaemic cells in vivo. Bone marrow pH was measured by a needle electrode. Nitroimidazol‐theophylline (NITP) was used to identify hypoxic cells, and we applied bromodeoxyuridine (BrdUrd) to identify DNA replicating cells.  The leukaemia progressed slowly until day 27 after which a rapid deterioration could be observed leading to severe changes over the following 5 d. In whole blood there was evidence of progressing metabolic acidosis. In bone marrow the fraction of leukaemic cells increased to > 90% and the pH dropped to about 6.5. The fraction of NITP+ cells increased to > 80% in bone marrow and to about 40% in blood. The fraction of BrdUrd+ cells was unchanged in blood, but decreased in bone marrow both for normal cells (from about 20% to 5%), and for leukaemic cells (from about 45% to 25%), evidently as a result of the severely changed micro‐ environment. In this study we have demonstrated in vivo the development of an acidic and hypoxic bone marrow hampering normal haemopoiesis during leukaemic growth. Our data support the notion of BNML as a valuable tool for studying leukaemogenesis.
Quantitation of minimal residual disease in Philadelphia chromosome positive chronic myeloid leukaemia patients using real‐time quantitative RT‐PCR
Authors: Menskin  , Van De Locht  , Schattenberg  , Linders  , Schaap  , Geurts Van Kessel  , De Witte
Published: March 2003

We used a recently developed system for real‐time quantitative polymerase chain reaction (PCR) to determine residual disease in patients with chronic myeloid leukaemia. The expression of the Bcr‐Abl hybrid oncogene was determined and normalized by using the PBGD housekeeping gene product as endogenous reference. The sensitivity and reproducibility of the assay was tested on cell line K562. A dilution of Bcr‐Abl‐positive cell line K562 remained positive at up to 250 fg of RNA. 10 copies of Bcr‐Abl DNA in water could still be detected. The dynamic range of the method spanned six orders of magnitude. Analysis of 10 identical assays on K562 RNA resulted in a variation of 15%. To test the feasibility of normalization of Bcr‐Abl dosage by the PBGD product, we compared the efficiencies of the RT‐PCRs in 150 patient analyses. We concluded that PBGD was a suitable and stringent quality control standard. Three patients who were treated with donor leucocyte infusions for chronic myeloid leukaemia who had relapsed after bone marrow transplantation were followed over time. The normalized Bcr‐Abl dosage was compared to the results of cytogenetics. Cytogenetic analysis was negative below a normalized Bcr‐Abl dose of about 3 × 10−2. This semi‐automated method is fast, sensitive and accurate and enables a high throughput of samples.
Chimaeric anti‐interleukin 6 monoclonal antibodies in the treatment of advanced multiple myeloma: a phase I dose‐escalating study
Authors: Van Zaanen  , Lokhorst  , Aarden  , Rensink  , Warnaar  , Van Der Lelie  , Van Oers
Published: March 2003

Interleukin 6 plays a key role in the pathogenesis of multiple myeloma (MM). Therefore we conducted a phase I dose‐escalating study with chimaeric monoclonal anti‐IL6 antibodies (cMab) in MM patients resistant to second‐line chemotherapy. The cMab (CLB IL6/8; Kd 6.25 × 10−12 M) was given in two cycles of 14 daily infusions, starting on day 1 and day 28, repectively, with a daily dose of 5 mg in patients 1–3, 10 mg in patients 4–6, 20 mg in patients 7–9 and 40 mg in patients 10–12 (total dose 140 mg, 280 mg, 560 mg and 1120 mg of anti‐IL6, respectively). 11/12 patients had elevated pretreatment IL6 levels.

Except for transient thrombocytopenia in two patients there was no toxicity. There were no changes in haemoglobin levels, granulocyte count, liver enzymes or renal function. No human anti‐chimaeric antibodies were induced. This was also reflected in a long half‐life time of the cMab (median 17.8 d), resulting in accumulation of the anti‐IL6 cMab and high levels of circulating IL6. However, this was in the form of biologically inactive IL6/cMab complexes and did not result in acceleration of the disease. Although C‐reactive protein (CRP) levels were decreased to below detection level in 11/12 patients, indicating effective IL6 blocking, none of the patients achieved a response according to the standard criteria. We conclude that this chimaeric anti‐IL6 Mab has a low toxicity, low immunogenicity and a long T1/2. A dose of 40 mg/d for 14 d can safely be used in future phase II studies.
Expression of the leptin receptor in human leukaemic blast cells
Authors: Nakao Takafumi , Hino Masayuki , Yamane Takahisa , Nishizawa Yoshiki , Morii Hirotoshi , Tatsumi Noriyuki
Published: March 2003

The leptin receptor is a member of the cytokine receptor superfamily, and is expressed in CD34 haemopoietic stem cells. We examined expression of the leptin receptor in fresh human leukaemia cells. Northern blot analysis showed the leptin receptor was expressed in leukaemic cells from patients with acute myeloblastic leukaemia, acute lymphoblastic leukaemia and chronic myeloid leukaemia (CML). In CML, higher expression was observed in blast crisis than in chronic phase. The expression of leptin receptor decreased during in vitro differentiation of leukaemic blast cells. It appeared that expression of the leptin receptor was associated with immature leukaemic blast cells. Our findings may indicate the possibility that leptin has some role in leukaemia.
Double heterozygosity of the GPIIb gene in a Swiss patient with Glanzmann's thrombasthenia
Authors: Ruan Jian , Peyruchaud Olivier , Alberio Lorenzo , Valles GÉraldine , Clemetson Kenneth , Bourre FranÇois , Nurden Alan T.
Published: March 2003

Glanzmann's thrombasthenia (GT) results from a qualitative or quantitative defect of GPIIb–IIIa complexes (integrin αIIbβ3), the fibrinogen receptor on platelets. This integrin plays a critical role in platelet aggregation. In this report we describe the molecular abnormalities of a patient with clinical and laboratory findings typical of type I Glanzmann's thrombasthenia. SDS‐PAGE with Western blotting revealed an absence of GPIIb but small amounts of normally migrating GPIIIa in his platelets. A non‐radioactive PCR‐SSCP procedure and direct sequence analysis of PCR‐amplified DNA fragments showed the patient to be a compound heterozygote for mutations in the GPIIb gene. A single point mutation (G to A) at nucleotide 1064 of the cDNA derived from the mother's allele led to a Glu324 to Lys amino acid substitution in GPIIb. It was responsible for a MscI restriction site in exon 12 of the GPIIb gene. This amino acid substitution changes the electric charge between the second and third Ca++‐binding domains of GPIIb. The second mutation was inherited from his father and is in exon 18 of the GPIIb gene. It was a T → C base transition at position 1787 of GPIIb cDNA and results in a Ile565 to Thr substitution. The two GPIIb mutations identified in this study will provide new information on GPIIb–IIIa structure and biosynthesis.
Review of computerized decision support systems for oral anticoagulation management
Authors: Fitzmaurice  , Hobbs  , Delaney  , Wilson  , Mcmanus
Published: March 2003

Computerized decision support systems (CDSS) are available to assist clinicians in the therapeutic management of oral anticoagulation. We report the findings relating to CDSS for oral anticoagulation management of a primary‐care‐based systematic review which largely focused on near‐patient testing. Seven papers were reviewed which covered four different systems. The methodology of these papers was generally poor, although one randomized controlled trial showed improved therapeutic control associated with computerized management compared with human performance.
Establishment and characterization of EBV‐positive and EBV‐negative primary effusion lymphoma cell lines harbouring human herpesvirus type‐8
Authors: Carbone Antonino , Cilia Anna Maria , Gloghini Annunziata , Capello Daniela , Todesco Michela , Quattrone Silvia , Volpe Rachele , Gaidano Gianluca
Published: March 2003

In this study we report on the establishment and characterization of two novel lymphoma cell lines (CRO‐AP/3 and CRO‐AP/5) which carry infection by human herpesvirus type‐8 (HHV‐8) and have derived from AIDS‐related primary effusion lymphoma (PEL). These two cell lines are representative of different virologic subtypes of PEL, i.e. HHV‐8+/EBV− PEL in the case of CRO‐AP/3 and HHV‐8+/EBV+ PEL in the case of CRO‐AP/5. Consistent with the diagnosis of PEL, both CRO‐AP/3 and CRO‐AP/5 expressed indeterminate (i.e. non‐B, non‐T) phenotypes although immunogenotypic studies documented their B‐cell origin. Both cell lines are devoid of genetic lesions of c‐MYC, BCL‐2 and p53 as well as gross rearrangements of BCL‐6. Detailed histogenetic characterization of these novel PEL cell lines suggests that PEL may derive from a post‐germinal centre B cell which has undergone pre‐terminal differentiation. The CRO‐AP/3 and CRO‐AP/5 cell lines may provide a valuable model for clarifying the pathogenesis of PEL. In particular, these cell lines may help understand the relative contribution of HHV‐8 and EBV to PEL growth and development and may facilitate the identification of recurrent cytogenetic abnormalities highlighting putative novel cancer related loci relevant to PEL.
Functional, phenotypic and molecular characterization of cytokine low‐responding circulating CD34+ haemopoietic progenitors
Authors: Pierelli Luca , Scambia Giovanni , Fattorossi Andrea , Bonanno Giuseppina , Battaglia Alessandra , Rumi Carlo , Marone Maria , Mozzetti Simona , Rutella Sergio , Menichella Giacomo , Romeo Virgilio , Mancuso Salvatore , Leone Giuseppe
Published: March 2003

Circulating CD34+ cell populations characterized by a low rate (up to five) or high rate (more than five) of cell divisions were isolated from 8 d cultures in the presence of stem cell factor (SCF), interleukin‐3 (IL‐3), granulocyte‐macrophage colony stimulating factor (GM‐CSF), granulocyte colony stimulating factor (G‐CSF), erythropoietin (EPO), Flt3 ligand and Peg‐rHu megakaryocyte growth and development factor (Peg‐rHuMGDF) using the fluorescent dye 5,6‐carboxyfluorescein diacetate succinimidyl ester (CFDA‐SE) and flow cytometric cell sorting. Phenotypic characterization of cells which had experienced up to five divisions (CFDA‐SEbright) showed a similar surface antigen expression to starting, freshly isolated CD34+ cells. Conversely, cells which experienced more than five divisions (CFDA‐SEdim) showed a differentiating behaviour, down‐regulating CD34 antigen and acquiring differentiation markers. CFDA‐SEbright cells were significantly enriched in CD105 (endoglin) positive precursors as compared to both freshly isolated CD34+ and CFDA‐SEdim cells. Functional analysis indicated that CFDA‐SEbright had a 3‐fold and 10‐fold greater cumulative cloning efficiency as compared to freshly isolated CD34+ cells and CFDA‐SEdim cells, respectively. CFDA‐SEbright cells retained the vast majority of LTC‐IC and showed a LTC‐IC frequency 2.8‐fold higher than that found in freshly isolated CD34+ cells. RT‐PCR and Western blot analyses showed significantly higher bcl‐2 RNA and protein levels in CFDA‐SEbright cells as compared to freshly isolated CD34+ and CFDA‐SEdim cells. This study indicates that cytokine low‐responding circulating CD34+ cells (CFDA‐SEbright cells) represent a functionally, phenotypically and molecularly distinct multipotent progenitor population with biological properties associated with primitive precursors.
Establishment of a novel human myeloid leukaemia cell line (FKH‐1) with t(6;9)(p23;q34) and the expression of dek‐can chimaeric transcript
Authors: Hamaguchi Hiroyuki , Nagata Kaoru , Yamamoto Katsuya , Fujikawa Izumi , Kobayashi Masaru , Eguchi Mitsuoki
Published: March 2003

Translocation t(6;9)(p23;q34), resulting in a dek‐can gene fusion, is a recurrent chromosomal abnormality mainly associated with specific subtypes of acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS). Patients with this type of chromosomal change are usually young and their prognosis is poor. The role of fusion protein generated from dek‐can chimaeric transcript on the leukaemogenesis of t(6;9) AML or MDS is as yet unknown. We have established the first permanent cell line (FKH‐1) with t(6;9), derived from the peripheral blood of a patient with t(6;9) AML transformed from Philadelphia chromosome (Ph1)‐negative chronic myelocytic leukaemia (CML). The FKH‐1 expressed myelomonocytic markers and dek‐can chimaeric transcript. In the presence of 10 ng/ml recombinant human granulocyte colony‐stimulating factor (G‐CSF), the cells doubled every 54 h and showed multilineage myeloid differentiation, resulting in heterogenous morphologies such as macrophages, basophils, eosinophils and neutrophils. Thus, this cell line may be derived from a pluripotent myeloid stem cell and should be a useful tool for biomolecular studies on the pathogenesis of t(6;9) myeloid malignancies which have rarely been investigated because of the lack of continuously proliferating cells.
Elevated SCF levels in the serum of patients with chronic renal failure
Authors: Kitoh Takashi , Ishikawa Hitoshi , Ishii Tatsuo , Nakagawa Shigeki
Published: March 2003

Serum stem cell factor (SCF) and soluble KIT (sKIT) levels were estimated in patients with chronic renal failure (CRF) and anaemia, and compared with clinical parameters of blood cells and renal function. Serum SCF levels in CRF patients were 5‐fold higher than those in healthy controls. However, serum sKIT levels in haemodialysis (HD)‐CRF patients were only slightly higher than those of healthy controls. In untreated CRF patients and healthy controls, serum SCF levels were significantly correlated with blood urea nitrogen (BUN), creatinine, haemoglobin, red blood cell (RBC) count and sKIT. In untreated CRF patients, serum SCF levels were significantly correlated with BUN, creatinine, and sKIT. These results suggest that serum SCF levels increased with the deterioration of renal function and might be related to erythropoiesis.
Detection of karyotypic aberrations in acute myeloblastic leukaemia: a prospective comparison between PCR/FISH and standard cytogenetics in 140 patients with de novo AML
Authors: Krauter Jürgen , Peter Wolfgang , Pascheberg Ulrich , Heinze Barbara , Bergmann Lothar , Hoelzer Dieter , Lübbert Michael , Schlimok Günther , Arnold Renate , Kirchner Hartmut , Port Maria , Ganser Arnold , Heil Gerhard
Published: March 2003

In 140 patients with de novo acute myeloid leukaemia (AML) standard cytogenetics were compared with RT‐PCR for the detection of t(8;21), t(15;17) and inv(16) and fluorescence in situ hybridization (FISH) for numerical aberrations of chromosomes 7, 8, X and Y. RT‐PCR detected 18 cases with t(8;21), 12 with t(15;17) and seven with inv(16). In two cases with t(8;21), two with t(15;17) and four with inv(16) these aberrations had not been detected by standard cytogenetics. There were no false negative PCR results. In 12 patients with these chromosomal changes, standard cytogenetics revealed additional chromosomal aberrations. In 16 patients sole numerical aberrations of the chromosomes 7, 8, X or Y were found by FISH. In these patients the sensitivity of FISH and standard cytogenetics was comparable. In 87 patients no aberrations could be found by PCR and FISH whereas in 24 of these patients standard cytogenetics revealed an abnormal karyotype. These data recommend the combination of standard cytogenetics and molecular techniques to improve the sensitivity for the detection of genetic lesions in AML. Once chromosomal markers have been identified by combined analysis these markers could be used to monitor residual disease during/after chemotherapy, by RT‐PCR and/or FISH.
pH is a potent modulator of erythroid differentiation
Authors: Mcadams Todd A. , Miller William M. , Papoutsakis E. Terry
Published: March 2003

Physiological parameters such as pH and oxygen tension probably play significant roles in the regulation of haemopoiesis in the bone marrow microenvironment, but these roles have yet to be characterized in detail. We have found that changes in culture pH (0.2 units) can cause significant changes in the culture composition of mature cells and colony‐forming cells (CFCs), especially in the presence of erythropoietin (Epo). Peripheral blood (PB) CD34+ cells cultured at different pH values (7.15–7.6) were characterized using total cell counts, colony assays, morphological analysis, haemoglobin staining, flow cytometry, immunocytochemical staining, and Western blots. Cultures performed at high (7.6) pH contained greater numbers of haemoglobin‐positive and band‐3‐positive cells, and acquired these erythroid differentiation markers sooner than standard (7.35) and low (7.1) pH cultures. Flow cytometry using CD71 and CD45RA antigens also indicated that erythroid differentiation proceeds faster at high pH and is blocked at an intermediate stage by low pH. Morphological data confirmed that high pH cultures had been shifted towards late‐stage erythroid compartments as compared to low and standard pH cultures. These findings have important implications both in elucidating the regulatory role of pH in the bone marrow microenvironment and for the design of in vitro systems to study the development of erythroid cells.
Indolent course as a relatively frequent presentation in T‐prolymphocytic leukaemia
Authors: Garand Richard , Goasguen Jean , Brizard AndrÉ , Buisine Jacqueline , Charpentier AgnÈs , FranÇois Claisse Jean , Duchayne Eliane , Lagrange Monique , Segonds Christine , Troussard Xavier , Flandrin Georges ,   the Groupe Français d'Hématologie Cellulaire
Published: March 2003

T‐prolymphocytic leukaemia (T‐PLL) is a rare disorder with a poor outcome. Presentation features were studied in 78 T‐PLL cases. Although 53 patients (group A) presented with typical progressive disease including rapidly increasing leucocytosis, 25 patients (group B) experienced an initial indolent clinical course with stable moderate leucocytosis. The morphology and antigenic profile of abnormal cells were similar in both groups, except for a lower incidence of CD45RO+ CD45RA− pattern in group B. A high incidence of inv(14)(q11;q32), t(14;14)(q11;q32) and i(8)(q10) chromosomal abnormalities were found in both groups. After an initial indolent phase (median 33 months; 6–103 months), 16 group B patients progressed to an aggressive stage with clinical and laboratory features similar to group A. Moreover, median survival after progression was short in both groups. In conclusion, T‐PLL may start as an indolent disease similar to that reported in ataxia telangectasia. In this rare genetic disorder, some patients develop stable T‐cell clones which progress toward T‐PLL‐like leukaemia. Moreover, ATM gene mutations have been reported in T‐PLL. Thus, both diseases are likely to be closely related.
Ifosfamide and vinorelbine: an active regimen for patients with relapsed or refractory Hodgkin's disease
Authors: Bonfante  , Viviani  , Santoro  , Devizzi  , Di Russo  , Zanini  , Soncini  , Soto Parra  , Valagussa  , Bonadonna
Published: March 2003

Twenty‐six patients with relapsed or refractory Hodgkin's disease (HD) were treated with an intensive salvage regimen combining ifosfamide (3000 mg/m2/d, days 1–4 through continuous intravenous infusion) and vinorelbine (25 mg/m2, i.v. days 1 and 5) with mesna uroprotection and G‐CSF support. Courses were given at 3‐week intervals.

Ten patients achieved a complete and 10 patients a partial response, yielding an overall response rate of 77%. The main toxic effect was neutropenia and the combination was well tolerated.
Haemopoietic cell transplantation in children with juvenile myelomonocytic leukaemia
Authors: Macmillan  , Davies  , Orchard  , Ramsay  , Wagner
Published: March 2003

Seven children, 11 months to 5.9 years of age, with juvenile myelomonocytic leukaemia (JMML) underwent allogeneic haemopoietic cell transplantation (HCT) using related or unrelated donor bone marrow or umbilical cord blood. All patients had active disease at time of transplant despite chemotherapy in five patients and chemotherapy and splenectomy in one patient prior to HCT conditioning. All patients received cyclophosphamide and TBI, with the addition of busulphan in two cases. Engraftment was documented in all cases. Notably, six of seven patients relapsed after allogeneic HCT with one achieving a return to full donor chimaerism after cyclosporin A (CSA) withdrawal. Two patients are alive in remission, 23+ and 30+ months after transplant. The role of allogeneic HCT in patients with JMML is discussed. A cooperative multicentre trial is needed to establish the optimal therapy for these patients.
Splenic irradiation for symptomatic splenomegaly associated with myelofibrosis with myeloid metaplasia
Authors: Elliott Michelle A. , Chen Michael G. , Silverstein Murray N. , Tefferi Ayalew
Published: March 2003

Twenty‐three patients who had myelofibrosis with myeloid metaplasia (MMM) were treated at our institution with 50 courses of splenic irradiation (SI) for symptomatic splenomegaly. The median dose of radiation per course was 277.5 cGy, administered in a median of 7.5 fractions. 8/23 patients received multiple courses of SI. Of 49 evaluable courses of SI, 46 (93.9%) resulted in an objective decrease in spleen size. The median duration of response was 6 months (range 1–41). Reduction in spleen size was associated with symptomatic relief in all patients. Overall median survival after SI was 22 months. Significant cytopenia occurred in 10 (43.5%) patients, or 16 (32%) of the 50 courses of SI. Prolonged, life‐threatening pancytopenia after a single course of SI occurred in six patients (26%), resulting in fatal sepsis or haemorrhage in three (13%). Nine patients underwent subsequent splenectomy; the perioperative mortality rate was 11%. One third of patients experienced postoperative intra‐abdominal haemorrhage necessitating surgical re‐exploration. SI can provide symptomatic relief and a reduction in spleen size in most MMM patients. The increased risk of postoperative bleeding in patients requiring subsequent splenectomy dictates against considering SI as an alternative to splenectomy for patients who are otherwise good surgical candidates.
Salvage of patients with acute promyelocytic leukaemia with residual disease following ABMT performed in second CR using all‐trans retinoic acid
Authors: Grimwade David , Jamal Rahman , Goulden Nicholas , Kempski Helena , Mastrangelo Stefano , Veys Paul
Published: March 2003

Detection of residual disease after completion of therapy or following bone marrow transplantation (BMT) in patients with acute promyelocytic leukaemia (APL) predicts relapse and is associated with a poor prognosis. Here we describe the successful treatment of residual disease post‐transplant in APL using prolonged all‐trans retinoic acid (ATRA) therapy in two children in whom autologous BMT (ABMT) had been performed in second complete remission (CR). ATRA treatment was well tolerated and found to be beneficial despite its prior use as a component of the initial induction protocol. ATRA therapy post‐transplant led to molecular remission as determined by fluorescence in situ hybridization (FISH) as well as reverse transcriptase‐polymerase chain reaction (RT‐PCR) analyses and remission now exceeds 3.5 years in both patients. Overall, this study not only demonstrates that ATRA may successfully salvage APL patients with residual disease post‐transplant, but also suggests a potential role for retinoids post‐consolidation as a means of eliminating residual disease which could be beneficial even in patients previously exposed to ATRA as a component of the induction protocol.
No treatment for low‐risk thrombocythaemia:results from a prospective study
Authors: Ruggeri  , Finazzi  , Tosetto  , Riva  , Rodeghiero  , Barbui
Published: March 2003

Essential thrombocythaemia (ET) is a chronic myeloproliferative disorder characterized by the occurrence of thromboembolic episodes, particularly in patients aged > 60 years or with a previous history of thrombosis, and/or by haemorrhages in patients with an exceedingly high platelet count. In these subgroups of patients the use of cytoreductive therapy is beneficial in terms of risk/benefit ratio. Only limited anecdotal data are available on the thrombotic or haemorrhagic risk and survival in young asymptomatic ET patients with a platelet count < 1500 × 109/l. Therefore the optimal management of these patients is unknown.

To assess the incidence of thrombosis and haemorrhages in this group of patients we carried out a prospective observational study in a cohort of 65 patients with ET, aged < 60 years, with no history of thrombosis or haemorrhage and platelet count < 1500 × 109/l, and in 65 age‐ and sex‐matched controls. Patients were not treated with cytoreductive therapy until the occurrence of thrombosis or haemorrhage. Arterial or venous thrombotic events were objectively documented both in cases and in controls.

The median follow‐up was 4.1 years, with an incidence of thrombosis in patients and controls of 1.91 and 1.50 cases/100 patient‐years, respectively. The age‐ and sex‐adjusted risk rate ratio was 1.43 (95% CI 0.37–5.4). Only three minor haemorrhagic episodes occurred in patients, with an incidence of 1.12 cases/100 patient‐years.

Pregnancy and surgery were not associated with thrombosis in these patients.

We conclude that the thrombotic risk in young ET patients, with no thrombotic history and a platelet count < 1500 × 109/l, is not increased compared to the normal population and that a conservative therapeutic approach should therefore be considered in these patients.
Additional genetic risk factors for venous thromboembolismin carriers of the factor V Leiden mutation
Authors: Tosetto Alberto , Rodeghiero Francesco , Martinelli Ida , De Stefano Valerio , Missiaglia Edoardo , Chiusolo Patrizia , Mannucci Pier Mannuccio
Published: March 2003

Only a minority of subjects with factor V (FV) Leiden mutation develop venous thromboembolism (VTE), suggesting that additional genetic risk factors may be present in symptomatic carriers. We screened 157 unrelated carriers of the FV Leiden mutation with a first episode of VTE and 291 unrelated asymptomatic FV carriers for the presence of two frequent mutations, i.e. G20210A of the prothrombin gene and C677T of the methylenetetrahydrofolate reductase gene. Carriers with other inherited or acquired thrombophilia‐associated abnormalities were excluded from analysis. Heterozygotes for the G20210A mutation were more prevalent among symptomatic carriers than in asymptomatic carriers (10.8% v 2.7%, P < 0.0001); homozygotes for the C677T mutation were also more prevalent in symptomatic carriers (21.6% v 14.4%, P = 0.05). Factor V Leiden carriers who had had a VTE episode during oral contraceptive intake were more frequently carriers of the G20210A mutation (14.3%, P = 0.03). These results further support the idea that VTE in carriers of FV Leiden results from interaction with additional genetic or circumstantial risk factors, and that an accurate search for such factors is required to identify carriers at risk.
Chromosomal abnormalities in systemic amyloidosis
Authors: Fonseca Rafael , Ahmann Gregory J. , Jalal Syed M. , Dewald Gordon W. , Larson Dirk R. , Therneau Terry M. , Gertz Morie A. , Kyle Robert A. , Greipp Philip R.
Published: March 2003

Primary systemic amyloidosis (AL) is a plasma cell disorder characterized by deposition of monoclonal light chains in different organ systems. Although multiple and complex numerical chromosomal abnormalities have been described in patients with multiple myeloma, it is currently unknown whether such changes occur in systemic amyloidosis.

Bone marrow samples from 21 patients with AL were studied by standard cytogenetics and interphase fluorescencein situhybridization (FISH) for the presence of numerical chromosomal abnormalities. We tested for six chromosomes (7, 11, 9, 15, 18 and X) using centromere‐specific probes. The monoclonal plasma cells were identified by simultaneous fluorescent staining of the monotypic cytoplasmic immunoglobulin. We compared these results with those obtained from 19 patients with monoclonal gammopathy of undetermined significance (MGUS) and normal controls.

Multiple numerical chromosomal abnormalities were detected in AL by interphase FISH, including trisomy of chromosomes 7 (42%), 9 (52%), 11 (47%), 15 (39%), 18 (33%) and X (13% in women and 54% in men). Monosomy of chromosome 18 was seen in 72% of cases. Previous exposure to alkylator therapy did not appear to correlate with these abnormalities. No significant difference was observed in the prevalence of these abnormalities between AL and MGUS.

Multiple chromosomal numerical abnormalities were detected by interphase FISH analysis in patients with AL, especially monosomy of chromosome 18. Aneuploidy in the monotypic plasma supports a neoplastic nature for the disorder.
Evidence of cytogenetic and molecular remission by allogeneic cells after immunosuppressive therapy alone
Authors: Carella Angelo M. , Lerma Enrica , Corsetti Maria T. , Dejana Anna , Celesti Lidia , Casarino Lucia , De Stefano Francesco , Frassoni Francesco
Published: March 2003

An immunosuppressive but not myeloablative regimen followed by HLA‐matched donor mobilized haemopoietic stem cell transplantation was employed in two high‐risk patients. The first patient had refractory anaemia with excess blasts (RAEB) and cytogenetic evidence of translocation 1;3(p36;q21). The second patient had Philadelphia‐negative but p190 BCR‐ABL chimaeric gene positive chronic myelogenous leukaemia in accelerated phase (AP‐CML). The conditioning regimen consisted of fludarabine (30 mg/m2/d, days 1–3) with cyclophosphamide (300 mg/m2/d, days 1–3). Cyclosporine and methotrexate were employed for acute graft‐versus‐host disease (aGVHD) prophylaxis. In both cases the engraftment of donor cells was demonstrated by cytogenetics and short tandem repeat polymorphisms via PCR. Both patients are alive with normal cytogenetic (RAEB) and molecular (AP‐CML) remissions, 100 and 150 d after allografting, respectively. In particular, in the AP‐CML patient, the BCR‐ABL became undetectable and the BCR‐ABL/ABL ratio was <0.0001.
Differential effects of three radiographic contrast media on platelet aggregation and degranulation: implications for clinical practice?
Authors: Heptinstall S. , White A. , Edwards N. , Pascoe J. , Sanderson H. M. , Fox S. C. , Henderson R. A.
Published: March 2003

We have determined the effects of three radiographic contrast media on platelet aggregation and degranulation in vitro. Aggregation was measured as loss of single platelets, and degranulation was measured as P‐selectin expression using flow cytometry. Iopamidol added to hirudinized blood induced aggregation directly and also potentiated that induced by weak platelet agonists such as adenosine diphosphate (ADP). Iodixanol also potentiated platelet aggregation, but ioxaglate inhibited it. Iopamidol also caused marked platelet degranulation. The pro‐aggregatory effect of iopamidol was evident in non‐anticoagulated blood as well as in hirudinized blood, but not in citrated blood. In platelet‐rich plasma (PRP) prepared from hirudinized blood neither iopamidol nor iodixanol directly induced platelet aggregation, but they rendered platelets hypersensitive to ADP. ADP antagonists inhibited the platelet aggregation and degranulation induced by iopamidol in whole blood, whereas aspirin, an inhibitor of thromborane A2 synthesis, did not. These data are consistent with clinical reports of increased thromboembolic risk with non‐ionic low‐osmolar media, and raise concerns about the routine use of these contrast media during diagnostic and interventional arteriographic procedures. Routine use of citrate in previous experiments may have masked a pro‐aggregatory effect of some contrast media.
Possible presence of enhancing antibodies in idiopathic thrombocytopenic purpura
Authors: Yang Yi‐Yuan , Fischer Peter , Leu Sy‐Jye , Zhu Min , Woods, Jr Virgil L , Chen Pojen P
Published: March 2003

It is difficult to detect IgG anti‐platelet autoantibodies in idiopathic thrombocytopenic purpura (ITP). Recently, it was reported that reactivity with glycoprotein IIb/IIIa was lost when IgG anti‐GPIIb/IIIa antibodies from seven ITP patients were digested with pepsin to yield F(ab′)2 fragments. These findings suggested that some IgG anti‐platelet autoantibodies in ITP may be of low affinity and thus require the presence of ‘enhancing’ anti‐IgG antibodies (i.e. rheumatoid factors, RFs) for detection. To test this hypothesis, we used a phage display technique to isolate five IgG RFs from an ITP patient (patient 1). Sequence analysis revealed that these RFs consisted of two clones, represented by GG3 and GG48. Both representative RFs bound specifically to IgG Fc fragments with apparent dissociation constants of 8.2 × 10−8 m and 8.8 × 10−7 m, respectively. Moreover, IgG RFs were subsequently found in a serum sample from patient 1. Combined, these results suggest that IgG RFs may occur in ITP, and may be required for the detection of some IgG anti‐platelet autoantibodies and for the corresponding antibody‐mediated platelet destruction in autoimmune ITP.
Associations of blood rheology and interleukin‐6 with cardiovascular risk factors and prevalent cardiovascular disease
Authors: Woodward Mark , Rumley Ann , Tunstall‐Pedoe Hugh , Lowe Gordon D. O.
Published: March 2003

Haemorheological variables (whole‐blood, plasma and relative blood viscosity, haematocrit, red cell aggregation, white cell count and fibrinogen) were measured in 753 men and 821 women aged 25–74 years, and related to cardiovascular risk factors and prevalent cardiovascular disease (CVD). Men had higher levels than women of blood viscosity, haematocrit, corrected viscosity and relative viscosity. Post‐menopausal women had higher levels than pre‐menopausal women of blood viscosity, haematocrit, corrected blood viscosity, plasma viscosity and fibrinogen: each of these differences was completely or partly abolished by use of hormone replacement therapy. Serum total cholesterol, triglycerides, diastolic blood pressure, body mass index and smoking markers showed positive associations with most rheological variables, whereas HDL‐cholesterol, plasma vitamin C and social class showed inverse associations. Rheological variables were associated with prevalent CVD after age‐adjustment. However, after multiple risk factor adjustment only plasma viscosity and red cell aggregation showed significant (P < 0.04) associations in both men and women (comparing top to bottom quarters). Plasma interleukin‐6 (measured in a 25% subsample of 196 men and 221 women) correlated significantly with age, fibrinogen, white cell count, plasma and blood viscosity, current smoking, and (in men) with low serum vitamin C levels; but not with other major risk factors or with prevalent cardiovascular disease.
Economic impact of using clodronate in the management of patients with multiple myeloma
Authors: Bruce N. J. , McCloskey E. V. , Kanis J. A. , Guest J. F.
Published: March 2003

The economic impact of using prophylactic clodronate as an adjunct to chemotherapy in the management of multiple myeloma for the first 4 years following diagnosis was established from the perspective of the National Health Service (NHS). A state‐transition model of the course of multiple myeloma was constructed using the MRC VI myelomatosis trial results and information on patient management obtained retrospectively from clinical trialists. Data were collected on resource use and corresponding costs for standard management and managing severe hypercalcaemia, vertebral and non‐vertebral fractures. Managing patients with prophylactic clodronate cost the NHS a mean £22 934 per patient; comprising £16 697 for standard management, £4862 for clodronate therapy and £1376 for adverse events. Managing patients without prophylactic clodronate cost a mean £19 557 (£16 697 and £2860 for standard management and adverse events respectively). Therefore prophylactic clodronate therapy increased the cost by £3377, or 17% per patient. Hospitalization accounted for 32% of the total cost, whereas chemotherapy accounted for 5%. The results were robust to sensitivity analyses (range £2605–£4150). Further studies are required to assess the impact of prophylactic clodronate on quality of life to enable the clinical benefits and additional cost of this treatment to be compared with other healthcare interventions.