bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell density‐dependent VP‐16 sensitivity of leukaemic cells is accompanied by the translocation of topoisomerase IIα from the nucleus to the cytoplasm
Authors: Valkov  , Gump  , Engel  , Sullivan
Published: March 2003

The resistance of several leukaemic and myeloma cell lines (CCRF, L1210, HL‐60, KG‐1a and RPMI 8226) to VP‐16 was found to increase with cell density and to be maximal (3.5‐ to 39‐fold) in plateau phase cell cultures, as measured by clonogenic and MTT assays. Non‐transformed confluent Flow 2000 human fibroblasts and Chinese hamster ovary (CHO) cells were also five‐ and 15‐fold resistant to VP‐16 respectively. The transition from log to plateau phase was accompanied by a drastic decrease in topoisomerase (topo) IIα content in CHO cells and human fibroblasts, while the leukaemic cells maintained constant cellular levels of topo IIα and topo IIβ. However, the nuclear topo IIα content was found to decrease as a result of translocation of the enzyme to the cytoplasmic compartment in the leukaemic cells. This was confirmed by subcellular fractionation experiments, Western blotting analyses and immunocytochemistry studies. The quantity of topo IIα in plateau phase cytoplasmic fractions ranged from 18% in L1210 cells to 50% in HL‐60 and 8226 cells, as measured by both immunoblotting and quantification of the label in immunofluorescent images. The cytoplasmic fraction from plateau phase cells retained topo II catalytic activity, as measured by the decatenation of kinetoplast DNA. The nuclear–cytoplasmic ratio of topo IIα may be critical in determining the sensitivity of leukaemic cells to topo II inhibitors. Cytoplasmic trafficking of topo IIα was observed in plasma cells obtained from patients with multiple myeloma, and perhaps contributes to drug resistance in this disease.
Comparative analyses of megakaryocytes derived from cord blood and bone marrow
Authors: Miyazaki Rika , Ogata Hajime , Iguchi Tomoko , Sogo Sinji , Kushida Taketoshi , Ito Tomoki , Inaba Muneo , Ikehara Susumu , Kobayashi Yohnosuke
Published: March 2003

Thrombocytopenia is typically observed in patients undergoing cord blood transplantation. We hypothesized that delayed recovery of the platelet count might be caused by defects in the megakaryocytic differentiation pathway of cord blood progenitors. To test this hypothesis, we compared the features of in vitro megakaryocytopoiesis between cord blood progenitors and those in bone marrow cells after isolation of CD34+ cells as progenitors. The proliferative responses of the progenitors in cord blood are higher than those in bone marrow cells in the presence of interleukin (IL)‐3, stem cell factor (SCF) and thrombopoietin (TPO). However, the ability to generate mature megakaryocytes was higher in bone marrow progenitors than in cord blood in the same in vitro culture system, when examined by the expression of CD41, polyploidy and proplatelet formation. Furthermore, an earlier induction of c‐mpl protein, a receptor for TPO, was observed in the progenitors from bone marrow than in those from cord blood in the presence of SCF and IL‐3. Therefore, the ability to generate mature megakaryocytes in bone marrow progenitors is superior to that in cord blood, and the delayed engraftment of platelets after cord blood transplantation might be attributed to the features of cord blood megakaryocyte progenitors.
Transfusion of platelet concentrates cryopreserved with ThromboSol plus low‐dose dimethylsulphoxide in patients with severe thrombocytopenia: a pilot study
Authors: Pedrazzoli Paolo , Noris Patrizia , Perotti Cesare , Schiavo Roberta , Ponchio Luisa , Belletti Simona , Da Prada Gian Antonio , Balduini Carlo Luigi , Salvaneschi Laura , Robustelli della Cuna Gioacchino , Siena Salvatore
Published: March 2003

We have recently reported the possibility of supporting the phase of severe thrombocytopenia after high‐dose chemotherapy (HDC) and stem cell transplantation using 5% dimethylsulphoxide (DMSO)‐cryopreserved autologous platelet concentrates (PCs). The aim of the present study was to evaluate the therapeutic potential of ThromboSol (a recently developed platelet storage solution) plus PCs cryopreserved in 2% DMSO in patients undergoing myeloablative chemotherapy and autologous transplantation. PCs were collected from 14 women with breast cancer by a single plateletapheresis and cryopreserved in ThromboSol/2% DMSO by either direct insertion in a −80°C freezer or in liquid nitrogen after computer‐controlled rate (CR) freezing. When required, PCs were thawed, centrifuged to remove the cryoprotectants and transfused. In vitro studies on thawed platelets showed loss of epitopes of surface glycoproteins and a marked reduction of functional activity compared with fresh platelets. Transfusion of CR‐frozen PCs was associated with a mean 1 h corrected count increment (CCI) of 9.2 ± 5.4 × 109/l and only one allogeneic PC was required in this group. In contrast, six out of seven patients required additional allogeneic transfusions in the −80°C group (CCI = 2.7 ± 1.4 × 109/l). ThromboSol‐treated PCs have the ability to overcome thrombocytopenia if processed by a CR freezing protocol, but appear ineffective when frozen by direct placing at −80°C.
Oral magnesium pidolate: effects of long‐term administration in patients with sickle cell disease
Authors: De Franceschi  , Bachir  , Galacteros  , Tchernia  , Cynober  , Neuberg  , Beuzard  , Brugnara
Published: March 2003

Prevention of erythrocyte dehydration by specific blockade of the transport pathways promoting loss of potassium (K) is a potential therapeutic strategy for sickle cell (SS) disease. Dietary magnesium (Mg) pidolate supplementation over a 4‐week period has been shown to inhibit K–Cl co‐transport and reduce dehydration. We report here the results in 17 of 20 patients with SS disease treated in an open‐label unblinded study of the effects of long‐term (6 months) oral Mg pidolate administration (540 mg Mg/d). A significant decrease (P < 0.0025) was observed with Mg therapy in the distribution widths for red cell mean cell haemoglobin concentration (MCHC) (haemoglobin distribution width; HDW), reticulocyte mean cell volume (red cell distribution width of reticulocytes; RDWr) and MCHC (reticulocyte HDW; HDWr), activity of red cell K–Cl co‐transport, Na/Mg exchanger and Ca2+‐activated (Gardos) K+ channel, whereas red cell K and Mg contents were significantly increased. Hb levels and absolute reticulocyte counts did not change with Mg therapy. Two patients did not complete the trial because of diarrhoea and one did not complete the trial for unrelated reasons. Although the median number of painful days in a 6‐month period decreased from 15 (range 0–60) in the year before the trial to 1 (range 0–18; P < 0.0005) during the period of Mg therapy, no firm conclusion on therapeutic efficacy could be drawn from this unblinded open‐label trial.
Safety profile of the oral iron chelator deferiprone: a multicentre study
Authors: Cohen  , Galanello  , Piga  , DiPalma  , Vullo  , Tricta
Published: March 2003

In previous trials, the orally active iron chelator deferiprone (L1) has been associated with sporadic agranulocytosis, milder forms of neutropenia and other side‐effects. To determine the incidence of these events, we performed a multicentre prospective study of the chelator. Blood counts were performed weekly, and confirmed neutropenia mandated discontinuation of therapy. Among 187 patients with thalassaemia major, the incidence of agranulocytosis (neutrophils < 0.5 × 109/l) was 0.6/100 patient–years, and the incidence of milder forms of neutropenia (neutrophils 0.5–1.5 × 109/l) was 5.4/100 patient–years. All cases of neutropenia resolved after interruption of therapy. Neutropenia occurred predominantly in non‐splenectomized patients. Nausea and/or vomiting occurred early in therapy, was usually transient and caused discontinuation of deferiprone in three patients. Mild to moderate joint pain and/or swelling did not require permanent cessation of deferiprone and occurred more commonly in patients with higher ferritin levels. Mean alanine transaminase (ALT) levels rose during therapy. Increased ALT levels were generally transient and occurred more commonly in patients with hepatitis C. Persistent changes in immunological studies were infrequent, although sporadic abnormalities occurred commonly. Mean zinc levels decreased during therapy. Ferritin levels did not change in the overall group but decreased in those patients with baseline levels > 2500 μg/l. This study characterized the safety profile of deferiprone, and, under the specific conditions of monitoring, demonstrated that agranulocytosis is less common than previously predicted.
Comparing near misses with actual mistransfusion events: a more accurate reflection of transfusion errors
Authors: Ibojie  , Urbaniak
Published: March 2003

In a retrospective review of transfusion errors in a large teaching hospital, we found the true incidence of errors to be at least four times the actual mistransfusion events detected. Seventy‐five per cent of the errors were detected as near misses. The mistransfusions equated to 1/8610 compatibility procedures, and 1/27 007 units of blood issued, whereas the number of true transfusion errors equates to 1/2153 compatibility procedures and 1/6752 units of blood issued. The major error‐prone activities included patient identification at phlebotomy and the final infusion of the blood product at the bedside. Of the cases, 95.2% were due to non‐compliance with existing guidelines. Potential disasters were avoided only by the vigilance of the blood bank staff and the systems in place to detect errors.
IgA myeloma of donor origin arising 7 years after allogeneic renal transplant
Authors: Grey Mark , Townsend Neal , Lappin David , Roberts Ian , Stanford Rachel , Sheldon Melanie , Tate David , Short Colin , Lucas Guy
Published: March 2003

We report the case of a 38‐year‐old man who developed IgA myeloma of donor origin 7 years after allogeneic renal transplant. The diagnosis of multiple myeloma was unequivocal and based on positive results from serum electrophoresis, skeletal survey and bone marrow investigations. The donor origin of the myeloma cells was confirmed by DNA fingerprinting. We believe this is the first reported case of disseminated multiple myeloma of donor origin developing after allogeneic renal transplant and, as such, gives some insight into the natural history and biology of the disease.
Investigation of the effect of marathon running on leucocyte counts of subjects of different ethnic origins: relevance to the aetiology of ethnic neutropenia
Authors: Bain Barbara J. , Phillips Doraline , Thomson Kirsty , Richardson David , Gabriel Ian
Published: March 2003

Healthy subjects of African ancestry, including Afro‐Caribbeans, have been observed to have lower total white cell counts and neutrophil counts than healthy Caucasian subjects. The cause of this ethnic neutropenia is unclear. We have previously found no evidence that increased margination of neutrophils is responsible. In this study, we have investigated mobilization of neutrophils from the bone marrow granulocyte reserve by endurance exercise. We investigated subjects of different ethnic origins before and after they had competed in a marathon race. Before the race, the neutrophil counts of Africans/Afro‐Caribbeans were significantly lower than those of Caucasians (means 2.49 and 3.21 × 109/l respectively; P = 0.043). After the race, the difference was greater, as was the degree of significance (means 10.21 and 12.33 respectively; P = 0.017). The percentage increment was almost identical whereas the absolute increment was considerably less, although of marginal statistical significance (increment 7.72 and 9.12 × 109/l respectively; P = 0.10). Similar changes were observed in the monocyte count. Before the race, the difference in the means was of marginal statistical significance (0.35 and 0.41 × 109/l respectively; P = 0.105), whereas after the race the difference was greater and was highly significant (means 0.75 and 1.05 × 109/l respectively; P = 0.001). These observations support the results of our earlier study, both of which suggested that ethnic neutropenia is likely to result from a diminished bone marrow reserve rather than being consequent on altered distribution of neutrophils within the blood stream.
Early haemorrhagic morbidity and mortality during remission induction with or without all‐trans retinoic acid in acute promyelocytic leukaemia
Authors: Di Bona Eros , Avvisati Giuseppe , Castaman Giancarlo , Luce Vegna Maria , De Sanctis Vitaliana , Rodeghiero Francesco , Mandelli Franco ,   for the Gruppo Italiano per le Malattie Ematologiche dell'Adulto
Published: March 2003

A total of 622 consecutive patients with acute promyelocytic leukaemia (APL) treated within the Gruppo Italiano per le Malattie Ematologiche dell'Adulto (GIMEMA) group during 1989–97 have been reviewed to assess the clinical effectiveness of all‐trans retinoic acid (ATRA) on the incidence of early haemorrhagic deaths and on APL‐associated coagulopathy. Of them, 499 were treated with idarubicin plus ATRA (study A) and 123 with Idarubicin alone (study B). In both studies, similar guidelines for supportive treatment were used. Haemorrhagic symptoms were evaluated according to a reproducible score system. Deaths occurring within 10 d of starting treatment were 19 (3.8%) in study A and nine (7.3%) in study B (P = 0.09), with 15 (3%) and five (4.1%) (P not significant) due to haemorrhage. Overall, induction mortality was 7.6% and 16.2% respectively (P < 0.003). In study A, days with platelet counts ≤ 20 × 109/l or with fibrinogen ≤ 1 g/L were reduced by about 30%, the haemorrhagic score by 50% and the consumption of blood products by about 40%, and fewer patients were treated with antihaemorrhagic drugs (39% vs. 61%; P < 0.001). On multivariate analysis, early deaths were influenced by blast count at diagnosis > 30 × 109/l (P < 0.001) in both studies, and by a haemorrhagic score of 3 in study A (P < 0.001).

Although the reduction of early fatal haemorrhages was not significant, a substantial clinical improvement was evident in terms of reduction of the severity of bleeding symptoms, blood product consumption and overall induction mortality when ATRA was combined with idarubicin.
The G1691 → A mutation of factor V, but not the G20210 → A mutation of factor II or the C677 → T mutation of methylenetetrahydrofolate reductase genes, is associated with venous thrombosis in patients with lupus anticoagulants
Authors: Galli Monica , Finazzi Guido , Duca Francesca , Norbis Francesca , Moia Marco
Published: March 2003

Arterial and venous thrombosis are the most common manifestations of antiphospholipid syndrome. To investigate whether genetic determinants contribute to their thrombotic risk, we studied the prevalence of the G1691 → A mutation in the gene coding for factor V, the G20210 → A mutation in the prothrombin gene and the C677 → T mutation in the methylenetetrahydrofolate reductase gene in 152 patients with lupus anticoagulants. One hundred and twenty‐eight cases (84%) also had increased titres of anticardiolipin antibodies. History of thrombosis was present in 96 patients (63%); 67 suffered from venous thrombosis only, 23 cases had arterial thrombosis only, six patients had both venous and arterial thrombosis. Five patients were heterozygous for the G1691 → A mutation in the factor V gene (3%). All of them (100%) suffered from venous thrombosis compared with 68 out of the 147 cases without the mutation (46%) (P = 0.0474). The prevalence of the G20210 → A mutation in the prothrombin gene was evaluated in 145 patients; eight of these patients were heterozygous (5%). Four of these patients (50%) experienced venous thrombosis compared with 65 out of the 137 patients without the mutation (47%) (P = ns). Neither mutation was associated with arterial thrombotic events. No patient carried both mutations. The C677 → T mutation in the methylenetetrahydrofolate reductase gene was assessed in 83 patients; 15 of them (18%) were homozygous and 37 (44%) were heterozygous. There was no significant association between the status of the mutation and history of venous and arterial thrombosis. No significant correlation was found among the three groups. In conclusion, only the G1691 → A mutation in the factor V gene was associated with the thrombotic risk of patients with lupus anticoagulants.
Performance of the OptiMAL malaria antigen capture dipstick for malaria diagnosis and treatment monitoring at the Hospital for Tropical Diseases, London
Authors: Moody Anthony , Hunt‐Cooke Angela , Gabbett Elisabeth , Chiodini Peter
Published: March 2003

We report here the sensitivity and specificity of OptiMAL for the diagnosis of acute malaria in patients presenting to the Hospital for Tropical Diseases (HTD), a tertiary referral centre for Tropical and Infectious diseases. A sensitivity of 95.3% and a specificity of 100% for Plasmodium falciparum and a sensitivity of 96% and a specificity of 100% for Plasmodium vivax was obtained. The ability to follow the course of the parasitaemia using OptiMAL during treatment and its significance for use in areas where expert microscopy is not available is discussed.
Resveratrol induces Fas signalling‐independent apoptosis in THP‐1 human monocytic leukaemia cells
Authors: Tsan Min‐Fu , White Julie E. , Maheshwari Jewraj G. , Bremner Theodore A. , Sacco Joseph
Published: March 2003

Resveratrol, a natural product present in wine, has recently been shown to inhibit the growth of a number of cancer cell lines in vitro. In the current study, we have demonstrated that resveratrol inhibits the growth of THP‐1 human monocytic leukaemia cells in a dose‐dependent manner with a median effective dose of 12 μm. It did not induce differentiation of THP‐1 cells and had no toxic effect on THP‐1 cells that had been induced to differentiate into monocytes/macrophages by phorbol myristate acetate. A significant fraction of resveratrol‐treated cells underwent apoptosis as judged by flow cytometric analysis of DNA content, DNA fragmentation and caspase‐specific cleavage of poly(ADP‐ribosyl) polymerase. Resveratrol treatment had no effect on the expression of Fas receptor or Fas ligand (FasL) in THP‐1 cells, nor did it induce clustering of Fas receptors. In addition, THP‐1 cells were resistant to activating anti‐Fas antibody, and neutralizing anti‐Fas and/or anti‐FasL antibodies had no protective effect against resveratrol‐induced inhibition of THP‐1 cell growth. The effect of resveratrol on THP‐1 cells was reversible after its removal from the culture medium. These results suggest that (1) resveratrol inhibits the growth of THP‐1 cells, at least in part, by inducing apoptosis, (2) resveratrol‐induced apoptosis of THP‐1 cells is independent of the Fas/FasL signalling pathway and (3) resveratrol does not induce differentation of THP‐1 cells and has no toxic effect on differentiated THP‐1 cells. Thus, resveratrol may be a potential chemotherapeutic agent for the control of acute monocytic leukaemia.
Prenatal and peripartum management of congenital afibrinogenaemia
Authors: Kobayashi Takao , Kanayama Naohiro , Tokunaga Naoki , Asahina Toshihiko , Terao Toshihiko
Published: March 2003

We experienced three cases and four successful deliveries with congenital afibrinogenaemia and propose the following guidelines for the prenatal and peripartum management: (i) genital bleeding usually begins at 5 weeks' gestation and spontaneous abortion always occurs at 6–8 weeks' gestation without fibrinogen infusion; (ii) the fibrinogen level must be at least 0·60 g/l and, if possible, higher than 1·0 g/l during the pregnancy; (iii) the necessary amounts of fibrinogen increase as the pregnancy progresses and the preterm labour occurs; (iv) the fibrinogen level under the continuous infusion of fibrinogen during labour must be at least 1·5 g/l and, if possible, higher than 2·0 g/l to prevent placental abruption; (v) the puerperium is usually uneventful with a reduced dose of fibrinogen infusion.
A single nucleotide polymorphism at nucleotide −1793 in the von Willebrand factor (VWF) regulatory region is associated with plasma VWF:Ag levels
Authors: Harvey Philip J. , Keightley Angela M. , Lam Y. Miu , Cameron Cherie , Lillicrap David
Published: March 2003

von Willebrand Factor (VWF) is a large multimeric glycoprotein involved in the transport and protection of factor VIII and in mediating platelet–subendothelium and platelet–platelet interactions. We have documented the presence of a single nucleotide polymorphism (SNP) at nucleotide (nt) −1793 (G 0·36 or C 0·64) in the VWF 5′‐flanking sequence. This polymorphism is in strong linkage disequilibrium with the previously reported SNPs at nts −1234, −1185 and −1051 and, like this other group of polymorphisms, shows a significant association with plasma VWF levels. This association is more marked in subjects who are more than 40 years of age. Further, circumstantial evidence to support a role for the −1793 sequence in regulating VWF expression comes from our demonstration of differential binding of endothelial cell nuclear proteins, including the transcription factor NFκB, by this sequence. In summary, the association of the −1793 SNP with plasma VWF levels provides additional evidence for the role of the VWF regulatory region between nts −1793 and −1051 in controlling VWF expression.
Fetal bone marrow as a source of stem cells for in utero or postnatal transplantation
Authors: Golfier Fran´ois , Bárcena Alicia , Harrison Michael R. , Muench Marcus O.
Published: March 2003

We examined the potential of human fetal bone marrow (FBM) as a source of haematopoietic stem cells for transplantation. The median number of cells obtained between 20 and 24 weeks' gestation was 1·9 × 109 and a median 1·17 × 108 of these cells expressed CD34. Flow cytometry was also used to estimate the content of three different candidate stem cell populations in the tissues older than 20 weeks' gestation. A median 8·8 × 105 CD34++CD38− cells, 1·37 × 106 CD34++CD4+ cells and 2·20 × 106 CD34++CD90+ cells were detected. The content of colony‐forming units culture (CFU‐C) in the FBM ranged from 2·8 × 104 to 6·0 × 106 per fetus. The CFU‐C content could be expanded 50‐fold by culture for 1 week in serum‐deprived medium and the growth factors kit ligand and granulocyte–macrophage colony‐stimulating factor. Positive selection of FBM CD34+/++ cells was achieved using the Baxter Isolex 50 device. An average purity of 82% and yield of up to 19% of CD34+/++ cells was achieved. T cells were depleted by 99·84%. Analysis of candidate stem cell populations and primitive CFU‐C suggested a preferential enrichment of these cells over the total population of CD34+/++ cells. All FBM samples were found to be free of microbial contamination at the time of harvest and after selection of CD34+/++ cells. Thus, FBM is a safe source of stem cells. The large number of progenitors and candidate stem cells that can be obtained from FBM makes it suitable for in utero and possibly postnatal transplantation.
Generation of HLA‐DRB1*1501‐restricted p190 minor bcr–abl (e1a2)‐specific CD4+ T lymphocytes
Authors: Tanaka Yuji , Takahashi Tsuyoshi , Nieda Mie , Masuda Shigeo , Kashiwase Koichi , Ogawa Seishi , Chiba Shigeru , Juji Takeo , Hirai Hisamaru
Published: March 2003

A small population of cells in acute lymphoblastic leukaemia is characterized by a specific translocation of the c‐abl oncogene on chromosome 9 to the break point cluster lesion (bcr) on chromosome 22, t(9; 22)(q34; q11) (e1a2). Theoretically, the junction‐spanning sequences of oncogene fusion proteins might be ideal targets for immunotherapy because these are not present in normal cells. In this study, we show for the first time that in vitro immunization with a 17‐mer e1a2 peptide representing the p190 minor bcr–abl fusion protein resulted in HLA‐DRB1*1501‐restricted peptide‐specific proliferative CD4+ T lymphocytes, using peptide‐pulsed monocyte‐derived dendritic cells as the antigen‐presenting cells.
Monitoring of minimal residual disease in patients with MLL–AF6‐positive acute myeloid leukaemia by reverse transcriptase polymerase chain reaction
Authors: Mitterbauer Gerlinde , Zimmer Christine , Pirc‐Danoewinata Hendrati , Haas Oskar A. , Hojas Sabine , Schwarzinger Ilse , Greinix Hildegard , Jäger Ulrich , Lechner Klaus , Mannhalter Christine
Published: March 2003

We studied 210 unselected patients with acute myeloid leukaemia (AML) for MLL abnormalities. Twenty‐seven patients (13%) with rearranged MLL genes were identified by means of Southern blot analysis. An MLL–AF6 fusion transcript was detected in six patients by a reverse transcriptase polymerase chain reaction (RT‐PCR) for the MLL–AF6 translocation. Sequence analysis showed fusion of MLL exon 7 as well as exon 6 (two patients) or MLL exon 6 as well as exon 5 (four patients) to AF6 exon 2. In only three patients could the t(6;11) also be identified by cytogenetic and/or fluorescence in situ hybridization (FISH) analysis. The MLL–AF6‐positive patients were monitored by RT‐PCR for a period of 6–33 months. Complete haematological remission (CR) was achieved in all six cases, but was short in 5/6 patients (range 2·6–8·3 months). In these five patients, the MLL–AF6 transcripts were detected in every sample tested after induction and consolidation chemotherapy. One patient received autologous bone marrow transplantation (BMT) which also did not lead to PCR negativity. Intensive salvage therapy was unable to induce a second remission in the relapsed patients. One of the six MLL–AF6‐positive patients achieved a molecular CR. He is still in CR at 33 months after diagnosis. Survival analysis indicates a poor prognosis in MLL–AF6‐positive patients. The median event‐free survival was 6·8 months, the median overall survival 15 months. Persistent PCR positivity was consistently associated with relapse. Thus, RT‐PCR provides a valuable and sensitive tool for the identification of t(6;11)‐positive AML and the monitoring of response to treatment in these patients. The results of RT‐PCR may be useful to evaluate therapeutic procedures and to make treatment decisions, which will enable molecular remissions to be achieved and improve the clinical outcome in this group of patients.
The polyneuropathy associated with Waldenström's macroglobulinaemia can be treated effectively with chemotherapy and the anti‐CD20 monoclonal antibody rituximab
Authors: Weide R. , Heymanns J. , Köppler H.
Published: March 2003

A 50‐year‐old male developed headache, impaired balance, visual defects and severe deafness. Ten months later he presented with markedly reduced power and tremor of his right arm. Waldenström's macroglobulinaemia (WM) with accompanying polyneuropathy was diagnosed. The patient received chemotherapy, which resulted in a partial improvement of the arm tremor. Subsequently, he was treated with rituximab (4 × 375 mg/m2), leading to complete resolution of the tremor and the paresis of his arm. Additionally, his headache and imbalance disappeared. Fifteen months after rituximab therapy he remained free of any neurological symptoms. This is the first report showing that WM‐associated polyneuropathy can be treated effectively with a combination of chemotherapy and the anti‐CD20 monoclonal antibody rituximab.
t(4;11)(q21;p15) translocation involving NUP98 and RAP1GDS1 genes: characterization of a new subset of T acute lymphoblastic leukaemia
Authors: Mecucci Cristina , La Starza Roberta , Negrini Massimo , Sabbioni Silvia , Crescenzi Barbara , Leoni Pietro , Di Raimondo Francesco , Krampera Mauro , Cimino Giuseppe , Tafuri Agostino , Cuneo Antonio , Vitale Antonella , Foà Robin
Published: March 2003

Two cases of T acute lymphoblastic leukaemia (T‐ALL) with an identical t(4;11)(q21;p15) translocation were identified within a prospective study on the biological and clinical features of adult ALL patients enrolled into the therapeutic protocol ALL0496 of the GIMEMA Italian Group. In both cases, the molecular characterization showed an involvement of the NUP98 gene on 11p15 which rearranges with the RAP1GDS1 gene on 4q21. The morphological and immunological features of the leukaemic cells, as well as the clinical behaviour and response to induction therapy, were the same in both patients. Based on the available data, the t(4;11)(q21;p15) translocation involving the NUP98–RAP1GDS1 fusion gene emerges as a new highly specific genetic abnormality that characterizes a subset of T‐ALL.
In vitro and in vivo effects of 2′‐deoxycoformycin (Pentostatin) on tumour cells from human γδ+ T‐cell malignancies
Authors: Aldinucci Donatella , Poletto Dalisa , Zagonel Vittorina , Rupolo Maurizio , Degan Massimo , Nanni Paola , Gattei Valter , Pinto Antonio
Published: March 2003

Hepatosplenic γδ+ T‐cell lymphoma represents a rare neoplasm of post‐thymic phenotype, characterized by an aggressive clinical course and a poor response to conventional chemotherapy. In the present study, we have examined the cytotoxic effects of the purine analogue 2′‐deoxycoformycin (dCF) on cultured mononuclear cells and purified γδ+ tumour cells from bone marrow or peripheral blood of four patients with hepatosplenic γδ+ T‐cell lymphoma. At a concentration of 10 µm, dCF, in the presence of 2′‐deoxyadenosine (dAdo), displayed an early and selective cytotoxic effect on γδ+ tumour T cells. After 48 h of in vitro exposure to dCF, the absolute number of viable CD3+/γδ+ tumour T cells was reduced by more than 90% in all samples with respect to control cultures, with absolute counts of viable CD3+/αβ+ lymphocytes being reduced only by 6–40% of the initial cell input. Analysis of cultures after 5 d of exposure to dCF plus dAdo revealed the persistence of normal CD3+/αβ+ T cells, which accounted, however, for only 20–25% of the initial cell input. Accordingly, the combination of dCF (10–100 µm) plus dAdo was able to induce a dose‐dependent inhibition of clonogenic growth and [3H]‐thymidine incorporation in purified CD3+/CD4−/CD8−γδ+ tumour cells. We also report that one patient with hepatosplenic γδ+ T‐cell lymphoma in terminal leukaemic phase showed a striking haematological response to single‐agent dCF given as fourth‐line treatment. In particular, the selective clearance of γδ+ tumour T cells in peripheral blood and bone marrow was observed starting after the second course of treatment. Our results suggest that dCF may represent a potentially active drug for the management of this aggressive form of T‐cell lymphoma.
Amifostine can reduce mucosal damage after high‐dose melphalan conditioning for peripheral blood progenitor cellautotransplant: a retrospective study
Authors: Capelli D. , Santini G. , De Souza C. , Poloni A. , Marino G. , Montanari M. , Lucesole M. , Brunori M. , Massidda D. , Offidani M. , Leoni P. , Olivieri A.
Published: March 2003

Amifostine (WR‐2721; Ethyol) is a well‐known cytoprotector, but a possible role in preventing extrahaematological toxicity after high‐dose therapy (HDT) has never been investigated. We compared two historical groups of patients who either received (group A, n = 35) or did not receive (group B, n = 33) amifostine (740 mg/m2) before high‐dose (HD) melphalan, followed by autologous infusion of peripheral blood progenitor cells (PBPCs). Amifostine was well tolerated at this dose level. Emesis grade 1–2 was the most important side‐effect, but the interruption of infusion was never required. The incidence and median duration of severe mucositis (grade 3–4) was 21% and 0 d (range 0–11 d) in group A and 53% and 7 d (range 0–11 d) in group B. The duration of analgesic therapy was also significantly lower in group A (0 d; range 0–12) than in group B (6 d, range 0–20) (P = 0·0001). Severe diarrhoea (3% vs. 25%; P = 0·01) and emesis (9% vs. 34%; P = 0·01) were also reduced in group A in comparison with group B. No differences were observed between the two groups for haematological recovery. This retrospective study strongly suggests that amifostine can reduce severe mucositis and the use of analgesic drugs in this setting. A randomized study is warranted to confirm these preliminary results.
Ferritin expression in maturing normal human erythroid precursors
Authors: Vaisman Boris , Meyron‐Holtz Esther G. , Fibach Eitan , Krichevsky Anna M. , Konijn Abraham M.
Published: March 2003

We studied the expression of H‐ and L‐ferritin subunits at sequential stages of maturation of normal human erythroid precursors. The erythroid cells developed in liquid culture and were purified immunomagnetically before analysis. It was found that the content of both ferritin subunits decreased exponentially with maturation: the decrease was rapid when cellular haemoglobin was low, and it slowed down when the haemoglobin was increased. This mode of decline was especially pronounced for the L‐subunits. The H‐/L‐subunit ratio did not change significantly during the investigated period. The synthesis of both subunits was equal at each given developmental stage, and declined significantly with maturation. However, this decline was just slightly faster than that of total protein synthesis. The data indicated that the degradation of H‐ and L‐ferritin also declined as maturation proceeded. No decrease was observed in mRNA levels of either ferritin subunit. Thus, the ferritin content and turnover were maximal at the beginning of haemoglobin accumulation and diminished later. As the rate of ferritin turnover determines the rate of incorporation and release of its iron, the results presented suggest that ferritin mediates cellular iron transport and donates iron for haem synthesis, mainly at the beginning of haemoglobin accumulation. The synthesis of both ferritin subunits is regulated during erythroid maturation at the post‐transcriptional level.
No impact of high‐dose cytarabine on the outcome of patients transplanted for acute myeloblastic leukaemia in first remission
Authors: Cahn Jean‐Yves , Labopin Myriam , Sierra Jorge , Blaise Didier , Reiffers Josy , Ferrant Augustin , Bergmann Lothar , Visani Giuseppe , Cornelissen Jan , De Witte Theo , Bosi Alberto , Frassoni Francesco , Gorin Norbert‐Claude ,   for the Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT)
Published: March 2003

High‐dose cytarabine is currently used in combination with anthracycline in the treatment of acute myeloblastic leukaemia (AML). Moreover, high‐dose cytarabine has been reported to produce long‐term disease‐free survival in a proportion of patients, especially in certain subtypes of AML. However, it remains unknown whether the outcome of patients undergoing allogeneic or autologous stem cell transplantation is influenced by previous treatment with high‐dose cytarabine. To this end, 1672 patients with AML in first remission who were reported to the Acute Leukaemia Working Party registry of the European Group for Blood and Marrow Transplantation (EBMT) and who were transplanted between 1980 and 1995 were analysed according to the dose intensity of cytarabine given at induction and/or consolidation. Autologous stem cell transplantation (ABMT) was performed in 846 patients and allogeneic bone marrow transplantation (BMT) in 826 patients. This study shows that the dose of cytarabine (Ara‐C) given at induction and/or consolidation did not influence the relapse incidence in patients subsequently allografted or autografted. In addition, it did not give any advantage in terms of overall outcome. Therefore, high‐dose (HD) Ara‐C may not be needed for patients who have a planned stem cell transplantation (SCT) as post‐remission therapy. Nevertheless, HD Ara‐C may be utilized in certain subtypes of AML that are believed to be curable by chemotherapy alone.
A comparison of molecular and enzyme‐based assays for the detection of thiopurine methyltransferase mutations
Authors: Coulthard Sally A. , Rabello Celia , Robson Jill , Howell Christopher , Minto Lynne , Middleton Peter G. , Gandhi Maher K. , Jackson Graham , McLelland Janet , O'brien Hugh , Smith Stephen , Reid Michael M. , Pearson Andrew D. J. , Hall Andrew G.
Published: March 2003

S‐Methylation by thiopurine methyltransferase (TPMT) is an important route of metabolism for the thiopurine drugs. About one in 300 individuals are homozygous for a TPMT mutation associated with very low enzyme activity and severe myelosuppression if treated with standard doses of drug. To validate the use of molecular genetic techniques for the detection of TPMT deficiency, we have determined red blood cell TPMT activity in 240 adult blood donors and 55 normal children. Genotype was determined by restriction fragment length analysis of polymerase chain reaction products in a cohort of 79 of the blood donors and five cases of azathioprine‐induced myelosupression, and this confirmed a close relationship between genotype and phenotype. In 17 of the 24 cases in which mutations were found, DNA was also available from remission bone marrow. In one of these cases, DNA from the remission marrow sample indicated the presence of a non‐mutated allele that had not been seen in the blast DNA sample obtained at presentation. These results indicate that polymerase chain reaction‐based assays give reliable and robust results for the detection of TPMT deficiency, but that caution should be exercised in relying exclusively on DNA obtained from lymphoblasts in childhood leukaemia.
T‐cell prolymphocytic leukaemia: antigen receptor gene rearrangement and a novel mode of MTCP1 B1 activation
Authors: De Schouwer P. J. J. C. , Dyer M. J. S. , Brito‐Babapulle V. B. , Matutes E. , Catovsky D. , Yuille M. R.
Published: March 2003

T‐cell prolymphocytic leukaemia (T‐PLL) is a sporadic, mature T‐cell disorder in which there is usually an aberrant T‐cell receptor alpha (TCRA) rearrangement that activates the TCL1 or MTCP1‐B1 oncogenes. As mutations of the Ataxia Telangiectasia (A‐T) gene, ATM, are frequent in T‐PLL and as ATM seems to act as a tumour suppressor through a mechanism involving V(D)J recombination, we examined V(D)J recombination in T‐PLL. Using Southern blotting and the polymerase chain reaction, two of 60 TCRG coding joints were abnormal. In all cases, both TCRD alleles were deleted, IGH was germline, and patterns of TCRB and TCRA rearrangement were normal. However, in a case harbouring t(X;7)(q28;q35), we identified TCRB segment Jβ2·7 juxtaposed to MTCP1 exon 1. This is the first time that TCRB has been implicated in MTCP1 B1 activation. The structure of the breakpoint supports a model in which translocation activates a cryptic MTCP1 promoter. This analysis of V(D)J recombination is consistent with it being a variable that is independent of ATM in T‐PLL.
Acute leukaemia in acromegaly patients
Authors: Au W. Y. , Chow W. S. , Lam K. S. L. , Ko G. T. C. , Cockram C. S. , Kwong Y. L.
Published: March 2003

Acromegaly patients are known to have an increased risk of malignancies, especially colonic adenocarcinoma. This may be as a result of the growth‐stimulating effect of growth hormone (GH). The clustering of leukaemia in children treated with GH has also caused concern. There have been a few reports of leukaemia in acromegaly patients. We report two patients with acute lymphoblastic leukaemia and one patient with acute myeloid leukaemia among 106 acromegaly patients treated over a 15‐year period. Two of the cases received radiotherapy as part of their treatment. Adjusted for age and follow‐up years, the incidence of leukaemia in this cohort is significantly higher than the general population. The incidence is also higher than would be expected as a result of radiotherapy alone, suggesting that GH may play a synergistic role.
Defective platelet response to arachidonic acid and thromboxane A2 in subjects with PlA2 polymorphism of β3 subunit (glycoprotein IIIa)
Authors: Andrioli Giuseppe , Minuz Pietro , Solero Pietro , Pincelli Silver , Ortolani Riccardo , Lussignoli Sabrina , Bellavite Paolo
Published: March 2003

The membrane complex αIIbβ3 is the major receptor for fibrinogen and is involved in platelet adhesion and aggregation. Evidence has been presented that the PlA2 allele of the β3 PlA1/A2 gene polymorphism might be an independent risk factor for coronary thrombosis, but the matter is still controversial. We investigated the relationship between this polymorphism and possible alterations of platelet functions in vitro. The platelet adhesion to fibrinogen‐coated microplate wells and the aggregation induced by several different agonists were tested in 63 healthy volunteers, among them, 49 subjects with PlA1/A1 polymorphism, 12 subjects with PlA1/A2 polymorphism and two subjects with PlA2/A2 polymorphism. Subjects with PlA1/A2 polymorphism or with PlA2/A2 polymorphism showed significantly lower platelet responses as compared with PlA1/A1 subjects when either arachidonic acid or the thromboxane A2 analogue, U46619, were used as agonists. In resting condition and after thrombin or ADP stimulation, platelet function was normal in all the subjects. An increased sensitivity to the anti‐aggregatory effect of acetylsalicylic acid was observed in platelets from subjects with the PlA2 allele. Finally, using a flow‐cytometric evaluation and determining the β‐thromboglobulin plasma levels, we did not find any evidence of a PlA2 platelet hyper‐reactivity ex vivo. Our findings are not consistent with the hypothesis that the purported increase of cardiovascular risk in these subjects may be as a result of platelet hyperactivation. On the contrary, the PlA2 allele is associated with a platelet functional deficiency, specifically linked to the activation of the fibrinogen receptor by thromboxane A2.
Early induction of apoptosis in B‐chronic lymphocytic leukaemia cells by hydroxychloroquine: activation of caspase‐3 and no protection by survival factors
Authors: Lagneaux L. , Delforge A. , Carlier S. , Massy M. , Bernier M. , Bron D.
Published: March 2003

We have investigated the effect of hydroxychloroquine (HCQ), an anti‐rheumatic drug, on malignant B cells from 20 patients with B‐chronic lymphocytic leukaemia (B‐CLL). HCQ induced a decrease in cell viability in a dose‐ and time‐dependent manner. The mean IC50 was 32 ± 7 μg/ml (range, 10‐75 μg/ml) for 24 h of exposure. This cytotoxic effect was owing to apoptosis, as demonstrated by morphological changes, annexin V binding capacity and DNA fragmentation (28 ± 4% of apoptotic cells as early as 5 h post incubation, increasing to 82 ± 4% at 18 h post treatment). The apoptosis was associated with caspase‐3 activation because the cleavage and activity of caspase‐3 were increased by HCQ. The amount of bcl‐2 protein was reduced during apoptosis, evidenced using quantitative flow cytometry. As early as 1 h post‐HCQ treatment, a reduction of the mitochondrial transmembrane potential was measured by 3,3’‐dihexyloxacarbocyanine iodide. Interestingly, the HCQ effect was not affected by exposure to interleukin‐4 or co‐culture with bone marrow stromal cells. Our observations suggest that HCQ may offer a new therapeutic tool in the treatment of B‐CLL patients.
Familial pseudohyperkalaemia Chiswick: a novel congenital thermotropic variant of K and Na transport across the human red cell membrane
Authors: Haines Philip G. , Crawley Charles , Chetty Margaret C. , Jarvis Helen , Coles Suzanne E. , Fisher Julie , Nicolaou Anna , Stewart Gordon W.
Published: March 2003

Two families with inherited abnormalities in Na and K transport across the red cell membrane are described. Both presented with ‘pseudohyperkalaemia’ as a result of loss of K from the red cells on storage at room temperature. Routine haematology was essentially normal, except for macrocytosis in one family. Studies of the temperature dependence of the passive leak to K showed a novel shoulder pattern with a minimum at 25°C, a maximum at 10°C, followed by a further fall. As in other cases of red cell‐based pseudohyperkalaemia, the abnormal temperature dependence of this ‘leak’ flux could be held to account for the loss of K from the cells at room temperature. These cases represent a novel variant of the temperature dependence of the passive leak of K and Na across the red cell membrane, and can be classified as a mild, non‐haemolytic form of the group known as the hereditary stomatocytosis and allied disorders’.
Long‐lasting complete remission of hepatitis C virus (HCV) infection and HCV‐associated immunocytoma with alpha‐interferon treatment
Authors: Patriarca Francesca , Silvestri Federico , Fanin Renato , Zaja Francesco , Sperotto Alessandra , Baccarani Michele
Published: March 2003

Several epidemiological data suggest the involvement of hepatitis C virus (HCV) in the pathogenesis of some histotypes of B‐cell non‐Hodgkin's lymphomas, in particular immunocytoma. We report a patient with HCV‐associated immunocytoma, first treated with six courses of fludarabine. A partial response was achieved and subsequent therapy with alpha‐interferon resulted in the clearance of the virus and a long‐lasting complete clinical and histological remission of the lymphoproliferative disease.
Interferon α extends the survival of human myeloma cells through an upregulation of the Mcl‐1 anti‐apoptotic molecule
Authors: Puthier Denis , Thabard Wilfrid , Rapp Marie‐José , Etrillard Marguerite , Harousseau Jean‐Luc , Bataille Régis , Amiot Martine
Published: March 2003

We have recently reported that Mcl‐1, an anti‐apoptotic member of the Bcl‐2 family, is upregulated by interleukin (IL)‐6 in human myeloma cells through the janus kinase/signal transducers and activators of transduction (JAK/STAT) pathway. In the current study, we have explored the effects of interferon (IFN)‐α, a cytokine which has been shown to increase myeloma cell survival. Our results demonstrate that IFN‐α potently upregulates Mcl‐1 on both myeloma cell lines and purified native myeloma cells. Of note, this upregulation is not due to an induction of an IL‐6 autocrine loop. Furthermore, we showed that IL‐6 and IFN‐α had no additive effect on Mcl‐1 upregulation, suggesting that both cytokines act through a common mechanism. Finally, the analysis of signalling transduction pathways strongly suggests that Mcl‐1 upregulation induced by IFN‐α depends on STAT3 activation. Altogether, our data show that IFN‐α has an IL‐6‐like effect on human myeloma cells and suggest that it could be deleterious in some patients.
Epstein–Barr virus‐positive aggressive lymphoma as a consequence of immunosuppression after multiple salvage treatments for follicular lymphoma
Authors: Orlandi Ester , Paulli Marco , Viglio Alessandra , Pagnucco Guido , Riboni Roberta , Baldanti Fausto , Lazzarino Mario
Published: March 2003

We report on a patient with follicular non‐Hodgkin's lymphoma (NHL) who developed a fatal high‐grade Epstein–Barr virus (EBV)‐positive NHL after conventional chemotherapies. The sudden onset of the high‐grade lymphoma was accompanied by increasing circulating EBV genome copies and was complicated by spontaneous rupture of the spleen. Splenic tissue was diffusely infiltrated by large B cells. In situ hybridization for Epstein–Barr‐encoded RNA (EBER) 1–2 was positive in 70% of cells, and molecular analysis revealed the presence of EBV DNA and a monoclonal IgH gene rearrangement. This case shows that the immunosuppression of multiple treatments may induce uncontrolled reactivation of a latent EBV infection, contributing to high‐grade transformation in heavily treated lymphoma patients.