bluuebunny/crossref_metadata_2025 huggingface dataset:
Tropism of human herpesvirus 8 for peripheral blood lymphocytes in patients with Castleman's disease
Authors: Kikuta Hideaki , Itakura Osamu , Taneichi Koji , Kohno Michifumi
Published: November 2003

Multicentric Castleman's disease (MCD), also called multicentric angiofollicular lymphoid hyperplasia, is a systemic lymphoproliferative disorder causing fever, lymphadenopathy and splenomegaly. Recently, Kaposi's sarcoma‐associated herpesvirus/human herpesvirus 8 (KSHV/HHV‐8) DNA sequences have been detected in cases of MCD. We examined HHV‐8 DNA sequences in the peripheral blood mononuclear cells (PBMCs) of two HIV‐negative patients with MCD and in PBMCs and the lymph node of a HIV‐negative patient with localized Castleman's disease (LCD) by the polymerase chain reaction. The novel sequences were detected in all DNA samples. Furthermore, the sequences were detected in only the CD19+ B‐lymphocyte fraction of the patient with LCD as previously reported. However, the sequences were detected in CD19+ B‐lymphocyte and CD2+ T‐lymphocyte fractions of two patients with MCD. These results suggest that HHV‐8 has tropisms for both B lymphocytes and T lymphocytes in Castleman's disease.
G‐CSF after peripheral blood stem cell transplantation in lymphoma patients significantly accelerated neutrophil recovery and shortened time in hospital: results of a randomized BNLI trial
Authors: Linch David C. , Milligan Donald W. , Winfield David A. , Kelsey Stephen M. , Johnson Stephen A. , Littlewood Timothy J. , Smith Graeme M. , Hutchinson Robert M. , Goldstone Anthony H. , Fielding Adele K. , Vaughan Hudson Gillian
Published: November 2003

We have undertaken a prospective randomized study in 90 patients with relapsed or resistant lymphomas to assess the value of G‐CSF (lenograstim) in the acceleration of myeloid recovery after peripheral blood stem cell transplantation (PBSCT). A common regimen of cyclophosphamide 1.5 g/m2 on day 1 and lenograstim 263 μg s.c. on days 2–10 with two aphereses on days 10 and 11 was used for stem cell mobilization. 77% of patients achieved an adequate PBSC collection in two harvests (>2 × 108 MNC/kg or >2 × 106 CD34+ cells/kg). 65 patients went on to receive high‐dose BEAM chemotherapy and engraftment data was available for 62. 34 patients had been randomized to receive lenograstim 263 μg/d s.c. and 28 to no growth factor. The median time to ANC > 0.5 × 109/l was 9 d in the lenograstim arm versus 12.5 d in the no‐lenograstim arm (P = 0.0001). This was associated with a median duration of time in hospital post PBSCT of 13 d in the lenograstim arm versus 15.5 d in the no‐lenograstim arm (P = 0.0002). Median days to platelet independence, platelet transfusions, incidence of infection and red cell transfusion were the same in both arms. These data indicate that lenograstim significantly accelerated myeloid recovery after PBSCT and shortened the duration of hospital stay.
FK506 inhibits anti‐IgM antibody‐induced apoptosis and 17 kD endonuclease activity in the human B‐cell line, MBC‐1, established from Burkitt's lymphoma
Authors: Higashigawa Masamune , Komada Yoshihiro , Shimono Yoshiki , Nagata Tsunehisa , Inamochi Hideki , Mao Xiao‐Yan , M'soka Tendai , Hori Hiroki , Kawasaki Hajime , Sakurai Minoru
Published: November 2003

The endonuclease which causes antibody‐induced apoptotic cell death in B cells is not completely understood. We previously established a B‐cell line (MBC‐1) from a patient with Burkitt's lymphoma at the leukaemic stage which demonstrated the typical morphology and internucleosomal DNA fragmentation of apoptotic cell death when treated with anti‐IgM antibody. FK506, an immunosuppressive agent and calcineurin inhibitor, partially rescued the anti‐IgM antibody‐induced cell death in these MBC‐1 cells. DNA SDS‐PAGE nuclease activity assay demonstrated that a 17 kD protein exhibited endonuclease activity. Active gel assay showed nuclease activity in the cellular nuclear extract not treated with anti‐IgM antibody. This nuclease activity was inhibited by FK506 at concentrations of 10–200 ng/ml in the active gel assay. These results raise the possibility that the 17 kD endonuclease is one of the nuclear members of the immunophilin family, which may function as an endogenous endonuclease in MBC‐1 cells.
Cyclic AMP phosphodiesterases in human lymphocytes
Authors: Sheth S. B. , Chaganti K. , Bastepe M. , Ajuria J. , Brennan K. , Biradavolu R. , Colman R. W.
Published: November 2003

The function of lymphocytes, like platelets, has been shown to be inhibited by agents which increase intracellular cyclic AMP. Two high‐affinity cAMP phosphodiesterases (PDEs), the cyclic GMP‐inhibited cAMP phosphodiesterase, PDE3, and the cAMP‐specific phosphodiesterase PDE4, are known to regulate cAMP concentration in haemopoietic cells by degrading cAMP to AMP. We characterized the relative contribution of the two PDEs to total lymphocyte PDE activity. We then determined which of the different gene products, PDE3A, typical of myocardium and platelets, or PDE3B, typical of adipocytes, were present in lymphocytes. The PDE3‐specific inhibitor, milrinone, and the PDE4 inhibitor, rolipram, suppressed hydrolysis by 70% and 30% respectively, which indicated that both PDE4 and PDE3 were present, and that PDE3 was predominant. RT‐PCR yields the expected size fragment for the primer pair PDE3B and not for PDE3A. The DNA sequence obtained had > 95% identity with PDE3B. PDE3B appears to be the major cAMP PDE in lymphocytes. In contrast to human platelets, human lymphocytes appear to contain the PDE3B subtype. Since PDE3B in adipocytes is subject to hormonal regulation, lymphocytes may be similarly modulated. Understanding the role of cAMP regulation and the involvement of cAMP in lymphocyte function may have important implications in drug development.
Heart transplantation for end‐stage heart failure caused by iron overload
Authors: Koerner Michael M. , Tenderich Gero , Minami Kazutomo , Zu Knyphausen Edzard , Mannebach Hermann , Kleesiek Knut , Meyer Hans , Koerfer Reiner
Published: November 2003

Few reports exist concerning heart transplantation in recipients with end‐stage myocardiopathy‐associated heart failure caused by iron overload occurring with β‐thalassaemia, Diamond‐Blackfan syndrome or haemochromatosis. Seven potential transplant candidates (six male, one female, mean age 26 years) with such heart failure, following desferrioxamine application subcutaneously over a number of years, and intravenously during their hospitalization before transplantation, were retrospectively analysed. Five were New York Heart Association (NYHA) class IV, three experienced one or more resuscitations immediately before transplantation could be performed. Continuous, high‐volume, veno‐venous haemofiltration was necessary in two patients. One of these two candidates additionally had to be bridged, first with a right ventricular, then with a biventricular assist device. Five of the seven patients survived, two with haemochromatosis, one with β‐thalassaemia major and one with Diamond‐Blackfan syndrome following transplantation. One non‐transplanted candidate with β‐thalassaemia major has been recompensated for 5 years. Survival was 14–74 months. Our results demonstrate the feasibility and indication of transplantation in patients with such heart failure and the satisfying outcome of immunosuppression is described.
The role of transforming growth factor‐β in PEG‐rHuMGDF‐induced reversible myelofibrosis in rats
Authors: Yanagida Makoto , Ide Youichi , Imai Atsushi , Toriyama Mie , Aoki Toshiko , Harada Katsuhiko , Izumi Hideakira , Uzumaki Hiroya , Kusaka Masaru , Tokiwa Tomonobu
Published: November 2003

Pegylated recombinant human megakaryocyte growth and development factor (PEG‐rHuMGDF) injected at a suprapharmacologic dose (100 μg/kg) daily for 5 d in normal rats caused marked increases in marrow megakaryocytes and platelet counts at 6–8 d followed by gradual decreases to control levels at 10–20 d. Interestingly, in addition to the expected thrombopoiesis, PEG‐rHuMGDF was associated with myelofibrosis with a predominance of reticulin fibres at day 10 followed by complete normalization by day 20. At 6–8 d, the levels of transforming growth factor‐β1 (TGF‐β1) in the extracellular fluid of the marrow, the platelet poor plasma, and the platelet extract were increased 23‐, 7‐ and 2‐fold, respectively. The elevated levels of TGF‐β1 were gradually reduced to baseline levels at 13–20 d in accordance with the normalization of myelofibrosis and thrombopoiesis. An ultrastructural analysis showed that large fragments of megakaryocytes were deposited in the marrow parenchyma of PEG‐rHuMGDF‐treated rats at day 6. PEG‐rHuMGDF administration at pharmacologic doses (1 and 10 μg/kg) did not induce the deposition of reticulin fibres in the marrow. These findings suggest that TGF‐β1 leaked from megakaryocytes is involved in the development of the PEG‐rHuMGDF‐induced myelofibrosis and that this is a reversible process related to the regulation of the excess production of platelets.
A population‐based registry on paraproteinaemia in the Netherlands
Authors: Ong F. , Hermans J. , Noordijk E. M. , Wijermans P. W. , Seelen P. J , De Kieviet W , Gerrits W. B. J , Kluin Ph. M , Kluin‐Nelemans J. C. ,   for the Comprehensive Cancer Centre West, Leiden, The Netherlands
Published: November 2003

Patients with monoclonal gammopathies comprise a heterogenous group. The few studies on incidence and follow‐up are single‐centre‐based and may reflect referral bias. To avoid this, all patients (n = 1275) in mid‐western Netherlands with a newly discovered paraproteinaemia in 1991, 1992 and 1993 were included in a population‐based registry and divided into four major diagnostic groups: multiple myeloma and plasmacytoma (n = 230, 18%), other haematological diseases (n = 141, 11%), paraprotein‐related internal diseases (n = 191, 15%) and monoclonal gammopathy of undetermined significance (MGUS, n = 713, 56%). To avoid a possibly erroneous diagnosis, patients who were classified as having MGUS but who did not undergo confirmatory bone marrow examination were included in a separate group ‘provisional MGUS’ (n = 524, 41%), whereas patients who did were classified as having ‘definite MGUS’ (n = 189, 15%). The ‘provisional MGUS’ patients were relatively older and had more often a poor performance status, but differences between this and the ‘definite MGUS’ group were otherwise small. Patients complaining of general malaise more often had a full work‐up of their paraproteinaemia. Bone pain, hypercalcaemia, high total protein, and high ESR occurred predominantly in the myeloma group, whereas fever or infection was less often seen in these patients. This registry of patients with paraproteinaemias provided valuable data related to all different diseases associated with paraproteinaemia.
Interdonor variability of platelet response to thrombin receptor activation: influence of PlA2 polymorphism
Authors: Lasne Dominique , Krenn Maria , Pingault Véronique , Arnaud Emmanuel , Fiessinger Jean‐Noël , Aiach Martine , Rendu Francine
Published: November 2003

Considering that platelet response to thrombin receptor activation might be critical for the development of arterial thrombosis, we measured the dense granule release under stimulation by the thrombin receptor activating peptide (TRAP) in a series of 102 healthy volunteers. The threshold TRAP concentration which initiated a secretion ranged from 3 to 20 μm. A good concordance (79%, k = 0.677) between two tests performed at a 1 month interval indicated that platelet response to thrombin receptor activation was characteristic of each individual donor. Since the threshold concentration required to initiate secretion corresponded to the threshold concentration which induced a biphasic aggregation, all volunteers were genotyped for the PlA2 polymorphism, the Pro33 variant of GPIIIa. Platelets from subjects with the PlA2 polymorphism required higher TRAP concentrations to aggregate than those from subjects with no PlA2 allele (P = 0.0012). However, they also required a higher ADP concentration to aggregate. In order to exclude any influence of GPIIIa polymorphism on TRAP‐induced secretion, we studied the variability of platelet response to TRAP among the 77 individuals with no PlA2 allele, and found the same interdonor variability with the same distribution of threshold TRAP concentrations as for the 102 individuals. The results suggest that (i) platelet secretion in response to thrombin receptor activation could be a genetically controlled phenotype independent of the GPIIIa polymorphism; (ii) the PlA2 polymorphism is associated with platelet hypoaggregability.
Expression of the Evi‐1 gene in haemopoietic cells of children with juvenile myelomonocytic leukaemia and normal donors
Authors: Gerhardt Thomas M. , Schmahl Gerd E. , Flotho Christian , Rath Armin V. , Niemeyer Charlotte M.
Published: November 2003

Activation of the Evi‐1 gene was first described to be associated with the transformation of murine myeloid leukaemias and has previously been detected in cases of human acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML) in blast crises and in myelodysplastic syndromes. In this study we determined the frequency and the level of Evi‐1 expression in juvenile myelomonocytic leukaemia (JMML) and in normal haemopoiesis. Using RT‐PCR and Southern blot hybridization mRNA of Evi‐1 could be detected in bone marrow (BM) and peripheral blood (PB) mononuclear cells (MNC) of normal donors. In JMML 12/20 patients examined expressed elevated levels of Evi‐1 compared to normal controls. In these samples over‐expression of the gene was correlated with a higher percentage of blasts (P = 0.02). Expression levels in BFU‐E and CFU‐GM derived colonies from BM of JMML patients were lower than those in the corresponding MNC samples. Analysis of CD34+ and CD34− cells demonstrated that Evi‐1 is primarily expressed in the CD34+ cell population of both JMML and normal donors. These findings suggest that Evi‐1 expression is linked to the early stages of haemopoiesis. Studies on the regulation of Evi‐1 expression in CD34+ cells will elucidate its function in progenitor cells and clarify its possible role in the pathogenesis of JMML.
Aminophospholipid exposure, microvesiculation and abnormal protein tyrosine phosphorylation in the platelets of a patient with Scott syndrome: a study using physiologic agonists and local anaesthetics
Authors: Dachary‐Prigent Jeanne , Pasquet Jean‐Max , Fressinaud Edith , Toti Florence , Freyssinet Jean‐Marie , Nurden Alan T.
Published: November 2003

The Scott syndrome is a rare inherited haemorrhagic disorder characterized by the inability of blood cells to expose aminophospholipids and to shed microparticles. We have had the opportunity to study a recently reported French patient with this syndrome and have confirmed by means of a fluorescence assay for transbilayer lipid movement a reduced aminophospholipid exposure when platelets were stimulated with the calcium‐ionophore ionomycin, in spite of a normal elevation of intracellular Ca2+. Secretion and calpain activation were also shown to be normal. Significantly, the level of phosphotyrosine‐labelled proteins in platelets treated with thrombin or a thrombin + collagen mixture and in particular the phosphorylation of a 40 kD band were severely reduced. Furthermore, inhibition of thiol‐containing enzymes, including tyrosine‐phosphatases, by N‐ethyl maleimide did not lead to aminophospholipid exposure in the patient's platelets, in spite of increased tyrosine protein phosphorylation. In contrast, amphiphilic membrane drugs such as tetracaine and propranolol induced both surface aminophospholipid exposure in Scott platelets and the shedding of microparticles, thereby showing that membrane perturbation can lead to loss of phospholipid asymmetry in this syndrome. Our results provide the first insight that the lack of expression of procoagulant phospholipids and microparticle formation in Scott syndrome platelets is associated with a defect of intracellular signalling.
Donor lymphocyte infusion for childhood acute lymphoblastic leukaemia relapsing after bone marrow transplantation
Authors: ATRA A. , MILLAR B. , SHEPHERD V. , SHANKAR A. , WILSON K. , TRELEAVEN J. , PRITCHARD‐JONES K. , MELLER S. T. , PINKERTON C. R.
Published: November 2003

Four children with acute lymphoblastic leukaemia (ALL) who relapsed after allogeneic bone marrow transplantation (BMT) were treated with donor lymphocyte infusion (DLI) without prior conditioning. Three patients had previously received a non‐T‐cell‐depleted matched sibling BMT and the fourth had a T‐cell‐depleted matched unrelated BMT. Two patients developed grade III–IV acute graft‐versus‐host‐disease (GVHD) of the skin, which required intervention. Both are alive in complete haematological remission 7 and 10 months from DLI with chronic GVHD of the skin requiring immunosuppressive therapy. A third patient went into haematological remission 6 weeks after DLI, but with no clinical evidence of GVHD. His bone marrow remained in remission 11 months post‐DLI despite the disease (ALL) relapsing in extramedullary sites. The fourth patient showed no clinical or haematological response to three consecutive doses of DLI given at 4‐weekly intervals and died from progressive disease 11 months after relapse. These preliminary observations indicate that in constrast to experience in adult ALL, DLI may be effective in inducing sustained remission in children with ALL relapsing after BMT, and a response may occur even in the absence of clinical evidence of GVHD.
Clonal chromosomal abnormalities as direct evidence for clonality in nasal T/natural killer cell lymphomas
Authors: TIEN HWEI‐FANG , SU IH‐JEN , TANG JIH‐LUH , LIU MING‐CHI , LEE FEN‐YU , CHEN YAO‐CHANG , CHUANG SOU‐MING
Published: November 2003

Nasal T/natural killer (NK) cell lymphoma is a distinct clinicopathologic entity which is more prevalent in Asia than in America and Europe. The clonal nature of the infiltrating lymphoid cells is difficult to demonstrate because of the lack of immunologic markers for clonality and the absence of clonal T‐cell receptor gene rearrangement in most cases. In this study, clonal chromosomal abnormalities were detected in the tumour cells from four patients with nasal T/NK cell lymphoma. This finding provided direct evidence for clonality of the disease. Moreover, nonrandom cytogenetic abnormalities, including isochromosome for the short arm (p) of chromosome 6, isochromosome for the long arm (q) of chromosome 1, partial deletion of 6q, and aberrations at 11q, were disclosed. Isochromosome 6p was the sole structural abnormality in one patient, which may be a pathognomonic change in nasal lymphoma.
Heterogenous response of B lymphocytes to transforming growth factor‐beta in B‐cell chronic lymphocytic leukaemia: correlation with the expression of TGF‐β receptors
Authors: Lagneaux Laurence , Delforge Alain , Bron Dominique , Massy Martine , Bernier Michel , Stryckmans Pierre
Published: November 2003

We investigated the potential role of transforming growth factor‐beta (TGF‐β) on spontaneous and cytokine‐induced proliferation of B‐cell chronic lymphocytic leukaemia (B‐CLL) cells in vitro. Purified B lymphocytes from 21 B‐CLL patients were cultured for 5 d in the presence of medium alone, IL‐2 and/or IL‐10, in the presence or absence of TGF‐β, and proliferation was measured by 3H‐thymidine incorporation. TGF‐β inhibited B‐cell proliferation in the majority of patients (15/21) but no inhibition was detected in 6/21 patients whatever the type of stimulant used. Addition of neutralizing antibodies to TGF‐β increased spontaneous and cytokine‐induced proliferation; this effect was dose dependent and specific because addition of an irrelevant chicken IgG had no effect on B‐CLL proliferation. In resistant patients, neutralizing antibodies to TGF‐β did not increase the proliferation. The expression of TGF‐β receptors on B‐CLL cells was significantly lower than the one observed on normal CD5+ B lymphocytes for which the sensitivity to TGF‐β inhibition was more marked than in CLL. In addition, we found a strong correlation between the response of leukaemic B cells to TGF‐β inhibitory action and the expression of TGF‐β receptors on these cells. In summary, TGF‐β appears to function in CLL as a negative regulator of B lymphocytes but loss of responsiveness to this factor accompanied by a decrease of TGF‐β receptor expression, might provide a selective advantage to B‐CLL lymphocytes.
The site of destruction of autologous 111In‐labelled platelets and the efficiency of splenectomy in children and adults with idiopathic thrombocytopenic purpura: a study of 578 patients with 268 splenectomies
Authors: Najean Y. , Rain J‐D. , Billotey C.
Published: November 2003

The indication for splenectomy in chronic idiopathic thrombocytopenic purpura (ITP) remains a controversial subject. The mortality rate of persistent thrombocytopenia is very low, except in severe cases. Conversely, the risks of splenectomy are significant (in the present series, morbidity: 4.1% mortality: 1.4%), with a success rate of only 60–75%. It is therefore useful to define a parameter able to predict the efficacy or failure of splenectomy.  An analysis of 578 cases of chronic ITP, where the site of platelet destruction has been determined, is presented. 268 of these cases had been splenectomized. When platelet destruction was splenic, 96% of subjects aged 5–30 years and 91% of cases over the age of 30 years obtained a remission. Conversely, when platelet destruction was hepatic or diffuse, failure or incomplete results were observed in 92% of cases. The site of platelet destruction therefore constitutes a parameter which can help the clinician to make the decision to perform splenectomy.
TRANSFER OF ANTI‐D ANTIBODIES ACROSS THE ISOLATED PERFUSED HUMAN PLACENTAL LOBULE AND INHIBITION BY HIGH‐DOSE INTRAVENOUS IMMUNOGLOBULIN: A POSSIBLE MECHANISM OF ACTION
Authors: Urbaniak S. J. , Duncan J. I. , Armstrong‐Fisher S. S. , Abramovich D. R. , Page K. R.
Published: November 2003

Using an in vitro perfusion model, therapeutic intravenous immunoglobulin (IVIgG) and IgG anti‐D have been shown to cross the placenta from the maternal circuit to the fetal circuit. The transfer of all IgG species was linear with respect to time, and the amount of IgG transferred was proportional to the concentration of IgG in the maternal circuit ([IgG]m), but reached saturation at upper limits. With total [IgG]m at 6.5 g/l, 11.1 g/l or 26.2 g/l the increase in the fetal concentration of total IgG was 4.6 mg/l/h, 8.9 mg/l/h and 9.9 mg/l/h respectively. The rate of transfer of specific anti‐D antibody to the fetal circuit was 0.026 IU/ml/h at a concentration of 38 IU/ml in the maternal circuit ([anti‐D]m). High‐dose therapeutic IVIgG added to the maternal circuit (total [IgG]m 29.2 g/l) significantly inhibited (P < 0.001) anti‐D transfer to 0.004 IU/ml/h. Addition of the same IVIgG at a lower concentration (total [IgG]m 11.1 g/l) also reduced anti‐D transfer, but only to 0.015 IU/l/h. The inhibitory effect of IVIgG does not appear to be mediated by anti‐idiotypic or non‐specific complexing with the anti‐D, but may be the result of competition with IgG anti‐D for placental Fcγ receptors involved in the endocytotic uptake of IgG. The efficacy of IVIgG in this model suggests that it may be clinically useful in preventing HDN and other immune cytopenias, provided a sufficiently high dose is given.
Activated protein C resistance can be associated with recurrent fetal loss
Authors: Brenner Benjamin , Mandel Hanna , Lanir Naomi , Younis Johnny , Rothbart Haim , Ohel Gonen , Blumenfeld Zeev
Published: November 2003

As thrombosis of placental vessels may result in recurrent fetal loss, we analysed 39 consecutive women with recurrent fetal loss of unknown cause for activated protein C resistance. Factor V Leiden (FVL) mutation (19 cases) or APC resistance without FVL (nine cases) were found among these 39 women.  Evaluation of 128 pregnancies in 19 patients with factor V Leiden mutation and 56 gestations in nine women with acquired APC resistance, revealed over 50% first‐trimester abortions and 17% late abortions. Intra‐uterine fetal death occurred in nine out of 19 FVL patients (47%). Only 34 of 184 gestations (18%) in hereditary or acquired APC‐resistance women resulted in a live birth, with 11 of the 34 (32%) being premature deliveries. These data suggest that, in some patients with recurrent fetal loss, hereditary and acquired APC resistance are potential causes of vascular placental insufficiency.
Tobacco smoking and risk of haematological malignancies in adults: a case–control study
Authors: PASQUALETTI PAOLO , FESTUCCIA VITTORIO , ACITELLI PIERLUIGI , COLLACCIANI ANTONIO , GIUSTI ALDO , CASALE RAFFAELE
Published: November 2003

A retrospective study was conducted in 1216 cases to investigate the possible association between tobacco smoking and the risk of haematological malignancies. A small, but not significant, increase in malignancy was observed in smokers. Significant association was demonstrated between tobacco smoking and acute non‐lymphoblastic leukaemia, and myelodysplastic syndromes. The duration and amount smoked increased the risk; heavy smokers presented significant positive associations with overall malignancies, acute nonlymphoblastic leukaemia, myelodysplastic syndromes, and monoclonal gammopathy of undetermined significance, whereas light smokers did not present any significant association. These data support a causal relationship between certain haematological malignancies and tobacco smoking. Further research is needed to examine the risk according to dose–response effect, and the variation in risk according to the histological subtype of the malignancy.
Prognostic factors in elderly acute lymphoblastic leukaemia
Authors: LEGRAND OLLIVIER , MARIE JEAN‐PIERRE , MARJANOVIC ZORA , CADIOU MONIQUE , BLANC CLAUDE , RAMOND SYLVIE , VIGUIÉ FRANCK , PERROT JEAN‐YVES , ZITTOUN ROBERT
Published: November 2003

A retrospective study was performed on 46 unselected acute lymphoblastic leukaemia (ALL) elderly patients aged 60 years or more. Only 50% of these patients were included in the EORTC cooperative clinical trials, thus confirming the important selection bias in most of the published series on elderly ALL patients. 43% of the elderly patients achieved a complete remission (CR). The median survival was 10 months and the 5‐year overall survival was only 7.6±4%. In multivariate analysis, W.H.O. performance status and peripheral blast counts at day 7 were found to significantly influence achievement of CR and survival. In patients with W.H.O. performance status 2, 35% died during induction treatment versus 4% in patients with W.H.O. performance status <2. Patients >70 years old showed a marked drop of the CR rate (27%) compared to those aged 60–69 (67%), and a very high death rate during the induction period (38% versus 4%). This suggests that ALL protocol treatments should be proposed until 70 years in patients with good‐performance status, whereas less intensive treatment should be offered to elderly patients with performance status 2 and/or age 70. Peripheral blast counts at day 7 may help to adjust the treatment during induction phase.
Polymorphism of the tumour necrosis factor‐alpha and lymphotoxin‐alpha genes in chronic lymphocytic leukaemia
Authors: Demeter Judith , Porzsolt Franz , Rämisch Stephanie , Schmidt Dorothee , Schmid Mathias , Messer Gerald
Published: November 2003

The neoplastic cells of CLL are able to produce TNF which is known to stimulate the proliferation of CLL cells in an autocrine and paracrine manner. Genetic polymorphism of molecules of the TNF ligand superfamily has been described and certain alleles were suspected to predispose to variant biological responses. Previously, the rare allele TNFB*1 of the TNF‐β/lymphotoxin (LT)‐α gene (NcoI, asparagine at amino acid position 26) was found to be associated with a stronger LT‐α response of PBMC in vitro.

We now report on a significant increase of the allele TNF1 (TNFA −308 G) of the TNF‐α promoter/enhancer polymorphism in a group of 73 CLL patients when compared to healthy individuals (RR=3.18, 95% confidence interval 1.57–8.3; P=0.006). The allelic distribution of the TNF‐β/LT‐α NcoI polymorphism did not differ significantly from randomized healthy controls. On the other hand, the frequency of the allele TNFB*2 was increased in CLL patients with advanced clinical stage (P=0.0004). These findings indicate immunogenetic associations involving polymorphisms of cytokine genes serving as paracrine and autocrine growth factors, which thus can contribute to the pathogenesis of the TNF/LT‐sensitive haematological malignancy CLL.
PTHrP‐mediated hypercalcaemia in a case of CML blast crisis
Authors: KUBONISHI ICHIRO , ASAHI YASUTOMO , MACHIDA HISANORI , UEMURA YOSHIKI , HATAKEYAMA NOBUO , KUBOTA TETSUYA , OHTSUKI YUJI , MIYOSHI ISAO
Published: November 2003

We report a 52‐year‐old man with chronic myelogenous leukaemia (CML) who developed hypercalcaemia in blast crisis. There was a high serum level of parathyroid hormone‐related protein (PTHrP). The leukaemia cells secreted PTHrP into culture medium when cultured in vitro and were shown to express PTHrP mRNA by reverse transcription‐polymerase chain reaction. Transplantation of the leukaemia cells into nude mice resulted in the production of Ph1 chromosome‐positive tumours which caused increased levels of calcium and PTHrP in the blood. These results indicated that the hypercalcaemic event in the patient was induced by PTHrP of CML cell origin.
High intensity of oral anticoagulant therapy in patients with cerebral haemorrhage: cause or consequence of the bleeding?
Authors: Cannegieter S. C. , Rosendaal F. R. , Bollen E. L. E. M. , Brie¨t E.
Published: November 2003

In assessing the optimal intensity of anticoagulant therapy, the International Normalized Ratio (INR) at admission is used as a basis for INR‐specific incidence rates. In 47 patients suffering a haemorrhagic stroke we tested the assumption that the INR at admission is an acceptable measure for the INR that preceded the haemorrhage. We found high D‐dimer levels in 70% of the patients, which indicated activated coagulation and fibrinolysis. This was not of such an extent that it could also be measured with other routine coagulation tests, with the possible exception of two patients. We found normal INRs in 33 non‐anticoagulated patients, and only a mildly prolonged INR of 1.9 in one patient, which was most probably caused by a vitamin K deficiency. We concluded that the INR at admission can be used in studies to assess the optimal level of anticoagulation.
Cyclosporin for thrombotic thrombocytopenic purpura after autologous bone marrow transplantation
Authors: Van Ojik Heidi , Biesma Douwe H. , Fijnheer Rob , Hené Ronald J. , Lokhorst Henk M. , Jan Sebo , Rooda Eelkman , Verdonck Leo F.
Published: November 2003

Thrombotic thrombocytopenic purpura (TTP) after bone marrow transplantation (BMT) is a serious complication that, in contrast to classic TTP, is generally not responsive to therapeutic plasma exchange. We describe the remission of TTP after autologous BMT by administration of cyclosporin in four patients, either after failure of plasmapheresis (two patients) or as primary treatment modality (two patients). Cyclosporin may be an attractive drug for post autologous BMT‐TTP.
Cytogenetics adds independent prognostic information in adults with acute lymphoblastic leukaemia on MRC trial UKALL XA
Authors: Secker‐Walker L. M. , Prentice H. G. , Durrant J. , Rchards S. , Hall E. , Harrison G.
Published: November 2003

Cytogenetic classification of 350 adults with acute lymphoblastic leukaemia on MRC UKALL XA trial showed the following statistically significant associations: t(9;22) (11%) increased with increasing age and leucocyte counts (WBC) and most had a C/pre‐B immunophenotype. t(4;11) (3%) was associated with higher WBCs, increasing age and null immunophenotype. Other abnormalities of 11q (abn11q) (4%) were associated with male sex and T‐cell ALL. High hyperdiploidy (7%) and abn9p (5%) decreased with increasing WBC. High hyperdiploid patients were younger and tended to have C/pre‐B ALL. Triploidy/tetraploidy (3%) decreased and pseudodiploidy (11%) increased with increasing WBC. Cytogenetic classification was prognostically important (chi‐square for heterogeneity of classification = 53.36; P < 0.0001) and added significance to age, sex and WBC. A poor prognosis for patients classed as t(9;22) (13% disease‐free survival at 3 years), as t(4;11) 24% at 3 years) and hypodiploid (11% at 3 years), and good prognosis for abn12p (4% of subjects) and high hyperdiploidy (74% and 59% at 3 years respectively) were statistically significant, but the 54% 3‐year disease‐free survival for patients with t(1;19) was not. The prognosis of patients classed as t(9;22) was independent of other single variables. Abn12p, abnormalities of 11q (including t(4;11) cases) and hypodiploidy added prognostic significance to all other variables combined.
A severe transfusion‐dependent congenital dyserythropoietic anaemia presenting as hydrops fetalis
Authors: Cantù‐Rajnoldi A. , Zanella A. , Conter U. , Faccini P. , Soligo D. , Gornati G. , Vegni C. , Nicolini U.
Published: November 2003

We describe a case of congenital dyserythropoietic anaemia (CDA) presenting at week 20 of pregnancy with hydrops fetalis and very severe anaemia. Fetal blood examination showed marked erythroblastosis with morphologic alterations while the basic haematological and biochemical tests were in the normal range. The fetus received intravascular red blood cells transfusion at 20 and 25 weeks of gestation. After a caesarean section at week 33 the child became transfusion dependent. Haematological investigations demonstrated that the child is affected by a CDA‐like picture which, although morphologically similar to CDA II, is Ham test negative and does not show the typical membrane proteins alterations.
Characterization of a novel bleeding disorder with isolated prolonged bleeding time and deficiency of platelet microvesicle generation
Authors: Castaman Giancarlo , Yu‐Feng Li , Battistin Eliseo , Rodeghiero Francesco
Published: November 2003

Platelet prothrombinase activity and microvesicle (MV) generation were measured in four patients from three unrelated families with a life‐long bleeding disorder associated with slightly prolonged bleeding time and isolated defective serum prothrombin consumption, without platelet function abnormality or von Willebrand factor defect. MV generation was reduced in all the patients either after thrombin plus collagen or A23187 calcium ionophore stimulation, whereas, at variance with Scott syndrome, prothrombinase activity was normal. This abnormality constitutes a new bleeding disorder, which provides new insights into the possible role of platelet microvesicles in health and disease. Furthermore, the results of this study suggest that MV generation should be investigated in patients with a bleeding history and apparently isolated prolonged bleeding time when prothrombin comsumption in serum is defective and all other investigations are normal.
Evaluation of DNA analysis for evidence of apoptosis in megaloblastic anaemia
Authors: Ingram C. F. , Davidoff A. N. , Marais E. , Sherman G. G. , Mendelow B. V.
Published: November 2003

This study involved DNA analysis of bone marrow cells of 15 patients with megaloblastic anaemia. The diagnosis was based on the morphological changes seen in the bone marrow, associated with either a low red cell folate or serum vitamin B12 level and an adequate response to appropriate therapy as confirmation of the diagnosis. Flow cytometric DNA analysis showed an increase in the S and G2 phases of the cell cycle, but conventional agarose gel electrophoretic DNA analysis did not confirm the characteristic ‘ladder pattern’ which might have been expected in classic apoptosis. In addition, cells showing morphological changes suggestive of apoptosis, such as nuclear condensation and fragmentation, did not show evidence of DNA fragmentation using the ApopTagTMin situ digoxigenin nucleotide labelled, peroxidase detection system. Further studies using annexin V flow cytometric analysis and pulsed field gel electrophoresis were also unable to detect evidence of apoptosis as a significant cause of cell death in megaloblastic anaemia.
Reliability of delayed INR determination: implications for decentralized anticoagulant care with off‐site blood sampling
Authors: Baglin Trevor , Luddington Roger
Published: November 2003

In order to develop decentralized anticoagulant care by off‐site blood sampling and transport of samples to a centralized laboratory for International Normalized Ratio (INR) determination we have performed a direct comparative study of INR stability. Analysis was performed daily for 5 d using nine thromboplastins. The overall mean difference of INR after 3 d was only 0.05 INR units for samples with a therapeutic INR. After 5 d there was a mean difference of 0.11 INR units with ‘non‐Manchester’ reagents and 0.44 INR units with ‘Manchester’ reagents. With over‐anticoagulated samples mean differences of 0.55–0.72 INR units were observed after 3 d and 1.16–2.46 INR units after 5 d. Although there was some variation in stability of results with different thromboplastins, the difference over time with each thromboplastin was much less than the difference between thromboplastins.

In conclusion, there is no clinically significant change in INR when analysis is delayed for up to 3 d. Off‐site blood sampling can accommodate a large increase in patient workload without a major revenue increase in primary care and with continued total quality management and central expert advice.
A simplified statistical method for local INR using linear regression
Authors: Poller L. , Barrowcliffe T. W. , Van den Besselaar A. M. H. P. , Jespersen J. , Tripodi A. , Houghton D.
Published: November 2003

A simplified method of International Normalized Ratio (INR) derivation using linear regression of certified INR plotted against local prothrombin time (PT) results has been compared with INR from conventional orthogonal regression. Linear regression assumes error only with the local PT results whereas orthogonal regression assumes error with both reference and local results. The reliability of local INR derivation using lyophilized plasmas has been assessed in a collaborative study. INR from conventional fresh plasma International Sensitivity Index (ISI) calibrations have been compared with INR from calibrations with two types of lyophilized plasma, artificially depleted and coumarin.

Although calibration slopes differed with the two types of analysis and the different lyophilized plasmas, both gave reasonable approximations to fresh plasma ISI calibrations. With orthogonal regression the overall percentage INR deviation was 5.25% with the artificially depleted plasmas and 6.85% for the results with lyophilized coumarins. With the linear regression, deviation was 8.40% for the artificially depleted plasmas and 5.05% for coumarin‐treated patients' lyophilized plasmas. The simpler regression method appears to be worthy of further study as the present report has demonstrated that if the calibrant plasmas are accurately certified with the thromboplastin International Reference Plasma (IRP) results approximate to the conventionally determined INR using the manual PT technique. Coagulometers require further assessment.
A method for canine nasal mucosa superfusion, in vivo; evidence that pseudomonas‐induced neutrophil migration is inhibited by α2‐adrenoceptor agonist
Authors: STJÄRNE P. , AGUSTI C. , CARDELL L. O. , WAGSTAFF S. , NADEL J. A.
Published: November 2003

Methods and Results
A method for isolating and superfusing a segment of the nasal cavity in the anaesthetized dog was developed. Introduction of Pseudomonas aeruginosa supernatant (PA), which is known to induce IL‐8 release, resulted in a marked time‐dependent exponential increase in neutrophil recruitment to the nasal chamber. When oxymetazoline (final concentration. 10‐5 M), was added to the superfusate, the PA induced‐neutrophil migration was inhibited; the addition of benzalchoniul chloride to the superfusate had no effect on the PA induced‐neutrophil migration.

Conclusion
We conclude that a α2 adrenoceptor agonist profoundly inhibits neutrophil migration in response to bacterial products. Our novel method allows continuous introduction of stimuli and monitoring of responses in the nasal mucosa.
Predictability of early atopy by cord blood‐IgE and parental history
Authors: BERGMANN R. L. , EDENHARTER G. , BERGMANN K. E. , GUGGENMOOS‐HOLZMANN I. , EORSTER J. , BAUERl C. P. , WAHN V. , ZEPP F. , WAHN U.
Published: November 2003

Background
Atopic family history and cord blood IgE have been used as predictors of atopic disease in newborns for about 20 years, but at least for cord blood IgE the sensitivity has been shown to be very low. The objective of this paper was to evaluate whether parental history and cord blood‐IgE were more accurate predictors for the appropriate atopic phenotypes in the infants rather than for any atopy.

Methods
A total of 1314 newborn infants was recruited in six German obstetric departments in 1990 and followed‐up for 2 years. Four hundred and ninty‐ninc (38%) were at high risk for atopy with at least two first degree atopic family members and/or elevated cord‐blood IgE concentrations.

Results
The cumulative incidence of atopic dermatitis over the first 2 years of life (AD24) amounted to 20. 1%, and there was a significant association with AD history of the mother (OR 2.5, 95%‐Cl 1.46–4.26) and of the father (OR 3.53, 95%cC1 1.90–6.54). The cumulative incidence of recurrent wheezing in the first 2 years of life (RW24) amounted to 16.1%, and was positively associated with asthma history (OR 2.11, 95%CI 1.33–3.60) and sensitization history (OR 1.64, 95%C1 1.34–2.36) of the mother, but with neither for the father. RW24 was less prevalent in girls than in boys (OR 0.64. 95%Cl 0.47–0.89). Thirty‐one per cent of infants were sensitized (CAP test value > 0.35 kU/L) against at least one of nine food or inhalative allergens (S24) and this was signilicantly associatcd with cord blood‐IgE value (OR 2.43, 95%C1 1.69–3.49). and sensitization history of the mother (OR 1.64, 95%CI 1.18–2.41). Using multiple logistic regression analysis, the prediction of AD24 by AD of parents, of RW24 by asthma of parents, and of sensitization by cord blood IgE was of low accuracy.

Conclusion
The predictive capacity of parental history and cord blood IgE is not high enough to recommend them as screening instruments for primary prevention. The majority of atopic manifestations and of sensitization occur in infants with no demonstrable risk at birth.
Allergen‐induced CD30 expression on T cells of atopic asthmatics
Authors: LEONARD C. , TORMEY V. , FAUL J. , BURKE C. M. , POULTER L. W.
Published: November 2003

Background
The importance of TH2‐type T cell cytokines in atopic disease is widely accepted. CD30, a member of the TNF/NGF receptor superfamily, is expressed on a proportion of activated CD45RO+ T cells and has been proposed as a marker for TH2 phenotype. CD30 ligand‐CD30 interaction has been shown to positively influence development of the TH2 phenotype, and serum levels of soluble CD30 (sCD30) have been used as prognostic markers in HIV, SLE, Epstein‐Barr Virus infection and Hodgkin's Lymphoma but not as yet in allergic disease.

Objectives
To establish if serum levels of sCD30 are elevated in atopic asthma and determine whether allergen‐induced proliferation/activation of PBMCs from atopic asthmatics promotes CD30 expression on CD4+ T lymphocytes. Further, to determine if expression of CD30 and sCD30 correlate with disease severity.

Methods
Eighteen atopic asthmatics were each assigned a symptomatic disease score based on symptoms and bronchodilator rescue usage. Serum sCD30 was measured in peripheral blood by ELISA. PBMCs from atopic asthmatics were analysed with flow cytometry to obtain the proportions of CD4 T cells expressing CD45RO and CD30. The cells were then cultured for 10 days with IL‐2 with or without house dust mite antigen. A proliferation index was recorded and expression of CD30 and CD45RO retested. As a control, stimulation with PHA was used. Results with patients’ PBMCs were compared with results of a parallel analysis of PBMCs from non‐atopic healthy controls.

Results
Serum sCD30 was elevated in the 18 atopic asthmatics compared with a group of normal subjects but levels did not correlate with symptomatic disease activity. CD4CD45RO expression was low (14%) in atopic asthmatic peripheral blood but increased to 41% after 10 days culture with allergen. The CD4:CD8 ratio increased after Der p stimulation. A significant rise in the percentage of CD44 T cells expressing surface CD30 (29%) was seen along with increased mean fluorescence intensity. Both these results correlated with symptomatic disease severity score. Non‐specific PHA stimulation failed to significantly affect CD30 expression.

Conclusions
There is a specific response to allergen in atopic asthma which causes significant increases in CD30 expression. This may correlate with disease severity.
Comparison of inflammatory cell counts in asthma: induced sputum vs bronchoalveolar lavage and bronchial biopsies
Authors: GROOTENDORST D. C. , SONT J. K. , WILLEMS L. N. A. , KLUIN‐NELEMANS J. C. , VAN KRIEKEN J. H. J. M. , VESELIC‐CHARVAT M. , STERK P. J.
Published: November 2003

Background
Induced sputum potentially allows monitoring of airway inflammation in patients with asthma in a non‐invasive way. However, the relationship between the cellular content in sputum and airway tissue has not been fully clarified.

Objective
We compared the cellular compositions of hypertonic saline‐induced sputum, bronchoalveolar lavage fluid (BAL) and bronchial biopsies in 18 clinically stable patients with mild to moderate atopic asthma (baseline FEV1: range 61–114%pred, PC20 methacholine: 0.04–4.7 mg/mL). They were treated with inhaled short‐acting bronchodilators on demand, with (n = 8) or without (n= 10) regular inhaled steroids.

Methods
Each patient underwent sputum induction and fiberoptic bronchoscopy on separate days in random order. Differential cell counts of induced sputum, bronchoalveolar lavage and bronchial wash were determined on May‐Grünwald‐Giemsa stained cytospins. Flow cytometry was performed on sputum and BAL samples. Immunohistochemical techniques were used to stain inflammatory cells in 6 μm cryostat sections of bronchial biopsies.

Results
Sputum cell differentials were not different between the patients with and without inhaled steroids, and showed a median value of 19.4% squamous cells, with 1.0% eosinophils, 3.3% lymphocytes, 28.7% neutrophils, 49.4% macrophages and 6.9% cylindric epithelial cells (in percentage non‐squamous cells). The percentage eosinophils in sputum was significantly correlated with their percentage in bronchial wash(Rs= 0.52, P = 0.03) and in BAL (Rs= 0.55,P = 0.02), whilst there was a trend towards such a correlation between the number of eosinophils/mL sputum and the number of EG2+ eosinophils/mm2 lamina propria in bronchial biopsies (Rs= 0.44,P= 0.07). In addition, the percentage of CD4+ lymphocytes correlated between sputum and BAL (Rs= 0.55, P= 0.03).

Conclusion
We conclude that the eosinophil counts in hypertonic saline‐induced sputum from patients with asthma are related to those in bronchial wash and BAL and, to a lesser extent, with the counts in bronchial biopsies. This suggests that induced sputum can be used to monitor the presence and severity of airway inflammation in asthma.