bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytomegalovirus (CMV) antigenaemia for rapid diagnosis and monitoring of CMV‐associated disease after bone marrow transplantation
Authors: Gondo Hisashi , Minematsu Toshio , Harada Mine , Akashi Koichi , Hayashi Shin , Taniguchi Shuichi , Yamasaki Kazuo , Shibuya Tsunefumi , Takamatsu Yasushi , Teshima Takanori , Eto Tetsuya , Nagafuji Koji , Mizuno Shin‐ichi , Hosoda Kenji , Mori Ryoichi , Minamishima Yoichi , Niho Yoshiyuki 
Published: March 2008

A technique for the rapid detection of cytomegalovirus (CMV) antigen‐positive blood leucocytes (CMV antigenaemia) was evaluated in 15 marrow transplant patients as a means of diagnosis and for monitoring CMV‐associated disease. CMV antigenaemia was determined by direct immunoperoxidase staining of leucocytes with a peroxidase‐labelled monoclonal antibody, HRP‐C7, which binds an immediate‐early antigen of human CMV.CMV antigenaemia occurred in 7/15 marrow transplant patients (47%) and was initially detected between 4 and 6 weeks after transplantation. CMV‐associated diseases developed in 3/15 patients (20%). All patients with CMV‐associated disease had a relatively large number of CMV antigen‐positive leucocytes, exceeding 10 per 50000 white blood cells (WBCs). In the remaining 12 patients, CMV antigen‐positive leucocytes were less than 10 per 50000 WBCs or were undetectable. CMV‐associated disease did not develop in these patients during the period of monitoring. CMV antigen‐positive leucocytes were detected more frequently in patients who developed acute graft‐versus‐host disease (GVHD) or haemorrhagic cystitis than in those without such complications. CMV antigens were detectable from 1 to 4 weeks before the onset of CMV‐associated disease which allowed initiation of ganciclovir treatment at an early stage. The degree of CMV antigenaemia paralleled the clinical symptoms and signs, higher degrees of antigenaemia being associated with more significant disease. Thus, the detection of CMV antigen‐positive blood leucocytes is useful for the diagnosis and monitoring of CMV‐associated disease following bone marrow transplantation.
Abnormal cAMP‐induced phosphorylation of rap 1, protein in grey platelet syndrome platelets
Authors: Enouf Jocelyne , Corvazier Elisabeth , Papp Béla , Quarck Rozenn , Magnier Clarice , Kovàcs Tünde , Bredoux Raymonde , Lévy‐Tolédano Sylviane , de Gunzburg Jean , Wuytack Frank , Caen Jacques 
Published: March 2008

Summary We previously demonstrated abnormal Ca2+ transport by microsomes in platelets from a grey platelet syndrome patient. Here, we investigated the platelet Ca2+ ATPases that mediate this transport, as well as its possible regulation by rap 1 protein. We showed that grey platelet syndrome platelets expressed the same two distinct Ca2+ ATPases as those recently described in normal platelets; the 100 kD SERCA2‐b isoform (Sarco/Endoplasmic Reticulum Ca2+ATPase) and a new 97 kD SERCA isoform. The two Ca2+ ATPases formed similar amounts of transient phosphorylated intermediates. The expression of these two Ca2+ ATPases was compared by Western blotting using specific antibodies, which again emerged in similar amounts in normal and grey platelet syndrome platelets. As regards the protein phosphorylated by cAMP, it was found to be identical to rap 1 protein when it was immunoprecipitated with an antibody raised against a synthetic peptide specific for rap 1 protein. Although the expression of rap 1 protein was similar in membranes isolated from grey platelet syndrome and normal platelets, its exogenous phosphorylation by cAMP was abnormal, with a concentration (10 μg/ml) of the catalytic subunits of the cAMP‐dependent protein kinase (C.Sub.), as it decreased to half the control level.It is concluded that the abnormal Ca2+ transport found in grey platelet syndrome platelets is not due to the abnormal expression of the Ca2+ ATPases, but is associated with an abnormality of rap 1 protein phosphorylation by cAMP.
Lack of restricted T‐cell receptor β‐chain variable region (Vβ) usage of reactive T‐lymphocytes in Hodgkin's disease
Authors: Meggetto Fabienne , Al Saati Talal , Rubin Bent , Delsol Georges 
Published: March 2008

Summary. T‐cell response against tumour‐associated antigens is mediated by the TCR complex. To determine a possibly restricted TCR‐Vβ repertoire in reactive T‐lymphocytes in Hodgkin's disease (HD), 20 cases (of which 10 were EBV‐positive cases) were investigated using 14 monoclonal antibodies (MoAbs) recognizing 11 different TCR‐Vβ region family products and Northern blot analysis with cDNA probes specific for mRNA transcripts of 11 Vβ families that were not detectable by MoAbs. Four Vβ families (Vβ5, Vβ6, Vβ8, Vβ19) were investigated using both immunohistochemistry (IHC) with anti‐Vβ MoAbs and Northern blot analysis. Immunohistochemical and Northern blot findings were correlated with the detection of the Epstein‐Barr virus (EBV) genome in Hodgkin's and Reed‐Sternberg cells (H‐RS).The non‐neoplastic lymphocytes in HD were predominantly of T‐phenotype (CD3+). Most of these cells were TCR‐αβ+ (βF1+) and only a few T‐cells were reactive for TCR‐δ1 antibody (TCR‐γδ+). In the majority of cases helper/inducer T‐cells (CD4+) outnumbered suppressor/cytotoxic T‐cells (CD8+). Labelling of these samples with the panel of 14 anti‐Vβ MoAbs showed that only a small percentage (0·2–5·5%) of βF1+ lymphocytes were positive with each of these MoAbs. The proportion of these cells was comparable to that seen in normal tissues. Most TCR Vβ+ cells were randomly distributed, but in virtually all cases occasional Vβ+ cells pertaining to the various Vβ families were seen in close contact to H‐RS cells.Using total RNA extracted from malignant and normal tissues, no visible band was detected with the various Vβ probes. As determined in the present study, the percentage of T‐cells expressing a given Vβ family must be 10% to be detected with Northern blot. Thus, the percentage of Vβ+ cells expressing Vβ families which were explored only with Northern blot were within the same range as those of the 11 different TCR‐Vβ region families assessed with IHC, i.e. 1–10% of lymphoid cells. The results of the present study show that in HD there is no restricted T‐cell Vβ repertoire usage regardless of the detection of EBV. In addition, since the various Vβ families are represented in T‐cell subpopulations forming rosettes around H‐RS cells, we conclude that the T‐cells attracted by H‐RS cells constitute a polyclonal population.
Molecular defects in CRM+ factor VII deficiencies: modelling of missense mutations in the catalytic domain of FVII
Authors: Bernardi F. , Liney D. L. , Patracchini P. , Gemmati D. , Legnani C. , Arcieri P. , Pinotti M. , Redaelli R. , Ballerini G. , Pemberton S. , Wacey A. I. , Mariani G. , Tuddenham E. G. D. , Marchetti G. 
Published: March 2008

Summary. The molecular defects causing CRM+ factor VII deficiency were investigated in seven unrelated subjects and several members of their families.Four missense mutations located in the catalytic domain of factor VII were found. The previously reported 304ArgGln substitution was present in the homozygous and heterozygous forms, with different polymorphic haplotypes, thus demonstrating that it is recurrent and frequent in the Italian population. The 310Cys Phe substitution was found in the homozygous form and in the compound heterozygous condition with the nonsense mutation 356Trpstop. Two missense mutations, 298MetIle and 342GlyArg, were found in the homozygous and in the heterozygous condition respectively.Molecular heterogeneity was further increased by finding of the 353ArgGln polymorphism in the doubly heterozygous condition with the 304 and 342 mutations.Plausible explanations for loss of FVII function were found by inspecting a model of the serine protease domain of factor VIIa. Inefficient activation of the catalytic site is predicted for 298MetIle. 342GlyArg would directly distort the geometry of the ‘oxyanion hole’preventing formation of a substrate enzyme intermediate. 310Cyshe is predicted to have an adverse effect on tissue factor interaction. These mutations point to important regions of the factor VII molecule.
Marked and reproducible increase in trilineage blood cell counts by administration of granulocyte colony‐stimulating factor in a patient with refractory anaemia with excess blasts in transformation
Authors: Chiba Shigeru , Inamori Ken , Mitani Kinuko , Hirai Hisamaru , Yazaki Yoshio 
Published: March 2008

Summary. We report a patient with refractory anaemia with excess blasts in transformation (RAEB‐t) who presented with severe pancytopenia and received four intermittent series of granulocyte colony‐stimulating factor (G‐CSF) treatment over 1·5 years. In addition to the increase in mature neutrophils, platelet count and haemoglobin level were dramatically increased. These haematological improvements were dependent on G‐CSF during these treatment series. Bone marrow colony assay revealed that G‐CSF increased both CFU‐E‐ and BFU‐E‐derived colonies in vitro.Clinical usage of G‐CSF in myelodysplastic syndrome (MDS) is discussed, with particular emphasis on mechanisms of trilineage response.
Identification of a subgroup of myelodysplastic patients with a neutrophil stimulation‐signalling defect
Authors: Lowe G. M. , Dang Y. , Watson F. , Edwards S. W. , Galvani D. W. 
Published: March 2008

Summary. f‐Met‐Leu‐Phe‐stimulated luminol‐enhanced chemiluminescence was found to be repeatedly defective in some MDS patients. This defect was not attributed to myeloperoxidase deficiency, nor to a defect in NADPH oxidase function, because PMA chemiluminescence was found to be normal in these individuals. An arbitrary value of 7 mV (half the mean control value) was chosen to subdivide the group: MDS patients with values <7 mV had a mean f‐Met‐Leu‐Phe chemiluminescence response of 2·5±0·5 compared to MDS patients with values <7 mV who had a mean response of 15·6±1·6mV, P<0·01 (healthy controls 14±2 mV). The characteristics of the f‐Met‐Leu‐Phe receptor and initial calcium flux results suggested that the receptor itself was normal in number and function in low f‐Met‐Leu‐Phe responders. The rate of superoxide generation, which is calcium‐dependent, was also found to be in the normal range in low f‐Met‐Leu‐Phe responders, although total superoxide production was reduced in some of these patients.When MDS neutrophils with a low f‐Met‐Leu‐Phe response were stimulated with PMA, chemiluminescence was normal, suggesting normal activity of the NADPH‐oxidase complex. Furthermore, myeloperoxidase activity was reduced in only three out of the 11 low f‐Met‐Leu‐Phe responders.Following priming with GM‐CSF, f‐Met‐Leu‐Phe chemiluminescence was 27·1±6 mV in low f‐Met‐Leu‐Phe responders compared to controls (87·7±11 mV, P<0·005). Thus, although responses were improved, they were not as marked as in control neutrophils.These data suggest that a subgroup of MDS patients have a low f‐Met‐Leu‐Phe chemiluminescence response which is not due to a defect in the f‐Met‐Leu‐Phe receptor or oxidase activity, and in the majority of cases MPO activity is normal. Initial patient survival data suggest that these patients may have an increased risk of infective mortality. It is proposed that defective f‐Met‐Leu‐Phe chemiluminescence results from a putative defect in cell‐signalling mechanism upstream of PKC, and GM‐CSF priming only partially improves responsiveness.
Trisomy 8q due to i(8q) or der(8) t(8;8) is a frequent lesion in T‐prolymphocytic leukaemia: four new cases and a review of the literature
Authors: Mossafa Hossain , Brizard André , Huret Jean‐Loup , Brizard Françoise , Lessard Michel , Guilhot François , Tanzer Joseph 
Published: March 2008

Summary.Cytogenetic abnormalities found in four cases of T‐cell prolymphocytic leukaemia (T‐PLL) are described. An isochromosome 8q was found in three patients and a t(8;8) in one. In the four cases, karyotypes were complex and showed a high degree of instability. In addition, we reviewed 27 published cases of cytogenetically studied T‐PLL. On the whole, the most frequently recurring anomalies in T‐PLL are 14q lesions with nonrandom breakpoints, inversion (14)(q11q32) or tandem translocations (14;14) (not seen in any of our cases) and trisomy for 8q, mainly due to i(8q), found in more than 40% of patients each.Similar structural anomalies were found almost as frequently among the 23 cytogenetically studied cases of so‐called T‐chronic lymphocytic leukaemia (T‐CLL) reported prior to 1989. It is now accepted that the T‐cell counterpart of B‐CLL either does not exist or is exceedingly rare and thus previously reported cases of T‐CLL sharing the chromosomal characteristics of T‐PLL may well have been misdiagnosed examples of T‐PLL.Isochromosomes 8q are exceptionally found in other types of haematological malignancies. However, i(8q) could not be shown to be the primary lesion in any case in T‐PLL and the role of trisomy for 8q, as well of the associated monosomy 8p, is entirely unknown.
Rapid onset of chronic granulocytic leukaemia after treatment for Hodgkin's disease
Authors: Jackson G. H. , Latham J. A. , Lennard A. L. , Taylor P. R. A. , Proctor S. J. 
Published: March 2008

Summary. A 17‐year‐old white female presented with stage IVB Hodgkin's disease. After chemotherapy and radiotherapy she achieved a complete clinical remission and underwent an autologous bone marrow transplant (ABMT). 22 months later she developed chronic granulocytic leukaemia (CGL).Polymerase chain reaction (PCR) analysis of bone marrow harvested at the time of ABMT did not show any evidence of the bcr‐abl sequence that was detectable at the diagnosis of CGL. This case provides further information on the kinetics of the development of CGL and adds to the small pool of data on CGL developing after treatment for Hodgkin's disease.
Establishment and characterization of a novel myeloid cell line from the bone marrow of a patient with the myelodysplastic syndrome
Authors: Tohyama Kaoru , Tsutani Hiroshi , Ueda Takanori , Nakamura Toru , Yoshida Yataro 
Published: March 2008

SUMMARY. A novel long‐term cultured myeloid cell line was established from the bone marrow of a patient with myelodysplastic syndrome (MDS). This cell line, designated MDS92. proliferated in the presence of interleukin‐3 or granulocyte‐macrophage colony‐stimulating factor and transiently in the presence of Steel factor, with a tendency for gradual maturation, and formed myeloid colonies in the semi‐solid culture condition.In addition, the MDS92 cell line represented rather complicated karyotypic abnormalities including the deletion of fifth and seventh chromosomes and a point mutation at codon 12 of the N‐ras oncogene.These characteristics of the MDS92 cell line are exclusively compatible with the property of preleukaemia.
Dissociation between the phospholipases C and A2 activities in stimulated platelets and their involvement in the arachidonic acid liberation
Authors: Faili Ahmad , Emadi Sharareh , Vargaftig B. Boris , Hatmi Mohamed 
Published: March 2008

Summary. In previous work we have demonstrated that platelets depleted from secretory phospholipase A2 (sPLA2) produced similar amounts of thromboxane (Tx)B2 as control platelets upon stimulation by thrombin. However, since depletion of sPLA2 was not total, this sole finding only suggested the non‐involvement of sPLA2 in arachidonic acid release.In the present study we provide further evidence for the non‐involvement of sPLA2 in arachidonic acid liberation during platelet activation. Thus, rabbit platelets exposed to thrombin secreted sPLA2, released free arachidonic acid and formed TxB2 and inositol phosphates. In contrast, U46619, a stable prostaglandin (PG)H2 analogue, activates phospholipase C (PLC) and induces release of sPLA2 without TXB2 generation nor arachidonic acid liberation. At each concentration tested of both agonists, stimulation of sPLA2 activity paralleled the production of inositol phosphates. These data suggest that sPLA2 is dependent on phosphoinositide hydrolysis and on the release reaction and that it is not involved in the liberation of arachidonic acid from stimulated platelets. In addition, a dissociation was observed between sPLA2 and the enzyme involved in the arachidonic acid mobilization, suggesting that the liberation of this fatty acid from membrane phospholipids was mediated by cytosolic phospholipase A2 (cPLA2). Finally, PLC does not play a major role in arachidonic acid liberation, since U46619, which induced the breakdown of inositol phospholipids, failed to release arachidonic acid. In confirmation, neomycin, which inhibits PLC activity, failed to inhibit ATP, sPLA2 and arachidonic acid release upon stimulation of platelets by fluoroaluminate. These data demonstrate that sPLA2 is not involved in the arachidonic acid release by stimulated platelets and indicate that the activations of PLC, sPLA2 and cPLA2 are independent events.
Isolation and culture of endothelial cells from human bone marrow
Authors: Masek Lisa C. , Sweetenham John W. 
Published: March 2008

Summary : Adhesive interactions between haemopoietic progenitor cells and bone marrow sinusoidal endothelium are potentially important in the homing of these cells back to re‐establish haemopoiesis following stem cell transplantation. A simple method for the isolation and culture of human bone marrow endothelial cells is described using bone marrow aspirates obtained from patients undergoing bone marrow harvests for autologous or syngeneic bone marrow transplantation. The method is based on the selective binding of the lectin Ulex europaeus agglutinin‐1 (UEA‐1) to endothelial cells. Magnetic Dynabeads coupled with UEA‐1 were incubated with single cell suspensions of bone marrow following red cell lysis, and bound cells were isolated with a magnet. The isolated cells demonstrated positive immunofluorescence staining for von Willebrand factor. Cells were plated onto tissue culture flasks coated with extracellular matrix derived from human umbilical vein endothelial cells in an endothelial serum‐free medium together with 5% fetal calf serum for 24h. Cells were then cultured in endothelial serum‐free growth medium supplemented with 5% fetal calf serum, endothelial cell growth supplemented with 5% fetal calf serum, endothlial cell growth supplement and heparin. After 2–4 weeks in culture, two morphologically different cell populations can be identified. One has a polygonal spindle‐shaped morphology with a rapid growth rat, the other a rounded morphology and a slow growth rate. Both population have a vasiculated cytoplasm. Positive immunostaining of the cells was demonstrated with a number of endothelial cell markers including von Willebrand factor, and antibodies to ICAM‐1, VCAM‐1, E‐wlectin, CD31 and BMA120. Weibel‐Palade bodies were observed by electron microscopy.Culture of these cells will allow detailed in vitro studies of adhesion mechanisms in the homing of haemopoietic progenitor cells.
Phosphorylation of 2‐chlorodeoxyadenosine (CdA) in extracts of peripheral blood mononuclear cells of leukaemic patients
Authors: Arner Elias S. J. , Spasokoukotskaja Tatjana , Juliusson Gunnar , Liliemark Jan , Eriksson Staffan 
Published: March 2008

Summary2‐Chlorodeoxyadenosine (CdA) is an antileukaemic agent used in treatment of hairy cell leukaemia (HCL) and chronic lymphocytic leukaemia of B‐ and T‐cell type (B‐CLL and T‐CLL). The aim of this study was to elucidate the interpatient variability of CdA phosphorylation and its relation to response to CdA treatment. In extracts of peripheral blood mononuclear cells of patients with B‐CLL (n= 39), CdA phosphorylation was significantly higher than in HCL (n= 19) when calculated per protein (391 ± 155 pmol CdA phosphorylated/mg protein/min versus 288 ± 166 pmol/mg/min, P < 0.001), but was the same when calculated per cell (12 ± 5.9 pmol/106 cells/min versus 14 ± 5.9 pmol/106 cells/min) due to a larger cell volume in HCL. In T‐CLL (n= 6), CdA phosphorylation was significantly lower than in B‐CLL, both when calculated per protein (128 ± 68 pmol/mg/min, P < 0.001) or per cell (5.7 ± 2.7 pmol/106 cells/min, P < 0.05). This low CdA phosphorylation in T‐CLL was unexpected because normal B‐ and T‐lymphocytes contain equal amounts of CdA phosphorylation. With B‐CLL, 21 patients who responded (complete and partial response) to CdA treatment showed a significantly higher CdA phosphorylation than 13 patients not responding to CdA treatment (456 ± 170 pmol/mg/min versus 309 ± 97 pmol/mg/min, P < 0.01). We conclude that the level of CdA phosphorylation is correlated with the response of leukaemias to CdA treatment.
Adult T‐cell leukaemia with various abnormalities in endocrine and metabolic systems
Authors: Adachi Mitsunobu , Shiomura Tadahiko , Shimada Hideto , Kishimoto Norihumi , Iida Kei , Ikeno Humiaki , Nishimura Haruo , Inoue Tatsuhide , Sako Yoshiyasu 
Published: March 2008

SummaryAdult T‐cell leukaemia (ATL) is a unique type of T‐cell malignancy closely associated with human T‐cell leukaemia virus‐1 (HTLV‐1). Despite frequent descriptions of hypercalcaemia, cases accompanied by diabetes insipidus or syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in ATL patients have rarely been reported. We present an unusual case of ATL with various abnormalities in his endocrine and metabolic systems involving anterior pituitary function, thyroid function, lipid metabolism and Ca metabolism. Some of these abnormalities were considered to arise from infiltration or leukaemic cells into systemic organs after elimination of the above symptoms. Clinical and haematological data showing improvement following chemotherapy are also presented.
The proliferative activity of myelopoiesis in myelodysplasia evaluated by multiparameter flow cytometry
Authors: Jensen Inger Marie , Hokland Peter 
Published: March 2008

SummaryDouble staining of bone marrow cells for CD13 and CD33 leucocyte differentiatian antigens and for DNA content has allowed us to evaluate the proliferative capacity of myelopoiesis in patients with myelodysplastic syndromes (MDS) using flow cytometry. By analysing 39 patients (15 RA/RAS, 14 RAEB and 10 RAEB‐t) and eight normal controls, we found significant differences in both the percentage of cells positive for these immature myeloid antigens between the FAB groups as well as in the fractions of CD13 and CD33 positive cells in S or S‐G2M phase of the cell cycle. Moreover, a clear decrease in the immature myeloid cell proliferative activity upon progression within the FAB groups was evident. Finally, we found a significant negative association between the percentage of myeloblasts in the bone marrow and the proliferative activity of the immature myeloid cells. indicating that the block in differentiation in MDS patients might be coupled to a simultaneous block in proliferation, especially in advanced stages. These data suggest that the use of double parameter assays in the longitudinal follow‐up of MDS patients might yield new information about the biology of MDS.