bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional studies of bone marrow haemopoietic and stromal cells in the myelodysplastic syndrome (MDS)
Authors: Coutinho L. H. , Geary C. G. , Chang J. , Harrison C. , Testa N. G. 
Published: March 2008

SummaryLong‐term bone marrow culture (LTBMC) was used to investigate the proliferative behaviour of marrow cells from a spectrum of cases of the myelodysplastic syndrome (MDS), and the results compared with those obtained in the conventional short‐term clonal assay. Two broad patterns of growth were revealed in LTBMC. In one group the incidence of haemopoietic progenitor cells steadily declined to abnormally low levels at 4 weeks, while in a second group they were maintained near normal levels for periods of up to 7 weeks. These growth patterns, which were not predictable from clonogenic assays on the marrow cells prior to LTBMC, or from the morphology of the bone marrow, may reflect the stage of evolution of the disease. Further studies of clonality are required to establish whether or not patients exhibiting the second pattern have a potentiality to harbour residual normal haemopoiesis.LTBMC was also used to study the function of MDS marrow stroma in terms of its ability to sustain the growth of normal haemopoietic progenitor cells. Although the phenotype of the cultured adherent cell layer, obtained from some patients, was atypical, no consistent functional defect of MDS stroma could be identified by studying the level of haemopoiesis reached by normal cells seeded into MDS stroma.
Bone marrow biopsy in myelodysplastic syndromes: morphological characteristics and contribution to the study of prognostic factors
Authors: Ríos A. , Cańtizo M. C. , Sanz M. A. , Vallespí T. , Sanz G. , Torrabadella M. , Gomis F. , Ruiz† C. , San Miguel J. F. 
Published: March 2008

SummaryTen characteristics of bone marrow (BM) biopsies in paraffin sections, obtained at diagnosis from patients with myelodysplastic syndromes (MDS) classified according to the FAB criteria, were analysed to identify both the most relevant morphologic data and any possible influence on survival.Agreement between two observers was obtained for 94% of the data. BM cellularity was increased in 63% of the cases and was higher in refractory anaemia with excess of blasts (RAEB), RAEB in transformation (RAEB‐t) and chronic myelomonocytic leukaemia (CMML) (P= 0.001). Dysmegakaryopoiesis and dyserythropoiesis were present respectively in 83% and 72% of the cases, with slight differences among the FAB subtypes. Abnormal localization of immature precursors (ALIP) was found in more than half of the cases and somewhat more frequently seen in the RAEB + RAEB‐t + CMML group (P= 0.07). Eosinophilia, plasmacytosis and reticulin fibrosis were evident in 26%, 18% and 47% of the cases respectively. Cellularity (P= 0.006), eosinophilia (P=0.009) and, to some extent, dysmegakaryopoiesis (P=0.07) bore a certain relationship with survival on univariate analysis. The presence of ALIP was not seen to affect the outcome. Multivariate analysis showed that the cellularity and presence of dysmegakaryopoiesis, in BM biopsy, added significant independent prognostic information to that achieved with age, platelet count and proportion of blast cells in BM aspirate, three variables with proven prognostic value in MDS patients. Using a regression model including these five characteristics we have stratified the patients into low, intermediate and high‐risk groups with different survivals (P=0.00001). The present findings show that BM biopsy is able to provide both morphological characteristics and information about the prognosis of survival, and should thus be included in the initial evaluation of MDS.
The effects of rhGM‐CSF on the neutrophil respiratory burst when studied in whole blood
Authors: Jaswon Mervyn S. , Khwaja Asim , Roberts Pamela J. , Jones H. Mark , Linch David C. 
Published: March 2008

SummaryA simple semiquantitative cytofluorometric method has been developed for measuring neutrophil respiratory burst activity in whole blood samples. This technique avoids the introduction of laboratory artefacts which modulate neutrophil function. In addition, flow cytometric analysis allows the response to be studied in individual cells. We show here that neutrophils examined freshly ex vivo, exhibit only weak respiratory burst activity in response to stimulation with the chemotactic peptide FMLP (10−6m). Prior incubation with rhGM‐CSF results in an increase in the number of responding cells from 13.5±2.36% (mean ±SEM) to 46.7±6.3% (P<0.0001) with an increase in total respiratory burst activity of 567% (P=0.001). The majority of neutrophils in whole blood (67.1±8.1%) exhibit respiratory burst activity in response to stimulation with phorbol ester (1 μg/ml of TPA), and this response is also significantly primed by rhGM‐CSF (P=0.004). The enhancement of respiratory burst activity induced by rhGM‐CSF is due to both recruitment of previously unresponsive neutrophils, and to intensification of the response of the responding cells. In vivo administration of rhGM‐CSF also results in priming of the respiratory burst in response to FMLP, although the enhancement of activity is not as great as that obtained when pre‐infusion blood samples are incubated with rhGM‐CSF in vitro.
Clinical and biochemical characterization of antithrombin III Franconville, a variant with Pro 41 Leu mutation
Authors: Roux N. de , Chadeuf G. , Molho‐Sabatier P. , Plouin P.‐F. , Aiach M. 
Published: March 2008

SummaryWe describe a familial study of AT III, a type III antithrombin III variant which was identified in the proposit‐us by gene analysis as Pro 41 Leu heterozygous mutation. None of the four members of the family who presented with defective heparin cofactor (hep‐cofactor) activity, and therefore probably carried the mutation, had experienced deep venous thrombosis. The abnormal AT III was purified from the propositus’plasma, taking advantage of the difference in NaCl concentrations required to elute variant and normal AT III from heparin–Sepharose. The antithrombin and anti‐Xa activities of the purified variant AT III were comparable to those observed for normal AT III, but hep‐cofactor activity was strikingly reduced. The enhancement by heparin of thrombin and F Xa inhibition by normal and variant AT III was compared in the absence of NaCl and in the presence of normal NaCl concentrations. The difference between the degrees of inhibition by normal and variant AT III was maximal at physiological ionic strength (i.e. at a concentration of 0.15 m). The quantification of heparin AT III interaction with both normal and variant purified proteins in a double reciprocal plot yielded similar dissociation constants but a 9‐fold decrease in the maximal pseudo‐first order constant. This suggests that Pro 41 is more involved in the molecular changes induced by heparin than in the primary binding of the activator.
Subcutaneous administration of recombinant human erythropoietin to subjects on continuous ambulatory peritoneal dialysis: an erythrokinetic assessment
Authors: Hughes R. T. , Cotes P. Mary , Pippard M. J. , Stevens J. M. , Oliver D. O. , Winearls C. G. , Royston J. P. 
Published: March 2008

SummaryErythrokinetic studies were performed in subjects on continuous ambulatory peritoneal dialysis, during a trial examining the effectiveness of subcutaneous administration of recombinant human erythropoietin (r‐HuEPO) in correcting the anaemia associated with end stage renal disease. 15 subjects (mean haemoglobin concentration 6.9 g/dl, SD 1.1) entered the study, and during treatment 9 were restudied at a haemoglobin concentration of 11–11.5 g/dl and six under‐went a third study at haemoglobin 13–13.5 g/dl. By adjusting the dose of r‐HuEPO, a stepwise increase in haemoglobin concentration was achieved, and this was accompanied by increases in total red cell volume and erythron transferrin uptake. Plasma volume decreased as red cell volume increased, leaving total blood volume essentially unchanged. Red cell survival, modestly reduced before treatment (mean 64, range 44–96 d, n= 6) tended to increase during treatment and when subjects were retested at a haemoglobin concentration of 13–13.5 g/dl (after 38–62 weeks treatment), the mean increase in red cell survival was 20 d (95% confidence interval 1–39 d).Thus subcutaneous r‐HuEPO is effective in correcting the anaemia of end stage renal disease when administered thrice weekly to subjects on continuous ambulatory peritoneal dialysis. It produces an increase in haemoglobin concentration primarily by expanding the erythron, and may have a secondary effect, seen after several months of treatment, of increasing red cell survival.
Rearrangements of immunoglobulin and TCR genes in lymphoid blast crisis of Ph + chronic myeloid leukaemia
Authors: Kerim Simonetta , Stul Michel , Mecucci Cristina , Vandenberghe Elisabeth , Cuneo Antonio , Cin Paola Dal , Michaux Jean‐Louis , Louwagie Andries , Cassiman Jean‐Jacques , Berghe Herman Van den 
Published: March 2008

SummaryClonal rearrangements of immunoglobulin heavy chain genes as well as both T cell receptor (TCR) δ and γ genes were found in four cases of blast crisis of Ph + chronic myeloid leukaemia with unequivocal B cell precursor (common) immunophenotype. In one case, the TCR β chain gene was also rearranged. Although the developmental sequence of TCR δ, γ and β rearrangements in T lymphocytes appeared to be respected, a full phenotypic effect, characteristic of T cell was not observed in these otherwise typical ‘common’ blast cells. Cytogenetic analysis ruled out the occurrence of TCR rearrangement due to structural chromosome changes. A high incidence of unexpected TCR gene rearrangements has been previously reported in the de novo‘common’ acute lymphoblastic leukaemias (ALL). Our cases of chronic myeloid leukaemia (CML) in lymphoid blast crisis show that genotypic similarities may exist between these two haematological entities.
Severe recessive poikilocytic anaemia with a new spectrin α chain variant
Authors: Lecomte M. C. , Feo C. , Gautero H. , Bournier O. , Galand C. , Garbarz M. , Boivin P. , Dhermy D. 
Published: March 2008

SummaryA severe neonatal haemolytic anaemia was observed in a propositus from the West Indies. Frequent blood transfusions were required until complete splenectomy was carried out. Blood smears showed predominant poikilocytosis with spherocytes and microcytes as observed in hereditary pyropoikilocytosis. Erythrocytes were completely fragmented after incubation at 45°C. The two asymptomatic parents had normal haematological profiles. The erythrocyte membrane electrophoretic patterns of the splenectomized propositus and her parents were normal. The propositus had a moderate defect in the spectrin (Sp) dimer self‐association. Limited tryptic digestion of the propositus’ Sp under standard conditions (0°C, 20 h, enzyme‐substrate ratio of 1/100) revealed an increased sensitivity to tryptic digestion. The major features detected by one‐ and two‐dimensional electrophoresis gels of Sp tryptic digests were the absence of high molecular weight peptides from the SpαII (48 kDa and 35 kDa peptides) and SpαIII (52 kDa peptide) domains with increased amounts of the lower molecular weight peptides from the SpαII and SpαIII (29 kDa peptide and 47 kDa peptide) domains respectively. Kinetic studies of Sp tryptic digestion (10 min to 36 h) confirmed the increased tryptic susceptibility of Sp. Immunodetection with specific anti‐α‐chain domain antibodies showed that the highest molecular weight peptides from the αII and αIII domains are released early in digestion, but disappear quickly, with an increase in their corresponding smaller peptides. The molecular weights of the peptides corresponding to the 48 kDa and 35 kDa peptides from the αII domain are slightly modified. Peptides from the αI and αIV domains showed no significant abnormalities. The Sps of both parents were normal. These results indicate that the patient has a novel Spα chain defect, which is probably located in the region of the αII domain which adjoins the αIII domain.
Identification of mRNA for PDGF B‐chain in human megakaryocytes isolated using a novel immunomagnetic separation method
Authors: Gladwin A. M. , Carrier M. J. , Beesley J. E. , Lelchuk R. , Hancock V. , Martin J. F. 
Published: March 2008

SummaryA rapid and simple method has been developed for the separation of pure populations of intact human megakaryocytes from whole bone marrow. Megakaryocytes were specifically recognized using monoclonal antibodies coupled to magnetizable articles. Cells labelled with the magnetizable probe were separated from unlabelled cells by introduction of a magnetic field. The technique yields megakaryocyte suspensions with a purity of > 98%. Electron microscope examination showed that the ultrastructure of the isolated megakaryocytes was well preserved. Using this method of cell purification, we have investigated expression of the gene for platelet‐derived growth factor (PDGF). RNA was isolated from cells harvested from either ribs or posterior iliac crest. The RNA was spotted onto nitrocellulose, and then hybridized using a c‐sis riboprobe specific for PDGF B chain mRNA. We demonstrate that mRNA for PDGF B‐chain is identifiable in samples of 50000 cells. We conclude that PDGF is synthesized by the megakaryocyte.
Haemophiliacs with factor VIII inhibitors in Finland: prevalence, incidence and outcome
Authors: Rasi V. , Ikkala E. 
Published: March 2008

SummaryTwenty‐five of the 139 Finnish patients (18·0%) with severe haemophilia A alive in 1960 or born later have or have had an inhibitor against factor VIII. 19 of the 110 patients alive have an inhibitor and the current prevalence is 17·3%. The incidence of new inhibitors was 10·3 per thousand patient years for the observation period starting from 1960, median 16 years. The age dependent cumulative risk of developing an inhibitor was 22% at the age of 10.There have been no deaths from bleeding since 1976, which has resulted in a marked decrease in mortality. The annual death rate of patients with inhibitors was 5·8 per thousand years of life in 1980–89 compared to 41·7 in the previous decade. The recent progress in the modalities for treatment of bleedings has markedly improved the outcome of patients with factor VIII inhibitors.
Urinary autoantibodies against intrinsic factor in pernicious anaemia patients
Authors: Wahlstedt Virve 
Published: March 2008

SummaryIn radioimmunoassay seven concentrated urines of penicious anaemia (PA) patients were positive for intrinsic factor (IF). Four were studied by gel filtration. Two contained both binding and blocking antibodies against IF, one had only blocking antibodies and one lacked both types of antibodies. The antibodies were mainly of the IgG‐type. No such antibodies were found in the urine of a healthy person. None of the urines studied contained enough protein to be classified as proteinuric. Not until the interferences of the autoantibodies in the IF assay can be eliminated is the assay of value in the diagnosis of PA.
Haemophilia protects against ischaemic heart disease: a study of risk factors
Authors: Rosendaal F. R. , Briét E. , Stibbe J. , Herpen G. van , Leuven J. A. Gevers , Hofman A. , Vandenbroucke J. P. 
Published: March 2008

Summary. We previously reported that mortality due to ischaemic heart disease was lower in haemophilia patients than in the general male population. To support the hypothesis that this could be attributed to a protective effect of the clotting defect in haemophilia and not to differences in cardiovascular risk factors, we performed a second study. We examined 95 haemophilia patients for the presence of major risk factors for ischaemic heart disease and compared their risk factor profile with the data of epidemiologic surveys of the general Dutch population.Haemophilia patients had on average higher blood pressures than the comparison population, were more often hypertensive and used antihypertensive drugs twice as often.The mean serum cholesterol level of the patients was markedly lower than in the comparison population (4.8 versus 5.6 mmol/1. 9 5% confidence interval of the difference: 0.5–1.1 Mmmol/l).The risk factors were weighted into one theoretical risk ratio for ischaemic heart disease by application of logistic regression coefficients. The theoretical risk ratio based on the risk factor profile was 0.78. This risk ratio can only explain a moderate reduction in the, incidence of ischaemic heart disease, much smaller than the mortality ratio of 0.20 we reported previously. Therefore these data support the hypothesis of a direct protective effect of haemophilia on the development of ischaemic heart disease.
Natural history of stage A chronic lymphocytic leukaemia untreated patients
Authors:   French Cooperative Group on Chronic Lymphocytic Leukaemia (see appendix for list of members)
Published: March 2008

Summary. In chronic lymphocytic leukaemia (CLL), stage A was defined in 1979 as the best prognostic group of the three‐stage (A, B, C) classification which was thereafter adopted by an International Workshop on CLL. Recently, the question of whether or not all stage A patients should be treated was raised by authors who described potential harmful effects of treatment among stage A patients and heterogeneity of stage A with respect to survival and disease progression.In this work, we studied the natural history of stage A‐CLL in order to determine on which grounds a decision to treat should be made. We performed a prognostic study on 309 untreated stage A‐CLL patients enrolled in a randomized clinical trial from 1980 to 1985. Our aim was to segregate a subgroup with a low probability of disease progression and death. We used three endpoints: overall survival (50 deaths), disease progression to stage B or C (78 events) and disease progression to stage C (39 events). Multivariate analyses, using Cox's model, selected nine variables as having a prognostic value for at least one endpoint, namely age, sex, general symptoms, involved axillary lymph node, splenomegaly, haemoglobin, platelets, lymphocyte count and bone‐marrow infiltration. By considering the three endpoints jointly, we obtained a proposed definition of smouldering CLL based on four variables, namely haemoglobin level, lymphocyte count, the number of enlarged areas and bone‐marrow infiltration. However, we did not succeed in better defining smouldering CLL than in a previous proposal, based on haemoglobin level and lymphocyte count. Moreover, whatever the definition used, including other previous works, disease progression within 5 years was still observed for about 10% of patients with smouldering CLL. Future attempts to define smouldering CLL could perhaps benefit from using biological markers.
Induction of patching and its reversal on surface‐activated human platelets
Authors: White James G. 
Published: March 2008

Summary. Fibrinogen coupled to colloidal gold (Fgn/Au) has been used to follow GPIIb‐IIIa receptors on surface‐activated platelets and to evaluate the effects of the anti‐actin agent, cytochalasin B (CB), on the movement of receptor‐ligand complexes. Fgn/Au binds diffusely to spread platelets, but is translocated rapidly from the peripheral margin to cell centres and to channels of the open canalicular system (OCS).CB added after spreading and incubation with Fgn/Au reverses the centripetal movement of receptor‐ligand complexes. Fgn/Au particles move back toward the margins of spread cells and form patches. The patches of receptor‐ligand complexes overlie segments of the OCS filled with Fgn/Au. Washing out the CB reverses the process and receptor‐ligand complexes are again translocated toward platelet centres.
Interleultin‐2 treatment‐associated eosinophilia is mediated by interleultin‐5 production
Authors: Macdonald D. , Gordon A. A. , Kajitani H. , Enokihara H. , Barrett A. J. 
Published: March 2008

Summary During a trial using recombinant human interleukin‐2 (rhIL‐2) immunotherapy for acute myeloblastic leukaemia (AML) in remission, eosinophilia was observed in all patients. We used in‐vitro clonogenic assays to investigate the mechanism of the eosinophilia in five patients.The mean eosinophil count increased from 0.05 ± 109/1 before rhTL‐2 to 0.98 ± 109/1 within 48 h of stopping the infusion, and an exponential correlation between the pre‐treatment lymphocyte CD4:CD8 ratio and the maximum eosinophil count was observed. KhIL‐2 did not stimulate eosinophil colony formation by normal bone marrow. However, serum collected from patients during rhIL‐2 infusion was a potent stimulator of eosinophil colony forming units (CFU‐Eo), but had no significant stimulatory effect on granulocyte‐macrophage colony forming units (CFU‐GM). The CFU‐Eo stimulation by pre‐treatment serum was 2.8‐fold higher than control serum. Serum collected during treatment stimulated CFIJ‐Eo 12 times more than control serum (P<045).By pre‐incubating patient serum, collected during rhIL‐2 treatment, with monoclonal antibodies to murine IL‐5. or human granulocyte‐macrophage colony stimulating factor (GM‐CSF). a reduction of 80% and 38% respectively in eosinophil and GM colony production was found. The CFU‐Eo stimulating effect of patient serum was in the range of the CFU‐Eo stimulating effect of normal serum, after the addition of 5 u/ml of recombinant murine IL‐5.The results suggest that eosinophilia was caused by IL‐5 and GM‐CSF production by rhIL‐2 stimulated CD4 positive lymphocytes. The location on chromosomes 5 of the genes for IL‐5. GM‐CSF and IL‐3 may be associated with regulation of expression. by a common mechanism. of all the factors known to be involved in eosinophil production. This mechanism may be activated by IL‐2 stimulation. The separate location on chromosome 17 of the G‐CSF gene may explain the ability of IL‐2 to produce a distinct stimulus to eosinophil but not neutrophil production.
Expression of the multiple drug resistance gene (mdr‐1) and epitope masking in chronic lymphatic leukaemia
Authors: Cumber Peter M. , Jacobs Allan , Hoy Terry , Fisher Janet , Whittaker John A. , Tsuruo Takashi , Padua Rose Ann 
Published: March 2008

Summary Resistance to cytotoxic agents is a common clinical problem in the treatment of chronic lymphatic leukaemia (CLL). The multidrug resistant (MDR) phenotype characterized by increased levels of a specific cell membrane p‐glycoprotein. confers cross resistance to a wide range of structurally dissimilar antineoplastic drugs. We have studied the expression of this p‐glycoprotein in chronic lymphatic leukaemia measured by immunofluorescence using a monoclonal antibody MRK 16 by flow cytometry.Initial results showed that only 12% of lymphocyte samples from CLL patients showed increased p‐glycoprotein, conflicting with a previous observation that 53% of CLL patients had an increased level of mdr‐1 mRNA. Treatment of the cells with neuraminidase to remove sialic acid residues increased the proportion of patients showing increased p‐glycoprotein to 52%. This suggests that in a subset of CLL patients post translational modification of the protein occurs masking the epitope recognized by MRK 16. Abnormal sialylation patterns associated with malignancy are a well‐recognized phenomenon.