bluuebunny/crossref_metadata_2025 huggingface dataset:
Children in long‐term remission after treatment for acute lymphoblastic leukaemia show persisting haemopoietic injury in clonal and long‐term cultures
Authors: Bhavnani M. , Jones P. H. Morris , Testa N. G.
Published: March 2008

Twenty children who were in unmaintained full haematological remission after treatment for acute lymphoblastic leukaemia (ALL) showed a significantly lower incidence of granulocyte‐macrophage progenitor cells (GM‐CFC) in the bone marrow compared to controls. This low incidence lasted for up to at least 3 years after the cessation of chemotherapy. There was no tendency to higher values with longer times after treatment, and the low incidence was not predictive of relapse.

Long‐term cultures from ALL bone marrows and from controls achieved similar levels of production of mature cells through the whole period of culture (6 weeks). However, cultures from patients’ bone marrow had on average about 5 times lower numbers of GM‐CFC, indicating that the level of mature cell production was achieved by a higher level of post‐GM‐CFC amplification than needed in the controls. This is taken to be due to compensatory mechanisms operative during stressed haemopoiesis which appears to be a long‐lasting effect after current chemotherapy of ALL.
Diagnostic differentiation of chronic B‐cell malignancies using monoclonal antibody L161 (CD1c)
Authors: Jones Richard A. , Master Peter S. , Child J. A. , Roberts Bryon E. , Scott Colin Stephen
Published: March 2008

The expression of membrane CD1c, as defined by monoclonal antibody L161, was examined on malignant lymphoid cells from 191 cases of chronic lymphoproliferative disease and on eight ‘normal’ enriched tonsil B‐cell extracts. Of 79 cases of chronic lymphocytic leukaemia (CLL) studied, 77 showed low (<20% positive cells) CD1c expression whereas 63/71 (89%) cases of B‐PLL, HCL and B‐NHL showed increased CD1c+ (but not CD1a or CD1b) components. In contrast, malignancies corresponding to terminal stages of B‐cell differentiation (immunocytoma and myeloma) generally showed low CD1c expression as did lymphoid cells from 10 cases of post‐thymic malignancy. Although there was some correlation between the expression of membrane CD1c and immunoglobulin (SIg) light chain densities (P<0.001), it is relevant in diagnostic terms that seven cases of B‐NHL, with low SIg staining intensities more typically associated with CLL were CD1c+. CD1c expression was not, however, correlated with the presence of CD23 or FMC7 determinants but did show a similar pattern of expression to that previously reported for beta‐2 microglobulin. Determination of cellular CD1c by APAAP immunocyto‐chemistry confirmed the presence of higher antigen densities in malignant B‐cells at intermediate/late stages of differentiation and this interpretation was further supported by the finding that the majority of phenotypically mature tonsil B‐cells were also CD1c+. The determination of CD1c expression by malignant B‐cells may therefore be of particular value in the diagnostic differentiation of chronic lymphoproliferative disorders.
Heterogenous response of B cell chronic lymphocytic leukaemia (B‐CLL) cells to anti‐human IgM antibody (anti‐μ) and B cell stimulatory factor 1 (BSF‐1)
Authors: Kawano Michio , Iwato Koji , Asaoku Hideki , Tanabe Osamu , Tanaka Hideo , Ishikawa Hideaki , Imamura Nobutaka , Kuramoto Atsushi
Published: March 2008

B cell chronic lymphocytic leukaemia (B‐CLL) cells from seven patients were examined for 3H‐TdR uptake with anti‐human IgM antibody (anti‐μ) and B cell stimulatory factor 1 (BSF‐1)/IL‐4. B‐CLL cells from one patient could proliferate with anti‐μ plus BSF‐1, but not with BSF‐1 alone. B‐CLL cells from two patients responded to BSF‐1 alone; one could proliferate more by adding anti‐μ, but the proliferation of the other cells was inhibited by adding anti‐μ. On the other hand, B‐CLL cells from the other four patients responded neither to anti‐μ nor to anti‐μ plus BSF‐1. Surface densities of membrane IgM of B‐CLL cells were also analysed by fluorescence activated cell sorter (FACS), but there was no correlation between proliferative response (DNA synthesis) of B‐CLL cells to anti‐μ and densities of surface membrane IgM. These results show that there is a functional heterogeneity of B‐CLL cells with regard to proliferative response to anti‐μ and BSF‐1.
Plasma D‐dimer levels and their relationship to serum fibrinogen/fibrin degradation products in hypercoagulable states
Authors: Wilde J. T. , Kitchen S. , Kinsey S. , Greaves M. , Preston F. E.
Published: March 2008

Plasma D‐dimer was measured and compared with serum fibrinogen/fibrin degradation product levels (FDPs) in patients with disseminated intravascular coagulation (DIC) and other conditions associated with a hypercoagulable state. D‐dimer (N<200 ng/ml) was elevated in all 43 patients with DIC, in 48 of 59 patients with liver disease, in 22 of 27 patients with acute leukaemia at presentation, in 17 of 23 patients with malignant disease, in 29 of 39 women in the third trimester of a complicated pregnancy, in 17 of 18 patients with deep venous thrombosis and in only four of 27 patients with acute myocardial infarction. There was a significant correlation between plasma D‐dimer and serum FDP levels(P<0.01) as follows; DIC: r =r=0.58, liver disease:r=0.57, acute leukaemia:r=0.84, malignancy:r=0.87. The frequent elevation of D‐dimer observed in liver disease, acute leukaemia, malignancy and complicated pregnancy indicates that a hypercoagulable state is a common occurrence in these conditions although in liver disease elevated levels resulting from a failure of normal clearance mechanisms cannot be excluded. The close relationship between D‐dimer and FDP levels suggests that serum FDPs predominantly arise from the interaction of plasmin with crosslinked fibrin rather than with fibrinogen in the conditions in which these were compared.
Immunochemical characterization of an autoantigen on platelet glycoprotein IIb in chronic ITP: comparison with the Baka alloantigen
Authors: Tomiyama Y. , Kurata Y. , Shibata Y. , Honda S. , Furubayashi T. , Mizutani H. , Tsubakio T. , Yonezawa T. , Tarui S.
Published: March 2008

Employing an immunoblotting procedure, we have identified and characterized an autoantigen carried on glycoprotein (GP) IIb in a patient with chronic idiopathic thrombocytopenic purpura (ITP), and have compared the location of the autoantigen with that of the platelet‐specific alloantigen Baka. Immunoblots, using the partially purified GP IIb/IIIa complex as the target antigen, indicated that GP IIIα carried both the ITP autoantigen and the Baka alloantigen. The ITP plasma contained another antibody against a 100 kD protein (P100), a trace contaminant in the GP IIb/ IIIa sample, which is probably a proteolytic fragment of an internal 124 kD protein. After chymotrypsin treatment, the auto‐ and alloantigen were found to be located on 65 kD fragments detectable under reducing conditions. In addition, immunoblots made after two‐dimensional nonreduced‐reduced SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE) directly demonstrated that both 65 kD fragments had a molecular weight of 80 kD under nonreducing conditions; this provides evidence that these fragments were one and the same, and were derived from GP IIbα. Immunoblots of platelets digested in situ with chymotrypsin indicated that the 65 kD fragment of GP IIbα was retained by the platelet membrane. We conclude, therefore, that a 65 kD fragment, which represents the membrane side of the chymotrypsin cleavage site on GP IIbα, carries a clinically important determinant(s) recognized not only by the anti‐Baka alloantibody, but also by the ITP autoantibody.
Maternal alloimmunization to HLA, platelet and granulocyte‐specific antigens during pregnancy: its influence on cord blood granulocyte and platelet counts
Authors: Skacel Patricia O. , Stacey T. E. , Elizabeth C. , Tidmarsh F. , Contreras Marcela
Published: March 2008

In order to obtain an estimate of the frequency of platelet‐specific and granulocyte‐specific antibodies and of the effect of such antibodies on the platelet count and granulocyte count of the newborn infant, serum from 147 women in their second or subsequent pregnancies was tested. No platelet‐specific antibodies were found but 29 of the women had granulocyte‐specific antibodies and the corresponding infants had granulocyte counts which were significantly lower than those of infants without antibodies. HLA antibodies were found in the sera of 57 women but were not associated with diminished platelet or granulocyte counts in the corresponding infants. Maternal granulocyte antibodies may be an underestimated contributory factor in the pathogenesis of neonatal neutropenia.
Spectrin autoantibodies in normal human serum and in polyclonal blood grouping sera
Authors: Ballas S. K.
Published: March 2008

Naturally occurring spectrin autoantibody was detected in normal human sera and in polyclonal blood grouping sera by the immunoblotting technique. The autoantibody seems to be IgG, stable, of high titre and low affinity. It was detected in all sera tested. Preparations of monoclonal antibodies directed against some blood group antigens and anti‐A1 lectin reagent were devoid of spectrin autoantibody as expected. This autoantibody may be instrumental in clearing red cell membrane components from the circulation during haemolysis. Care must be exercised in studies designed to determine the association of some blood group antigens with the red cell membrane skeleton, when using polyclonal sera which contain spectrin autoantibodies in addition to the antibodies against the blood group antigen in question.
Haematological changes during acute mental stress
Authors: Jern Christina , Wadenvik Hans , Mark Hans , Hallgren Jonas , Jern Sverker
Published: March 2008

To study haematological effects of emotional stress, blood samples were obtained from 29 healthy, normotensive, non‐smoking males aged 20–34 years before, during and after 10 min of mental arithmetic. There were significant increases in pheripheral blood cell count, haemoglobin concentration, and haematocrit in response to mental stress. Parallel to these changes significant increases in heart rate, and systolic and diastolic blood pressure were observed. The relative increments of leucocyte (8%) and platelet (3·5%) count were significantly higher than the increase in haemoglobin concentration (2%). There was a significant positive correlation between the blood pressure increase and the mobilization of leucocytes, whereas the increase in erythrocyte count, haemoglobin concentration, and haematocrit showed significant positive correlations with heart rate reactivity. It is concluded that mental stress causes an increase in leucocyte and platelet count that could not solely be accounted for by the concurrent haemoconcentration.
Marrow transplantation following busulfan and cyclophosphamide for chronic myelogenous leukaemia in accelerated or blastic phase
Authors: Copelan Edward A. , Grever Michael R. , Kapoor Neena , Tutschka Peter J.
Published: March 2008

Summary. Between July 1984 and October 1987, 21 patients with chronic myelogenous leukaemia in an accelerated or blastic phase were treated with 16 mg/kg of busulfan and 120 mg/kg of cyclophosphamide followed by infusion of bone marrow from an HLA‐identical sibling donor. The regimen was well tolerated. Except for one individual with severe marrow fibrosis all patients achieved a complete remission. Only one patient relapsed. Seven of 13 patients transplanted in an accelerated phase and five of eight transplanted in a blastic phase are alive without Philadelphia chromosomes between 8 and 48 months (median 29 months) following transplantation. The estimated probability of relapse‐free 3‐year survival is 55%. These results indicate that busulfan and cyclophosphamide combined with allogeneic marrow transplantation exert a potent anti‐leukaemic effect in patients in the accelerated or blastic phase of CML.
The bleeding disorder in acute promyelocytic leukaemia: fibrinolysis due to u‐PA rather than defibrination
Authors: Bennett B. , Booth N. A. , Croll A. , Dawson A. A.
Published: March 2008

Summary. Three consecutive patients with acute promyelocytic leukaemia who presented with severe haemorrhagic syndromes were studied and the findings contrasted with those of two patients with classical defibrination after electroshock or complicated labour. The leukaemic patients showed no depletion of fibrinogen. There was no evidence of disordered thrombin generation by either intrinsic or extrinsic pathway sufficient to account for their haemorrhage. All, however, showed strikingly enhanced fibrinolytic activity, which could have accounted for bleeding. This fibrinolytic disorder was characterized by free u‐PA in the plasma and differed from that seen after classical defibrination, where free t‐PA was observed. U‐PA was found also in malignant promyelocytes, which may be the source of u‐PA activity in the patients' plasma. Bleeding in promyelocytic leukaemia may be primarily a fibrinolytic disorder.
Menstrual cyclic thrombocytopenia
Authors: Tomer Aaron , Schreiber Alan D. , McMillan Robert , Cines Douglas B. , Burstein Samuel A. , Thiessen A. Robert , Harker Laurence A.
Published: March 2008

Summary. We studied three patients with cyclic thrombocytopenia which occurred in phase with the menstrual cycle. The platelet count in each patient reached a nadir of 5–20 × 109/1 at the onset of menses. Thrombocytopenia was associated with bruising, epistaxis and menorrhagia and was followed 5–14 d later by normal or elevated platelet counts (up to 900 × 109/1). Repeat bone marrow examinations performed at the time of reduced platelet counts showed megakaryocytic hyperplasia. 111In‐platelet‐disappearance from the circulation was measured in one patient: T50, time to half activity, was shortened to 0.7 d during the period of thrombocytopenia and was prolonged to 3.2 d when the platelet count increased (normal platelet T50 is about 4.8 d). In two of three patients platelet‐associated anti‐glycoprotein Ib autoantibodies were present and remained elevated despite normalization of the platelet count. In both of two patients the decrease in platelet count at the onset of menses was associated with an increase in the expression of monocyte Fcy receptors. Based on the reported capacity of oestrogenic hormones to modulate macrophage Fcy receptor expression, we propose that hormonal changes during the menstrual cycle may alter the Fcy receptor‐mediated clearance of antibody‐coated platelets by macrophages, modulate platelet survival, and cause cyclic thrombocytopenia.
Decreased frequency of bone marrow NK progenitors in aplastic anaemia
Authors: Yoda Yasuhiro , Kawakami Zenji , Abe Tsukasa
Published: March 2008

Summary. Twelve patients with aplastic anaemia were studied with regard to the frequency of NK progenitors in the bone marrow (BM) to investigate the mechanism of depressed NK activity and low NK cell count in the peripheral blood. NK cell (CD16+ cell) count and NK (K562) activity were significantly decreased (P < 0.02 and P < 0.001, respectively) in the patients as compared to eight healthy control subjects. Nylon wool non‐adherent (NW‐NA) BM mononuclear cells (MNC) of each patient and control were prepared. Mature T and NK cells were extensively depleted by sheep red cell rosette formation followed by a centrifugation on Ficoll‐Hypaque and monoclonal antibody mediated complement dependent cytolysis. Those selected BM cells were cultured in the presence of recombinant interleukin 2 (rIL2). Generation of NK activity was significantly decreased (P < 0.01) in the patients with aplastic anaemia. Frequency of NK progenitors in the selected BM cells assayed by the limiting dilution method was significantly decreased in those patients (P < 0.05). The frequency of BM NK progenitors relative to NW‐NA BM cells were related to NK cell count (P < 0.01). Those results indicate that depressed NK activity in aplastic anaemia is closely related to decreased NK cell count which is probably due to decreased production of NK cells in the BM.
The recovery of circulating progenitor cells after chemotherapy in AML and ALL and its relation to the rate of bone marrow regeneration after aplasia
Authors: Reid C. D. L. , Kirk A. , And J. Muir , Chanarin I.
Published: March 2008

Summary. Peripheral blood levels of BFU‐e, CFU‐GM and CFU‐mix were studied serially in nine patients with acute leukaemia in remission during the period of recovery that followed induction or consolidation chemotherapy. Following 23 courses of treatment in the nine patients, the values for all three classes of progenitor were found to be higher in ALL than in AML (mean peak CFU‐GM levels 5.8 × 103/ml and 0.8 × 103/ml respectively) and the highest levels were observed in patients recovering the most rapidly from bone marrow aplasia. Peak levels of these progenitors correlated best with the rate and extent of platelet recovery and all patients achieving blood levels of > 1.0 × 105 CFU‐GM/ml had recovered > 100 × 109/1 platelets by 20 d and had peak platelet counts of > 400 × 109/l within 40 d following chemotherapy. The peak values for circulating progenitors fell markedly after repeated courses of treatment in three of the five AML patients studied and this is likely to limit useful harvesting of such cells during later consolidation courses in this disease.
Antioxidant enzymes in lymphocytes from normal subjects and patients with chronic lymphocytic leukaemia: increased glutathione peroxidase activity in CLL B lymphocytes
Authors: Farber Charles M. , Kanganis Denise N. , Liebes Leonard F. , Silber Robert
Published: March 2008

Summary. The activities of several enzymes that protect against oxidative injury were determined in blood lymphocytes from patients with B chronic lymphocytic leukaemia (CLL) and from normal subjects. Similar glutathione reductase (GR), catalase and glucose‐6‐phosphate dehydrogenase (G6PD) activities were found in normal and CLL lymphocytes. Higher glutathione peroxidase (GP) activity was found in CLL lymphocytes. This activity in CLL B lymphocytes was 2‐fold higher than that of normal B lymphocytes, and 3‐fold higher than that of T lymphocytes from either source. Several disease processes have been associated with decreased glutathione peroxidase activity. Our finding with CLL B lymphocytes is believed to be the first example of an increased GP activity in a disease. It may reflect either the expansion of a rare type of B cell population or be an expression of the malignant process.
Human lymphoblastoid interferon as initial therapy in hairy cell leukaemia: a multicentre study in non‐splenectomized patients
Authors: Federico M. , Chisesi T. , Lauria F. , Bernasconi C. , Capnist G. , Castoldi G. , Damasio E. , Pagnucco G. , Frassoldati A. , Resegotti L. , Silingardi V.
Published: March 2008

Summary. The Italian Cooperative Group for Hairy Cell Leukaemia (ICGHCL), between April 1985 and June 1987, conducted a multicentre study using human lymphoblastoid alpha‐interferon as primary therapy as an alternative to splenectomy. Forty‐eight evaluable patients with HCL entered the study, 38 of them had splenomegaly, in five patients the spleen was not palpable and five were unfit for surgery because of age and general condition. Daily dose of 3 MU s.c. alpha‐IFN was given for 12 weeks, or until a satisfactory and stable response was obtained.

Among these 48 patients the response rate after 3 months of therapy was 63%, with seven patients (1 5%) achieving complete remission and 23 (48%) partial remission; 13 (27%) patients had a minor response. In five patients no response was observed and they died within 2 months of treatment. Five other patients, after an initial response, presented a re‐expansion of the disease.

Actuarial survival at 30 months was 88.8% for the entire group of 48 patients and 92% for the 38 patients who would normally be treated by splenectomy.

Thus, alpha‐IFN as primary treatment in HCL offered a reasonable therapy for splenomegalic patients. The timing and validity of splenectomy still remains an open question.
Comparison of allogeneic or autologous bone marrow transplantation and chemotherapy in patients with acute myeloid leukaemia in first remission: a prospective controlled trial
Authors: Reiffers J. , Gaspard M. H. , Maraninchi D. , Michallet M. , Marit G. , Stoppa A. M. , Corront B. , David B. , Gastaut J. A. , Sotto J. J. , Broustet A. , Carcassonne Y. , HOLLARD D.
Published: March 2008

Summary. Eighty‐five adult patients under the age of 50 years with acute myeloid leukaemia (AML) were entered into a prospective controlled study conducted to compare the effectiveness of allogeneic or autologous bone marrow transplantation and intensive chemotherapy for patients in first complete remission. Sixty‐one patients (72%) achieved complete remission then received a consolidation treatment. After consolidation, 58 patients who were still in remission were assigned to three different therapeutic modalities. Fifty‐two patients were evaluable: 20 patients who had an HLA‐identical sibling donor underwent allogeneic bone marrow transplantation within 3 months after achievment of complete remission; the other 32 patients were randomized to receive autologous bone marrow transplantation or intensive sequential chemotherapy. The actuarial risk of relapse at 3 years was 18% for the allogeneic patients, 50% for the autologous patients and 83% in the chemotherapy group. The difference was highly significant (P<0.0002). The disease‐free survival was respectively 66% (95% confidence interval 41–85%), 41% (95% confidence interval 16–66%) and 16% (95% confidence interval 0–31%) (P<0.004). We conclude that allogeneic bone marrow transplantation is presently the best therapeutic approach for patients with AML in first complete remission.
Spontaneous and interleukin 2 induced secretion of tumour necrosis factor and gamma interferon following autologous marrow transplantation or chemotherapy
Authors: Heslop Helen E. , Gottlieb David J. , Reittie Joyce E. , Bello‐Fernandez Concha , Meager Anthony , Prentice H. Grant , Brenner Malcolm K.
Published: March 2008

Summary. Culture of lymphocytes from allograft recipients or from normal donors with Interleukin 2 (IL2) induces high levels of gamma‐interferon (gamma IFN) and tumour necrosis factor (TNF) secretion. We now show that production of these two cytokines can also be increased after autologous bone marrow transplantation (BMT) and induced following chemotherapy for haematological malignancy. These effects occur even though the regenerating IL2 responsive lymphocytes in this context have previously been extensively exposed to cytotoxic agents. IL2 induced secretion of gamma‐IFN and TNF is higher in autograft recipients than in patients treated by chemotherapy alone. This effect may be related to differences in the phenotypic profile between recovering lymphocytes in the two groups and to an increased degree of prior in vivo lymphocyte activation in the autologous bone marrow transplant recipients. In both groups of patients, IL2 acts on CD3 + T cells and on CD16 + NK cells so that depletion of either subset incompletely abrogates IL2 dependent gamma‐IFN secretion. As both TNF and gamma‐IFN possess anti‐leukaemic and anti‐infective activity, enhancement of their secretion by IL2 infusion after autologous bone marrow transplant and induction after chemotherapy may be of therapeutic benefit.
Immune‐mediated agranulocytosis related to drugs and their metabolites: mode of sensitization and heterogeneity of antibodies
Authors: Salama A. , Schütz B. , Kiefel V. , Breithaupt H. , Mueller‐Eckhardt C.
Published: March 2008

Summary. Two major unresolved problems in drug‐related immune agranulocytosis are understanding the mechanism by which sensitization takes place in vivo, and verification of the diagnosis. Using a sensitive, competitive enzyme‐linked immunoassay (ELISA) we were able to characterize the causative antibodies in 13 patients with drug‐related agranulocytosis [metamizole (n= 5), penicillin (n= 5), dimethylaminophenazone (n=1), propyphenazone (n=1) and diclofenac (n= 1)]. Irrespective of the causative drug, the majority of patients appear to have developed autoantibodies (aab) in addition to drug‐dependent antibodies (ddab) of the IgG and/or IgM classes. In all cases related to metamizole, and in the single case related to diclofenac, the ddab appeared to recognize only metabolites of the drug since they were reactive in the presence of ex vivo antigens (urine from individuals receiving therapeutic levels of the drugs), but not the native drugs. Only a few ddab were reactive with granulocytes pretreated with the drug (cell‐drug complexes); the majority of ddab could not be detected unless the drug or ex vivo antigen was added to the incubation mixture as well as the solution used for subsequent washes.

Our results indicate that drugs and/or their metabolites interact with target cells and thereby directly function as immunogenic haptens, even when the drugs do not bind tightly to the cells.
Rapid identification of donor and recipient cells after allogeneic bone marrow transplantation using specific genetic markers
Authors: Hutchinson R. M. , Pringle J. H. , Potter L. , Patel I. , Jeffreys A. J.
Published: March 2008

Summary. The fate of the donor graft in allogeneic bone marrow transplantation, the presence of chimaerism and early relapse can be monitored by identification of the donor or recipient origin of haemopoietic cells in peripheral blood and bone marrow. We have done this by the use of highly informative locus‐specific hypervariable DNA probes in two sex and blood group matched transplants. In four sex mismatched transplants the Y chromosome was identified by in situ hybridization using a biotinylated Y probe.

Early engraftment by day 13 was detected in five of the six patients. Transient chimaerism occurred in half of the cases and was accompanied by the temporary appearance of the Philadelphia chromosome in one patient with chronic myeloid leukaemia without subsequent relapse. Persistence of the graft in the face of falling peripheral counts was documented in four of the six patients studied. The morphology as well as origin of the haemopoietic cells could be characterized by the Y probe analysis. In one patient, recipient lymphocytes were shown to co‐exist with myeloid series of donor origin.

We conclude that both techniques are highly specific, sensitive and can provide information within 24–48 h. Thus they are of value in guiding early therapeutic intervention in allogeneic transplantation.
Host and donor erythrocyte repopulation patterns after allogeneic bone marrow transplantation analysed with antibody‐coated fluorescent microspheres
Authors: Bär B. M. A. M. , Schattenberg A. , Van Dijk B. A. , De Man A. J. M. , Kunst V. A. J. M. , De Witte T.
Published: March 2008

Summary. Analysis of erythrocyte populations, using red blood cell antigen differences between host and donor as marker, was performed with a sensitive fluorescent microsphere assay to monitor marrow engraftment and mixed red cell chimaerism after allogeneic bone marrow transplantation (BMT). An adapted transfusion policy, using marker negative erythrocyte transfusions, was required for this analysis. In all patients the marrow graft was depleted of lymphocytes by counterflow centrifugation. Thirty‐seven patients were evaluable for donor repopulation at one or more points in the first 6 months after BMT. At 0·5 month donor erythrocytes were detectable in 19 out of 22 patients. At 6 months donor erythrocytes were detectable in 100% of the evaluable patients. In the first 3 months after BMT the average donor erythrocyte repopulation in recipients of major ABO mismatched transplantations was delayed. Thirty‐eight patients were evaluable for chimaerism at 6 months or later after BMT. A high incidence of mixed red cell chimaerism was observed varying from 50% to 71% at different points of analysis. Mixed red cell chimaerism with low percentages (<1%) of host cells was not related with relapse, nor did high percentages (>10%) of host cells necessarily indicate relapse.
Chromosomes and other prognostic factors in acute lymphoblastic leukaemia: a long‐term follow‐up
Authors: Secker‐Walker Lorna M. , Chessells Judith M. , Stewart Elizabeth L. , Swansbury G. J. , Richards Susan , Lawler Sylvia D.
Published: March 2008

Summary. Cytogenetic, clinical and laboratory features at diagnosis were examined in a group of 80 children with acute lymphoblastic leukaemia (ALL) who had been followed up for a minimum of 5 1/2 years. The 17 (21%) with high hyperdiploidy tended to have low leucocyte counts and common ALL, but their favourable outcome (75% event‐free survival) was independent of these factors. No patient with hypodiploidy survives while the pseudodiploid and normal groups have an intermediate prognosis. Cytogenetic analysis showed examples of patients with the well‐recognized translocations and a number with apparently unique ones. Among the latter were some long‐term survivors. We conclude that Cytogenetic analysis identifies a good risk group of patients who remain well on long‐term follow‐up, but that the presence of a translocation does not necessarily imply a poor outcome.
T‐cell receptor genes expression in B‐cell leukaemias
Authors: Narni Franco , Mariano Maria Teresa , Colò Alessandro , Grantini Marcello , Merli Francesco , Donelli Amedea , Montagnani Giuliano , Di Prisco Alfredo Ubaldo , Torelli Giuseppe , Torelli Umberto
Published: March 2008

Summary. Using Northern‐blot analysis we have studied the expression of T‐cell receptor (TCR) α, β and γ chain genes in primary cells obtained from 36 leukaemic patients. Fifteen patients had myeloid leukaemias, two had T‐cell leukaemias, and 19 leukaemias corresponding to various stages of B‐lymphocyte differentiation. We observed that truncated TCR β mRNAs were produced in B‐cells at relatively high levels even in the absence of detectable gene rearrangements. Ti α mRNAs of abnormal size were also frequently found. Such transcripts were not detectable in total RNA extracted from leukaemic myeloid cells. Factors allowing Ti α and β gene transcription must be active in leukaemic pre‐B and B cells but not in myeloid cells. Neither B‐lineage nor myeloid leukaemias expressed TCR γ gene at detectable levels.
Increased in vivo biosynthesis of prostacyclin and thromboxane A2 in chronic idiopathic thrombocytopenic purpura
Authors: Rousson D. , Guichardant M. , Lagarde M. , Viala J. J. , Dechavanne M.
Published: March 2008

Summary. The production of thromboxane A2 (TxA2) and prostacyclin (PGI2) was studied in patients with chronic idiopathic thrombocytopenic purpura (10 patients) compared to central thrombocytopenia (five patients) and healthy subjects (10 subjects). This production was monitored by the assay of urinary 2,3‐dinor‐TxB2 and 2,3‐dinor‐6‐keto‐PGF1α as respective breakdown products of TxA2 and PGI2 by stable isotope dilution assays employing negative ion‐chemical gas‐chromatography‐mass‐spectrometry. Evidence is presented for the existence of an enhanced PGI2 and TxA2 urinary excretion in the group of idiopathic thrombocytopenic purpura (ITP) patients. Moreover, production of serum TxB2 per platelet was decreased in ITP group. These results provide arguments for an in vivo platelet cyclooxygenase hyperactivity during chronic ITP.
Diagnostic and prognostic significance of myelomonocytic cell surface antigens in acute myeloid leukaemia
Authors: Merle‐Beral Félène , Cong Duc Laurent Nguyen , Leblond Véronique , Boucheix Claude , Michel Ariane , Chastang Claude , Debre Patrice
Published: March 2008

Thirty‐six cases of acute myeloid leukaemia (AML) were tested with a large battery of monoclonal antibodies (moAbs) detecting surface markers normally expressed by myelomonocytic, T and B lymphoid, megakaryocytic and erythroid lineages. Differences in antigenic expression were observed among the various FAB subgroups: HLA‐class II molecules were found in almost all AML cases but not in the promyelocytic subgroup (M3); CD14 and CD36 antigens were detected in monocytic leukaemias (M4 and M5); the CD34 moAb (MY10) recognizing an epitope described on myeloid stem cells was positive in 88% of the M1 and 80% of the M3 cases. By a multivariate analysis, only the CD14b (MY4) discriminated significantly between M1–M2 and M4–M5 subgroups. Using Cox's model to assess the prognostic importance of variables including immunophenotyping on survival, we undertook a one by one analysis and found that the presence of CD17 antigen predicted for a shorter survival (P= 0.03). In addition this marker appeared more significant than other clinical and biological parameters.
The effect of phospholipase C on plasma factor VII
Authors: Hubbard Anthony R. , Parr Lynne J.
Published: March 2008

The proposed link between a circulating factor VII–phospholipid complex and the risk of cardiovascular disease (CVD) has stimulated us to investigate the effect of phospholipase C (PLC) on the factor VII (FVII) activity in plasma from healthy individuals. PLC caused a rapid fall in FVII activity which was larger with heparinized than with citrated plasma. EDTA inhibited the PLC effect so emphasizing the involvement of divalent cations. PLC dependent loss of FVII activity varied widely between individuals, showed a highly significant correlation with plasma triglyceride concentrations, and was always greater in post‐prandial compared to fasting plasma samples. Experiments using pure recombinant FVIIa and plasma depleted of FVII by adsorption indicated that loss of FVII activity only occurred in the simultaneous presence of absorbed plasma. FVIIa and PLC. Preincubation of PLC with adsorbed plasma before adding FVIIa did not lead to loss of FVII activity. It appears that PLC may act on lipoproteins already bound to FVII. in order to inhibit FVII activity. Other results indicated that competition between different plasma components (lipoproteins) in binding to FVII may govern the extent of the PLC dependent reduction in FVII activity.
Inhibition of cellular iron uptake by haem in mouse erythroleukaemia cells
Authors: Hradilek Antonin , Neuwirt Jan
Published: March 2008

Haemin inhibited iron uptake from transferrin (Tf) by mouse erythroleukaemia cells (MELC) induced for differentiation by hexamethylene bisacetamide (HMBA). The rate of 59Fe internalization was decreased, but the rate and the extent of 125I‐Tf endocytosis was unaffected by the addition of haemin. Haemin inhibited 59Fe incorporation into haem by a greater proportion than the overall uptake of 59Fe from Tf. The reduction of total cellular 59Fe uptake was more pronounced at 59Fe‐Tf concentrations closer to saturation. Exogenous 5‐aminolaevulinic acid stimulated 59Fe utilization for haem synthesis in MELC but did not revert the inhibition induced by haemin. Haem synthesis measured by 14C‐glycine incorporation into haem was maintained for at least 1 h without an external transferrin iron source and was inhibited by the addition of haemin equally over the whole range of Tf concentrations studied. Desferrioxamine (DFO) stimulated cellular uptake of 59Fe by the uninduced cells and reverted the inhibition of 59Fe transport into HMBA treated cells caused by haemin. Addition of DFO within a short‐term incubation had no effect on haem synthesis measured by 14C‐glycine incorporation into haem. No evidence for a direct effect of haem on the transferrin cycle or iron release was found. It was concluded that the reduction of iron uptake by haemin treated MELC is secondary to the decrease in iron utilization for haem synthesis.
Induction of eosinophilic colonies by interleukin‐5 on acute myeloblastic leukaemic cells
Authors: Motoji Toshiko , Okada Michiko , Takanashi Minoko , Masuda Michihiko , Tanaka Kumi , Oshimi Kazuo , Mizoguchi Hideaki
Published: March 2008

The effect of interleukin‐5 (IL‐5) on blast colony formation of acute myeloblastic leukaemia (AML) cells was studied. IL‐5 showed a weak but significant colony stimulatory effect in one out of six cases studied. IL‐5 induced eosinophilic colony growth in this case, and chromosomal and histochemical analyses strongly suggested that these eosinophilic colonies were derived from leukaemic blast progenitor cells. To our knowledge, this is the first report that IL‐5 induces eosinophilic colonies on fresh AML cells. A possible induction of differentiation of AML blast cells by IL‐5 is suggested.
Interleukin‐4 suppresses the interleukin‐3 dependent erythroid colony formation from normal human bone marrow cells
Authors: De Wolf Joost Th. M. , Beentjes John A. M. , Esselink Mariet T. , Smit Jan W. , Halie M. Ruud , Vellenga Edo
Published: March 2008

Human recombinant interleukin 4 (IL‐4) was studied for its effects on the erythroid burst forming unit (BFU‐E) from human bone marrow cells. IL‐4 alone neither supports nor suppresses the erythropoietin (Epo)‐dependent colony formation. Different results were obtained when IL‐4 was combined with interleukin‐3 (IL‐3) in the presence of Epo. IL‐4 suppressed the IL‐3 supported erythroid colony formation in all cases (an increase of 58 ± 8% with IL‐3 versus an increase of 14 ± 7% with IL‐3 plus IL‐4, n= 8). This antagonizing effect was dependent on the continuous presence of IL‐4 in the culture medium, but was independent of adherent cells. B‐, T‐cells, or the presence of serum in the culture medium. Finally, the effects of IL‐4 and IL‐3 were studied on the ‘Epo‐independent’BFU‐E by adding Epo on day 3. A decline of the IL‐3 supported BFU‐E was observed in the presence of IL‐4 but the degree of reduction was equivalent to the results obtained when Epo was supplied at day 0. These findings indicate that IL‐4 acts as suppressive growth factor for the IL‐3 supported erythroid colony formation from human bone marrow cells.
Studies on a patient with thrombocytopenia, giant platelets and a platelet membrane glycoprotein Ib with reduced amount of sialic acid
Authors: Aakhus A. M. , Stavem P. , Hovig T. , Pedersen T. M. , Solum N. O.
Published: March 2008

A 70‐year‐old patient with a life‐long bleeding tendency, giant platelets and thrombocytopenia (10‐40 x 109 platelets/1) has been studied. This is a condition often associated with lack of platelet membrane glycoprotein Ib (GP Ib). Electron microscopy of fixed platelets incubated with monoclonal antibodies to GP Ib (AN 51, AP 1) and goldlabelled goat anti‐mouse IgG, showed a distinct distribution of GP Ib on the patient's platelets, however. Crossed immunoelectrophoresis and SDS‐PAGE demonstrated a reduced mobility of the patient's GP Ib which could be explained by absence of sialic acid. Blotting with peroxidase‐conjugated peanut agglutinin confirmed this conclusion. This lectin binds to galactose‐N‐acetyl‐galactosamine residues exposed terminally when sialic acid is absent from the carbohydrate side‐chains. Such binding could be seen with normal GP Ib only after neuraminidase treatment. Fluorescence studies with FITC‐conjugated peanut agglutinin showed binding of the lectin to intact patient platelets, indicating that lack of sialic acid was not introduced during the platelet isolation procedure. Neither could the lack of sialic acid be attributed to increased neuraminidase activity as studied in vitro. Platelets treated with neuraminidase in vivo or in vitro are rapidly cleared from the circulation. Therefore the patient's thrombocytopenia may be associated with the reduced amount of GP Ib sialic acid. As far as we know, similar cases have not been described previously.
Formate metabolism in the cobalamin‐inactivated rat
Authors: Deacon Rosemary , Perry Janet , Lumb Michael , Chanarin Israel
Published: March 2008

Endogenous formate levels in blood and liver were assayed in rats both after inactivation of cobalamin (Cbl) by exposure to N2O as well as in air‐breathing controls. The uptake of [14C]formate by tetrahydrofolate (H4folate) in bone marrow cells and liver homogenate and the incorporation of [14C]formate into purine, pyrimidine, methionine, serine and choline, was measured. There was a significant accumulation of endogenous formate following Cbl inactivation. There was impaired utilization of [14C]formate for single unit carbon (C1 unit) transfers mediated by folate in Cbl‐inactivated tissues, other than for synthesis of adenine. The impairment was not accompanied by any accumulation of labelled methylH4folate indicating that methylfolate trapping played no part in impaired single carbon unit transfer. The effect of Cbl lack was a failure to form formylH4folate so that formate accumulated. The reason for this is not known.
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

Long‐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

Ten 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

A 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

We 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.