bluuebunny/crossref_metadata_2025 huggingface dataset:
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

Erythrokinetic 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.
Induction of B‐cell chronic lymphocytic leukaemia and hairy cell leukaemia like phenotypes by phorbol ester treatment of normal peripheral blood B‐cells
Authors: Visser Lydia , Poppema Sibrand
Published: March 2008

Summary. To investigate the relationship between normal B‐cells, B‐cell chronic lymphocytic leukaemia (B‐CLL) cells and hairy cell leukaemia (HCL) cells the three cell types were incubated with phorbol myristic acetate (PMA). The parameters studied were morphology, immunophenotype and tartrate resistant acid phosphatase (TRAP). PMA stimulation of B‐cells induced morphological changes as well as CD5, CD11c, B‐ly 7 and TRAP positivity. The cells formed small aggregates, the cell membranes were ruffled and frequently hairy and a small number of cells became plastic adherent and developed dendritic structures. CD5 and CD11c appeared on day 2, imitating a B‐CLL phenotype. On day 3 and 4 a decrease of CD5, an increase of CD11c and the appearance of B‐ly 7 could be seen mimicking an HCL phenotype. The changes in B‐CLL and HCL upon PMA stimulation were mainly morphological; the B‐CLL cells became ruffled and aggregated strongly, while the HCL cells developed dendritic features and became adherent. The immunophenotype of the PMA stimulated HCL cells did not change. The B‐CLL cells remained CD5 positive and did not become B‐ly 7 positive. The findings indicate that although PMA stimulation of normal peripheral blood cells results in CLL‐ as well as HCL‐like phenotypes, similar stimulation of B‐CLL cells does not result in an HCL‐like phenotype.
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

Clonal 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

A 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

A 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

Twenty‐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

In 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.
Contrasting patterns of neoplastic cell behaviour in long‐term culture of bone marrow from patients with acute leukaemia and myelodysplastic disorders
Authors: Firkin Frank C. , Birner Raquella , Russell Sue H. , Garson O. Margaret
Published: March 2008

Summary. Alterations in neoplastic cell behaviour responsible for increased production of terminally‐differentiated granulocytes during long‐term culture of bone marrow in different categories of acute leukaemia and myelodysplasia have been investigated. An increase in neutrophils associated with transition to a morphological picture identical to normal control cultures occurred in 15 of 25 studies on acute leukaemia in contrast to one of six studies on myelodysplastic disorders. An abnormal neoplastic karyotype was employed as a marker for monitoring the course of the neoplastic cell population in 11 studies in which there was progression towards a normal pattern of differentiation. An increase in differentiation was shown by this means to represent increased maturation of cells of the neoplastic process in one study on a myelodysplastic disorder, demonstrating domination of proliferative activity in culture by all of the myelodysplastic disorders examined. Transition towards normal differentiation in nine studies on acute leukaemia, however, correlated with partial or complete replacement of the acute leukaemic cells by normal haemopoietic series in de novo acute leukaemia, and by Ph positive cells in blast crisis of CML. Conversion to morphologically and cytogenetically normal cell populations in five studies on de novo acute leukaemia occurred in four cases which failed to respond to remission‐induction therapy, suggesting the selective toxic effect capable of purging acute leukaemic cells from bone marrow operated by a mechanism which lacked cross‐resistance to currently‐employed cytotoxic agents.
Testicular irradiation in childhood lymphoblastic leukaemia
Authors: Eden O. B. , Lilleyman J. S. , Richards S. ,   on behalf of the Medical Research Council Working Party on Leukaemia in Childhood
Published: March 2008

Summary. The effect of randomly allocated testicular irradiation on the subsequent incidence of testicular infiltration and disease‐free survival was assessed in two Medical Research Council Childhood Leukaemia Trials, UKALL VI and UKALL VII. None of the 83 boys who actually received testicular radiotherapy subsequently developed gonadal disease, whereas 18 of the 163 who were not irradiated did. Despite this there is no apparent difference in disease‐free survival for those randomized to receive testicular irradiation compared to those who were not. after a minimum of 8 years follow up. Although prophylactic testicular irradiation appears to prevent subsequent gonadal relapse there is no evidence that it improves overall prognosis when adequate systemic chemotherapy is used. As it has considerable long‐term side effects it cannot be recommended as routine therapy.
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.
Hereditary deficiency of all vitamin K‐dependent procoagulants and anticoagulants
Authors: Brenner B. , Tavori S. , Zivelin A. , Keller C. B. , Suttie J. W. , Tatarsky I. , Seligsohn U.
Published: March 2008

Summary. Hereditary combined deficiency of vitamin K‐dependent factors is a rare entity. We report a 7‐year‐old girl of Arab origin with hereditary deficiency of the procoagulants factors II, VII, IX and X and the natural anticoagulants proteins C and S. The patient is the tenth offspring of a consanguinous marriage and presented at 6 weeks with spontaneous intracerebral haemorrhage. Symptoms improved following plasma infusion. A sibling died at 5 d from uncontrollable umbilical bleeding. Blood coagulation work‐up at 6 years showed: factor II:C (activity) 12 U/dl, factor II: Ag (antigen) 40 U/dl; factor VII:C 12 U/dl; factor IX:C 36 U/dl, factor IX: Ag 57 U/dl; factor X:C 17 U/dl, factor X: Ag 54 U/dl; protein C activity 43 U/dl; protein C:Ag 45 U/dl; protein S:Ag 34 U/dl; levels of factors V:C and VIII:C were normal. Assays of coagulation factors in the parents and five of the siblings were within the normal range. Following acute infection and dilantin therapy procoagulant activity levels were reduced further and were partially increased after vitamin K infusion. Crossed immunoelectrophoresis of prothrombin in the presence of calcium lactate revealed a population of des‐carboxyprothrombin. Serum vitamin K epoxide levels were undetectable. The data suggest that the defect in our patient stems from abnormal carboxylation of the vitamin K‐dependent proteins and that the mode of inheritance is autosomal recessive.
Isolation and characterization of the translation product of a β‐globin gene nonsense mutation (β121 GAA→TAA)
Authors: Adams Junius G. , Steinberg Martin H. , Kazazian Haig H.
Published: March 2008

Summary. The β°‐thalassaemia gene of an individual who was a mixed heterozygote for this allele (GAA to TAA in codon 12) and β+‐thalassaemia (IVS‐1 position 110 G to A) was examined to determine if the β°‐thalassaemia allele directed the synthesis of any detectable protein product. This β°‐thalassaemia allele was of particular interest, because it is the only example of a premature chain termination codon in the third exon of the β‐globin gene that produces thalassaemia. A very small amount of an abnormal protein was found in the red blood cells of the proband and was purified by preparative column chromatography. This abnormal protein was digested with trypsin, the peptides were separated by reverse phase high performance liquid chromatography (HPLC), and the amino acid content of each peptide was determined. All of the soluble β‐globin peptides were found except for T‐13. T‐14 and T‐15 (residues 121‐146), indicating the presence of a truncated protein that corresponded to the translation product of the β121 Glu→Term mRNA. This truncated globin was estimated to comprise between 0.05% and 0.1% of the total non‐α‐globin protein. These results may explain the phenotype of inclusion body β‐thalassaemia in heterozygotes, which is atypical of heterozygous β°‐thalassaemia.
Antilymphocyte globulin therapy enhances impaired function of natural killer cells and lymphokine activated killer cells in aplastic anaemia
Authors: Myint A. A. , Malkovska V. , Morgan S. , Luckit J. , Wonke B. , Gordon‐Smith E. C.
Published: March 2008

Summary. MHC‐unrestricted cytotoxic lymphocytes, namely natural killer (NK) and lymphokine activated killer (LAK) cells, have been implicated in the regulation of haemopoiesis. To investigate the possible role of these lymphocytes in the pathogenesis of aplastic anaemia (AA), we studied their functions in the peripheral blood mononuclear cells (PBMC) and bone marrow mononuclear cells (BMMC) of patients with AA treated with antilymphocyte globulin (ALG). Before treatment, both NK and LAK activities in the PBMC of 25 patients were low (NK=1.9±2.1 × 103LU/l. LAK = 4.7±3.6 × 103 LU/l) compared to normal (NK=6.0±3.0 × 103 LU/l, LAK=10.0±3.5 × 103 LU/l) or multiply transfused (NK=7.8±6.6 × 103 LU/l, LAK = 25.2±13.6 × 103 LU/l) controls. The NK and LAK activities in the BMMC in AA patients were not significantly different from those in PBMC. In all patients with low LAK and NK activities pre ALG there was an increase in activity 2‐24 weeks after therapy which eventually reached normal levels and which was maintained for up to 2 years. Analysis of lymphocyte phenotypes in AA patients before treatment showed both significantly low mean proportion and absolute numbers of CD16+ cells compared to normals, which increased after therapy. Changes in MHC‐unrestricted cyto‐toxicity and lymphocyte phenotypes post therapy were not correlated with haemopoietic recovery. These data suggest that ALG treatment can enhance the functions of MHC‐unrestricted lymphocytes independently from haemopoiesis. It is unlikely that these cells play a role in the pathogenesis of AA.
Revised recommendations for the measurements of the serum iron in human blood
Authors:   Iron Panel of the International Committee for Standardization in Haematology
Published: March 2008

Summary. Certain modifications to the previously published ICSH recommendations (1978) have been approved by the Iron Panel of the International Committee for Standardization in Haematology (ICSH). They include substitution of the chromogen bathophenanthroline sulphonate with ferrozine or ferene, which are more sensitive and cheaper, and a reduction in the volume of the test sample from 2 ml to 0.5 ml.
A study of prognostic factors in blast crisis of Philadelphia chromosome‐positive chronic myelogenous leukaemia
Authors: Cervantes Francisco , Rozman María , Rosell Jordi , Urbano‐Ispizua Alvaro , Montserrat Emilio , Rozman Ciril
Published: March 2008

Summary. In 80 patients with Ph‐positive chronic myelogenous leukaemia the main clinical, haematological and cytogenetical data were recorded at diagnosis of blast crisis and evaluated for prognostic significance. At the time of the analysis 73 patients had died with a median survival of 4.8 months from diagnosis of blast crisis for the whole series. When analysed as a time‐dependent variable, the achievement of a favourable response to chemotherapy resulted in a longer patient's survival, On the other hand, the univariate analysis identified six pretreatment characteristics associated with a poorer prognosis: a longer chronic phase, presence of extramedullary blastic involvement, a platelet count below 200 × 109/1, a less marked leucocytosis, a blood blast cell percentage higher than 10%, and presence of trisomy 8. The latter parameters were included in a multiple regression model together with the blast cell phenotype (lymphoid versus non‐lymphoid), and only four of them (trisomy 8, duration of chronic phase, platelet count, and leucocyte count) retained their prognostic influence. When the therapeutical response was also included in the regression model, it proved to be the most important prognostic variable, followed by trisomy 8, length of chronic phase, extramedullary disease, and platelet count, whereas the leucocyte count lost its predictive value. Thus, in spite of the short overall survival of blast crisis patients, the identification of prognostic factors in such a haematological condition may be of interest, especially in the interpretation of new therapeutical approaches.
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.
Philadelphia chromosome positive B‐cell type malignant lymphoma expressing an aberrant 190 kDa bcr‐abl protein
Authors: Mitani Kinuko , Sato Yuko , Tojo Arinobu , Ishikawa Fuyuki , Kobayashi Yukio , Miura Yasusada , Miyazono Kohei , And Akio Urabe , Takaku Fumimaro
Published: March 2008

Summary The Philadelphia (Ph) chromosome translocation, t(9:22) (q34;q11) is found in some acute lymphoid leukaemias (ALL) and acute myeloid leukaemias (AML). Although cytogenetically all Ph chromosomes appear similar, the 22q11 breakpoints found in acute leukaemias are of two kinds, those within the major breakpoint cluster region (Mbcr‐1) of the BCR gene as found in chronic myelogenous leukaemia (CML), and those within the first intron of this gene. In the former group the molecular events are the same as those found in CML, p210 bcr‐abl, encoded by 8.5 kb mRNA; however, a new aberrant protein, p190 bcr‐abl, is found in the latter group. Ph translocation is also found in a few cases with malignant lymphoma, but it has not been characterized at the molecular level. We describe here a non‐Hodgkin's lymphoma case with primary splenic presentation, which showed a complex Ph translocation. Neoplastic cells were of a B‐cell origin (HLA‐DR+, sIgM+, sIgΔ+, CALLA‐). Molecular studies revealed the expression of p190 bcr‐abl with no Mbcr‐1 rearrangement. Our case indicates that the same Ph translocation as seen in acute leukaemias can be found in hematologic disorders other than leukaemias, suggesting that a c‐abl gene activating mechanism may be involved in the pathogenesis of wide spectrum of haematologic malignancies.
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.
The effect of high doses of recombinant human erythropoietin on megakaryocytopoiesis and platelet production in splenectomized mice
Authors: Tsukada Junichi , Misago Masahiro , Kikuchi Makoto , Sato Tadatsugu , Susumu Oda Rhyosuke Ogawa , And Shyozo Chiba , Eto Sumiya
Published: March 2008

Summary The effects of recombinant human erythropoietin (rHuEpo) on megakaryocytopoiesis and platelet production were investigated in splenectomized, sham‐operated and intact mice. When splenectomized mice were injected with 50 U of rHuEpo daily for 4 d, peripheral platelet counts, megakaryocyte (MK) size and the percentage of mature MK (stage IV MK) increased from 1 to 4 d after the initial rHuEpo injection. The total number of marrow MK colony forming units (CFU‐MK) increased from 2 to 4 d after the initial injection. Furthermore, from days 6 to 8, the total number of marrow MK also increased. In addition, when the dose of rHuEpo was increased to 20 ˜ 200 U per mouse, clear dose responses were detected in platelet counts and MK numbers. When sham‐operated and intact mice were injected with rHuEpo, no significant change in platelet numbers was detected. The number of MK, MK size and the number of CFU‐MK increased only in spleen, not in bone marrow. Our data indicate that Epo has stimulatory effects on platelet production in vivo, if used in sufficient quantities.
Total circulating red cells versus haematocrit as the primary descriptor of oxygen transport by the blood
Authors: Jones J. G. , Holland B. M. , Hudson I. R. B. , Wardrop C. A. J.
Published: March 2008

Summary Peripheral haematocrit (PCV) is the traditional target and monitor in many transfusion regimens. Without negating the importance of PCV as a determinant of whole blood viscosity, the present article outlines two important reasons why the red cell volume (RCV) should replace PCV in the central target role during blood transfusion in intensive care and other emergency situations:

1. PCV reflects both RCV and plasma volume (PV) and is therefore not directly proportional to the total blood oxygen carrying capacity. At best, the relationship between PCV and RCV is hyperbolic and this is often overlooked when relating the two parameters in practice. At worst, the hyperbolic relationship is unreliable because PV and RCV can vary independently and the PCV is a fluctuating ratio of variable numbers.

2. PCV is not a good indicator of blood volume (BV), which is another important determinant of oxygen delivery to tissues and a crucial parameter in intensively managed patients. BV is directly proportional to RCV and this relationship also is often overlooked in clinical practice.

The recommended values for RCV are 30 ml/kg in men, 25 ml/kg in women and between 30 ml/kg and 45 ml/kg in neonates within the first week of life.
Using teicoplanin for empiric therapy of febrile neutropenic patients with haematological malignancies
Authors: Menichetti F. , Favero A. Del , Bucaneve G. , Aversa F. , Fiorio M.
Published: March 2008

Summary The cumulative experience with teicoplanin in treating febrile neutropenic patients included in three different comparative clinical trials conducted at a single institution during a 3‐year period, is presented. 1 52 febrile episodes in 129 neutropenic patients were treated with iv. teicoplanin (6 mg/kg/d) combined with amikacin (15 mg/kg/d) plus ceftazidime (90 mg/kg/d). The study population comprised 75 patients with acute leukaemia and 77 marrow recipients: 53(% (81/152) had a central venous catheter in place and 68%) (103/152) had severe neutropenia (< 100/mm3) at the beginning of the febrile episode.

The overall response rate of the evaluiable febrile episodes was excellent: 88% (107/122) improved. Bacteraemias due to Gram‐positive cocci accounted for 75% of the total (42/56) and pathogens in the blood isolates were mostly staphylo‐cocci (coagulase‐negative 14, coagulase‐positive 13) and streptococci (131. The response rate of Gram‐positive bacteraemias was good: 88% (37/42) improved and 75% (9/12) of Gram‐positive bacteraemias having teicoplanin as the only antibiotic with in vitro activity against the infective strains were cured. Death due to infection accounted for 7% of total febrile episodes (11/152). Side effects were documented in 14(%, of the episodes. In a setting of high prevalence of Grampositive infections caused by strains with a high rate of resistance to aminoglycoside and betalactam antibiotics, there may be an advantage in including teicoplanin in the initial empiric antibiotic regimen for febrile neutropenic cancer patients.
Surface antigen expression of human neoplastic plasma cells includes molecules associated with lymphocyte recirculation and adhesion
Authors: Hamilton M. S. , Ball J. , Bromidge E. , Franklin I. M.
Published: March 2008

Summary The surface phenotype of neoplastic plasma cells from peripheral blood of plasma cell leukaemia patients and bone marrow of patients with myelomatosis was investigated with two monoclonal antibody panels including 50 selected from the B cell panel of the IVth International Workshop on Leucocyte Differentiation Antigens. The majority of myelomas expressed CD24 (HB8 epitope only), CD38, CD44, CD54, and the antigen recognized by the monoclonal antibody 8A. A range of other antigens may also be expressed including CD10, CD32 (FcR II), CD19, CD20 and MHC Class II. Antigens expressed by myeloma plasma cells can be considered in three groups: (a) antigens associated with lymphocyte and plasma cell differentiation: (b) antigens which are not lineage specific: and (c) molecules concerned with lymphocyte recirculation and intercellular adhesion (CD44 and CD54). The significance of CD44 and CD54 expression by plasma cells and the potential interaction of plasma cells with T lymphocytes and monocytes is discussed.
Occurrence of the αI 22 Arg→His (CGT→CAT) spectrin mutation in Tunisia: potential association with severe elliptopoikilocytosis
Authors: Baklouti F. , Marechal J. , Morle L. , Alloisio N. , Wilmotte R. , Pothier B. , Ducluzeau M.‐T. , Kastally R. , Delaunay J.
Published: March 2008

Summary A category of spectrin αI domain variants are manifested by the increase of the αI 74 kDa fragment at the expense of the parent 80 kDa fragment following partial tryptic digestion. We describe a particular case of αI/74 abnormality in a Tunisian family. The propositus was severely ill and had an elliptopoikilocytosis. To the contrary, his father, who carried the same αI/74 variant, displayed no clinical signs and a few elliptocytes. The increase of the αI 74 kDa fragment was more pronounced in the propositus than in his father. Unexpectedly, the spectrin content was reduced to similar (and limited) extents in both of them, and the father displayed nearly as pronounced an increase of the spectrin dimer percentage as the propositus following low ionic strength extraction. In vitro spectrin dimer reconstitution experiments indicated that the primary mutation was located in the α‐chain itself (not in the β‐chain as is the case in some αI/74 mutants). Following polymerase chain reaction (PCR) amplification, cloning and sequencing of exon 2 of spectrin α‐gene in the father, we found the G→A substitution at position 2 of codon 22 (CGT→CAT; Arg→His). This mutation has been recently discovered in a family of French descent. Dot blot hybridization confirmed that the substitution was transmitted with the αI/74 abnormality. As previously shown, the enhancement of its expression level in the propositus, with respect to the father, was accounted for by the presence of a factor carried by the α‐spectrin allele in trans and transmitted by the mother.
Treatment of refractory pure red cell aplasia with cyclosporine A: disappearance of IgG inhibitor associated with clinical response
Authors: Means Robert T. , Dessypris Emmanuel N. , Krantz Sanford B.
Published: March 2008

Summary Remissions were obtained in 6/9 evaluable patients with pure red cell asplasia (PRCA) refractory to other immunosuppresive agents who were treated with cyclosporine A (CsA). Four of these patients have remained in continuous remission off all treatment for 4–19 months. Another patient who stopped CsA abruptly relapsed, but responded to reinstitution of therapy. The sixth patient died of a cerebrovascular accident while in remission on a low dose of CsA. Acute side effects were minimal and were responsive to dose reduction. One patient developed a lymphoma while in an unmaintained remission, and one patient who did not respond to CsA was found to have a lymphoma approximately a year after stopping treatment.

In vitro studies of autologous erythroid progenitors in a patient with an IgG inhibitor of erythropoiesis showed a reduction of autoantibody associated with the response to CsA. The antigen to which this inhibitor is directed was expressed only during the marrow erythroid burst‐forming unit (BFU‐E) period of erythroid differentiation.

CsA can induce sustained remissions in cases of PRCA refractory to other multiple agents, and these remissions may be associated with a reduction in autoantibody to erythroid progenitor cells. Further studies of patients with PRCA who respond to CsA may lead to an improved understanding of this disorder.
Results of Medical Research Council Childhood Leukaemia Trial UKALL VIII (Report to the Medical Research Council on behalf of the Working Party on Leukaemia in Childhood)
Authors: Eden O. B. , Lilleyman J. S. , Richards S. , Shaw M. P. , Peto J.
Published: March 2008

Summary During the 1970s, despite apparently similar treatment, the prognosis for children with lymphoblastic leukaemia (ALL) improved more in some countries, notably the United States and West Germany, than in others. To find out why, the first phase of the United Kingdom (UK) Medical Research Council (MRC) childhood ALL trial, UKALL VIII, was designed to see whether similar results to the United States Children's Cancer Study Group (CCSG) could be obtained in the U.K. using an identical protocol (CCG 162). Protocol 162 was one of a series of regimens devised by the American Children's Cancer Study Group in the 1970s and was used specifically for their average risk patients (all children with ALL with an initial white cell count up to 50 × 109/l except those aged 3–6 years with white cell counts under 10 × 109/l). One arm (1A) of their study was adopted by the MRC for all children in the U.K. aged 0–14 years with confirmed ALL.

Eight hundred and twenty‐nine consecutive patients were entered between 1980 and 1984. The first 199 patients formed a single arm study as per the original protocol 162 (arm 1A), but the subsequent 630 children were randomized to receive or not two doses of daunorubicin on the first 2 d of induction. This randomization was an attempt to answer the important question as to whether event‐free survival was influenced by the use of four rather than three induction agents. A second randomization between 2 and 3 years continuing therapy was also introduced at this stage as it had been by the CCSG in their protocol. With a minimum follow up period of more than 5 years, disease‐free survival for the whole group is 55%, a considerable improvement on all previous UKALL trials. Results for patients directly comparable with those in CCSG 162 (‘average risk’patients) and their American counterparts were similar.

Daunorubicin was associated with more early deaths but improved disease‐free survival for those achieving remission. More children relapsed who stopped treatment after 2 years than those who continued for 3, but this was balanced by increased treatment mortality in the third year.

The fact that for UKALL VIII the results were similar to those of the CCSG suggests that previous MRC protocols were not sufficiently sustained and intensive, particularly during the maintenance phase of treatment. Daunorubicin may have provided better disease control, albeit with increased toxicity, and 3 years of maintenance therapy did not confer any advantage in long‐term survival over 2 even though it may have reduced the rate of early relapse.
Effects of recombinant human granulocyte colony‐stimulating factor on leucopenia in zidovudine‐treated patients with AIDS and AIDS related complex, a phase I/II study
Authors: Van Der Wouw Poll A. , Van Leeuwen Remko , Van Oers Rien H. J. , Lange Joep M. A. , Danner Sven A.
Published: March 2008

Summary. Twelve male patients, eight with the acquired immunodeficiency syndrome (AIDS) and four with AIDS related complex (ARC), who had zidovudine associated neutropenia (<1 x 109 neutrophils/l) were treated with recombinant human granulocyte colony‐stimulating factor (G‐CSF) in a phase I/II study. Treatment consisted of daily subcutaneous injections with G‐CSF in a weekly increasing dose of 0·4, 2, 5 or 10 μg/kg body weight until a neutrophil count of more than 3 x 109 neutrophils/l was observed. This effective dose was continued for up to 4 weeks, followed by 4 weeks observation period without G‐CSF treatment. Two patients (both with ARC) reached target neutrophil counts at the lowest G‐CSF dose, whereas nine patients needed 2 μg/kg. One patient discontinued treatment before he reached target neutrophil counts. Mean(±SD) neutrophil counts before and after 1 and 4 weeks of effective dose treatment were 0·65(±0·188) × 109, 6·016(±2·595) x 109 and 5·54(±4·237) x 109/l respectively (P<0·01). The number of monocytes increased from 0·171(±0·113) to 0·501(±0·274) and 0·474(±0·374) x 109/l after 1 and 4 weeks of treatment (P<0·01). Other haematologic parameters did not change significantly. Two weeks post‐treatment the numbers of neutrophils and monocytes had returned to pre‐treatment values. Mild side effects consisting of bone, joint or muscle pain were observed in three patients. Two patients (both with AIDS) did not complete the study. One patient stopped treatment because of fever and malaise, attributable to a generalized cytomegalovirus (CMV) infection and one patient had to stop zidovudine treatment because of severe thrombocytopenia. We conclude that G‐CSF increases the number of circulating neutrophilic granulocytes in zidovudine‐treated patients at relatively low doses and with few side‐effects.
Proposal for the recognition of minimally differentiated acute myeloid leukaemia (AML‐MO)
Authors: Bennett J. M. , Catovsky D. , Daniel M‐T. , Flandrin G. , Galton D. A. G. , Gralnick H. R. , Sultan C.
Published: March 2008

Summary. We describe a form of acute myeloid leukaemia (AML), designated AML‐MO, with minimal myeloid differentiation, not included previously in the FAB classification. AML‐MO cannot be diagnosed on morphological grounds alone as the blast cells are large and agranular, sometimes resembling L2 or, rarely, L1 lymphoblasts, and should be identified by the following features: negative myeloperoxidase (MPO) and Sudan Black B reaction (or positive in less than 3% of blasts), negative B and T lineage markers and expression of myeloid antigens recognized by at least one monoclonal antibody, CD13 or CD33. Other myeloid markers are also often positive and these include CDllb and the enzyme MPO demonstrated by immunocytochemistry and or electron microscopy analysis. The findings in a group of 10 cases satisfying the criteria for AML‐MO are described. AML‐MO represents 2–3% of all cases of AML and 1–1·5% of all acute leukaemias. Its clinical and biological significance is not yet apparent but its identification in a larger number of cases may achieve this aim.
T cell‐mediated inhibition of erythropoiesis in aplastic anaemia: the possible role of IFN‐γ and TNF‐α
Authors: Miura Akira , Endo Kazuyasu , Sugawara Tomohiro , Kameoka Jun‐ichi , Watanabe Norimichi , Meguro Kuniaki , Fukuhara Osamu , Sato Isao , Suzuki Chiyuki , Yoshinaga Kaoru
Published: March 2008

Summary. The inhibitory activity of T cells on autologous erythroid colony‐forming units (CFU‐E) (T cell inhibitory activity) in patients with aplastic anaemia (AA) was investigated. In 11 (32·4%) out of 34 AA cases, T cell inhibition on autologous CFU‐E growth was greater than that in normal individuals. In order to evaluate the mechanism of this inhibitory activity, T cell surface markers, interferon (IFN) production in peripheral blood mononuclear cell (PBMNC) liquid culture, and cytokine levels such as IFN and tumour necrosis factor‐α (TNF‐α) in CFU‐E clot cocultured with T cells, were measured in a portion of the patients. In five patients investigated for IFN production in PBMNC liquid culture, all produced statistically more IFN activity than normal individuals under phytohaemagglutinin (PHA‐P) stimulation (P<0·01) with no relation to T cell inhibitory activity. In only one patient whose T cells displayed increased CD8 and HLA‐DR antigen (CD8+HLA‐DR+) and inhibitory activity, a significant amount of IFN‐γ was observed in CFU‐E clot cocultured with T cells, and the addition of anti‐IFN‐γ antibody to the coculture resulted in recovered CFU‐E colony growth. These results suggest that IFN‐γ production by T cells may explain, at least in part, the pathogenesis of haematopoietic defects in AA. In other patients however, T cell inhibitory activity neither correlated to the T cell subpopulations (CD4+/CD8+, CD8+HLA‐DR+), IFN production in PBMNC liquid culture, nor to IFN and TNF‐α levels in CFU‐E clot culture. The roles played by cytokines other than IFN and TNF‐α on haematopoietic precursor cells require further evaluation in a larger sample of patients with AA.