bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA‐DA monoclonal antibodies inhibit the proliferation of normal and chronic granulocytic leukaemia myeloid progenitor cell
Authors: Goldman John M. , Hibbin Jill , Kearney Lyndal , Orchard Kim , Th'ng K. H.
Published: March 2008

Summary. We studied the capacity of murine monoclonal antibodies with HLA‐DR specificity to inhibit the proliferation in vitro of erythroid (BFU‐E and CFU‐E) and granulocyte‐macrophage (CFU‐GM) progenitor cells in normal bone marrow and the blood of patients with chronic granulocytic leukaemia (CGL). Two IgG2 antibodies (CA 2.06 and L243) inhibited the proliferation of normal BFU‐E and CFU‐GM at relatively high dilution; a third antibody, DA2, had no effect on either progenitor cell. A complement‐king monoclonal antibody with T‐cell activity (OKT3) produced only minor reduction in progenitor cell proliferation. Further studies with L243 showed that BFU‐E and CFU‐GM from the blood of patients with CGL were inhibited to the same degree as normal marrow progenitor cells. The inhibition of progenitor

cell proliferation by a given antibody was always complement dependent and was therefore presumed to be due to a direct cytotoxic effect. The inhibitory effect of monoclonal antibodies is a valuable approach to the characterization of antigenic determinants on myeloid progenitor cells and the differential cytotoxicity of selected monoclonal antibodies might be exploitable for therapy.
Quantitation of coagulant antigens and inhibition of activated protein C in combined factor V VIII deficiency
Authors: Giddings J. C. , Sugrue Angela , Bloom A. L.
Published: March 2008

Summary. Inhibition of activated human protein C was assessed in an amidolytic assay system using normal human plasma and samples from patients with hereditary coagulation abnormalities. In eight experiments normal plasma inhibited 63.5% (± 15.6%) protein C activity. Plasma from patients with haemophilia A or isolated factor V deficiency gave results which were not significantly different from normal. However, plasma from patients with combined factor V and factor VIII deficiencies inhibited an average of 24.5% (± 13.6%) of the amidolytic activity (P < 0.01). Two of these plasma samples failed to inhibit any protein C activity. The relationship between the level of inhibitor and those of factor V and factor VIII coagulant antigens (VCAg and VIIICAg) in the combined defect was investigated. There was no significant correlation between the level of inhibitor and any of the coagulation immunoassays on these stored samples but there was significant correlation between VCAg and VIIICAg in some assay systems. The levels of VCAg and VIIICAg was low in most samples from patients with the combined defect which was in contrast to the results obtained when normal plasma was incubated with activated protein C in vitro. The findings are consistent with the presence of biochemical similarities between factors V and VIIIC molecules, but the role of activated protein C and its inhibitor in hereditary combined factor V/VIII deficiency remains to be firmly established.
Deficient natural killer (NK) cell activity in paroxysmal nocturnal haemoglobinuria (PNH)
Authors: Yoda Yasuhiro , Abe Tsukasa , Mitamura Keiji , Saito Kenji , Kawada Kenichi , Onozawa Yasusuke , Adachi Yamao , Nomura Takeo
Published: March 2008

Summary. Natural killer (NK) cell activity was studied in nine patients with paroxysmal nocturnal haemoglobinuria (PNH) and 16 healthy subjects. NK cell activity was deficient in PNH and pretreatment of mononuclear cells with human leucocyte interferon in vitro failed to normalize this abnormality. The deficient NK cell activity may result from an abnormality of a lymphocyte subpopulation not described previously.
Secretion of plasminogen activator by the human macrophage‐like cell line, GCT: separation from colony‐stimulating and erythropoiesis‐enhancing activities
Authors: Barlow Grant H. , Brennan James K. , Abboud Camille N. , Francis Charles W. , Lichtman Andrew H. , Lichtman Marshall A.
Published: March 2008

Summary. The human macrophage‐like cell line, GCT, elaborates monokines such as colony‐stimulating activity (CSA) and erythropoiesis‐enhancing activity (EEA) which stimulate the growth of primitive blood progenitors in culture. These cells also secrete a fibrinolysis activator (FA), which can be identified if cells are cultured in serum‐free medium. FA was found to have a similar molecular weight to CSA and EEA by gel filtration but could be separated from them by ion exchange chromatography. Subcellular fractionation of GCT cells indicated that fibrinolytic activity was present in the cell membranes and cytosol, whereas CSA and EEA were present only in the cytosol. FA resembled urokinase in molecular weight and its strict requirement for plasminogen as a substrate. Double immunodiffusion of GCT activator and urokinase against anti‐urokinase antiserum resulted in a line of identity, and incubation of activator with antiserum resulted in loss of its fibrinolytic activity. Thus, GCT activator was similar, if not identical to the plasminogen activator, urokinase.
The in vitro effect of adenine on pyruvate kinase‐deficient red cells
Authors: Lappin T. R. J. , Elder G. E. , Savage G. A. , Cooke P. J. , Bridges J. M.
Published: March 2008

In pyruvate kinase (PK)‐deficient blood with high levels of reticulocytes, the degree of haemolysis after incubation at 37°C for 48 h was halved by the inclusion of adenine in the medium. The decrease in haemolysis was associated with a higher ATP level but was not related to the pH of the incubation mixture. The incorporation of adenine into nucleotides studied over a 3 h period was similar in normal and PK‐deficient blood. The observed increase in cellular ATP was equivalent to that shown by radioactive measurements to have been synthesized from added adenine. Inhibition of mitochondrial oxidative phosphorylation in the reticulocytes of the PK‐deficient blood by KCN reduced the amount of adenine taken up by the cells by a factor of 3 and altered the pattern of incorporation into the nucleotides. Only 25% of the adenine which entered the blood cells was converted into ATP compared with 85% in the absence of cyanide. Despite the synthesis of small amounts of ATP from labelled adenine, the overall ATP content fell to less than 50% of its original level. It is suggested that incubation with adenine increased the ATP level in the reticulocytes by virtue of mitochondrial oxidative phosphorylation thereby reducing the haemolysis of these cells.
Leukaemia of platelet precursors: diverse features in four cases
Authors: Bevan David , Rose Michael , Greaves Melvyn
Published: March 2008

Four patients are described with malignant cells of definite or probable megakaryocytic lineage in the bone marrow and blood. Megakaryocytic features were defined by morphological and cytochemical studies using light and electron microscopy, and in two cases by reaction with a monoclonal anti‐platelet antibody, AN51. All patients had increased bone‐marrow reticulin, which developed in less than 15 months in two cases. One of these fitted the clinical pattern of acute myelosclerosis of Lewis and Szur (now widely referred to as acute megakaryoblastic leukaemia). Three cases had unique clinical features which are described in detail. There may be several variants of megakaryocytic leukaemia, including chronic forms. One variant is associated with Down's syndrome (trisomy 21).
DEMONSTRATION OF VITAMIN B12 ANALOGUES IN HUMAN SERA NOT DETECTED BY MICROBIOLOGICAL ASSAY
Authors: Chanarin I. , Muir M.
Published: March 2008

Sera were absorbed with polyacrylamide beads to which purified human intrinsic factor was attached. This procedure removed the vitamin B12 analogues which are measured by microbiological assay with Lactobacillus leichmannii and Euglena gracilis and which are measured in an isotope dilution method using intrinsic factor. Such sera still contained B12 analogues that were assayed in an isotope dilution method using a non‐intrinsic factor vitamin B12 binder. Such vitamin B12 analogues make up approximately half of the total vitamin B12 analogues in human serum.
Treatment of aplastic anaemia with antilymphocyte globulin (ALG)
Authors: Fairhead S. M. , Chipping P. M. , Gordon‐Smith E. C.
Published: March 2008

Summary.

Thirty‐one patients with aplastic anaemia were treated with antilymphocyte globulin (ALG) followed by anabolic steroids. All patients were reviewed at least 1 year after receiving treatment. Twenty‐one patients were considered to have a severe form of aplasia. ALG from three sources was used during the trial. Sixteen patients were alive at the time of analysis, eight who had had severe aplastic anaemia (38%) and five of these no longer require blood support. No difference in response was found according to type of ALG used. Patients with a long history of aplasia before treatment fared worse than those with a short history. These results were better than those obtained with anabolic steroids alone in historical controls. It is concluded that ALG treatment is worth giving to patients with aplastic anaemia but that the optimum way of giving this immunosuppression may be different from that used in this presentation.
Studies of the heterogeneity of antithrombin III concentrates
Authors: Barrowcliffe T. W. , Eggleton C. Anne , Mahmoud M.
Published: March 2008

Summary.

The biological activities of antithrombin III (At III) concentrates, prepared by several manufacturers for clinical use, have been compared by three assay methods, and their heparin‐binding properties studied by crossed immunoelectrophoresis and heparin affinity chromatography. Concentrates from two of the four manufacturers showed discrepancies between assay methods, with concentrations by heparin co‐factor assays significantly lower than those by immunological and progressive antithrombin methods. Heterogeneity was also found by heparin binding studies, with about half the total At III antigen in these concentrates being unable to bind to heparin. These results confirm previous findings of heterogeneity in At III concentrates and show that some concentrates contain substantial amounts of altered At III molecules in which the heparin‐binding site has been denatured but the thrombin‐neutralizing site left largely intact.
Biosynthesis of ferritin subunits from different cell lines of HL‐60 human promyelocytic leukaemia cells and the release of acidic isoferritin‐inhibitory activity against normal granulocyte‐macrophage progenitor cells
Authors: Dörner Marianne H. , Broxmeyer Hal E. , Silverstone Allen , Andreeff Michael
Published: March 2008

Summary.

Biosynthesis of acidic isoferritins was investigated in human promyelocytic HL‐60 cells, characterized by diploid (2C), tetraploid (4C) and mixed diploid–tetraploid (2C–4C) DNA cell lines. The three cell lines were studied for the biosynthesis of ferritin and its subunits and for the release of acidic isoferritin‐inhibitory activity against normal CFU‐GM before and after addition of DMSO. While the tetraploid and mixed diploid–tetraploid cell lines synthesized more H‐ (Mr= 21) than L‐subunits (Mr= 19) after induction, the tetraploid line synthesized more H‐subunit before and after induction, compared to the diploid line. The release of acidic isoferritin‐inhibitory activity was greater before than after induction in both cell lines, but the tetraploid cell line released more acidic isoferritin‐inhibitory activity consistent with its greater production of Mr= 21 subunit. However, after induction no inhibitory activity could be detected from the diploid cells and much less activity was detected with the tetraploid cells, suggesting that differentiation caused a decrease in production of acidic isoferritin‐inhibitory activity.
Turnover of 131I‐human spleen ferritin in plasma
Authors: Cragg Susan J. , Covell Ann M. , Burch Anna , Owen Glynn M. , Jacobs Allan , Worwood Mark
Published: March 2008

Summary.

Human spleen ferritin was labelled with 131I and injected into two normal men. The labelled ferritin left the plasma rapidly. The experimental clearance curve could be fitted with accuracy into two single exponential functions. The first component, T1/2= 9 min, accounted for the clearance of about 90% of the labelled ferritin. Surface counting showed uptake of 131I by the liver but not by the spleen. Such a rapid plasma turnover is similar to that found after injection of tissue ferritins into experimental animals but contrasts with the slow turnover previously found for 131I‐labelled human plasma ferritin. Differential clearance of isoferritins from the plasma is an important factor explaining the biochemical and immunological differences between tissue and plasma ferritins.
Serum transcobalamin levels as an early indicator of bone marrow engraftment following transplantation
Authors: Naparstek E. , Rachmilewitz B. , Rachmilewitz M. , Fuks Z. , Slavin S.
Published: March 2008

Summary.

Three B12 binding proteins, the transcobalamins TCI, TCII and TCIII, were determined serially in the serum of five patients who underwent bone marrow transplantation. The increase in TCII, followed by the increase in TCI, proved to be an early indicator of bone marrow regeneration, reaching a peak of up to twice its normal levels at least 5 d prior to the rise of the peripheral white blood cell count.
Eosinophilic blast crisis in a case of chronic myeloid leukaemia
Authors: Marinone G. , Rossi G. , Verzura Paola
Published: March 2008

Summary.

A case of typical chronic myeloid leukaemia (CML) is described, of which the terminal blast crisis was characterized by an impressive proliferation of atypical eosinophils and by the simultaneous complete deficiency of neutrophilic myeloperoxidase. Since eosinophilic cells preserved a strong peroxidase positivity, a high number of immature non‐granular eosinophilic precursors could be recognized among the leukaemic blast cells, thus supporting the diagnosis of eosinophilic blast crisis of CML.
A good prognosis group in childhood acute lymphoblastic leukaemia
Authors: Stockley Robert J. , Ahlquist Patricia Y. , Mott Martin G.
Published: March 2008

Summary.

The records of 121 children presenting with acute lymphoblastic leukaemia between 1969 and 1982 were reviewed. The overall relapse free survival rate was 50%. However, girls presenting with a total white blood cell count of 20 × 109/1 or less (35% of all patients) had a particularly favourable prognosis with an 80% relapse‐free survival rate at 12 years. The difference in prognosis between the sexes is confirmed but this difference is confined to the low initial white cell count group. We suggest that girls in this group should be excluded from the more intensive arms of new treatment protocols.
The ultrastructural morphology of T lymphocytes in B‐chronic lymphocytic leukaemia: a study with monoclonal antibodies and the immunogold technique
Authors: Matutes E. , Crockard A. D. , Catovsky D.
Published: March 2008

Summary.

T‐lymphocytes from five patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) were analysed by light (LM) and electron microscopy (EM) by means of the immunogold technique with monoclonal antibodies combined with E rosettes. LM analysis confirmed the existence of a population of E+ lymphocytes unreactive with the OKT3 monoclonal antibody. The EM study showed that E + lymphocytes from B‐CLL can be distinguished morphologically from the leukaemic B‐cells which were identified by their labelling with FMC4 (anti HLA‐Dr). Within the E + fraction two cell types were seen which differed both in reactivity with OKT3 and ultrastructural morphology. T3 + lymphocytes are similar to normal T3 +, T4 + cells: they have high nucleocytoplasmic (N/C) ratio and few cytoplasmic organelles. Their reactivity with OKT3 is, however, considerably weaker than that of normal T3 + lymphocytes. T3 – (E +) lymphocytes, on the other hand, are characterized by low N/C ratio, active Golgi, lysosomal granules and parallel tubular arrays. These cells resemble normal Tγ lymphocytes which comprise cells with the membrane phenotypes: T3 +, T8 +, M1 – and T3 –, T8 –, M1 +. These results provide further evidence for a T‐cell imbalance in B‐CLL and help define better the cellular basis for this abnormality.
Phenotypic conversion of acute leukaemia from T‐lymphoblastic to myeloblastic induced by therapy with 2′‐deoxycoformycin
Authors: Murphy S. B. , Stass S. , Kalwinsky D. , Rivera G.
Published: March 2008

Summary.

A 6‐year‐old boy with T‐cell acute lymphoblastic leukaemia (ALL) in relapse was treated with the adenosine deaminase inhibitor, 2′‐deoxycoformycin (DCF). Remarkably, his residual leukaemia underwent an abrupt phenotypic shift, coincident with a massive anti‐leukaemic effect of DCF. Both at diagnosis and prior to therapy with DCF, blast cells had typical lymphoblastic morphology and T‐cell characteristics (terminal transferase +, T‐antigen +, Ia –, cALLa –, myeloperoxidase –, and high in adenosine deaminase content). After four courses of DCF by constant infusion, the blast cells were myeloid in appearance and reactivity to a variety of tests (terminal transferase –, myeloperoxidase +, Sudan black B +, esterase +, My‐1 +). We hypothesize that DCF therapy created a selection pressure, blocking pathways of T‐cell differentiation and proliferation, permitting the emergence of a newly dominant myeloid subclone of a multipotential leukaemic cell progenitor with the innate capacity for both T‐lymphocytic and myeloid differentiation.
E‐rosette positive acute lymphoblastic leukaemia in adolescents and adults
Authors: Baccarani Michele , Amadori Sergio , Willemze Roel , Haanen Clemens , Corbelli Giovanna , Gobbi Marco , Meloni Giovanna , Mandelli Franco , Tura Sante
Published: March 2008

Summary.

E‐rosetting of leukaemic blast cells is one of the markers of T‐cell acute lymphoblastic leukaemia (ALL). In children, E+ ALL has a bad prognosis. In adults, data are scarce. This report provides information on 25 E+ ALL adult patients who have a minimum follow‐up time of 36 months. Twenty‐two of 25 patients (88%) achieved complete remission (CR) (median duration 16 months), and six of them were alive, relapse‐free, and off therapy after 36–81 months, with a 26% projected 6‐year relapse‐free survival. In 97 patients with E−SmIg− ALL, who were treated at the same Institutions, over the same period of time, and by the same modalities, the outcome of therapy was almost identical: CR 80%, median duration of first CR 15 months, projected 6‐year relapse‐free survival 15%. The white blood cell (WBC) count at presentation influenced significantly and to the same degree first CR length in both E+ and E− cases. In this adult series, WBC count was not as high as in children. Moreover, a high Hb concentration, a very high WBC count, lymphadenomegaly, and mediastinal involvement, were found more frequently in adolescents and young adults than in adults. Based on these data, it is suggested that in adults E‐rosetting as such is not a marker of a poorer prognosis, that some of the typical features of children E+ ALL weaken with age, and that in adults the disease can have a less aggressive character.
Differential binding of IgG anti‐D and IgG autoantibodies to reticulocytes and red blood cells
Authors: Gray L. S. , Kleeman J. E. , Masouredis S. P.
Published: March 2008

Summary.

125I IgG anti‐D binding to reticulocytes obtained by density fractionation is reduced relative to that bound to all other red cell (RBC) fractions. Maximum D antigen reactivity occurs following reticulocyte maturation with no detectable change in D reactivity of mature RBC throughout their life span. Reticulocytes have in the range of about 60% of the content of mature RBC. Previously reported increased anti‐D agglutinability and binding to old RBC it is not due to an intrinsic increase in D antigen with age, but results from an ‘apparent’decrease in anti‐D binding to young RBC fractions due to reticulocyte enrichment.

IgG RBC autoantibodies obtained by elution from the RBC of eight Coombs‐positive blood donors, probably associated with α‐methyldopa (α‐MD) administration, showed decreased binding to reticulocytes as determined by 125I protein A (PA). Reticulocytes bound about 70% of the IgG bound to mature RBC, indicating that the membrane antigenic determinant defined by these autoantibodies was incompletely expressed in the reticulocyte. This difference in IgG autoantibody binding between reticulocytes and mature RBC is similar to the decreased D antigen content of reticulocytes and consistent with an autoantibody determinant associated with the Rh complex. Direct testing of density fractionated Coombs‐positive RBC in four out of five patients with autoimmune haemolytic anaemia (AIHA) showed reduced quantities of IgG on reticulocytes. The distribution of IgG between reticulocytes and mature RBC may be useful in serologically characterizing patients with AIHA and in identifying subpopulations of patients with this disorder.
Interaction between erythroblasts and macrophages in vitro: effect of neuraminidase‐treatment of erythroblasts and the role of serum factors
Authors: Wiener Edith , Wickramasinghe S. N.
Published: March 2008

Summary.

Some factors influencing the interaction in vitro between imethylsul‐phoxide‐induced Friend leukaemia erythroblasts (IFLE) and syngeneic mouse peritoneal macrophages (Mø) have been investigated. Desialation of erythroblasts by treatment with neuraminidase resulted in a significant increase in their association with and ingestion by Mø. In addition, the interaction with Mø of both neuramini‐dase‐treated and untreated IFLE was found to be influenced by a heat‐stable serum factor, γ‐globulin markedly enhanced the weak IFLE‐Mø interaction which occurred at low serum concentration suggesting that the heat‐stable factor may be an immunoglobulin. Desialation of the IFLE‐rendered them agglutinable by peanut lectin which was used as a probe for neuraminidase‐induced membrane changes. By contrast, cycloheximide treatment of IFLE which also enhances their interaction with Mø, did not result in the exposure of receptors for this lectin. It is proposed that both desialation and cycloheximide treatment of IFLE lead to secondary alterations in their membrane structure which renders them recognizable by Mø.
Cyclosporin A is associated with faster engraftment and less mucositis than methotrexate after allogeneic bone marrow transplantation
Authors: Atkinson K. , Biggs J. C. , Ting A. , Concannon A. J. , Dodds A. J. , Pun A.
Published: March 2008

Summary. Thirty‐eight patients with severe aplastic anaemia or acute leukaemia received HLA‐identical sibling marrow transplants. Seventeen were treated with methotrexate and 21 with cyclosporin A (CyA) to minimize graft‐versus‐host disease post transplant. Marrow engraftment was more rapid and oropharyngeal mucositis less severe in those receiving CyA. This resulted in a shorter period of neutropenia and thrombocytopenia, fewer platelet and granulocyte transfusions and less fever. These benefits have to be weighed against the increased nephrotoxicity of CyA.
Abnormal monoclonal antibody‐defined helper/suppressor T‐cell subpopulations in multiple myeloma: relationship to treatment and clinical stage
Authors: Mills K. H. G. , Cawley J. C.
Published: March 2008

Summary.Among peripheral blood T cells from 2 5 patients with myeloma, % 0KT8+suppressor cells were increased in all but two patients (mean 63 ±12%; normal 34 ±8%), but were present in normal absolute numbers (mean 0.5 ±0.3 × 109/1; normal range 0.2–0.6 × 109/1)‐ OKT4+ helper cells were reduced in relative and absolute numbers (mean 40 ±12%, 0.3 ±0.2 × 109/1; normal 64 ±8%, 0.6·1.4 × 1091). Similar results were obtained in treated and untreated cases, and in patients of different clinical stage. In the 12 patients tested, γ Fc‐rosetting T cells were present in normal numbers (mean 28 ±10%; normal 34±10%). In four patients with IgM disease, similar, but less marked, changes in monoclonal antibody‐defined T‐cell subsets were observed. The findings are discussed in relation to those in other B‐cell malignancies, and it is suggested that imbalances of OKT4+helper/OKT8+suppressor cells may be important in the immune suppression of many of these disorders.
Insulin stimulates erythroid colony formation independently of erythropoietin*
Authors: Kurtz Armin , Jelkmann Wolfgang , Bauer Christian
Published: March 2008

Summary. The effect of insulin on the proliferation of late stage erythroid precursor cells (CFU‐e) from fetal mouse liver and aduit bone marrow was studied in a serum‐free culture system. Insulin in supraphysiological concentrations (> 10 −8m) stimulated erythroid colony formation independently of Ep. The combined effect of Ep and insulin was smaller than the surn of their single effects. The number of colonies obtained with insulin was linearly related to the number of plated cells. These results suggest that insulin stimulates erythroid colony formation by a direct action on CFU‐e.
Solid‐phase vitamin B12 assays using polyacrylamide‐bound intrinsic factor and polyacrylamide‐bound R‐binder
Authors: Muir M. , Chanarin I.
Published: March 2008

Summary. A new solid phase vitamin B12 assay is described using intrinsic factor to measure microbiologically‐available B12 and R‐binder to measure total B12. The solid phase reagent consists of intrinsic factor coupled to polyacrylamide beads and salivary R‐binder coupled to polyacrylamide beads. The assay is simple to perform and separates completely sera from controls and patients with megaloblastic anaemia due to B12 deficiency.
The significance of pulmonary infiltrates developing in patients receiving granulocyte transfusions
Authors: Dana Bruce W. , Durie Brian G. M. , White Ruby F. , Huestis Douglas W. , Paplanus Samuel
Published: March 2008

Summary. We reviewed a series of 109 patients given granulocyte transfusions between 1974 and 1978, to determine if pulmonary infiltrates developing during granulocyte transfusion therapy carried any specific prognostic significance. Eighteen patients developed new infiltrates while receiving granulocytes. Six of these patients died during the acute episode, an overall mortality rate no different from patients not developing new infiltrates. However, the subgroup with pulmonary infiltrates who died had some special features including: infiltrates that developed between 7 and 21 d after transfusions were begun; infiltrates that progressed or persisted during the period of transfusion; and infiltrates that were local rather than diffuse. Autopsies on five of the six patients who died revealed disseminated fungal infection in three, pulmonary haemorrhage in one, and alveolar hyaline membranes in association with Pseudomonas sepsis in one. Besides this high‐risk group specific aetiologies could rarely be assigned for other more diffuse or earlier/later developing infiltrates, despite usual diagnostic studies.

We conclude that local infiltrates developing more than 1 week after transfusions are begun are life threatening; they are often caused by resistant infectious organisms and would seem to represent a failure of granulocyte therapy. Aggressive diagnostic and therapeutic interventions are warranted in patients developing this kind of infiltrate while receiving granulocytes. Conversely, pulmonary infiltrates occurring early or longer than 3 weeks after granulocyte tranfusions are begun are not clinically ominous and may be a manifestation of the beneficial effect of granulocyte therapy.
Serum erythropoietin titres in the anaemia of premature infants
Authors: Saito Tsunehiro , Urabe Akio , Uramoto Kyoko , Hashimoto Yasuo , Takaku Fumimaro
Published: March 2008

Summary. Erythropoietin (EPO) titres in the serum of premature infants were determined by an in vitro method using radioiron incorporation into haem in fetal mouse liver cells in culture. The serum EPO titres in 12 premature infants at birth were significantly lower than those of normal adults. In those premature infants who developed anaemia the serum EPO titres increased significantly by 4–6 weeks after birth, confirming that EPO production increases in premature infants in early days of life.
Penicillin‐G degradation products inhibit in vitro granulopoiesis
Authors: Neftel Klaus A. , Müller Marianne R. , Wälti Manfred , Erni Jana , Gugler Madeleine , Arrenbrecht Stefan
Published: March 2008

Summary. 38% of penicillin‐G in solution decays at 20°C within 24 h, 50% at 37°C and 66% within 3 h at 56°C. These degraded penicillin‐G solutions strongly inhibit growth and maturation of granulocytic stem cells in vitro. Inhibitory concentrations are in the range obtainable with high dose penicillin therapy in vivo. No cytotoxicity of degraded penicillin solutions on bone marrow cells was seen over 24 h. It is suggested that penicillin‐G degradation products are responsible for severe granulo‐cytopenia observed after high dose penicillin‐G therapy.
The production of colony stimulating activity by monocyte enriched fractions from murine continuous bone marrow culture adherent layers
Authors: Oblon David J. , Castro‐Malaspina Hugo , Broxmeyer Hal E.
Published: March 2008

Summary. The production of colony stimulating activity (GM‐CSA) within murine continuous bone marrow cultures was investigated by subjecting either the nonadherent or the adherent layer cells to separation by velocity sedimentation. The presence of GM‐CSA in conditioned medium was defined by the support of granulocyte/macrophage colony formation in soft agar culture. Cell free conditioned medium from weekly feedings of intact continuous marrow cultures and medium conditioned by each fraction of velocity sedimentation separated, nonadherent cells did not contain assayable GM‐CSA. However, medium conditioned by fractions of adherent layer cells with a modal sedimentation velocity of 8·8 mm/h (range 6·2–10·6 mm/h) contained GM‐CSA. Cytochemical studies with Wright's–Giemsa and non‐specific esterase stains in addition to immunofluorescent studies with anti‐collagen III, anti‐collagen IV and monoclonal anti‐Mac I antibodies to define fibroblasts, endothelial cells and monocytes, respectively, demonstrated that the cells within the GM‐CSA producing fractions were enriched with monocytes/macrophages. Fibroblasts and a small proportion of endothelial cells were also present. GM‐CSA is produced within the microenvironment (adherent layer) of murine continuous marrow cultures. Either adherent layer monocytes and/or a monocyte–endothelial cell interaction account for the GM‐CSA production.
Interferon production by human marrow stromal cells
Authors: Shah Girish , Dexter T. Michael , Lanotte Michel
Published: March 2008

Summary. We have demonstrated that normal human bone marrow stromal cells can be induced to produce high levels of β‐interferon (IFN). One of three randomly selected human stromal cell lines produced β‐IFN in similar amounts to one of the best β‐IFN producer cell lines (MG‐63 osteosarcoma cells). The marrow stromal cell lines did not produce γ‐IFN, though a low level of α‐IFN was apparently produced. The stromal cell lines differ from usual producer cell lines in that they can be subcultured for a much longer growth period and also maintain the ability to producer IFN. The cells are karyotypically normal and are not virus transformed. Such cell lines may be useful in the production of human IFN as well as allowing studies on the role of IFN in stromal cell‐haemopoietic cell interactions.
Receptor binding of lactoferrin by human monocytes
Authors: Birgens Henrik S. , Hansen Niels Ebbe , Karle Hans , Kristensen LarsØstergaard
Published: March 2008

Summary. The binding of iron‐saturated 125I‐lactoferrin to human monocytes was studied at pH 7·4 and 37°C. Monocytes in suspension bound 125I‐lactoferrin by a reversible, saturable and specific binding indicating the presence of a receptor. The dissociation constant (KD) of the binding was estimated at about 4·5 × 10−9 M and the number of receptors was about 1·6 × 106 per monocyte. The affinity of native lactoferrin (20% iron saturated) was only slightly below that of iron‐saturated lactoferrin (KD about 7·9 × 10−9 M). Human transferrin, horse cytochrome c and human immunoglobulin G were without inhibitory effect on the binding of 125I‐lactoferrin. The majority of cell‐bound 125I‐lactoferrin was dissociable. The dissociation rate was not affected by addition of unlabelled lactoferrin to the dissociation medium. The binding of 125I‐lactoferrin to adherent mononuclear blood cells showed an about 100‐fold lower affinity (KD about 2·5 × 10−7 M) than to cells in suspension, but the specificity of the binding was the same. These results are compatible with the idea that lactoferrin exerts a biological effect mediated by an interaction with cells of the monocyte/macrophage lineage.
ABH incompatible bone marrow transplantation: removal of erythrocytes by starch sedimentation
Authors: Dinsmore Robert E. , Reich Lilian M. , Kapoor Neena , Gulati Subhash , Kirkpatrick Dahlia , Flomenberg Neal , O'Reilly Richard J.
Published: March 2008

Summary. Nineteen patients with acute leukaemia underwent bone marrow transplantation despite major ABH incompatibility between donor and recipient. The marrow inoculum was prepared prior to infusion by admixture with hydroxy‐ethyl starch to sediment the incompatible erythrocytes, which were then discarded. The infusion was well tolerated with two patients developing transient haemoglobinuria, seven patients developing low grade fever and 10 experiencing no reaction. Durable haematopoietic engraftment was achieved in the 18 evaluable patients and was not influenced by pre‐transplant isohaemagglutinin titres. No difference in time to engraftment, incidence of GVHD or in overall survival was found, compared to ABH compatible transplants. Therefore, the presence of incompatibility did not appear to influence transplant outcome adversely. The technique described is a rapid and safe method for overcoming the ABH barrier in marrow transplantation.
Methotrexate‐induced immune haemolytic anaemia
Authors: Woolley Paul V. , Sacher Ronald A. , Priego Victor M. , Schanfield Moses S. , Bonnem Eric M.
Published: March 2008

Summary. A woman with metastatic breast carcinoma developed acute haemolytic anaemia on two occasions following treatment with methotrexate (MTX) and 5‐fluorouracil (5‐FU). The direct antiglobulin test was positive. Compatible control cells were pretreated with either MTX or 5‐FU and subsequently incubated with patient and control sera. Agglutination in the indirect antiglobulin test was observed only in the presence of patient serum and MTX, and did not occur following pretreatment with 5‐FU, folic acid, folinic acid or triamterene. There was no evidence of complement binding to the red cell surface. A macrophage assay showed increased phagocytic uptake of MTX‐treated cells in the presence of patient serum. This phagocytosis was not enhanced by the addition of complement. The antibody subclass was predominantly IgG‐3. The 51Cr red cell survival of autologous MTX cells was markedly shortened, indicating rapid in vivo destruction.

These studies demonstrate the occurrence of acute haemolytic anaemia due to an IgG‐3 antibody that reacts with erythrocytes in the presence of methotrexate. This antibody does not fix complement, but sensitizes red blood cells to phagocytosis by macrophages.
Separation of cobalamin analogues in human sera binding to intrinsic factor and to R‐type vitamin B12 binders
Authors: Muir M. , Chanarin I.
Published: March 2008

Summary. Intrinsic factor (IF) and cobalamin‐R‐binding protein (R‐binder) linked to polyacrylamide beads were used to absorb cobalamins from solutions and serum extracts. Both binding agents were equally effective in removing [57Co]B12 from aqueous solution. IF was more effective than R‐binder in removing [57Co]B12 added to a serum extract.

All endogenous cobalamins detectable in serum by saturation analysis assay were removed by absorption onto R‐binder. Absorption with IF removed microbiologically‐active cobalamins but left behind analogues assay able, with an R‐binder. However, when absorption with IF was continued the concentration of R‐binding cobalamins steadily declined indicating that IF bound both types of cobalamins though the binding was less avid for the microbiologically‐inactive analogues than for microbiologically‐active cobalamins. Finally, the R‐binding analogues in serum were carried on transcobalamin I and none was detectable on transcobalamin II.

The absorption studies establish the presence of two types of cobalamins one binding preferentially to IF and the other preferentially to R‐binder. Only the former is detected by microbiological assay.
Preservation of deformability (filterability) of sickle cells by BW12C during progressive deoxygenation
Authors: Kenny M. W. , Stuart J.
Published: March 2008

Summary. venous blood from patients with sickle‐cell disease in the steady state or in crisis was progressively deoxygenated in vitro to study the effect of BW12C, a new compound designed to stabilize haemoglobin in the oxy‐conformation, on the deformability (filterability) of washed erythrocytes. At a final concentration of 1·5 mM, BW12C significantly increased erythrocyte deformability, compared with no added compound, at all levels of deoxygenation below normal arterial PO2. At concentrations of 3·0 and 5·0 mM, BW12C prevented any significant reduction of erythrocyte deformability, or increase in sickled cells, with deoxygenation down to PO2 values below the normal venous level. These in vitro results demonstrate the considerable potential, as an anti‐sickling agent, of this novel compound.
Fractionated total body irradiation in marrow transplantation for leukaemia
Authors: Vitale V. , Bacigalupo A. , Lint M. T. van , Frassoni F. , Ricci G. , Siracusa G. , Marziano G. , Vitali M. L. , Scielzo G. , Tomassini A. , Bistolfi F. , Marmont A. M.
Published: March 2008

Summary. Thirty consecutive patients with leukaemia were prepared for bone marrow transplantation (BMT) with cyclophosphamide (CY) 120 mg/kg followed by total body irradiation (TBI). TBI was delivered in a single dose (sTBI) of 10 Gy, at a dose rate of 0·06–0·08 Gy/min, or in fractionated doses (fTBI) of 3·3 Gy/d, on each of 3 consecutive days, at the same dose rate. Lung shielding was adopted for all patients, in order to obtain a homogeneous dose delivered to the lung and at midline. The first 12 patients were prepared with sTBI and the following 18 with fTBI.

Several variables (remission status, mode of irradiation, prevention of graft versus host disease (GvHD) with methotrexate (MTX) or cyclosporin A(CyA) were then analysed for their influence on (a) incidence and mortality of GvHD, (b) incidence and mortality of interstitial pneumonia (IP), (c) 100‐d survival, and (d) relapses. Our data suggest that fTBI is the single most important factor associated with a lower incidence of IP (P= 0·002), a lower mortality from GvHD and IP (P= 0·03 and 0·001) and a better 100‐d survival (P= 0·03). Remission status had no significant influence on GvHD, IP and acute mortality. When compared to MTX, the use of CyA was associated with less GvHD and IP, but when only patients in remission given fTBI were analysed, this was no longer true. Relapses were mostly influenced by the remission status of the patient at transplant (P= 0·001).

Although this is not a randomized study, our data suggest that fTBI can reduce the acute risks of marrow transplantation in leukaemia.
Effects of dexamethasone on erythroid colony and burst formation from human fetal liver and adult marrow
Authors: Roodman G. D. , Lee J. , Gidari A. S.
Published: March 2008

Summary. Dexamethasone, a prototypic synthetic glucocorticoid, was added to cultures of human fetal liver and adult marrow cells to assess its effects on erythroid colony and burst formation. Dexamethasone decreased the number of fetal liver erythroid colonies and bursts formed in the presence of erythropoietin, and also decreased the number of cells per colony. The amount and type of haemoglobin produced per cell were unaffected by adding dexamethasone to the cultures. Dexamethasone inhibited the incorporation of 3H‐thymidine into DNA in fetal liver cells stimulated with erythropoietin, supporting the hypothesis that dexamethasone inhibits the proliferation but not the differentiation of fetal liver CFU‐E and BFU‐E. In contrast, addition of dexamethasone to adult bone marrow cultures had a variable effect on erythroid colony and burst formation.
Ferritin in the red cells of normal subjects and patients with iron deficiency and iron overload
Authors: Cazzola Mario , Arosio Paolo , Barosi Giovanni , Bergamaschi Gaetano , Dezza Laura , Ascari Edoardo
Published: March 2008

Summary. Red cell ferritin was measured in normal subjects and patients with iron deficiency and iron overload by means of radioimmunoassays with antibodies to liver (basic) and heart (acidic) ferritins. In most of the subjects examined, red cells were found to contain greater amounts of heart‐type than liver‐type ferritin. The basic ferritin content reflected the abnormal body iron status both in iron deficiency and iron overload while the acidic ferritin content was less closely related to the iron status. The two immunologically different red‐cell ferritins probably represent distinct ferritin molecules and may have different metabolic functions within haem‐synthesizing erythroid cells.
In vitro determination of red cell alloantibody significance using an assay of monocyte‐macrophage interaction with sensitized erythrocytes
Authors: Branch Donald R. , Gallagher Michael T. , Mison Angeles P. , Sy Siok Hian Anita L. , Petz Lawrence D.
Published: March 2008

Summary One hundred and forty‐eight red cell alloantibodies, of specificities generally considered to be of clinical significance, were studied in vitro for their ability to induce phagocytosis of sensitized red cells by allogeneic mononuclear phagocytes. Results indicate that only 53% of the alloantibodies studied mediated significant phagocytosis in vitro. The percentages for each blood group system were as follows: Kell, 73%; Jka, 32%; Jkb, 67%; D, 75%; E, 60%; Fya, 62%; Yta, 25%; Ge, 22%; and Vel, 25%. Significant phagocytosis was independent of the strength of the indirect antiglobulin test. The percentage of anti‐Jka and anti‐Fya mediating significant phagocytosis was increased when fresh complement was added during the sensitization procedure and/or red cells homozygous for the antigen in question were used. The in vivo clinical significance or lack of significance was documented for nine alloantibodies; five caused haemolysis and four did not. Those causing in vivo haemolysis mediated in vitro phagocytosis by monocyte‐macrophages whereas the antibodies that did not result in haemolysis showed no increased in vitro phagocytosis. Autologous monocytes were more reliable than random allogeneic monocytes in that phagocytosis was increased over that obtained using allogeneic monocyte‐macrophages with two of four alloantibodies having documented clinical significance. The use of target red cells homozygous for the antigen in question, the addition of fresh complement in the antibody sensitization procedure, and use of autologous and allogeneic monocyte‐macrophages appear necessary for optimal results.

Since 47% of those alloantibodies generally considered to be clinically significant failed to mediate phagocytosis in vitro, the monocyte‐macrophage assay should not be considered a predictive assay of a given alloantibody's in vivo significance or lack of significance until more extensive correlation of these assays with in vivo red blood cell survival is obtained.
Localization of factor VIIIC: antigen in guinea‐pig tissues and isolated liver cell fractions
Authors: Kelly D. A. , Summerfield J. A. , Tuddenham E. G. D.
Published: March 2008

Summary. Factor VIIIC: antigen (VIII: CAg) was estimated in guinea‐pig tissues by an immunoradiometric assay using a human inhibitor antibody. In homogenized guinea‐pig tissues, VIII: CAg was shown to be stable and to be predominantly located in the liver (9±1·2 units; mean ±SEM, n= 8). Lesser amounts were detected in spleen (1·3±0·02 units), lung (0·6±0·07) and kidney (0·4±0·06). In isolated liver cell fractions separated by centrifugal elutriation VIII: CAg was mainly detected in the hepatocyte fraction (0·3±0·07 units/108 cells; mean ± SEM, n= 5) and in lesser amounts in the endothelial (0·02±0·01 units/108 cells) and the Kupffer cell fractions (0·05±0·02 units/108 cells). The liver concentration of VIII:CAg was (0·17±0·02 units/g) which was 20% of the plasma concentration (0·96±0·01 units/ml, n= 8) suggesting that VIII: CAg may not be stored in the liver but is rapidly exported following synthesis.
The effect of spicules obtained from sickle red cells on clotting activity
Authors: Westerman M. P. , Cole E. R. , Wu K.
Published: March 2008

Summary. Spicules from sickle red cells were examined for their effects on the clotting activity of blood. The spicules were obtained from the sickle red cells after deoxygenation and oxygenation and were tested for clotting activity with Russell's viper venom assay. A marked increase in clotting activity was observed when spicules were added to the system. The increase was distinctly greater than that observed after the addition of sickle red cells while normal red cells had little effect. Vesicles prepared from sickle or normal red cells by incubation with the ionophore A‐23187 + Ca2+ also markedly increased clotting activity. The effect of spicules or vesicles on the clotting system may be related to reorganization of phospholipid in the spectrin‐poor membrane of the spicules or vesicles. Because of these effects, the spicules from the sickle red cells may contribute to the hypercoaguable state in these patients and possibly to their vaso‐occlusive crises since free spicules are present in their plasma. Vesicles from red cells from other types of anaemia with hypercoaguability may have a similar effect on coagulation.
In vitro and in vivo studies of iron delivery by human monoferric transferrins
Authors: van der Heul C. , Kroos M. J. , van Noort W. L. , van Eijk H. G.
Published: March 2008

Summary. According to the Fletcher‐Huehns hypothesis there exists a functional difference between the two iron‐binding sites of transferrin. In this study we present the results of an evaluation of this hypothesis in vitro and in vivo with human pure monoferric transferrins obtained by preparative isoelectric focusing in granulated gels. The uptake of iron from monoferric transferrins TfFec and FeNTf by erythroid bone marrow cells, hepatocytes and stimulated T‐lymphocytes in vitro was equal, even when both monoferric transferrins were present together in the incubation medium. Ferrokinetic studies in vivo, performed with both pure monoferric transferrins, showed that transferrin TfFec, as well as transferrin FeNTf, mainly deliver their iron to the erythron. As red cell 59Fe utilization, red cell iron turnover and other ferrokinetic parameters, obtained from this study, were identical too it is evident that both iron‐binding sites of transferrin are functionally homogeneous in vivo, with respect to iron delivery.