bluuebunny/crossref_metadata_2025 huggingface dataset:
Mitochondrial NADH dehydrogenase in iron‐deficient and iron‐repleted rat muscle: an EPR and work performance study
Authors: Ohira Yoshinobu , Hegenauer Jack , Strause Linda , Chen Chuan‐Show , Saltman Paul , Beinert Helmut
Published: March 2008

Summary.Iron may affect both respiratory O2transport and mitochondrial electron transport in the performance of muscle work. This study was designed to elucidate the molecular defect iron‐deficient work performance by identifying heretofore unmeasurable mitochondrial enzymes that are diminished by iron deficiency and may be restored by iron repletion. Female rats were made iron‐deficienty by dietary control and were repleted by oral iron. Iron deficiency reduced physical work capacity (treadmill running time), haemoglobin (Hb), and mitochondrial ironsulphur (Fe‐S) centres in heart and skeletal muscles; mitochondrial number was unaffected. Oral iron supplementation restored work capacity and Hb within 4 d to normal or near‐normal levels, but in general Fe‐S centrres of mitochondria due to NADH dehydrogenase remained at iron‐deficient levels. Subnormal concentrations of mitochondrial iron‐dependent NADH dehydrogenase in muscle are not by themselves rate‐limiting in work performance.
Uptake of transferrin by rat peritoneal macrophages
Authors: Nishisato Takuji , Aisen Philip
Published: March 2008

Summary. Activated rat peritoneal macrophages bind 125I‐apotransferrin in a time and temperature‐dependent process, the amount of transferrin taken up at 4°C amounting to only about 15% of that bound at physiological temperatures. Binding is reversible, saturable, and largely abolished by prior treatment of the cells with Pronase. A single class of high affinity binding sites is evidenced by Scatchard analysis, each cell binding about 110 000 apotransferrin molecules with an apparent affinity constant of 1.4 x 106 1 mol‐1. Macrophages are also capable of binding about one‐third as much iron‐saturated transferrin as iron‐free transferrin. Since binding of neither form of the protein is influenced by the presence of the other, separate and independent binding sites for apotransferrin and iron transferrin are presumed to exist on the macrophage.
Incidence of myeloperoxidase deficiency in an area of northern Italy: histochemical, biochemical and functional studies
Authors: Cramer R. , Soranzo M. R. , Dri P. , Rottini G. D. , Bramezza M. , Cirielli S. , Patriarca P.
Published: March 2008

Forty‐five subjects with a complete deficiency of myeloperoxidase were identified in an area of the region Friuli‐Venezia Giulia in north‐eastern Italy using the Hemalog D system as the screening technique. Histochemical and biochemical tests performed on the leucocytes of some of these subjects confirmed the defects shown by the Hemalog D system. The defect was of genetic origin in seven subjects. The genetic origin could be suspected in another eight subjects since more than two affected members were present in a given family. Eosinophil peroxidase, which is present in MPO deficient subjects, interfered with the guaiacol assay of MPO, and in several cases masked the genetic transmission. An assay was developed using o‐dianisidine as the electron donor which considerably reduced the interference by EPO. With this assay an autosomal recessive pattern of inheritance was found. The MPO deficient leucocytes had a higher respiratory burst than control cells and an impaired bactericidal activity, at early post‐phagocytic periods, which became comparable to that of control cells at later stages. Particle ingestion by the MPO‐deficient cells was normal.
In vivo stimulation and inhibition of granulopoiesis: the effect of an inflammatory reaction on murine diffusion chamber granulopoiesis
Authors: Symann M. , Anckaert M. A. , Huybrechts M. , Ninane J. , Canon J. L. , Sokal G.
Published: March 2008

Humoral factors influencing granulopoiesis have been evaluated using diffusion chambers (DC) implanted in the peritoneal cavity of mice challenged by an aseptic abcess produced by the subcutaneous implantation of copper rods. This resulted in an increase in peripheral blood neutrophils and an increase in tibial granulocytic elements. When DC loaded with bone‐marrow cells were implanted into mice stimulated the day before by an aseptic abcess significantly more CFU‐s, CFU‐c, proliferative and non‐proliferative granulocytes were produced, as compared to DC implanted into control hosts. When DC were implanted 4‐6 d after the induction of inflammation in mice a significant depression of DC granulopoiesis was observed. Levels of serum and DC fluid CSF and serum inhibitors of in vitro colony growth showed no correlation with DC myelopoiesis. The data show that mice undergoing an inflammatory reaction elaborate first humoral substance(s) enhancing CFU‐s and granulocytic growth in DC and next inhibitory factor(s) of DC granulopoiesis.
Basophils, histamine and gastric acid secretion in chronic myeloproliferative disorders
Authors: Gray A. G. , Boughton B. J. , Burt D. S. , Struthers G. R.
Published: March 2008

Fourteen patients with polycythaemia rubra vera (PRV) and four with chronic granulocytic leukaemia (CGL) were studied to examine the relationship between basophil counts, blood histamine, gastric acid secretion and dyspepsia or peptic ulceration. Basophil counts and blood histamine were markedly raised in untreated CGL but only marginally increased in other cases. Blood histamine and basophil counts were well correlated. Gastric acid secretion was normal in 10 out of 11 cases, and did not correlate with blood histamine values. The previously reported high incidence of dyspepsia and peptic ulceration is examined and its mechanisms are discussed.
The interaction between factor VIII clotting antigen (VIIICAg) and phospholipid
Authors: Yoshioka A. , Peake I. R. , Furlong B. L. , Furlong R. A. , Giddings J. C. , Bloom A. L.
Published: March 2008

Summary.

The interaction between factor VIII clotting antigen (VIIICAg) and phospholipid (PL) was studied using a two‐site solid phase immunoradiometric assay (IRMA) for VIII CAg. Incubation (2 h, 37°C) of normal plasma, cryoprecipitate or factor VIII concentrate with Diagen PL (0·5 mg/unit VIIICAg) resulted in 80–90% loss of IRMA‐measurable VIIICAg. No loss of factor VIII related antigen (VIIIRAg) or factor VIII clotting activity (VIIIC) was seen. Treatment of factor VIII concentrate with purified PLs showed greatest VIIICAg reduction with phosphatidylserine, less with phosphatidylethanolamine, and very little with phosphatidylcholine. The action of phospholipase‐C (PL‐C) on VIIICAg–PL complexes was investigated, with enzyme activity being monitored by thin‐layer chromatography. Treatment of normal plasma, cryoprecipitate or factor VIII concentrate with PL‐C (5 u/unit VIIICAg) resulted in 25%, 25% and 30% increases in VIIICAg. No increase in VIIIC or VIIIRAg was seen. Preincubation of factor VIII concentrate with PL, followed by PL‐C incubation, resulted in 70–80% recovery of measurable VIIICAg. Incubation of ‘activated’prothrombin complex with PL‐C increased VIIICAg by 44% (Autoplex) and 80% (FEIBA), indicating VIIICAg–PL complexes are present. Incubation of factor VIII concentrate with fresh platelet lysate led to a reduction in VIIICAg (100 u/dl to 21 u/dl). In fresh platelet lysate alone low VIIICAg levels were detectable (0·71 × 10−3 u/109 plt). After PL‐C incubation VIIICAg levels increased to 9·8 × 10−3 u/109 plt) (14‐fold increase). Thus VIIICAg in platelets may be hidden by VIIICAg–PL complexes.
Isoferritins in normal leucocytes
Authors: Jones B. M. , Worwood M. , Jacobs A.
Published: March 2008

Summary.

Monocytes, lymphocytes and polymorphs were separated from the peripheral blood of normal human subjects. Ferritin concentrations were determined with antibodies to both human spleen and heart ferritins. The heart type ferritin concentration in monocytes was 38·4±21·6 fg/cell (mean ±SD), in lymphocytes 8·6±6·6 fg/cell and in polymorphs 3·2±2·4 fg/cell. Spleen type ferritin concentrations (fg/cell) were 15·6±7·0 in monocytes, 6·6±5·7 in lymphocytes and 7·0±4·6 in polymorphs. The mean heart/spleen ferritin ratios were 2·8/1 for monocytes, 2·0/1 for lymphocytes and 0·6/1 for polymorphs. The cell extracts were also subjected to anion exchange chromatography. Heart type ferritin eluted at a higher chloride ion concentration than spleen type ferritin. Both H and L subunits were synthesized by mononuclear cells when incubated with 3H‐leucine. Human leucocytes contain a wide range of isoferritins and ferritin concentrations may be considerably underestimated in conventional assays for serum ferritin which employ antibodies to liver or spleen ferritin.
Demonstration of an immune‐mediated mechanism of penicillin‐induced neutropenia and thrombocytopenia
Authors: Murphy M. F. , Riordan T. , Minchinton R. M. , Chapman J. F. , Amess J. A. L. , Shaw E. J. , Waters A. H.
Published: March 2008

Summary.

Severe neutropenia may be a more common complication of high‐dose penicillin therapy than previously recognized. This report describes five such patients, one of whom also had thrombocytopenia. The neutrophil and platelet counts rapidly increased on stopping penicillin, and the bone‐marrow, which was hypocellular in some cases, became normal. Further studies on one of these patients, using a fluorescent antiglobulin technique with paraformaldehyde‐fixed cells, demonstrated a complement‐fixing IgG penicillin antibody reacting with the patient's granulocytes and platelets in the presence of the drug. This suggested an immune mechanism similar to the well‐recognized penicillin‐induced immune haemolytic anaemia. The associated bone‐marrow hypoplasia may also be due to antibody‐mediated suppression of penicillin‐coated precursor cells.
Oxygenation alters red cell folate levels
Authors: Nelson Nancy L. , Klausner Jeffrey S. , Branda Richard F.
Published: March 2008

Summary.

Folate activity is consistently higher in deoxygenated than oxygenated human blood as measured by both radioassay and Lactobacillus casei growth. Red cell, but not serum, folate levels are rapidly mutable: when fully oxygenated, the mean red cell folate level of 15 samples was 489 ng/ml; after deoxygenation the mean rose to 553 ng/ml; following re‐oxygenation the mean returned to 488 ng/ml. Transport of folate across the red cell membrane apparently does not contribute to these changes in folate levels, since agents known to inhibit permeation (N‐ethyl‐maleimide, p‐chlormercuriphenyl sulfonic acid and ethacrynic acid) do not affect the rise with deoxygenation. Moreover, new synthesis of folate compounds seems unlikely, since the changes are observed in energy‐depleted cells. Instead, direct binding of folate to haemoglobin is suggested by experiments in which the rise with deoxygenation was not seen in red cells exposed to carbon monoxide or treated with cyanate. Similar changes also occur in vivo: canine red cell, but not serum folate levels are significantly higher (P < 0.05) in samples collected from five different veins compared with blood from the paired artery (carotid‐jugular, renal, mesenteric, splenic, femoral) of four animals. Thus red cell folate levels are predictably influenced by the levels of oxygenation of the erythrocyte both in vitro and in vivo, possibly by reversibly binding to haemoglobin.
Incomplete glycosylation of erythrocyte membrane proteins in congenital dyserythropoietic anaemia Type II (CDA II)
Authors: Mawby W. J. , Tanner M. J. A. , Anstee D. J. , Clamp J. R.
Published: March 2008

Summary.

The alterations in the erythrocyte membrane proteins of individuals with congenital dyserythropoietic anaemia (CDA II) were studied. Alterations were observed in both the erythrocyte sialoglycoproteins and erythrocyte anion transport protein (Band 3). There was a decrease in the apparent molecular weight of the major sialoglycoprotein α (glycophorin A) as well as a general reduction in the intensity of staining of all the sialoglycoproteins by the PAS stain. Sialoglycoprotein α isolated from CDA II erythrocytes contained 30% less sialic acid than normal α. The anion transport protein of CDA II erythrocytes migrated as a band with a lower molecular weight than the normal protein on SDS‐gel electrophoresis. The CDA II anion transport protein had a substantially reduced content of N‐acetylglucosamine and galactose, which probably reflects a reduction in the number of N‐acetyl‐lactosamine units carried by the protein. Our results suggest that there is a general defect in glycosylation of the major membrane glycoproteins of CDA II erythrocytes. We suggest that this glycosylation defect is a consequence of bone marrow stress.
Cellular cooperation in endothelial cell thromboplastin synthesis
Authors: Lyberg T. , Galdal K. S. , Evensen S. A. , Prydz H.
Published: March 2008

Summary. Endothelial cells from human umbilical veins produce a procoagulant identified as thromboplastin (tissue factor, factor III) when stimulated with the phorbol ester 12‐0‐tetradecanoyl‐phorbol‐13‐acetate (TPA), phytohaemagglutinin (PHA) or endotoxin. Inducible thromboplastin synthesis (i.e. synthesis of the protein component of thromboplastin, apoprotein III) was totally inhibited by cycloheximide and actinomycin D, indicating that de novo protein and RNA syntheses are necessary. Serum enhanced the induced apoprotein synthesis.

Of the total thromboplastin activity in homogenates of stimulated endothelial cells, about 50–70% was available on the cell surface for interaction with other coagulation factors, inactivation by trypsin and neutralization with antiserum against apoprotein III.

Induced synthesis of thromboplastin in endothelial cells was 2–7‐fold enhanced by the presence of several other cell types in optimal ratio 4–10 cells per endothelial cell. Some of these cell types were themselves thromboplastin producers (U‐937, U‐937‐4), some were not inducible (lymphocytes, granulocytes and the lymphoblast lines Daudi and Molt 4). This enhancing effect was also seen with cell‐free culture supernatants, but these were generally somewhat less effective than the intact cells. Supernatants derived from cells cultured in the presence of TPA, PHA or endotoxin were in most cases more effective than supernatants from unstimulated cells.
Enrichment of erythroblasts from human bone marrow using complement‐mediated lysis: measurement of ferritin
Authors: Ali Faieza M. K. , May Alison , Jones Brian M. , Jacobs Allan
Published: March 2008

Summary. Sequential lysis of human bone marrow cells with a monoclonal antibody directed against myeloid cells (TG1) and a rabbit antiserum raised against peripheral blood mononuclear cells gave preparations in which 78–97% of the nucleated cells were erythroid, with a 24–77% recovery. Viability was high, morphology was good and the cells were able to divide and differentiate in culture. No metabolic experiments were carried out but the ferritin content of the erythroblasts was measured in four experiments and found to be about 200–2000 times higher than that found in normal erythrocytes. The H/S ratio was high in both erythroblasts and erythrocytes. Fractionation on the basis of density of two erythroblast preparations, one from a patient with sideroblastic anaemia and one from a patient with megaloblastic anaemia, showed that the most immature erythroblasts contained the highest content of ferritin and that this fell with maturation. The H/S ratio stayed the same or fell with maturation. It was concluded that this method would be valuable for the study of the role of erythroblast ferritin in normal and pathological situations.
Irreversible spectrin‐haemoglobin crosslinking in vivo: a marker for red cell senescence
Authors: Snyder L. M. , Leb L. , Piotrowski J. , Sauberman N. , Liu S. C. , Fortier N. L.
Published: March 2008

Summary. We have demonstrated that the in vivo presence of spectrin and haemoglobin aggregates occur in the oldest, dehydrated and most dense fraction of normal circulating red cells, following their separation by Percoll/hypaque density gradient. To our knowledge, this is the first evidence of irreducible in vivo oxidative damage to the red cell membrane cytoskeleton.
Influence of the tumour promoter TPA upon erythroid burst formation in vitro
Authors: Brockbank K. G. M.
Published: March 2008

Summary. A dose dependent effect of the tumour promoter TPA on burst formation by rabbit erythroid progenitors (BFU‐e) was demonstrated in cultures deficient in the early erythroid regulator burst‐promoting activity (BPA). In these culture conditions the burst number was highest (193% of controls) at 10 9m TPA and concentrations higher than 3 × 10 9m TPA were inhibitory. The degree of burst enhancement by TPA and bone marrow conditioned medium as a source of BPA was similar. The addition of optimal concentrations of both TPA and BPA simultaneously to cultures resulted in no further increase in burst number. Short‐term incubation of bone marrow cells with TPA failed to enhance the percentage of S‐phase BFU‐e under conditions in which BPA significantly increases the number of BFU‐e in the cell cycle.

These results indicate that the same population of BFU‐e responds to TPA and BPA, but TPA does not mimic the mitogenic effect of BPA upon BFU‐e.
Studies of the efficacy and potential hazards of methylene blue therapy in aniline‐induced methaemoglobinaemia
Authors: Harvey John W. , Keitt Alan S.
Published: March 2008

Summary. The similarity between poison and antidote was known to the ancient Greeks who used the same word, pharmakon, for both. This paper presents evidence that aniline (the toxin) and methylene blue (the therapy) are in fact remarkably similar and additive in some of their effects on erythrocytes. Studies were prompted by a case of aniline‐induced methaemoglobinaemia in which two injections of methylene blue did not rapidly eliminate cyanosis and were followed by severe, delayed haemolysis. Interactions between aniline and methylene blue were studied in cats which, although showing important differences from man in their haemoglobin and splenic vasculature, represent a useful model. Methylene blue potentiated the oxidative denaturation of haemoglobin by aniline as judged by the size and number of Heinz bodies and their turbidity in haemolysate. It also aggravated and prolonged the fall in erythrocyte reduced glutathione content which occurred at a time of maximum Heinz body production. While methylene blue in judicious dosage will reduce the content of methaemoglobin after aniline exposure, it may not eliminate visible cyanosis. Repeated injections of methylene blue can markedly aggravate subsequent haemolysis without further lowering methaemoglobin content.
Haematological reconstitution following high dose and supralethal chemo‐radiotherapy using stored, non‐cryopreserved autologous bone marrow
Authors: Burnett A. K. , Tansey P. , Hills C. , Alcorn M. J. , Sheehan T. , McDonald G. A. , Banham S. W.
Published: March 2008

Summary. Eighteen patients with small cell carcinoma of the lung received high dose cyclophosphamide (180–200 mg/kg) intensification following five pulses of ‘CHOP’ chemotherapy (cyclophosphamide 750 mg/m2 i.v., adriamycin 50 mg/m2 i.v., vincristine 1·4 mg/m2 i.v., prednisolone 40 mg orally for 5 d). They received infusions of autologous bone marrow which had been stored at 4°C for 34 h. Pancytopenia was predictable in onset and its duration acceptable. Recovery of neutrophils to greater than 1·0 × 109/l was achieved in 17·5 ± 0·9 d (mean ± SEM) and platelets to greater than 100 × 109/l in 17·5 ± 0·8 d. Four patients with acute myeloid leukaemia in complete remission received intensification with the supralethal combination of cyclophosphamide and total body irradiation followed by infusion of autologous marrow which had been stored at 4°C for 54 h. Haematological reconstitution in these patients was acceptable but slower (greater than 1·0 × 109/l neutrophils between days 26 and 40; greater than 20 × 109/l platelets between days 23 and 77). Except in one case, normal peripheral counts were attained in all patients.

It is concluded that bone marrow stored at 4°C for up to 54 h is a simple and practical source of viable stem cells which have the capacity for acceptable haematological reconstitution.
Normal equivalent cells of B cell malignancies: analysis with monoclonal antibodies
Authors: Gobbi M. , Caligaris‐Cappio F. , Janossy G.
Published: March 2008

Summary. Immunoglobulin heavy chain expression and reactivity of monoclonal antibodies RFA‐1, ‐2, ‐3, ‐4 and OKT10 discriminate between majority B lymphocyte populations in the bone marrow and in peripheral lymphoid organs. In this study normal tissues and various B cell malignancies were studied in cell suspensions and tissue sections. Pre‐B acute lymphoblastic leukaemia and multiple myeloma apparently reflect the phenotypes on normal B lineage cells of the bone marrow, while centroblastic‐centrocytic lymphoma, B chronic lymphoid leukaemia (CLL) and prolymphocytic leukaemia (PLL) show the characteristics of distinct peripheral B lymphoid subsets found at different sites in the lymphoid tissue. These ‘normal equivalent’ cells are centroblasts and centrocytes in the germinal centre, CLL‐like cells at the edge of the germinal centre (a minority population) and strongly Ig positive cells in the lymphocyte corona. Malignant cells in macroglobulinaemia are apparently more closely related to PLL and the corresponding normal peripheral B cells (in the corona) than to myeloma cells or the equivalent normal plasma cells in the bone marrow.
Sickle cell disease: the proportion of liganded haemoglobin needed to prevent crises
Authors: Franklin I. M. , Rosemeyer M. A. , Huehns E. R.
Published: March 2008

Summary. In an attempt to predict the likelihood of successfully treating sickle cell disease by increasing haemoglobin S (Hb S) oxygen affinity, two liganded derivatives of Hb S have been studied in an in vitro system that measures deoxy‐Hb S polymerization. The participation of these liganded forms in the polymers has been quantitated in terms of an exclusion factor that relates their behaviour to that of deoxy‐Hb S.

Carbonmonoxy‐Hb S has an oxy‐Hb‐like conformation and did not participate significantly in the polymerization. It was calculated that 30% carbonmonoxy‐Hb S would have to be maintained in vivo to prevent sickling. Met‐Hb S has a conformational equilibrium intermediate between oxy‐ (or carbonmonoxy‐) and deoxy‐Hb S and behaved in a similarly intermediate manner with regard to deoxy‐Hb S polymerization. 60% met‐Hb S would be needed to prevent in vivo sickling.

It is concluded that stabilizing the oxy(R)‐conformation is a potentially useful way of preventing sickling, and that a level of 30% R‐state Hb S would have to be maintained for this to be successful.
A trial of folate supplementation in children with homozygous sickle cell disease
Authors: Rabb L. M. , Grandison Yvonne , Mason Karlene , Hayes R. J. , Serjeant Beryl , Serjeant G. R.
Published: March 2008

Summary. A double blind controlled trial of supplementation with folic acid has been performed in 117 children with homozygous sickle cell (SS) disease aged 6 months to 4 years over a 1 year period. No megaloblastic change was observed in either group. At the end of the study period the folate supplemented group showed no significant differences in haemoglobin, growth characteristics, or in the proportion of children affected by major or minor infections, acute splenic sequestration, dactylitis or episodes of bone or abdominal pain. However, the folate supplemented group showed a significantly lower mean cell volume and the placebo group contained a significant excess of children experiencing multiple episodes of dactylitis.

The results are compatible with mild folate deficiency in some patients in the placebo group but the absence of striking effects on haematology or growth suggest that the policy of regular folate supplementation in children with SS disease should be critically reviewed.
In vitro culture of clonogenic leukaemic cells in acute myeloid leukaemia: growth pattern and drug sensitivity
Authors: Marie Jean‐Pierre , Zittoun Robert , Thevenin Danielle , Mathieu Monique , Viguie Franck
Published: March 2008

Summary. Bone marrow from 43 of 45 AML patients grew leukaemic colonies in culture with a technique using methyl‐cellulose semi‐solid medium and stimulation with PHA‐leucocyte conditioned medium. Plating efficiency was significantly greater in M4 FAB subtypes than in M1 or M2. The presence of Auer rods in cultured cells and the existence of cytogenetic abnormalities in both fresh and cultured blast cells in one patient confirmed the leukaemic origin of these colonies. These clonogenic cells were closely related to the growth fraction, as demonstrated by a high suicide index and a linear correlation between percentage of bone marrow blasts in S phase and plating efficiency. In vitro CFU‐L sensitivity to cytosine‐arabino‐side (ARA‐C) and to adriamycin (ADR) was tested in 22 patients treated with these two drugs. In the group sensitive in vitro to ARA‐C (10 patients), 70% entered complete remission. In the resistant group (12 patients), only 25% had complete remission while 75% had resistant disease. Eight of 14 patients sensitive to ADR in vitro achieved complete remission, while five were resistant to chemotherapy. On the other hand, six of eight patients resistant in vitro were resistant in vivo. When drug sensitivities to ARA‐C and ADR were cumulated, an excellent in vitro to in vivo correlation was found when the patient was sensitive or resistant to both drugs in vitro.
Fetal haemoglobin synthesis following fetal liver transplantation in man
Authors: Delfini C. , Saglio G. , Mazza U. , Muretto P. , Filippetti A. , Lucarelli G.
Published: March 2008

Summary. Transplantation of haematopoietic tissue from the fetus to the adult is of considerable clinical and theoretical interest. In particular it could help to understand whether the haemoglobin switch is under the influence of environmental factors or inherently programmed in the haemopoietic cell population. In the present study we have monitored haemoglobin synthesis following fetal liver transplantation (FLT) in a patient with acute leukaemia after intensive chemoradiotherapy. 28 days following FLT, the haemoglobin synthesis showed a predominantly fetal haemoglobin pattern, as determined by globin chain synthesis of bone marrow cells and by the peroxidase anti‐peroxidase (PAP) method applied to bone marrow sections treated with anti‐fetal haemoglobin antibodies. These findings demonstrate that, in man, fetal haemopoietic cells, transplanted into an adult environment, do not immediately switch to the production of adult haemoglobin.
Prognostic value of bone marrow plasma cell infiltration in stage I multiple myeloma
Authors: Cavo M. , Baccarani M. , Gobbi M. , Lipizer A. , Tura S.
Published: March 2008

Summary. We analysed for prognosis and response to chemotherapy a series of 48 consecutive stage I myeloma patients who were treated with melphalan and prednisone soon after diagnosis. It was found that the extent of bone marrow plasma cell infiltration had prognostic value, the survival of the patients with more than 50% plasma cells being significantly shorter (P<0·005) than that of the patients with less than 50% plasma cells. In the latter group, the observed number of deaths was so small (two cases) as to preclude any further analysis. In contrast, in the former group, a significant relationship between the response to chemotherapy and patients’ survival (P<0·001) was demonstrated. We conclude that a prospective randomized trial of treatment versus no treatment is warranted in stage I patients who have a high risk of death (i.e. with a high bone marrow plasma cell infiltration) in order to establish whether chemotherapy prolongs the length of survival.
Delayed release of an abnormal fibrinopeptide A from fibrinogen Manchester: effect of the Aα 16 Arg → His substitution upon fibrin monomer polymerization and the immunological crossreactivity of the peptide
Authors: Lane D. A. , Southan C. , Ireland H. , Thompson E. , Kehl M. , Henschen A.
Published: March 2008

Summary. Fibrinogen Manchester is an abnormal fibrinogen with an impaired release of fibrinopeptide A (FPA) and a polymerization abnormality. In the accompanying article we have identified the amino acid substitution in fibrinogen Manchester as Aα 16 Arg → His. When fibrinogen Manchester was digested with low thrombin concentrations approximately 40–50% of the total FPA content was released at a rate similar to FPA release from normal fibrinogen. The fibrin so formed exhibited an impaired polymerization of monomers. Digestion of fibrinogen Manchester with high concentrations of thrombin for prolonged times released the remaining FPA which had an abnormal retention time when studied by high performance liquid chromatography (HPLC). This fibrinopeptide has been shown previously to contain the Aα 16 Arg → His substitution. Fibrin resulting from this exhaustive digestion had normal polymerization of monomers. The normal and substituted FPAs were isolated by HPLC and compared in a double antibody competitive‐binding assay for normal FPA. The immunological cross‐reactivity of the abnormal peptide was reduced, so that approximately 5 times more abnormal peptide was required on a molar basis to displace labelled normal FPA. Normal intact fibrinogen was 10–fold less reactive (on a half molar basis) than free normal FPA and the crossreactivity of fibrinogen Manchester was measurably less than that of normal fibrinogen. It is concluded that immunological measurement alone of FPA released from abnormal fibrinogens may not give a complete description of the kinetics of peptide release if the amino acid substitution lies within the FPA sequence. The combination of radioimmunoassay and HPLC, however, provides a powerful analytical approach that should be useful in classifying and characterizing abnormal fibrinogens.
Mast‐cell precursors in various haematopoietic colonies of mice produced in vivo and in vitro
Authors: Sonoda T. , Kitamura Y. , Haku Y. , Hara H. , Mori K. J.
Published: March 2008

Summary. We have examined mast‐cell precursors in various in vivo and in vitro haematopoietic colonies of mice. Cells from haematopoietic colonies of (WB x C57BL/6)F1‐ +/+ mouse origin were injected into the skin of the congeneic W/Wv mice which are genetically depleted of tissue mast cells. The appearance of mast‐cell clusters at the injection site indicated the presence of mast‐cell precursors in the injected cell suspension. More than 40% of 12 d exogenous spleen colonies and 14 d in vitro mixed colonies contained mast‐cell precursors, but only a small proportion of 7 d exogenous spleen colonies, 9 d in vitro erythroid bursts or 14 d large in vitro neutrophil‐macrophage colonies contained mast‐cell precursors. Neither transient endogenous erythroid spleen colonies examined at the fifth day nor 7 d in vitro neutrophil‐macrophage colonies contained mast‐cell precursors. Among 12 d spleen colonies mast‐cell precursors were more frequent in the predominantly neutrophil than the predominantly erythroid colonies but 14 d in vitro mixed colonies, in which erythroid cells were predominant, contained mast‐cell precursors more frequently than 14 d in vitro neutrophil‐macrophage colonies. Therefore, differentiation of mast cells seems to be independent of either the neutrophil‐macrophage or erythroid cell lineages.
An electroblotting technique for the detection of factor VIII/von Willebrand factor multimers in plasma
Authors: Furlong B. L. , Peake I. R.
Published: March 2008

Summary. A new, relatively simple technique has been developed in order to study the multimers of factor VIII/von Willebrand factor (VIII/vWF). It involves electrophoresis on SDS agarose gels and electrophoretic transfer (electroblotting) of the separated protein bands onto nitrocellulose membranes, to which they are non‐covalently bound. VIII/vWF multimers are then detected by 125I‐labelled antibodies to VIII/vWF, and autoradiography. Optimum electrophoretic transfer occurred at 0·5 A, for 18 h, on 0·8% agarose gels, thus enabling detection of the multimeric profile of VIII/vWF in 5 μl of normal plasma. The multimeric profile for haemophilia A patients was identical to that for normal plasma. In plasma from patients with von Willebrand's disease (vWd), various patterns were seen, with a preponderance of smaller multimers in type II (atypical) vWd, similar to those seen in cryosupernatant. Heterogeneity within a particular type of vWd was also evident. Investigation of commercial factor VIII concentrates showed the presence of ‘doublets’ of VIII/vWF. Unlike other reported techniques, the rapid transfer and fixing of the protein bands to the nitrocellulose, minimizes loss of resolution, and handling of the paper is easier. It is possible to cut a sample electrophoresis strip into several areas, for incubation with different antibodies. Preliminary experiments also suggest that double antibody techniques are possible, or even removal of a first radiolabeled antibody by low pH, and then incubation of the separated proteins with a second, unrelated antibody.
Prospective platelet crossmatching for selection of compatible random donors
Authors: Freedman J. , Hooi C. , Garvey M. B.
Published: March 2008

Summary An indirect platelet radioactive antiglobulin test was used in a prospective crossmatch study in 29 patients for selecting compatible random donors for platelet transfusion. Patients received crossmatch‐compatible, crossmatch‐incompatible and uncrossmatched platelets. A successful outcome for transfusion was defined as a 24 h post‐transfusion corrected platelet increment of greater than 4·5 × 109/I. Of 47 predictions based on crossmatch results, the technique successfully predicted the outcome of the transfusion in 90% of cases. In all patients, transfusion of crossmatch‐compatible platelets resulted in a significantly higher 24 h corrected platelet increment than was obtained following transfusion of incompatible platelets. The mean±SEM 24 h corrected post‐transfusion increment (× 109/1) was 17·79±2·01 in 26 patients who received crossmatch‐compatible pooled random donor platelets; in 21 patients who received crossmatch‐incompatible platelets, it was 1·19 ±0·56; and in 25 patients who received uncrossmatched platelets, 4·42 ±0·97. The differences were highly significant. There was an 83% correlation of results with those obtained using an indirect platelet suspension immunofluorescence technique.
Accumulation and release of isoferritins during incubation in vitro of human peripheral blood mononuclear cells
Authors: Worwood M. , Hourahane D. , Jones B. M.
Published: March 2008

Summary Ferritin concentration has been measured in peripheral blood mononuclear cells and in the incubation medium following in vitro culture. Antibodies to both heart and spleen ferritin were used. Mononuclear cells cultured in medium containing about 12 μmol Fe/1 accumulate ferritin rapidly with an increase in the heart:spleen ferritin ratio from 3:1 to about 10:1. Higher concentrations of iron (100 μmol/1) produce an even greater effect. The accumulation of ferritin is prevented by the addition of desferrioxamine (2 mmol/1) to the incubation medium. Accumulation of ferritin appears to take place largely in monocytes. Phagocytosis of red blood cells also causes rapid accumulation of ferritin but without any change in the heart: spleen ratio. Small amounts of both spleen and heart type ferritin are released during incubation in an iron containing medium and following phagocytosis of red blood cells. Some concanavalin A binding ferritin is also released suggesting that phagocytic cells may be a source of the concanavalin A binding ferritin found in normal plasma.
Erythroid colony formation in primary thrombocythaemia: evidence of hypersensitivity to erythropoietin
Authors: Eridani S. , Batten Elaine , Sawyer Barbara
Published: March 2008

Summary In a group of 14 patients with primary thrombocythaemia (PT) the study of erythropoietic colony formation in vitro showed the development of so‐called ‘endogenous’colonies, namely colonies with no added erythropoietin (Ep), in all but one case. In the presence of added Ep the colony formation increased slightly in the PT group, but the rise in the control group was so pronounced that any statistical difference between the two groups disappeared. No ‘endogenous’colonies were observed in cases of secondary thrombocytosis. This finding reinforces the view that PT is a myeloproliferative disorder and establishes that some erythroid progenitors show the same high sensitivity to Ep as that found in polycythaemia vera; it is of interest that, in the patients studied, there was no apparent involvement of erythropoiesis.
Plasmin‐a2‐antiplasmin complexes in bleeding disorders characterized by primary or secondary fibrinolysis
Authors: Booth N. A. , Bennett B.
Published: March 2008

Summary. A two‐dimensional immunoelectrophoresis (2DIEP) method detects plasmin complexed to its major inhibitor, α2‐antiplasmin, in plasma in the blood of patients during (a) thrombolytic therapy with urokinase, (b) episodes of disseminated intravascular coagulation (DIC) with active fibrinolysis, and (c) episodes of fibrinolytic haemorrhage without evidence of DIC. Clearance of the complexes from the blood is rapid and their detection thus implies active plasmin generation at the time of blood sampling or within the preceding 24 h. Abolition of the complexes using tranexamic acid therapy allowed surgery without bleeding in two previously grossly haemorrhagic patients in group (c). Antithrombin III complexed with activated procoagulants was detected using a similar 2DIEP method in only two of fofour patients with DIC. Abnormalities of α2‐macroglobuin were detected on 2DIEP of plasma in the patients studied with proteolytic disorders; these did not appear to reflect complex formation.
Immunological study of in vitro maturation of human megakaryocytes
Authors: Vinci G. , Tabilio A. , Deschamps J. F. , Van Haeke D. , Henri A. , Guichard J. , Tetteroo P. , Lansdorp P. M. , Hercend T. , Vainchenker W. , Breton‐Gorius J.
Published: March 2008

Summary. Human megakaryocyte colonies were grown from the bone marrow in plasma clot or methyl cellulose cultures. Maturation of the megakaryocytic cells was sequentially studied from day 5 to day 16 of culture by fluorescent labelling with a panel of monoclonal and polyclonal antibodies against different platelet glycopro‐teins (Gp), PI AI antigen, factor VIII RAg platelet factor 4 (PF 4), fibrinogen and platelet‐derived growth factor (PDGF). Expression of Gp Ib was also studied by immunogold technique at electron microscopy. The first cells identifiable by these antibodies were found at day 5 of culture. They had the size of a lymphocyte. These small megakaryocyte precursors already expressed all the platelet antigens, HLA‐DR and transferrin receptors and were devoid of erythroid or myeloid markers. Among the platelet antigens, Gp IIIa was the most sensitive marker for the identification of these precursors. However, double‐fluorescent labelling demonstrated that the different platelet markers were coexpressed in a large majority of cells. Interestingly, cytoplasmic markers demonstrated that these small megakaryocyte precursors were themselves heterogenous by morphological criteria. During maturation, expression of Gps, particularly of Gp Ib, increased while the labelling pattern of anti factor VIII RAg and anti PF 4 antibodies switched from diffuse to granular staining. PDGF could also be detected in the megakaryocytes grown in culture.
Cytogenetic studies of early myeloid progenitor compartments in Ph1‐positive chronic myeloid leukaemia (CML) I. PERSISTENCE OF Ph1‐NEGATIVE COMMITTED PROGENITORS THAT ARE SUPPRESSED FROM DIFFERENTIATING IN VIVO
Authors: Dubé I. D. , Gupta C. M. , Kalousek D. K. , Eaves C. J. , Eaves A. C.
Published: March 2008

Summary. We have cytogenetically analysed individual haemopoietic colonies to investigate the level and extent of normal stem cell suppression that occurs in patients with Philadelphia chromosome (Ph1‐positive CML. Seventeen patients were studied at diagnosis prior to the initiation of chemotherapy and five of these were studied again 1–16 months later. Another nine patients were studied for the first time 2–96 months after diagnosis and initiation of chemotherapy. No chromosomally normal metaphases were found in either direct marrow preparations or in haemopoietic colonies obtained from simultaneous assays of marrow and/or blood samples from 20 of the 26 patients studied. In the other six, chromosomally normal haemopoietic progenitors (BFU‐E, CFU‐C and CFU‐G/E) were readily demonstrable even though in five of these patients all dividing cells in the bone marrow appeared to belong to the Ph1‐positive clone at the time of study. These results indicate that the suppressive effects of clonal expansion on normal haemopoiesis are more pronounced, and apparent sooner, in the more differentiated compartments. In addition, they support the view that the original population of normal stem cells does not disappear rapidly, although their numbers may be diluted to undetectable levels depending upon the extent of clonal expansion at the stem cell level by the time of diagnosis.
Bone marrow transplantation for acute nonlymphoblastic leukaemia: a survey of the European Group for Bone Marrow Transplantation (E.G.B.M.T.)
Authors: Zwaan F. E. , Hermans J. , Barrett A. J. , Speck B.
Published: March 2008

Summary. Between 1979 and 1982, 229 patients with acute nonlymphoblastic leukaemia in remission were given allogeneic bone marrow transplants from HLA‐identical siblings. Data on 183 patients transplanted in first complete remission were compared with results of 46 patients transplanted in second or subsequent remission. The 2‐year actuarial survival rate was 53% for patients transplanted in first remission and 36% for patients transplanted in second or subsequent remission (P= 0·02). The probability of survival was related to the age of the patient (P=0·02) and of the donor (P= 0·0002). The 2‐year actuarial leukaemia recurrence rate was 12% for patients transplanted in first remission and 44% for patients in a subsequent remission (P= 0·10). Other factors correlating with a higher probability of relapse after transplantation were the leukaemia subtypes M4 and M5 (P=0·007), cyclosporin A used as prophylaxis of graft‐versus‐host disease (P= 0·008), and the absence of chronic (but not of acute) graft‐versus‐host disease (P= 0·003). These data indicate that prolonged survival can be achieved in approximately 50% of patients with acute nonlymphoblastic leukaemia who receive bone marrow transplants during their first complete remission. But modifications of pre‐ and post‐transplant treatment should be considered to reduce the rate of relapse.
B‐chronic lymphocytic leukaemia patients with stable benign disease show a distinctive membrane phenotype
Authors: Caligaris‐Cappio F. , Gobbi M. , Bergui L. , Campana D. , Lauria F. , Fierro M. T. , Foa R.
Published: March 2008

Summary. The circulating B cells of 98 patients with B‐CLL were analysed with a panel of conventional markers and monoclonal antibodies including RFA‐1 and RFA‐4. A subgroup of patients was defined with a distinctive double membrane marker phenotype (RFA‐1+, RFA‐4+). These patients showed features of clinical stability. Their physical signs, haematological features and lymphocyte counts remained unchanged for 5–10 years: therefore, the RFA‐1+, RFA‐4+ membrane phenotype appears to identify a subgroup of B‐CLL patients with a particularly non‐progressive disease.
Low dose cytosine arabinoside: partial remission of acute myeloid leukaemia without evidence of differentiation induction
Authors: Leyden Michael , Manoharan Arumigum , Boyd Andrew , Cheng Zhao Ming , Sullivan John
Published: March 2008

Summary. Thirteen patients with acute myeloid leukaemia aged from 19 to 81 were treated with low dose cytosine arabinoside (ARA‐C) in a dose of 10–15 mg/m2 twice daily subcutaneously. Three complete remissions were obtained. Partial responses were observed in a further two patients. To analyse the action of low dose ARA‐C freshly isolated leukaemic cells and cells from the cloned promyelocytic leukaemia cell line (HL60) were cultured in vitro in the presence of cytosine arabinoside. Minimal evidence of differentiation induction was observed when compared with the cytotoxic effects of the drug. These results suggest that ARA‐C does not exert its anti‐leukaemic effects by halting proliferation through differentiation induction. Rather, it appeared that the capacity of this agent to kill cells in S‐phase produced a progressive depletion of the cycling leukaemic cells. This resulted in a corresponding steady decline in the total leukaemic cell population.
Activity of divicine in Plasmodium vinckei‐infected mice has implications for treatment of favism and epidemiology of G‐6‐PD deficiency
Authors: Clark I. A. , Cowden W. B. , Hunt N. H. , Maxwell L. E. , Mackie E. J.
Published: March 2008

Summary. Intravenous injection of divicine into mice infected with Plasmodium vinckei rapidly killed the parasites and caused haemolysis. Degenerating parasites were observed frequently inside intact circulating erythrocytes, implying that parasite death was not a passive consequence of haemolysis. Both parasite death and haemolysis were prevented by the iron chelator desferrioxamine. In vitro, divicine caused the accumulation of malonyldialdehyde and the depletion of reduced glutathione in normal mouse erythrocytes. Desferrioxamine inhibited the former event, but not the latter. These observations support the hypothesis advanced by Huheey & Martin (Experientia, 31, 1145, 1975) to explain the patchy geographical distribution of glucose‐6‐phosphate dehydrogenase deficiency in historic malarial areas and also suggest that desferrioxamine, a drug already in clinical use, is a potential treatment for favism and other examples of oxidative haemolysis.
Myeloid progenitor cells in the circulation of patients with myelofibrosis and other myeloproliferative disorders
Authors: Hibbin J. A. , Njoku O. S. , Matutes E. , Lewis S. M. , Goldman J. M.
Published: March 2008

Summary. We have measured the numbers of myeloid progenitor cells in the circulation of patients with myelofibrosis (MF) and other myeloproliferative disorders. In general, progenitor cell numbers were increased in the circulation of patients with MF compared with controls. The mean increases were 9‐fold for the multilineage progenitor cells (CFU‐GEMM), 13‐fold for the erythroid progenitor cells (BFU‐E), 37‐fold for the granulocyte‐macrophage progenitor cells (CFU‐GM) and 167‐fold for the megakaryocyte progenitors (CFU‐MK). Splenectomized patients generally had reduced numbers of circulating progenitor cells. In the CFU‐MK assay, mature megakaryocytes cultured from patients with MF regularly showed large vacuoles in the nucleus and cytoplasm, unlike control cells. The increased colony formation in patients with MK, involving especially CFU‐MK colonies, is consistent with the hypothesis that MF is a primary myeloproliferative disorder in which a megakaryocyte‐derived factor predisposes to the formation of marrow fibrosis.