bluuebunny/crossref_metadata_2025 huggingface dataset:
Ferritin and iron uptake by reticulocytes
Authors: Blight G. D. , Morgan E. H.
Published: March 2008

Summary.

The uptake of liver ferritin labelled with 125I or 59Fe by guinea‐pig reticulocytes has been studied to investigate the characteristics of the uptake process, compare it with transferrin uptake and determine whether ferritin‐iron is utilized by the cells in haem synthesis. The results confirmed that guinea‐pig reticulocytes, but not mature erythrocytes, take up liver ferritin by a saturable, time‐ and temperature‐dependent process. Up to 70% of the iron taken up by the cells was utilized in haem synthesis and competed directly with iron derived from transferrin. Scatchard analysis of the binding parameters indicated that 30–130 × 103 ferritin molecules were bound per cell to high affinity specific membrane receptors (Ka: 1·77 × 107 M−1). In contrast, rat took up much less ferritin than guinea‐pig reticulocytes and the process was entirely non‐specific. Release experiments with guinea‐pig reticulocytes at 37°C showed that a maximum of about 70% of the cell‐associated 125I‐ferritin was released from the cells of which up to 15% was trichloroacetic acid‐soluble.

We suggest that ferritin uptake by guinea‐pig reticulocytes involves receptor‐mediated endocytosis. The endocytotic vesicle fuses with a lysosome, iron is removed from the protein and enters a cytosolic pool in which it competes directly with transferrin‐derived iron to provide iron for mitochondrial haem synthesis. Some of the ferritin is catabolized and the rest is returned to the extracellular medium during membrane recycling.
Deposition of the terminal C5b‐9 complement complex on erythrocytes by human red cell autoantibodies
Authors: Salama A. , Bhakdi S. , Mueller‐Eckhardt C. , Kayser W.
Published: March 2008

Summary.

A radioassay for the detection of complement activating autoantibodies (autohaemolysins) in patients with autoimmune haemolytic anaemia (AIHA) is described. The method is based on immunoradiometric quantitation of the cytolytic C5b‐9 complement complex following its antibody‐dependent deposition on red blood cells (RBC). The use of affinity‐purified, radiolabelled antibodies directed against the neoantigens of the C5b‐9 complex ensured specificity of the test which proved more sensitive than conventional haemolytic assays and was not subject to disturbances by haemolytic sera.

The results obtained in 70 patients with various forms of AIHA (warm type (N= 45); cold type (N= 22); Donath‐Landsteiner type (N= 3)) support the prevailing assumption that autohaemolysins can trigger the complement cascade to completion. Since intact RBC from patients with detectable autohaemolysins carried C3/C4 components but never C5b‐9, it is inferred that regulatory mechanisms operate in vivo at the level of C4b/C3b inactivation to arrest the cascade and rescue the autologous cells.
Tissue ferritin in scorbutic guinea‐pigs
Authors: Roeser H. P. , Sizemore D. J. , Nikles A. , Cham B. E.
Published: March 2008

Summary.

Tissue ferritin metabolism was compared in control and ascorbic acid (AA) deficient guinea‐pigs. Concentrations of ferritin protein in the liver (0·98±0·61 mg/g wet weight) and spleen (0·48±0·23 mg/g) of control animals did not change after tissue depletion of AA. Iron dextran (75 mg/kg weight, i.m.) caused a 4–5‐fold increase in tissue ferritin concentrations in controls whereas no increase in tissue ferritin occurred in scorbutic animals. The rise in tissue total iron concentration was similar in the two groups. Liver ferritin synthesis was similar in control and scorbutic animals. After stimulation with iron (8 mg/kg iron dextran i.v.), ferritin synthesis rose in both groups of animals. However, the pattern of response differed. At 24 h after iron dextran, ferritin synthesis in controls was still significantly elevated (P< 0 · 001) and liver ferritin protein continued to rise, whereas in scorbutic animals, ferritin synthesis had declined to pre‐iron injection levels, and no rise in ferritin protein values occurred. It is concluded that the ferritin synthetic apparatus in AA deficient tissues remained intact and capable of responding to added iron. The absence of a sustained elevation in tissue ferritin protein after an iron load appeared to be due to inadequate stimulation of ferritin synthesis by intracellular iron. It is suggested that AA has a physiological role in the reduction of intracellular iron and that it is the reduced form of iron which stimulates ferritin synthesis. Abnormalities of iron metabolism occur in AA depleted tissues when the quantity of Fe3±us;entering cells exceeds the residual reducing capacity of those cells.
Five leucocyte labelling techniques: a comparative in‐vitro study
Authors: Colas‐Llnhart N. , Barbu M. , Gougerot M. A. , Bok B.
Published: March 2008

Summary. During the past years, several authors have used labelled leucocytes to detect inflammatory foci. However, before routine use in man, it is necessary to control the viability of labelled cells. Five leucocyte labelling techniques (111In‐oxine, 111In‐oxine without extraction, 99mTc oxine, pyrophosphate 99mTc, 51Cr) were compared using the same separation methods, conservation medium, viability assays and migration studies. Electron microscopic studies allowed the assessment of cellular damage induced by the labelling techniques as well as the calculation of the percentage of cells disrupted during preparation. Results obtained in vitro using 111In‐oxine were not satisfactory and in fact appeared contradictory to those published by the authors using this technique in vivo. Even the best method, pyrophosphate 49mTc labelling, was not completely atoxic, but the functional behaviour of the leucocytes did not seem affected. While in vitro studies offer much information concerning labelled cells, they cannot predict the in vivo behaviour of these cells.
Concurrent development of preleukaemic, lymphoproliferative and plasma cell disorders
Authors: Greenberg Bernard R. , Miller Claramae , Cardiff Robert D. , Mackenzie Malcolm R. , Walling Paula
Published: March 2008

Summary. A patient is described who presented with a combined preleukaemic, lymphoproliferative and plasma cell disorder. These disorders were not related to cytotoxic therapy. The clonal nature of the lymphoid component was demonstrated by lymphocyte surface marker studies and the clonal nature of the plasma cell component by immunoperoxidase studies. Since the involved lymphoid and plasma cells contained different heavy and light chains, they were felt to originate from two separate B cell clones not related to the clone responsible for the preleukaemic component.
Relationship of post‐transfusion viability to deformability of stored red cells
Authors: Card R. T. , Mohandas N. , Mollison P. L.
Published: March 2008

Summary. During storage in the liquid state, red cells progressively lose their ability to survive in vivo and they become less deformable. An opportunity to test the hypothesis that these two changes are directly related arose when a donor was encountered whose red cells survived particularly badly after storage. It was found that red cells from this donor lost deformability abnormally rapidly during storage. Measurement of red cell deformability after storage may be of value in predicting the viability of the cells.
Neonatal hyperbilirubinaemia in heterozygous giucose‐6‐phosphate dehydrogenase deficient females
Authors: Meloni Tullio , Forteleoni Gavino , Dore Angelo , Cutillo Stefano
Published: March 2008

Summary. Glucose‐6‐phosphate dehydrogenase (G6PD, D‐glucose 6‐phosphate: NADP oxidoreductase, E.C. 1.1.1.49) activity and the percentage of G6PD deficient erythrocytes was determined in 50 girls heterozygous for G6PD deficiency, 25 of whom had had hyperbilirubinaemia at birth and 25 who had normal bilirubin levels. The enzymatic activity was 2 32 ±0·87 l.U./g Hb in the first group and 3·31 ±0·92 l.U./g Hb in the second group. The percentage of G6PD deficient erythrocytes was 54.1 ±15.3 and 65.3 ±0.87, respectively. The level of enzymatic activity exceeded 4 I.U./g Hb and the percentage of G6PD deficient cells fell below 40% in only one of the subjects who had developed hyperbilirubinaemia. Levels of enzymatic activity below 4 I.U./g Hb, or percentages of G6PD deficient erythrocytes higher than 40% can therefore be considered to be associated with a high risk of developing neonatal hyperbilirubinaemia. In our opinion, these babies should receive prophylactic treatment with phenobarbital, as do G6PD deficient Mediterranean males.
Skin infiltration associated with chronic myelomonocytic leukaemia
Authors: Duguid J. K. M. , Mackie M.J. , McVerry B. A.
Published: March 2008

Summary. Chronic myelomonocytic leukaemia (CMML) is typically associated with a prolonged clinical course and is not usually very responsive to chemotherapeutic intervention. Skin infiltration has not been recognized previously as a feature of this illness. We have seen four patients recently with CMML, who during the the course of their illness developed marked skin infiltration. Whilst sensitivity to chemotherapy could be demonstrated in the peripheral blood cell population, skin infiltration was quite resistant to treatment. Skin infiltration heralded a more aggressive phase of the disease although no discernible change in morphology, cytochemistry or membrane marker analysis of the leukaemic cell population could be demonstrated in three of the patients studied; one patient, however, transformed to an acute leukaemia shortly thereafter.
Coexistent myelodysplasia and plasma cell neoplasia
Authors: Mufti G. J. , Hamblin T. J. , Clein G. P. , Race C.
Published: March 2008

Summary.The oncogenic effect of alkylating agents, particularly that of melphalan, has been postulated as the most likely cause of myelodysplastic syndrome (MDS) and acute leukaemia in patients with plasma cell neoplasia. Five patients aged between 69 and 79 years were found to have features of MDS at the time of diagnosis of multiple myeloma. In three of these cases MDS transformed into acute leukaemia. These patients had not received any chemotherapy prior to their presentation. The possible importance of these features is discussed.
The use of dichloromethylene diphosphonate for the management of hypercalcaemia in multiple myeloma
Authors: Paterson A. D. , Kanis J. A. , Cameron E. C. , Douglas D. L. , Beard D. J. , Preston F. E. , Russell R. G. G.
Published: March 2008

Summary. We have assessed the effects of the diphosphonate, dichloromethylene diphosphonate (Cl2MDP), in 19 patients with hypercalcaemia and increased bone resorption due to myeloma. Cl2MDP (800–3200 mg daily by mouth or 300 mg daily by intravenous infusion) decreased plasma calcium and biochemical indices of increased bone resorption in 16 of 19 patients. This effect persisted for the duration of treatment (up to 14 weeks). Prolonged treatment was associated with a progressive rise in serum alkaline phosphatase and only a transient fall in hydroxyproline suggesting the stimulation of bone repair. Since myeloma is associated with significant morbidity and mortality due to progressive bone loss, these results suggest that long‐term treatment of myeloma with Cl2MDP is worthy of further study.
Hypercalcaemia complicating acute myelogenous leukaemia: a syndrome of multiple aetiologies
Authors: Gewirtz Alan M. , Stewart Andrew F. , Vignery Agnes , Hoffman Ronald
Published: March 2008

Summary. Hypercalcaemia complicating acute myelogenous leukaemia is a rare but well‐recognized phenomenon. In most cases the pathogenetic mechanism causing the hypercalcaemia remains poorly understood. We recently studied in detail two patients with acute myelogenous leukaemia who developed hypercalcaemia during the course of their illness. The results of these studies conclusively excluded primary hyperparathyroidism or ectopic parathyroid hormone production as causes of the patients’ hypercalcaemia. In vitro studies carried out on short‐term suspension cultures of one patient's peripheral blood blast cells demonstrated production of a factor with potent bone resorbing activity, distinct from parathyroid hormone (iPTH) and prostaglandin E2 (PGE2). Further characterization of the bone resorbing factor suggested that it bore some similarity to osteoclast activating factor (OAF). Hypercalcaemia in the other case appeared to be due to a combination of skeletal invasion by malignant cells, and to ectopic secretion of an unidentified humoral factor with bone resorbing activity. These two cases demonstrate that the hypercalcaemia complicating acute myelogenous leukaemia may be due to a variety of mechanisms distinct from parathyroid hormone production.
Erythrocyte membrane skeleton abnormalities in hereditary spherocytosis
Authors: Burke Brian E. , Shotton David M.
Published: March 2008

Summary. Erythrocyte ghosts from eight individuals with hereditary spherocytosis have been compared with respect to their protein compositions as judged by SDS gel electrophoresis, their ease of spectrin extractability, and their freeze‐etch electron microscopic appearance after incubation in conditions designed to promote aggregation of the intramembrane particles. Four of these HS cases were unrelated, while the other four represented two generations from a single family, including a pair of identical twins, one of whom had not undergone splenectomy when this investigation was initiated.

Of the four unrelated cases, one showed no departures from normal under the conditions of this investigation, whereas the other three exhibited features which suggested a membrane skeleton lesion. In one of these there was a reduced proportion of spectrin tetramers relative to dimers in 4°C extracts, while the two remaining cases exhibited abnormal intramembrane particle aggregation. The four related cases had almost identical variations from normal. Spectrin was not extractable from their ghost membranes during a mild extraction incubation which removed spectrin from normal control ghosts. However, the intramembrane particle aggregation subsequently induced in these ghosts was of a degree unobtainable in normal ghosts without such spectrin extraction. In addition the ghosts from one twin, the only one of these patients who had not undergone splenectomy at the start of this investigation, showed a reduced amount of band 4.2. However, when this patient's blood was re‐tested after splenectomy, this protein was found to be at normal levels.

Our results support the view that hereditary spherocytosis is not a single disease, but is rather a term used to describe a variety of different molecular lesions of the erythrocyte membrane skeleton with similar clinical manifestations.
Blood group A antigen site distribution and immunoglobulin binding in relation to red cell age
Authors: Smalley C. E. , Tucker E. M.
Published: March 2008

Summary. Heterogeneity of A antigen site density was demonstrated on individual human A1 and A2 type red cells by immunoelectronmicroscopy using haemocyanin‐conjugated antiglobulin. Examination of red cells fractionated on a Percoll density gradient showed that the heterogeneity was not age‐related. The presence of autologous immunoglobulin on senescent red cells was also confirmed by the same technique.
Transplantation of bone marrow fibroblastoid stromal cells in mice via the intravenous route
Authors: Piersma A. H. , Ploemacher R. E. , Brockbank K. G. M.
Published: March 2008

Summary. Kinetics of fibroblastic colony‐forming cells (CFU‐F) were studied in mouse bone marrow after lethal total body irradiation and intravenous bone marrow transplantation. After an initial decrease, CFU‐F numbers recovered, and plateaued 5 weeks post‐treatment at 10% of normal values.

Using chromosome‐marked donor bone marrow cells we found that 1 day after transplantation 72% of donor CFU‐F had reached the recipient's bone marrow, indicating a highly specific lodgment of CFU‐F. Three months after transplantation donor CFU‐F were still detectable and comprised about half of the femoral CFU‐F population.

It is concluded that CFU‐F, a component of the haemopoietic microenvironment, are transplantable via the intravenous route.
Factor VIII inhibitor and raised platelet IgG levels associated with methyldopa therapy
Authors: Devereux S. , Fisher D. M. , Roter B. L. T. , Hegde U. M.
Published: March 2008

Summary. An inhibitor to the plasma coagulation factor VIII and abnormally high levels of platelet associated IgG (PAIgG) were found in a patient whilst on methyldopa. Both these parameters fell on initial withdrawal, but on rechallenge with the drug only the PAIgG rose to the high presenting levels. No inhibitory activity to factor VIIIc could be detected in a concentrated eluate prepared from the patient's platelets. These results may imply that two distinct antibodies were provoked by the administration of methyldopa, but could also be explained by the action of anti VIIIc alone on the patient's platelets.
Prothrombin Habana: a new dysfunctional molecule of human prothrombin associated with a true prothrombin deficiency
Authors: Rubio R. , Almagro D. , Cruz A. , Corral J. F.
Published: March 2008

Summary. A Cuban family with a new congenital dysprothrombinaemia is described. The propositus was a 5‐year‐old female who presented with umbilical bleeding after birth followed by easy bruising and bleeding tendency throughout her life. The main laboratory features of the defect included prolongation of prothrombin time and partial thromboplastin time. Prothrombin activity was less than 10% in several one‐ and two‐stage systems. However, the staphylocoagulase‐complexed prothrombin level and immunologic methods yielded levels of about 50%. The migration of the abnormal prothrombin was more anodic in single and bidimensional immunoelectrophoresis system and did not change by the addition of calcium. Family studies revealed that the father had approximately 50% prothrombin activity and antigen, whereas the mother had 45% prothrombin activity but about 100% prothrombin antigen. We suggest that the propositus is heterozygous for an abnormal prothrombin and heterozygous for true prothrombin deficiency.
Immunoenzymatic staining of haematological samples with monoclonal antibodies
Authors: Moir D. J. , Ghosh Anna K. , Abdulaziz Z. , Knight Patricia M. , Mason D. Y.
Published: March 2008

Summary. This paper describes the use of immunoenzymatic techniques (and in particular a recently developed immuno‐alkaline phosphatase procedure) for labelling haematological samples with monoclonal antibodies. Since cells are smeared and fixed before staining it is possible to combine optimal preservation of cellular detail with visualization of positive labelling. Additional advantages over conventional immunofluorescent procedures for detecting cellular antigens include the fact that samples may be stored for long periods both before and after staining, and that double labelling may readily be performed (either by combining immunoenzymatic staining with T cell resetting or by performing immunoperoxidase and immuno‐alkaline phosphatase techniques sequentially). Furthermore, these methods may be applied to samples containing too few cells for conventional examination (e.g. samples of cerebrospinal fluid). A total of 16 different antigens (including HLA‐DR, common ALL antigen and antigens associated with T cells, B cells, erythroid cells and megakaryocytes) were demonstrated by immuno‐enzymatic staining on a range of normal and neoplastic haematological samples. It is concluded that this approach to cellular antigen labelling is of potential value not only in routine haematological diagnosis, but also for research in many immunological and haematological fields.
Influence of heat‐treatment on rabbit liver‐ferritin kinetics
Authors: Frenkel Erik J. , Beld Bep van den , Marx Joannes J. M.
Published: March 2008

Summary. Rabbit liver‐ferritin was purified by heat‐treatment and immuno‐affinity chromatography or by immuno‐affinity chromatography alone. The ferritin was labelled with 125I and injected into rabbits. Significant differences between the two preparations were observed in the rate of clearance from plasma. The biological half‐life in vivo of the heat‐treated samples was 5·3·0·5 min, and of non‐heated preparations 8·5·0·5 min, thereby showing a significant (P < 0·001) affect on the final preparation of heat‐treatment during the isolation procedure. After the first 10–15 min period the clearance rate of a slower disappearing fraction was not dependent on the purification method used. The smaller amount of that fraction present in the heat‐treated preparation, however, implies a selective loss of ferritin forms with a slow plasma clearance during the isolation procedure that includes heat‐treatment.
Effect of alpha thalassaemia on the rheology of homozygous sickle cell disease
Authors: Serjeant B. E. , Mason K. P. , Kenny M. W. , Stuart J. , Higgs D. R. , Weatherall D. J. , Hayes R. J. , Serjeant G. R.
Published: March 2008

Summary. A study of rheological determinants (plasma viscosity, whole‐blood viscosity, and erythrocyte deformability) was made in 24 matched pairs of patients with homozygous sickle cell disease, with and without homozygous α‐thalassaemia 2. Patients with coexisting α‐thalassaemia showed a significant increase in erythrocyte deformability measured as filtration of washed erythrocytes through 5 μm diameter pores and also as viscosity of whole blood at high shear rate (230 s−1) and standard haematocrit (0·45). This rheological advantage may explain the beneficial effect of α‐thalassaemia 2 on haematological parameters and clinical events in homozygous sickle cell disease.
Allogeneic bone marrow transplantation: the monitoring of granulocyte macrophage colonies following the collection of bone marrow mononuclear cells and after the subsequent in‐vitro cytolysis of OKT3 positive lymphocytes
Authors: Gilmore M. J. M. L. , Prentice H. G. , Jones E. Price , Blacklock H. A. , Tidman N. , Schey S. , Goldstein G. , Janossy G. , Hoffbrand A. V.
Published: March 2008

Summary. Marrow nucleated cells from eight normal allogeneic donors was layered on Ficoll‐Metrizoate to isolate the mononuclear cell fraction. The cells were then washed to remove Ficoll‐Metrizoate and coagulation factors prior to resuspension in a balanced salt solution and the addition of the murine anti‐human T‐lymphocyte monoclonal antibody OKT3 and rabbit complement. The procedures were assessed for their effect on mononuclear cell viability (mean recovery 84·4%); the ability of the cells to proliferate granulocyte‐macrophage colonies (mean recovery 57·4%); the in‐vitro T‐lymphocytolysis (mean 75·7%) and the removal of rabbit complement (> 99%). Following marrow transplantation with this treated mononuclear fraction the mean day to recovery of ≥ 1·0 × 109/1 leucocytes was 20 d, with three patients developing ≥ Grade II acute graft versus host disease (GvHD). Thus, treatment of donor marrow with OKT3 and complement in a large volume system was not detrimental to subsequent engraftment, nor effective in complete T‐lymphocytolysis, nor in prevention of severe GvHD.
Inhibition of malarial invasion of red cells by chemical and immunochemical linking of spectrin molecules
Authors: Dluzewski A. R. , Rangachari K. , Gratzer W. B. , Wilson R. J. M.
Published: March 2008

Summary. The invasion of resealed human red cell ghosts by Plasmodium falciparum, and those from monkey cells by P. knowlesi, was strongly inhibited by anti‐spectrin antibodies introduced into their cytoplasm. Univalent F(ab)1 fragments gave no such effect, but a combination of these fragments and goat anti‐rabbit immunoglobulin, to restore bifunctionality, caused perceptible inhibition of invasion. Disulphide crosslinks introduced between spectrin molecules in intact red cells by the membrane‐permeant oxidizing agent, diamide, again led to inhibition of invasion. This effect was largely reversed by reduction of the cross‐links. Gel electrophoresis was used to confirm that cross‐linking was essentially confined to spectrin, and that extended covalent networks were not formed. It follows that local formation of bridges, whether by antibodies or oxidation of thiol groups, functions by inhibiting a local rearrangement of the cytoskeleton that forms a step in the invasion process.
The peroxidase activity and cellular content of granule proteins in PMN during pregnancy
Authors: ÖBerg G. , Lindmark G. , Moberg L. , Venge P.
Published: March 2008

Summary. The cellular content of myeloperoxidase (MPO), chymotrypsin‐like cationic protein (CCP) and lactoferrin (LF) of isolated polymorphonuclear leucocytes (PMNs) from pregnant women were analysed by immuno‐chemical methods. The cellular levels of MPO and CCP were unaltered during prenancy while the cellular levels of LF were increased during pregnancy and post partum (P<0·01). Peroxidase activity within isolated PMN was analysed by two methods, a Guaiacol method and a Chemiluminescence method. The activity was found to be reduced during pregnancy. The reduction was explained by the reduction in eosinophil peroxidase which in turn was a consequence of a reduced number of circulating eosinophils during pregnancy.
Platelet antibody levels do not correlate with response to therapy in idiopathic thrombocytopenic purpura
Authors: Kernoff L. M. , Malan E.
Published: March 2008

Summary. Platelet bound antibody was measured at presentation in 6 7 patients with idiopathic thrombocytopenic purpura by means of a quantitative antiglobulin consumption technique. The amounts of antibody present on the platelets of patients who failed steroid therapy were similar to those present on the platelets of patients who achieved a remission of their illness. It was also not possible on the basis of these data to identify those patients whose illness would remit after splenectomy. It is concluded that the pretreatment platelet antibody level does not assist in predicting the responses to treatment.
Heparin‐binding platelet proteins demonstrated by crossed affinity immunoelectrophoresis
Authors: Gogstad G. O. , Solum N. O. , Krutnes M. B.
Published: March 2008

Summary. Platelet proteins that interact with heparin were studied using crossed affinity immunoelectrophoresis. Platelet proteins solubilized in Triton X‐100 were applied to crossed immunoelectrophoresis against anti‐platelet antibodies, and an intermediate gel containing heparin covalently linked to Sepharose 4B was inserted. Six immunoprecipitates were absent or showed an altered position compared to control immunoplates, indicating that the corresponding antigens were bound to the immobilized heparin. These precipitates represented platelet factor 4, thrombospondin, glycoprotein lb, and three antigens termed G4,17 and 25. The subcellular location of the heparin‐binding proteins was either in the surface membrane (glycoprotein lb and the antigens 17 and 25), or in the a‐granules (platelet factor 4, thrombospondin and G4). Both forms of platelet factor 4 appearing after crossed immunoelectrophoresis, i.e. a line‐form and a peak‐form, bound strongly to the heparin. Glycoprotein lb showed a weak binding whereas its proteolytic split product glycocalicin did not significantly bind to the heparin in the present system. It is concluded that the platelets contain at least six heparin‐binding proteins which are present on the cellular surface or are able to be exposed to the extracellular medium after the release‐reaction has occurred.
Stimulation of fibrinogen biosynthesis by fibrinogen fragments D and E
Authors: Bell William R. , Kessler Craig M. , Townsend R. R.
Published: March 2008

Summary. Infusions of either fibrinogen fragment D or fibrinogen fragment E into rabbits were followed by increases in fibrinogen synthesis determined by the rate of incorporation of 75Se‐selenomethionine into circulating fibrinogen. The degree of stimulation was proportional to the amount of protein infused. When 4.5 mg of each fibrinogen fragment was administered separately to different groups of animals, fibrinogen fragment D was associated with a fourfold increase in fibrinogen synthesis above that in the control animals compared with 1‐5‐fold increase induced by fragment E. Fragments D and E were assayed for bound sialic acid, the absence of which facilitates binding, transport and catabolism of many circulating glycoproteins by the liver. Fibrinogen fragment D contained 1–3% sialic acid compared to 1–4% in fragment E. These data indicate conservation of sialic acid during plasmic digestion of fibrinogen. The capacity of these glycopolypeptide fragments to stimulate fibrinogen synthesis appears unrelated to the nearly identical quantities of N‐acetyl neuraminic acid found in each fragment.
The estimation of fetal haemoglobin in healthy adults by radioimmunoassay
Authors: Rutland P. C. , Pembrey M. E. , Davies T.
Published: March 2008

Summary. A radioimmunoassay (RIA) for the measurement of fetal haemoglobin (HbF) was established and used to determine the distribution of HbF in 2463 British adults between the ages of 18 and 65 years. Although a range in HbF values was similar in both genders a shift towards higher values is seen in females and the mean HbF% is higher, in females, at all ages between 18 and 55. A significant and different fall in HbF over the age range studied was noted between females (47%) and males (23%). The difference in mean HbF between the genders was less marked after age 40 years and a small rise in HbF production was measured in females between ages 50 and 54 years.
Hereditary persistence of fetal haemoglobin (HPFH) in conjunction with a chromosomal translocation involving the haemoglobin β locus
Authors: Jensen M. , Wirtz A. , Walther J.‐U. , Schemken E. M. , Laryea M. D. , Driesel A. J.
Published: March 2008

Summary An HPFH syndrome was found in a woman and her daughter who also carry a ‘balanced’cyclic translocation of chromosome segments involving four chromosomes, with one break point located in the region of the Hb β locus. This HPFH is characterized by 5% and 8% Hb F in peripheral blood, uneven distribution of Hb F in the red cells, and a Gγ/Gγ+Aγ ratio of 0·4. The mapping of the non alpha gene cluster shows no detectable deletion in the entire γ‐δ‐β‐globin gene region.
Mechanisms of deoxyadenosine toxicity in human lymphoid cells in vitro: relevance to the therapeutic use of inhibitors of adenosine deaminase
Authors: Lee N. , Russell N. , Ganeshaguru K. , Jackson B. F. A. , Piga A. , Prentice H. G. , Foa R. , Hoffbrand A. V.
Published: March 2008

Summary Deoxyadenosine (AdR) appears to be central to the molecular events mediating immunodeficiency in children born with adenosine deaminase (ADA) deficiency but it is still uncertain whether lymphotoxicity is due to AdR directly inhibiting transmethylation reactions in which S‐adenosylmethionine is the methyl group donor, or is due to phosphorylation of AdR to deoxyadenosine triphosphate (dATP) which then inhibits ribonucleotide reductase or is due to other mechanisms. Using AdR and the ADA inhibitor deoxycoformycin (dCF) and assessing cell viability, nucleoside incorporation into RNA and DNA, as well as measuring deoxyribonucleo‐side triphosphate (dNTP) concentrations and S‐adenosylhomocysteine (SAH) hydro‐lase activity, we have studied various types of human lymphoid cells and demonstrated in them the relative importance of the above two mechanisms of AdR toxicity. Treatment of normal resting peripheral blood lymphocytes in culture with AdR and dCF resulted in impaired viability. Although elevated dATP levels as well as decreased SAH hydrolase activities were both observed, the failure of a known inhibitor of ribonucleotide reductase (hydroxyurea) to produce toxicity, and the inability of deoxycytidine (CdR) to achieve a rescue effect, point to another mechanism, possibly inhibition of trans‐methylation or ATP depletion being the more likely causes of toxicity in resting lymphocytes. The same mechanism may well account for the rapid and severe lymphopenia in patients treated with dCF. On the other hand, in cultured lymphoblasts in the exponential phase of growth, AdR and dCF produced marked inhibition of growth and cell death both in a Thy‐Al,L line and in a c‐ALL line, in the absence of significant inhibition of SAH hydrolase, but with a substantial elevation in dATP concentrations and depressed levels of the other dNTP. Minor toxicity occurred in a proliferating B lymphoblast line despite almost complete inactivation of SAH hydrolase. These observations indicate inhibition of ribonucleotide reductase as the more likely mechanism of toxicity in rapidly proliferating lymphocytes. Other T‐cells actively synthesizing DNA, such as PHA‐stimulated or MIX activated lymphocytes and T‐lymphoid colony forming cells, are also likely to be affected by the same mechanism. Indeed in PHA‐stimulated lymphocytes, deoxycytidine caused significant although incomplete rescue from toxicity due to dCF and AdK. In patients with ADA deficiency or treated with ADA inhibitors, both mechanisms could be operative. These observations are also relevant to the possible use of dCF and AdK as immunosuppressive agents and for the removal of T‐cells or residual Thy‐ALL blasts from bone marrow in vitro prior to allogeneic or autologous bone marrow transplantation respectively.
Immunophenotyping of leukaemia: an immunoperoxidase method using air‐dried smears
Authors: Sandhaus Linda M. , Gajl‐Peczalska Kazimiera J. , Brunning Richard D.
Published: March 2008

Summary An immunoperoxidase (IP) method utilizing monoclonal antibodies to demonstrate the presence of the common acute lymphocytic leukaemia antigen (CALLA) on leukaemic cells using air‐dried blood and bone marrow smears is described. Two readily available CALLA‐specific monoclonal antibodies, BA‐3 and J5, and an avidin‐biotin immunoperoxidase methodology were utilized. Nineteen cases of acute leukaemia or blast crisis of chronic myelogenous leukaemia were studied. Immunofluorescent studies correlated closely with the results of the immunoperoxidase technique.
Extracellular cytolysis by leukaemic blast cells
Authors: Dallegri Franco , Patrone Franco , Frumento Guido , Ballestrero Alberto , Sacchetti Carlo
Published: March 2008

Summary Leukaemic blast cells from 20 patients with acute leukaemia were examined for their capacity to mediate cytotoxicity against ox red blood cells in the presence of phorbol myristate acetate (PMA), a system widely employed as an in vitro model of tissue damage by metabolically activated mature phagocytes. Blasts from certain patients with myelomonocytic and monocytic leukaemia behaved like efficient killer cells. Conversely, leukaemic myeloblasts and promyelocytes as well as leukaemic lymphoblasts were ineffective. Blast cells capable of inducing the target cell lysis were also capable of mounting an oxidative respiratory burst upon challenge with PMA, as detected by the superoxide anion release. N‐ethyl‐maleimide, superoxide dismutase and catalase completely inhibited the cytotoxicity by monocytoid blast cells, suggesting the involvement of oxygen reactive products in the lethal hit itself. The cytolytic potential of blasts committed to monocytic differentiation might be an additional factor contributing to the tissue damage in a subpopulation of leukaemic patients.
The ‘spent’phase of polycythaemia vera: hypersplenism in the absence of myelofibrosis
Authors: Najean Y. , Arrago J. P. , Rain J. D. , Dresch C.
Published: March 2008

Summary A clinical phase (spent phase) in the course of polycythaemia vera (PV) cases is described as enlargement of the spleen in spite of treatment, frequent cytopenia of one or several lines, persistent red cell hypervolaemia with considerable increase of plasma volume, persistence of myeloid hyperplasia with no collagen myelofibrosis or osteomyelosclerosis, absence of hepatosplenic erythro‐blastic metaplasia, as shown by radio‐iron kinetics and/or 111In‐transferrin scintigraphy.

The frequency of this phase was 5% in a study where it was not systematically sought, but it could in fact be greater. Its occurrence is not related to the clinical and biological parameters of PV. On the other hand, it is significantly more frequent and earlier in patients treated by phlebotomies than in those treated by myelosuppression (32P).

In four of the 12 cases, this phase was rapidly followed by an acute leukaemia. In eight cases there was a 1–5 year interval before a myelofibrosis with splenic myeloid metaplasia. This evolution could at this stage be delayed by chemotherapy. The efficacy of splenectomy should be studied.
Long‐term haemopoiesis in human fetal liver cell cultures
Authors: Cappellini M. D. , Potter C. G. , Wood W. G.
Published: March 2008

Summary. Haemopoiesis in human fetal liver is almost entirely restricted to the erythroid series but when fetal liver cells were cultured under conditions established for the long‐term maintenance of adult marrow haemopoiesis, a rapid switch to granulopoiesis was observed. Erythroid progenitor cells (BFU‐E) rapidly disappeared, even though no humoral or cellular inhibitors of erythropoiesis could be detected, while myeloid progenitors (CFU‐GM) increased in number. When the fetal liver cells were seeded onto stromal layers derived from adult marrow, in which endogenous haemopoiesis had ceased, granulopoiesis was established and maintained for more than a year, considerably longer than has previously been achieved with human haemopoietic cells.
Twenty‐five year survival of chronic granulocytic leukaemia with spontaneous karyotype conversion
Authors: Singer C. R. J. , McDonald G. A. , Douglas A. S.
Published: March 2008

Summary. Following busulphan‐induced bone marrow hypoplasia a woman with chronic granulocytic leukaemia has survived 25 years from diagnosis. Since the last course of busulphan therapy in 1959 she has remained in clinical and haematological remission. Repeated cytogenetic analysis of bone marrow showed Philadelphia chromosome mosaicism with a minority of abnormal metaphases till 1969. Analysis of 150 metaphases in 1982 revealed no cells containing the Philadelphia chromosome. The possible significance of this spontaneous karyotype conversion is discussed.
Differential inhibition of two molecular forms of melanoma cell plasminogen activator by a placental inhibitor
Authors: Lecander Ingegerd , Roblin R. , Astedt B.
Published: March 2008

Summary. The inhibitory effect of a placental urokinase inhibitor on the one‐ and two‐chain form of melanoma cell derived plasminogen activator (PA) was investigated. The melanoma cell PA has been shown to be similar to or identical with the PA found in normal tissue. With constant concentration of placental inhibitor the rate of inhibition of the two‐chain form was fast in contrast to that of the one‐chain form. With constant incubation time but increasing concentrations of the placental inhibitor the two‐chain form was inactivated to a greater extent at lower concentrations of placental inhibitor than was the one‐chain form. The increased reactivity of the two‐chain form of melanoma PA compared to the single‐chain form may explain the role of tissue PA in achieving high local concentrations of PA activity which facilitate selective fibrin clot lysis.
Identification of T lymphocyte subpopulations that regulate elaboration of granulocyte‐macrophage colony stimulating factor
Authors: Verma Dharmvir S. , Johnston Dennis A. , McCredie Kenneth B.
Published: March 2008

Summary. To determine the role of T lymphocytes (TL) that bear Fc receptors for IgG (Tγ+) and IgM (Tμ+) and of those that do not (Tγ‐ and Tμ‐) in the elaboration of granulocyte‐macrophage colony‐stimulating activity (GM‐CSA), we coincubated TL or the TL subpopulations in varying proportions with a constant concentration of autologous monocyte‐macrophages (MΦ) along with methanol extraction residue of BCG (MER). These conditioned media (CM) were assayed for GM‐CSA. MΦ and TL interacted maximally at 1:3–1:6 ratios, significantly less so at a 1:9 ratio. Coincubation of MΦ with Tγ‐ fraction markedly enhanced GM‐CSA elaboration at all MΦ:Tγ‐ ratios, progressively increasing colony‐stimulating activity as the proportion of Tγ‐ cells increased. MΦ interacted with the Tγ‐ fraction significantly better (P= 0.001) at all the ratios tested than it did with the Tγ‐μ+ or Tγ‐μ‐ subsets, suggesting that the Tγ‐μ+ and Tγ‐μ‐ subsets must interact. Coincubation of the Tγ+ fraction synergistically enhanced GM‐CSA elaboration only at MΦ:Tγ+ ratios of 1:1.5 and 1:3. Further increases in the proportion of Tγ+ cells rapidly and progressively decreased the capacity of the CM to stimulate granulocyte‐macrophage colony‐forming cells (GM‐CFC). Also, inclusion of Tγ+ cells in the coincubation mixtures of MΦ and Tγ‐ fraction significantly suppressed GM‐CSA elaboration. Dose‐response curves and CM‐mixing experiments revealed that the CM from coincubation of MΦ and Tγ+ not only contained smaller amounts of stimulating factors but also contained a GM‐CFC inhibitor. Experiments also demonstrated that the suppressor subpopulation enriched with the Tγ+ fraction was radiosensitive. These results suggest that the elaboration of GM‐CSA is controlled by helper and suppressor subpopulations that are enriched with Tγ‐ and Tγ+ fractions, respectively. Furthermore, the suppressor effect is radiosensitive.
Prophylactic antithrombotic therapy with stanozolol in patients with familial antithrombin III deficiency
Authors: Winter J. H. , Fenech A. , Bennett B. , Douglas A. S.
Published: March 2008

Summary. Three patients with familial antithrombin III deficiency underwent a trial of prophylactic antithrombotic therapy with stanozolol. An increase in plasma fibrinolytic activity and antithrombin III was seen. Acute venous thrombosis occurred in two patients; in the first patient thrombosis was precipitated by venography but in the second no precipitating factor was found. Our experience suggests that stanozolol is not a suitable prophylactic agent in patients with familial antithrombin III deficiency.
The coagulation factor VII in pregnancy
Authors: And K. Dalaker , Prydz H.
Published: March 2008

Summary. The hypercoagulable state in pregnancy is partly caused by the increased activity of factor VII in plasma. We demonstrate here that this activity is reduced to levels similar to those in plasma from non‐pregnant women by highly purified phospholipase C from Bacillus cereus, i.e. the activity increase is due to a circulating complex of factor VII and a phospholipase C‐sensitive compound. Phospholipase C had no effect on the levels of factor II and X in blood from pregnant women. This novel form of activated factor VII is not inhibited by an antiserum to the protein component of thromboplastin (apoprotein III). By gel filtration of plasma from pregnant women on Sephadex G‐100 the phospholipase C‐sensitive complex was partly separated from non‐phospholipase sensitive factor VII also present in the same plasma.
Radioimmunoassays of human high and low molecular weight kininogens in plasmas and platelets
Authors: Kerbiriou‐Nabias Daniegle M. , Garcia Francia O. , Larrieu Marie‐Josea
Published: March 2008

Summary. Radioimmunoassays of human high molecular weight kininogen (HMWK) and low molecular weight kininogen (LMWK) were developed using antibodies directed against the light and the heavy chains of kallikrein‐cleaved HMWK. With the anti‐light chain antibodies, the radioimmunoassay was specific for HMWK with a detection limit of 0.4 ng. The anti‐heavy chain antibodies were used to quantify the concentration of total kininogen antigens. In four different plasmas with a congenital deficiency in HMWK procoagulant activity, there was no detectable antigen in two cases and trace amounts, less than 1 μg/ml in the other plasmas (normal concentration: 72 ± 6 μg/ml). In the absence of HMWK, the radioimmunoassay performed with the anti‐heavy chain antibodies was specific for LMWK. The amount of LMWK was different in each of these patients’plasmas, ranging from no detectable antigen, i.e. less than 0.15 μg/ml, to a normal content.

Antigens immunologically indistinguishable from plasma kininogens were detected in lysates of five times washed platelets. HMWK antigen concentration was 3.17±0.87 μg per 1011 platelets (mean value in 11 donors). LMWK was also present in platelet lysates and the relative concentration versus HMWK was the same as in plasma.
Sequential effects of lithium on haematopoiesis
Authors: Joyce Robert A.
Published: March 2008

Summary. The administration of lithium salts to haematologically normal subjects is associated with increased blood neutrophil concentrations and marrow neutrophil production and with enhanced release of colony stimulating activity (CSA) required for growth of granulocyte‐macrophage progenitor cells (CFU‐GM) in vitro. To examine the haematopoietic changes associated with lithium salts, mice were given LiCl daily. Blood neutrophils and serum CSA levels increased as did blood platelet concentrations. These increments were preceded by expansion of marrow neutrophil production and increased concentrations of CFU‐GM as well as progenitor cells for megakaryocytes (CFU‐M) and erythrocytes (BFU‐E and CFU‐E). An earlier and sustained increase of transplantable pluripotential stem cells (CFU‐S) was detected beginning at day 2 of lithium administration. The sustained increase of CFU‐S with lithium was not associated with detectable changes of endogenous stem cells (E‐CFU) suggesting that a portion of the stem cell pool is resistant to the proliferative effects of lithium. These studies indicate that lithium acts initially to directly or indirectly increase marrow CFU‐S with later increments of progenitor cells. The more sustained increase of blood neutrophils with lithium administration may be the result of subsequent increments of CSA resulting in enhanced marrow granulocyte production.
Immunohistological analysis of human bone marrow trephine biopsies using monoclonal antibodies
Authors: Falini B. , Martelli M. F. , Tarallo F. , Moir D. J. , Cordell J. L. , Gatter K. C. , Loreti G. , And H. Stein , Mason D. Y.
Published: March 2008

Summary. This paper describes the use of a recently developed immuno‐alkaline phosphatase method (the ‘APAAP’technique) for labelling frozen sections of undecalcified bone marrow biopsies with monoclonal antibodies, including reagents reactive with T cells and their subsets, B cells, glycophorin, HLA‐DR antigen, common ALL antigen, epithelial cells and megakaryocytes. Use of an immuno‐alkaline phosphatase technique avoids problems due to endogenous enzyme activity encountered when staining bone marrow by immunoperoxidase procedures. Immunohistological labelling of frozen trephine biopsies is of particular value when it is impossible to aspirate marrow particles and for identifying cells which do not readily enter suspension (e.g. dendritic reticulum cells or stromal cells). Details are given of cases in which immunohistological analysis was used for the phenotyping of acute leukaemias, for the differential diagnosis of intramedullary T and B cell proliferations, and for identifying bone marrow metastases.
Evidence for clonal evolution in pre‐B‐cell leukaemia
Authors: Abromowitch Minnie , Williams Dorothy L. , Melvin Susan L. , Stass Sanford
Published: March 2008

Summary. This case study provides evidence for clonal evolution in pre‐B‐cell leukaemia. At diagnosis, the lymphoblasts from a 3‐year‐old boy were morphologically subtyped as L1 (French–American–British classification). Their immunophenotype was CALLA+, CIgM+, SIg‐, TdT+, and the karyotype was pseudodiploid with a 1;19 translocation. Striking shifts were apparent when the child relapsed 16 months later. The morphologic subtype had changed to L3, CALLA and TdT had disappeared, and a consistent karyotype was lacking. The modal chromosome number had increased through clonal evolution to 85, the 1;19 translocation was retained, and a new marker, a 14q+ (partial duplication) appeared and was present in a majority of cells. These cytogenetic findings are characteristic of a transforming state. However, despite the loss of TdT, the appearance of classic L3 morphology and the acquisition of a 14q+ marker, the cells retained a predominantly pre‐B phenotype.