bluuebunny/crossref_metadata_2025 huggingface dataset:
Observation of Echinocytosis in Eight Patients: a Phase Contrast and SEM Study
Authors: Feo C. J. , Tchernia G. , Subtil E. , Leblond P. F.
Published: March 2008

Summary. We report eight cases of echinocytosis discovered after screening of stained smears. The presence of echinocytes was exceptional in adults and children but was more frequent in neonates. In all cases we confirmed the presence of abnormal red cells by careful examination of the blood in wet preparations observed in phase contrast and after glutaraldehyde fixation and processing for SEM. No discernible common denominator was found to explain the presence of echinocytes in the blood of our patients. No echinocytogenic factor could be demonstrated in the fresh plasma. Echinocytes appeared quickly in patients who received exchange transfusion or transfusion of normal red cells. By contrast, in vitro incubation in the presence of substrates allowing constant ATP level never resulted in the formation of echinocytes when normal red cells were suspended in patient's plasma.
Hereditary Pyruvate Kinase Deficiency: Role of the Abnormal Enzyme in Red Cell Pathophysiology
Authors: Zanella A. , Rebulla P. , Vullo C. , Izzo C. , Tedesco F. , Sirchia G.
Published: March 2008

Summary. Two new mutant Pks, electrophoretically identical but kinetically slightly different, are reported. These two clinically innocuous PK variants, encountered in two non‐related subjects, have combined in their daughter to give a fully expressed haemolytic anaemia. The functional abnormalities of the daughter's PK (increased K0.5 PEP, abnormal response to FDP, increased urea and guanidine‐HCl stability, abnormal isoelectrofocusing and electrophoretic patterns) were like those of the parents but more pronounced, except for thermostability that was normal in the proband although markedly decreased in both parents. The family examined demonstrates that there is no relationship between in vitro properties of the variant and the severity of haemolysis. The hypothesis is put forward that the cause of haemolysis in PK deficiency may be associated with another defect located in the red cell membrane.
Some Features of Bone Marrow Macrophages in Patients with Homozygous β‐Thalassaemia
Authors: Wickramasinghe S. N. , Hughes Madeleine
Published: March 2008

Summary

The bone marrow macrophages of patients with homozygous β‐thalassaemia were frequently situated adjacent to collagen fibres and sometimes formed intrasinusoidal cytoplasmic protrusions. They also appeared to phagocytose processes of erythroblast cytoplasm (at times containing precipitated α‐chains) which projected into them from neighbouring erythroblasts. The cytoplasm of the macrophages included large numbers of heavily‐iron‐loaded secondary lysosomes of various sizes and shapes in addition to phagocytosed erythroblasts, erythrocytes and extruded erythroblast nuclei. Numerous ferritin molecules were found in the cytoplasmic matrix but there were hardly any in the mitochondria, endoplasmic reticulum or golgi saccules. A small number of ferritin molecules were present within the nucleus. Electron microscope autoradiographs of marrow fragments which had been incubated with [3H]leucine for 1 h revealed the presence of newly‐synthesized protein molecules in all types of secondary lysosomes. Light microscope autoradiographs showed that the [3H]thymidine labelling index of the bone marrow macrophages was less than 1% and suggested that only a very small proportion of these cells were actively preparing for division.
Autoimmune Antibodies in Chronic Lymphatic Leukaemia
Authors: Lewis C. M. , Pegrum G. D.
Published: March 2008

Summary

In chronic lymphocytic leukaemia a factor in patients' serum enhances the in vitro viability of the abnormal cells and this has been identified as an antibody. The activity of this factor can be removed by interaction with anti‐immunoglobulin and by ammonium sulphate precipitation with a degree of saturation in excess of 46%. Cohn fractionation and chromatography with A‐50 Sephadex show that the factor is not a complex but an immunoglobulin. No activity is removed after reaction of sera with 2‐mercapto‐ethanol and di‐thiothreitol. The evidence therefore suggests that a γ‐G immunoglobulin is involved. Concentrated washings from the leukaemic cells behave in exactly the same way as patients' sera and activity is retained in the same fraction during precipitation and purification procedure.

The extensive cross‐reactivity of the sera suggests a common chronic lymphatic leukaemic antibody and it is considered that an active autoimmune response may be an integral part of the disease.
Stereological Analyses of Leukaemic Cells
Authors: James V.
Published: March 2008

Summary. With the development of different treatment schedules for acute and chronic leukaemias it has become essential that a uniform, standardized classification of malignant cell type is achieved. Without a standardized diagnosis it is impossible to compare the results of differing treatment regimes. Present diagnosis is still based on clinical criteria and subjective assessment of cell morphology; objective and quantitative data are required. Stereological analyses are capable of providing quantitative data. Stereological techniques have been applied to the study of malignant cell types in different leukaemias for the first time and quantitative data has been obtained. The nuclear and mitochondrial volume fractions of the cells have been estimated for normal lymphocytes, the lymphocytes of chronic lymphocytic leukaemia, acute lymphoblastic leukaemia, the abnormal cells of hairy cell leukaemia and several other haematological malignancies.

The results clearly indicate that the cell types of different leukaemias possess measurably different properties and that stereological methods can be used to investigate the leukaemic cell quantitatively.
Ultrastructural Identification of Acid Complex Carbohydrate in Cytoplasmic Granules of Normal and Leukaemic Human Monocytes
Authors: Parmley R. T. , Spicer S. S. , O'Dell R. F.
Published: March 2008

Summary. Complex carbohydrate in granules of monocytes was compared with that in granules of neutrophils by ultrastructural cytochemical methods. The acid mucosubstance in granules of both cell types stained with dialysed iron after brief fixation with dilute glutaraldehyde, but that in monocyte granules differed in failing to stain after stronger fixation. Approximately 10% of granules in normal blood monocytes stained with this method, whereas more than 90% of granules in leukaemic monocytes from two of seven patients with acute myelomonocytic leukaemia stained intensely. This difference presumably results from unmasking of acid groups in immature granules or increased synthesis of granule mucosubstance in some leukaemic monocytes. Granules of monocytes differed further from those of neutrophils in failing after either type of fixation to stain with a high iron diamine technique for demonstration of sulphated mucosubstance. The absence of high iron diamine staining could reflect a lack of sulphate esters in monocyte granule mucosubstance, masking of the sulphate groups by other components, or extraction of the sulphated mucosubstance during specimen processing.
The Neonatal Platelet: Evaluation of Platelet Malonyl Dialdehyde Formation as an Indicator of Prostaglandin Synthesis
Authors: Stuart Marie J.
Published: March 2008

Summary. It is recognized that neonates exhibit a transient thrombocytopathy. Whether the cause of the platelet abnormality is due to a storage pool defect, or an‘aspirin‐like’abnormality with inhibition of platelet prostaglandin synthesis is unclear. In an attempt to assess neonatal platelet prostaglandin synthesis, platelet rich plasma (PRP) from 10 normal control maternal–neonatal pairs was obtained contemporaneously with PRP from donors who had ingested 600 mg aspirin within 12‐18 h prior to study. When platelet malonyl dialdehyde production in the presence of the sulphydryl inhibitor N‐ethyl maleimide (NEM), or thrombin, was measured as an indicator of platelet prostaglandin synthesis, a significant decrease was observed in neonatal platelets (2.46 ± 0.61 and 0.90 ± 0.19 nmol/109 platelets respectively) when compared to those of the paired maternal or adult controls (3. 23 ±0.31 and 1.30 ± 0.17 nmol/109 platelets). As expected, a marked decrease in platelet malonyl dialdehyde production in response to NEM or thrombin was also seen in the platelets from the donors who had ingested aspirin (0.25±0.14 and 0.10 ± 0.09 nmol/109 platelets). When compared to the normal maternal control PRP both the neonatal and aspirin donor PRPs demonstrated abnormalities in second phase platelet aggregation in response to ADP and epinephrine, and an abnormal response to collagen. However, mutual correction and secondary irreversible aggregation was obtained when both these abnormal PRPs were mixed in varying proportions. No mutually corrective effect was seen using neonatal PRP obtained from infants where maternal documentation of aspirin ingestion was present within 48 h prior to delivery, or when normal neonatal PRP was preincu‐bated with aspirin in vitro prior to mixing with PRP from donors who had recently ingested aspirin. These findings provide evidence that the neonatal platelet dysfunction cannot be attributed to an abnormality in prostaglandin synthesis or an‘aspirin‐like’defect. Although platelet malonyl dialdehyde formation in response to NEM and thrombin was decreased when compared to the adult the amount synthesized by the neonatal platelet in response to external stimuli appears to be
Oxidative Damage to Neutrophils in Glutathione Synthetase Deficiency
Authors: Spielberg Stephen P. , Boxer Laurence A. , Oliver Janet M. , Allen John M. , Schulman Joseph D.
Published: March 2008

Summary. Several episodes of neutropenia were observed in a child with glutathione synthetase deficiency (5‐oxoprolinuria). Studies of the patient's glutathione‐deficient neutrophils were undertaken to examine the responses of the cells to oxidative stress associated with phagocytosis. The patient's neutrophils contained 10–20% of normal glutathione content. Circulating neutrophils in infection‐free periods appeared less mature than normal by morphologic criteria, suggesting increased cell turnover. The cells ingested particles, responded to chemotactic stimuli, and oxidized 1‐14C glucose normally. However, following ingestion of particles, the cells accumulated excess hydrogen peroxide compared with normal cells, and showed impaired protein iodination and bacterial killing. Electron micrographs revealed damage to microtubules and membranous structures in the patient's neutrophils during phagocytosis. The level of glutathione in the cells appears inadequate to protect against peroxide generated during normal cell function, and the cells are thus damaged and rendered less effective in bacterial killing. The data provide evidence for a protective role of glutathione in normal neutrophil function.
Red Blood Cell Enzyme Abnormalities in Patients Treated with Chemotherapy
Authors: Etiemble J. , Bernard J. F. , Picat Ch. , Belpomme D. , Boivin P.
Published: March 2008

Summary. Sixteen red blood cell enzyme activities and fetal haemoglobin level have been assayed in 60 patients treated for haematologic or nonhaematologic malignant diseases with various combinations of cytostatic drugs. Acquired enzyme deficiency was found in 20 patients. The most frequently decreased activities were those of G6PD (12 cases), PK (seven cases), PFK (six cases) and AK (three cases). In many patients decreased activity of some enzymes contrasted with increased activity of others such as ALD, TPI, G3PD, PGK, ENOL and 6PGD. The number of abnormalities seems to be related to the duration of the treatment.

Incidence of enzyme deficiencies was similar in patients previously treated or not with radiotherapy. Enzyme abnormalities were correlated neither with haemoglobin level nor with reticulocyte count. They were very similar to those observed in primary acquired dyserythropoietic and preleukaemic states. Their prognostic value and mechanism are discussed.
Fatty Acid Composition of Erythrocytes in Hereditary Spherocytosis
Authors: Zail S. S. , Pickering A.
Published: March 2008

Summary. The fatty acid distribution of the four major phospholipids has been determined in the erythrocytes of five patients with hereditary spherocytosis and in five control subjects. In contrast to a report by other workers, we are unable to confirm their findings of a defect in fatty acid chain lengthening activity leading to disappearance of long chain fatty acid conjugates of more than 20 carbon atoms in some of the phospholipid fractions.
Coagulation Activation and Hyperviscosity in Infection
Authors: Richardson S. G. N. , Matthews K. B. , Cruickshank J. K. , Geddes A. M. , Stuart J.
Published: March 2008

Summary. A serial study of coagulation activation and whole‐blood viscosity was performed on 37 patients with local or systemic bacterial infection, malaria, or a viral infection. Thrombocytopenia, without consumption of coagulation factors, was the main feature of benign tertian malaria and viral infection, whereas in septicaemia and malignant tertian malaria it was associated with activation of coagulation and fibrinolysis. Patients with evidence of intravascular coagulation showed the highest levels of factor VIII related antigen which did not correlate with fibrinogen and probably reflected vascular endothelial cell damage rather than an acute‐phase protein reaction.

Hyperviscosity, which has been implicated in the pathogenesis of endotoxic shock and cerebral malaria, occurred in parallel with the acute‐phase rise in plasma fibrinogen. There was, however, no evidence to implicate hyperviscosity as a major causative factor in the pathogenesis of septic shock or severe infective illness.
Membrane Receptors of Human Leukaemic Myeloid Cells: Sequential Expression of the γFc Receptor
Authors: Burns G. F. , Cawley J. C.
Published: March 2008

Summary. An immunological surface marker study was performed on 13 patients with a variety of myeloid leukaemias. It was shown that expression of the receptor for the Fc of IgG (γFcR) starts to take place at the promyelocyte stage, and that the receptor is present on more mature granulocytic cells, but is absent from myeloblasts. Myeloblasts and promyelocytes are negative for the complement receptor CR2. Monoblasts, unlike myelobasts, were shown to express a γFcR and, to a lesser extent, CR2. Receptor expression therefore appears to be an earlier event in monocytic development. The possible diagnostic value of immunological marker studies in myeloid disorders is considered.
The Kinetics of Serum and Tissue Ferritins: Relation to Carbohydrate Content
Authors: Halliday J. W. , Mack U. , Powell L. W.
Published: March 2008

Summary. Significant differences were observed in the rate of disappearance from plasma of ferritins purified from rat serum and from different organs. Ferritin from all sources including purified serum ferritin was rapidly removed from plasma by the liver. No difference in biological half‐life was observed between apoferritin prepared by ultracentrifugation of liver ferritin and whole liver ferritin and ironloaded animals cleared injected serum ferritin from plasma at a comparable rate to normal rats. When amounts of 100 μg of ferritin were injected into rats the half‐life was significantly lengthened. The study confirmed the fact that ferritin iron and ferritin protein were removed from plasma at the same rate. No consistent effect of acidic or more basic isoferritin composition on biological half‐life was apparent. After chromatography on concanavalin A‐Sepharose 6B those ferritins which were predominantly bound to Con A‐Sepharose had a half‐life which was approximately twice that of ferritins which did not bind. It is concluded that the variation in plasma disappearance of ferritins of different tissue origin was explainable on the basis of carbohydrate content of the molecule.
Indium‐III: a New Radionuclide Label for Studying Human Platelet Kinetics
Authors: Heaton W. Andrew , Davis Harmon H. , Welch Michael J. , Mathias Carla J. , Joist J. Heinrich , Sherman Laurence A. , Siegel Barry A.
Published: March 2008

Summary. Indium‐III, when complexed with 8‐hydroxyquinoline (oxine), has been employed as a radioactive platelet label for thrombus imaging in animals and man. The short half‐life (2.8 d) and high yield of gamma photons of 111In make it ideal for in vitro counting and external imaging. To evaluate its suitability for studies of platelet turnover in man, platelet kinetic studies were carried out on 10 healthy volunteers using 111In‐and 51Cr‐platelets concurrently. For 111In labelling, platelets were harvested by differential centrifugation from 43 ml of whole blood drawn into acid‐citrate dextrose (ACD) solution. The platelets were washed and suspended in a mixture of ACD and isotonic saline and then incubated with 111In‐oxine, rewashed, and suspended in plasma for reinfusion. 51Cr labelling was performed using standard methods. Mean labelling efficiency was 73% with 111In and 6.5% with 51Cr. In vitro studies demonstrated minimal release, elution, and reutilization of the 111In label. There was no significant difference in the aggregation response of 111In‐ and 51Cr‐platelets to ADP and collagen. The in vivo recovery of 111In‐platelets was approximately 50% greater than that of 51Cr‐platelets whereas the platelet life spans were similar. These results indicate that 111In labelled platelets may be useful for thrombokinetic studies in man. The new method offers the advantages of reduced blood requirements, higher labelling efficiency, and the ability to perform external imaging of platelet distribution in vivo.
A Genetic Variant of Factor IX with Decreased Capacity for Ca2+ Binding
Authors: Bertina R. M. , Veltkamp J. J.
Published: March 2008

Summary. A genetic variant of factor IX is described that behaves identically to PIVKA IX (the precursor factor IX molecule induced by the absence of vitamin K or presence of vitamin K antagonists, acarboxy factor IX). It shows an increased electrophoretic mobility in the presence of Ca2+, a low affinity for adsorption to A1(OH)3 and a very low specific coagulant activity. This variant of factor IX has been demonstrated in the plasma of a patient with severe haemophilia B and in the plasmas of a number of possible carriers from the probands’ pedigree.
The Effect of Desferoxamine on Fibroblasts and Collagen Formation in Cell Cultures
Authors: Hunt J. , Richards R.J. , Harwood R. , Jacobs A.
Published: March 2008

Summary. Summary. Iron is essential for the activity of proline hydroxylase and is an important co‐factor in collagen synthesis. Fibroblast cultures exposed to desferrioxamine show impairment of DNA synthesis and reduced collagen formation, as measured by hydroxyproline synthesis and the deposition of hydroxyproline in the cell mat. In patients with transfusional iron overload long‐term treatment with desferrioxamine is said to result in the inhibition of hepatic fibrosis. It is suggested that this may be a direct effect on collagen synthesis rather than an effect of reduced iron stores.
Metabolic Changes in Red Blood Cells in Malignant Lymphomas
Authors: Bellis R. De , Boulard M. R. , Kasdorf H. , Rodriguez I. , Ferrando R. , Landro J. Di , Ferrari M. , Sanguinett C. M. , Tanzer J.
Published: March 2008

Serum copper levels (SCL) which are concomitantly related to red blood cell free copper are significantly increased in some malignant lymphomas in the phase of activity. This results in a profound inhibition of red cell key glycolytic enzymes, hexokinase (Hx) being the most sensitive. Fifteen patients (eight with Hodgkin's disease and seven with non‐Hodgkin's lymphoma) were studied for serum and red cell copper concentrations and Hx activity. The mean red cell life span was determined using 51Cr labelled red cells. The resulting data shows that in active disease an increase in SCL was associated with a decrease in Hx activity and a shortened red cell survival. In these cases there was no evidence of autoimmune phenomena or of direct bone marrow involvement by the disease. It is suggested that the increase in copper levels results in a shortened red cell life span through a copper‐induced inhibition of red cell Hx.
Membrane Abnormalities of Pyruvate Kinase Deficient Red Cells
Authors: Zanella A. , Brovelli A. , Mantovani A. , Izzo C. , Rebulla P. , Balduini C.
Published: March 2008

Two experimental systems have been applied to the study of PK‐deficient red cells in an attempt to reveal possible membrane abnormalities of these erythrocytes: (1) the glycoprotein self‐digestion of intact erythrocytes during in vitro incubation at 37°C; (2) the red cell sensitivity to the lytic action of mouse macro‐phages.

PK‐deficient erythrocytes display a more precocious than normal membrane glycoprotein self‐digestion and are much more susceptible than normal to the cytotoxic activity of mouse macrophages. This latter effect is more pronounced in young than in old PK‐deficient red cells. These observations indicate the existence of membrane abnormality in PK‐deficient red cells other than those so far suspected.
The Heterogeneity of Normal Hb A2‐β Thalassaemia in Greece
Authors: Kattamis C. , Metaxotou‐Mavromati A. , Wood W. G. , Nash J. R. , Weatherall D. J.
Published: March 2008

Nine patients have been observed with homozygous β thalassaemia in each of whom one parent has a normal level of Hb A2. On the basis of clinical, haematological and globin chain synthesis studies these families have been divided into two groups.

Group 1 (six families). Heterozygotes for normal Hb A2–β thalassaemia in this‐group showed minimal red cell abnormalities, normal osmotic fragility but imbalanced globin chain synthesis (α/β= 1.6), and appear to correspond to previous descriptions of ‘silent’β thalassaemia. Double heterozygotes with high Hb A2‐β thalassaemia have a clinical picture of mild β thalassaemia intermedia characterized by relatively low levels of Hb F (<20%) and γ chain synthesis.

Group 2 (three families), β Thalassaemia heterozygotes with normal Hb A2 levels in this group showed more marked red cell abnormalities, decreased osmotic fragility and more imbalanced globin chain synthesis (α/β=2.5) than those in group 1. Double heterozygotes with high Hb A2‐β thalassaemia are more severely affected and are transfusion dependent. Haemoglobin F and γ chain synthesis are high in these cases.

The frequency of normal Hb A2‐β thalassaemia in Greece may be as high as 10% of all β thalassaemia genes and this poses a significant problem for genetic counselling. Various molecular mechanisms are discussed which could account for the heterogeneity within these disorders.
Thrombin Receptors of Human Platelets: Thrombin Binding and Antithrombin Properties of Glycoprotein I
Authors: Ganguly P. , Gould N. L.
Published: March 2008

Washed human platelets were solubilized and the proteins were separated by preparative gel electrophoresis in the presence of sodium dodecyl sulphate. The gel was cut into slices and the effect of the eluted proteins on the clotting of fibrinogen by thrombin was evaluated. The isolate from only one gel slice strongly inhibited the clotting of fibrinogen. The prolongation of the clotting time was dependent on the concentration of the protein and reached a plateau around 5 μg. Gel electrophoresis of this isolate showed a prominent glycoprotein with an apparent Mr=150 000. Gel filtration studies with [125I]thrombin showed that the protein isolate bound a significant amount of thrombin which could be displaced with unlabelled thrombin. Another preparation from the same gel or purified γ‐globulin did not bind thrombin or prolong the clotting time of fibrinogen.

Glycoprotein I was isolated from human platelets by affinity chromatography on lectin–Sepharose columns. The isolated glycoprotein prolonged the clotting of fibrinogen and bound [125I]thrombin which could be displaced by unlabelled thrombin. It is proposed that the high affinity receptor of thrombin on human platelets is glycoprotein I. In addition, the antithrombin activity of intact platelets is due to binding of thrombin to this glycoprotein.
Immune Pancytopenia
Authors: Daneshbod‐Skibba Ghodsi , Shahidi Nasrollah T. , Becker Gary A. , Martin Jean
Published: March 2008

Summary. Circulating T‐lymphocytes from a 13‐year‐old boy with autoimmune anaemia, severe neutropenia and thrombocytopenia inhibited autologous and normal homologous bone marrow myeloid colony formation in vitro. This inhibition was abolished when the patient's antithymocyte globulin and complement‐treated T‐lymphocytes were used. T‐lymphocytes from normal individuals did not cause such an inhibition. The patient's lymphocytes showed no inhibitory effect on erythroid colony formation. Investigations of the patient's serum failed to disclose any leucoagglutinin, lymphocytotoxin or humoral factor against myeloid colony formation. These findings indicate that T‐lymphocytes may play a role in the pathogenesis of neutropenia in immune pancytopenia.
Splenic Irradiation in Myelofibrosis: Effect on Circulating Myeloid Progenitor Cells
Authors: Koeffler H. Phillip , Cline Martin J. , Golde David W.
Published: March 2008

Summary. We have investigated the mechanism of splenic irradiation‐induced granulocytopenia in two patients with myelofibrosis and marked splenomegaly. Serial assays were performed for circulating granulocyte‐monocyte progenitors capable of colony formation in vitro (CFU‐C). For comparison, similar studies were performed on two patients receiving whole brain irradiation for glioma. Splenic irradiation caused a significant decrease in circulating CFU‐C in the myelofibrosis patients. There was no decrease in circulating CFU‐C in the brain‐irradiated patients. No radiation‐induced humoral inhibitor of granulopoiesis and no increased CFU‐C radiosensitivity could be demonstrated in the myelofibrosis patients. These observations, taken together with previous data on splenic blood flow and pooling, suggest that the major mechanism of irradiation‐induced granulocytopenia in myelofibrosis is destruction of proliferating precursor cells in the splenic tissue and sinusoids.
A Modified Non‐radioisotope Method for Measurement of Platelet Production Time
Authors: De Haas H. A. , Clark S. E. , Zahavi J. , Kakkar V. V. , White A. M.
Published: March 2008

Summary. Platelet production time (PPT), based on the measurement of malondial‐dehyde (MDA) production prior to and after the intake of acetylsalicylic acid (ASA), was determined in 20 healthy subjects. High MDA levels were produced by stimulating platelet lipid peroxidation with arachidonic acid, resulting in a reliable, sensitive and accurate technique. PPT correlated well with platelet survival time measured by 51Cr autologous labelled platelets when both methods were used simultaneously in the same patients. This modified non‐radioisotope method might serve as a useful aid in the diagnosis of platelet disorders, thromboembolic diseases associated with increased platelet consumption and for the evaluation of drugs affecting platelet function.
Transitional Myeloproliferative Disorder
Authors: Pettit J. E. , Lewis S. M. , Nicholas A. W.
Published: March 2008

Summary. Eleven patients have been observed with clinical features of both polycythaemia vera and myelofibrosis. Detailed follow‐up and repeated haematological and isotopic investigations, including the assessment of erythropoietic distribution by 52Fe scanning, over a 10 year period, have indicated that patients who initially present with this syndrome may remain in a steady state for several years and that this transitional syndrome does not necessarily imply an active or irreversible transformation into classical myelofibrosis. Therapy with iron, folic acid, alkylating agents, splenectomy or splenic irradiation may reduce the extramedullary component of myeloproliferation and allow occasional patients to revert to more classical polycythaemia vera. Radioactive phosphorus (32P) therapy may be inappropriate in polycythaemic patients with dominant extramedullary erythropoiesis, as this form of therapy has a preferential medullary action and may selectively encourage extramedullary myeloproliferation.
Junctional Structures in Haemopoiesis: a Study of Bone Marrow using Freeze‐Fracture and Lanthanum Impregnation Techniques
Authors: Tavassoli Mehdi , Shaklai Mattiyahu
Published: March 2008

Summary. Intercellular regions of contact in the haemopoietic compartment of normal rat bone marrow were studied using freeze‐fracture and lanthanum tracer techniques. Small adhering junctions (like desmosomes and their variants) were found between haemopoietic and stromal cells but tight, gap or septate junctions could not be identified. These findings are in agreement with the concept that extensive junctional structures may be inconsistent with orderly development of this transient cell system, preventing the delivery of mature cells into the circulation and resulting in ineffective haemopoiesis. Occasionally ‘pinching off’of a portion of the cytoplasm of erythroid cells by stromal cells was seen, providing a means for intercellular communication. Structures similar to intercellular bridges responsible for direct intercellular communication were also seen.
Fluxes of Plasma Transcobalamins following Saturating Doses of Vitamin B12 in Man
Authors: Muir M. , Chanarin I.
Published: March 2008

Summary. The three vitamin B12 binding proteins in plasma were saturated with vitamin B12 by an intravenous injection of 57Co‐vitamin B12. All three unsaturated vitamin B12 binding proteins reappeared in plasma in detectable amounts within 15 min of the clearance of unbound vitamin B12 from plasma.
Generation of Factor VIII Coagulant Activity by Isolated, Perfused Neonatal Pig Livers and Adult Rat Livers
Authors: Owen Charles A. , Walter Bowie E. J. , Fass David N.
Published: March 2008

Summary. Isolated, normal, neonatal pig livers released VIII:C into perfusates of porcine von Willebrand blood lacking VIIIR:AG and VIII:RWF. Neonatal von Willebrand livers released VIII:C only when partially purified VIIIR:AG was added to the von Willebrand blood. Adult rat livers, perfused with frozen‐thawed erythrocytes suspended in Tyrode's solution containing 6% bovine serum albumin and hence no VIIIR:AG released VIII:C only when serum, adsorbed serum or cryoprecipitate from rat plasma was added to the perfusate. Cycloheximide blocked the release of VIII:C from rat liver.
Acute Myeloid Leukaemia Occurring in Untreated Chronic Lymphatic Leukaemia
Authors: Lawlor Emer , McCann S. R. , Whelan A. , Greally J. , Temperley I. J.
Published: March 2008

Summary. A case of acute myeloid leukaemia (AML) occurring in a patient with untreated chronic lymphatic leukaemia (CLL) is presented. The diagnosis of two simultaneous leukaemic processes is based on morphological, cytochemical and immunological findings. The significance of the development of AML in patients with CLL is discussed.
Rheological Properties of Erythrocytes in Heterozygous and Homozygous β Thalassaemia
Authors: Tillmann W. , Schröter W.
Published: March 2008

Summary. The rheological properties of erythrocytes were studied in 12 heterozygous and nine homozygous β thalassaemia patients, using filtration of erythrocytes through polycarbonate sieves with a pore diameter of 5 μm and viscosity measurements of erythrocyte and ghost suspensions. Erythrocyte deformability of all patients was decreased. In the heterozygous patients moderately diminished flexibility of cells was due to microcytosis of erythrocytes. Rigidity of erythrocytes in homozygous patients was due to altered cell shape, diminished fluidity of haemoglobin, and in splenectomized patients to the presence of inclusion bodies. Membrane flexibility of erythrocytes remained unaltered. We suggest that the decreased fluidity of haemoglobin as well as the presence of inclusion bodies result from the excess of α chains in erythrocytes of homozygous patients. The haemoglobin fluidity of erythrocytes of one patient with thalassaemia intermedia was unchanged. The flexibility of these cells was only moderately altered. In four patients who needed transfusions every 1–4 weeks deformability of transfused erythrocytes was likewise decreased when compared to controls. Erythrocyte suspensions obtained from the splenic artery and vein during splenectomy showed decreased flexibility of transfused cells after their passage through the spleen.
Hereditary Haemolytic Ovalocytosis with Defective Erythropoiesis
Authors: Torlontano G. , Fioritoni G. , Salvati A. M.
Published: March 2008

Summary. Patients belonging to four families with ‘atypical elliptocytosis’have been investigated. Clinical, haematological, erythrokinetic and enzymatic characteristics as well as the effect of splenectomy are discussed. These studies appear to define the fundamental features of a particular disorder or a variety of hereditary elliptocytosis; characterized by a genetic autosomal dominant character, moderate degree of RBC eccentricity, erythroid dysplasia with relative marrow failure and incomplete response to splenectomy.
Erythrocyte Membrane Lipid Reorganization during the Sickling Process
Authors: Chiu Danny , Lubin Bertram , Shohet Stephen B.
Published: March 2008

Summary. In order to study possible alterations in membrane lipids during sickling, we have measured the difference in susceptibility to lipid peroxidation, binding of trinitrobenzenesulfonic acid (TNBS) to aminophospholipids, and fatty acid uptake in cells containing sickle haemoglobin under aerobic and anaerobic conditions. We have also examined TNBS binding in irreversibly sickled cells in an attempt to evaluate the permanent effects of any such alterations.

We found that when erythrocytes were sickled by deoxygenation, the susceptibility to lipid peroxidation and binding of TNBS to aminophospholipids was markedly increased, while normal control cells showed no change. These effects appeared to be specific for the sickled state rather than a nonspecific consequence of cell age or the concentration of sickle haemoglobin within the cell. In contrast, fatty acid incorporation into membrane phospholipids, representing potential lipid renewal, was decreased in the sickle state. Cell fractions enriched in irreversibly sickled cells showed increased TNBS labelling in air and only modest rises with anoxia. Taken together, these data imply a rearrangement of membrane lipids during the sickling process and suggest a permanent reorganization of membrane lipids in the irreversibly sickled cell.
Ethanol Inhibition of Rabbit Reticulocyte Haem Synthesis at the Level of δ‐Aminolaevulinic Acid Synthetase
Authors: Ibrahim Nader G. , Spieler Paul J. , Freedman Michael L.
Published: March 2008

Summary. Ethanol inhibition of rabbit reticulocyte protein synthesis occurs as a result of a decrease in haem synthesis. The present study therefore was undertaken in order to localize the inhibitory site of ethanol on the haem biosynthetic pathway. Ethanol (0.05–0.15 m) inhibition of reticulocyte protein synthesis was prevented by simultaneous incubation with 0.025‐i mmδ‐aminolaevulinic acid (ALA). Ethanol inhibited both 14C‐glycine and 14C‐ALA incorporation into haem. However, the extent of haem formation with 14C‐ALA as substrate in the presence of ethanol was still equal to that when 14C‐glycine was used. These data suggest that ethanol inhibits the haem synthetic pathway at several loci, but that the decrease in haem synthesis, responsible for the decrease in protein synthesis, is due to the inhibition at the rate‐limiting enzyme, δ‐aminolaevulinic acid synthetase (ALA‐S). To confirm this, ALA‐S activity was then directly measured in intact reticulocytes, and it was shown that ethanol indeed inhibited its activity. The inhibition of ALA‐S was prevented by 10−4m dibutyryl cyclic AMP (db cAMP) or theophylline, agents which elevate intracellular cAMP and which have previously been shown to prevent and reverse ethanol inhibition of haem and protein synthesis. Thus, it appears that cAMP protects against ethanol toxicity by preventing inhibition of ALA‐S.
Evidence for the Platelet Specificity of β‐Thromboglobulin and Studies on its Plasma Concentration in Healthy Individuals
Authors: Ludlam C. A.
Published: March 2008

Summary. The concentration of normal human platelet β‐thromboglobulin (β‐TG) was measured in various washed organ samples by a radioimmunoassay. As only trace amounts were detected, β‐TG appears to be a platelet specific protein. Assay of β‐thromboglobulin in plasma samples from 180 normal individuals gave a range of 10–65 ng/ml. In the 10 subjects studied, plasma β‐TG concentration was related to platelet lifespan but not to turnover. The plasma β‐TG concentration rose with increasing age but did not correlate with the whole blood platelet count or the percentage of megathrombocytes. These results provide further substantial evidence that measurement of plasma β‐TG concentration is useful for assessing the participation of platelets in various disease processes.
The Separate Effects of H+ and 2,3‐DPG on the Oxygen Equilibrium Curve of Human Blood
Authors: Samaja M. , Winslow R. M.
Published: March 2008

Summary. Addition of non‐saturating amounts of 2,3‐DPG (2,3‐diphosphoglycerate) within the red cell (2,3‐DPG/haemoglobin less than I) intially reduces Hill's parameter, n. With increasing 2,3‐DPG/haemoglobin, n increases until a maximum is reached at 2,3‐DPG/haemoglobin greater than I. Thus, 2,3‐DPG influences the shape as well as the position of the whole blood oxygen equilibrium curve (OEC). The importance of this effect on the oxygen carrying capacity of the blood is considered. The effect of 2,3‐DPG on the position of the OEC (p50, the pO2 at one‐half maximal O2 saturation) is via its allosteric effect on haemoglobin at 2,3‐DPG/Haemoglobin less than I. Above that ratio, its effect is to reduce intracellular relative to the extracellular pH.
An Examination of Complement Proteins on Membranes of Paroxysmal Nocturnal Haemoglobinuria (PNH) and PNH‐like Red Cells
Authors: Burapakulsolsri Noppaporn , Yuthavong Yongyuth , Wilairat Prapon
Published: March 2008

Summary. Proteins obtained from membranes of PNH erythrocytes and of red cells treated with 2‐aminoethylisothiouronium bromide (AET) following lysis in acidified serum (Ham's test) were compared by sodium dodecylsulphate‐polyacrylamide gel electrophoresis. The peptide patterns were identical, providing support that AET‐treated red cells afford a suitable experimental substitute for PNH red cells in studies involving complement fixation.
Diagnosis of Haematological Disease using Anti‐i
Authors: Shumak Kenneth H. , Rachkewich and Rose A. , Beldotti Lorraine E.
Published: March 2008

Summary. Leukaemic blast cells were obtained from the blood of six patients with acute lymphoblastic leukaemia (ALL) and 15 patients with acute myeloblastic leukaemia (AML). The blasts were compared with lymphocytes from normal subjects in cytotoxicity and 125I‐labelled antibody binding tests using several examples of anti‐i. As much i antigen was detected on ALL blasts as on normal lymphocytes; much less i antigen was detected on AML blasts. Studies of three patients with morphologically undifferentiated acute leukaemia suggest that, in tests with anti‐i, blasts from such patients react either like lymphoblasts or myeloblasts despite the absence of the corresponding morphological features.
Further Studies on a Specific Platelet Antibody Found in Bernard‐Soulier Syndrome and its Effects on Normal Platelet Function
Authors: Tobelem G. , Levy‐Toledano S. , Nurden A. T. , Degos L. , Caen J. P. , Malmsten C. , Kindahl H.
Published: March 2008

Summary. An IgG antiplatelet antibody found in a multitransfused patient with Bernard‐Soulier syndrome (BSS), reacted with a normal platelet surface antigen of 150 000 daltons which was similar to the glycoprotein missing from BSS platelets. The BSS platelet antibody (BSS‐Pab) aggregated all control platelets which then released ADP and 5‐HT and synthesized thromboxane. When mixed with the antibody, BSS platelets did not aggregate, did not release ADP and 5‐HT and failed to synthesize thromboxane. The BSS‐Pab was not inactivated by incubation with BSS platelet stroma. While the antibody did not aggregate thrombasthenic platelets, its aggregating activity was lost after incubation with their stroma. The BSS‐Pab did not provoke ADP or 5‐HT release or thromboxane synthesis in thrombasthenic platelets or in the platelets of a patient with platelet cyclooxygenase deficiency or in normal platelets treated with indomethacin. The aggregating, release and synthetic responses of platelets after binding of BSS‐Pab to its membrane antigen (probably glycoprotein I) requires the presence of glycoprotein IIb and/or IIIa and the normal metabolism of arachidonic acid.
Heat‐Induced Erythrocyte Fragmentation in Neonatal Elliptocytosis
Authors: Zarkowsky Harold S.
Published: March 2008

Summary. Erythrocytes from neonates with elliptocytosis were studied for their pattern of heat‐induced fragmentation. Membrane alterations began at 44°C. There was a gradual progression in shape changes as the temperature was increased to 47°C, at which point frank fragmentation occurred. Normal erythrocytes show no morphologic changes until the critical temperature of fragmentation, 49°C is reached. Heat studies were repeated a few months later, at a time when the patients' erythrocyte morphology had become more typical of elliptocytosis. Morphologic changes occurred abruptly at 48°C with complete fragmentation. Increased thermal sensitivity of the red cell membrane has previously been demonstrated for pyropoikilocytes, and these studies suggest that some cases of elliptocytosis may be mild expressions of a similar membrane defect.
Stimulation of Human Haemopoietic Cells by Colony Stimulating Factors: Sensitivity of Leukaemic Cells
Authors: Francis G. E. , Berney J. J. , Chipping P. M. , Hoffbrand A.V.
Published: March 2008

Summary. The sensitivity of populations of human granulocyte precursors to factors with colony stimulating activity (CSA) was assessed in agar culture in vitro. The mean threshold of stimulation was estimated by analysis of dose‐response curves of clone growth against concentration of CSA. This method of assessment has the advantage that CSA production by cells contaminating the population under test does not affect results. Marrow cells from patients with acute myeloid leukaemia were less sensitive to CSA than marrow cells from normal individuals. Sensitivities of cells from chronic granulocytic leukaemia and from chronic myelomonocytic leukaemia were within the normal range, but this group also tended to require more CSA than controls. In addition, sensitivities of granulocyte precursors from patients with acute myeloid leukaemia were closely related to the culture pattern, and thus to the remission probability. The significance of this relationship is discussed.
Anticoagulant Activities of High and Low Molecular Weight Heparin Fractions
Authors: Barrowcliffe Trevor W. , Johnson Edward A. , Eggleton C. Anne , Kemball‐Cook Geoffry , Thomas Duncan P.
Published: March 2008

Summary. The anticoagulant activities of high and low molecular weight heparin fractions were measured by three assay methods, both in vitro, and after intravenous injection in volunteers. The low molecular weight (LMW) fraction had similar anti‐Xa activity in vitro to the high molecular weight (HMW) fraction, but in APTT assays the HMW fraction was about twice as potent. After intravenous injection, the two fractions gave equal heparin levels by anti‐Xa assays, but in APTT assays the LMW fraction had only half the activity of the HMW fraction. Anti‐Xa assays using synthetic substrate S‐222 gave about 20% lower levels than anti‐Xa clotting assays for both heparins. Complete protamine neutralization of the post‐injection heparin activity was found in APTT and synthetic substrate assays, but about 20% of the clotting anti‐Xa effect could not be neutralized. Complete neutralization of the fractions by protamine was shown by all three in vitro. This non‐neutralizable activity probably accounts for the difference between the anti‐Xa clotting and synthetic substrate assays. Studies by crossed immunoclectrophoresis and affinity chromatography indicated that the antithrombin III binding properties of the two fractions were similar.
The Adrenalin Binding Site on Human Platelets
Authors: McMillan R. , Bakich M. J. , Yelenosky R. J.
Published: March 2008

Summary. Using 3H‐adrenalin the quantity and characteristics of the adrenalin receptors on human platelets were evaluated. Maximum adrenalin binding by intact platelets from eight normal subjects averaged 3.2 ng adrenalin/108 platelets, equivalent to 105 000 binding sites per platelect. Bound3H‐adrenalin could be dissociated or binding could be inhibited by an excess of unlabelled adrenalin, noradrenalin, isoproterenol or dopamine; phentolamine or propanalol had no effect on 3H‐adrenalin binding, even in concentrations 10 000‐fold greater. These studies show that the epinephrine binding site can be detected and quantitated on human platelets and that its characteristics differ from those classically attributed to either alpha or beta adrenergic receptors.
Isozymes of Amino Acid Naphthylamidase in Human Haematopoietic Cell Lines
Authors: Slnna G. Afbu , Lundgren E. , Nllsson K. , Roos G.
Published: March 2008

Summary.

The occurrence of isozymes of the proteolytic enzyme amino acid naphthylamidase was investigated in a panel of 44 neoplastic and non‐neoplastic haematopoietic cell lines by polyacrylamide gel electrophoresis. Two isozymes (A and C) were common for all lines, whereas different forms of a third isozyme with intermediate electrophoretic mobility (B) appeared in nine lines. Isozyme A seemed to be associated with cell proliferation and was not a marker for malignancy. The appearance of different B isozymes not present in normal peripheral blood lymphocytes, occurred in some Burkitt lymphoma lines and in two non‐malignant lines maintained for a long time in culture. In three lines derived from myeloid leukaemia a variant of isozyme B seemed to be a marker for cell origin.
ABH Antigens and Bone Marrow Transplantation
Authors: Hershko Chaim , Gale Robert Peter , Ho Winston , Fitchen John
Published: March 2008

Summary.

We studied the role of ABH antigens in determining graft outcome in 104 patients who received HLA‐identical bone marrow transplants for aplastic anaemia and acute leukaemia. ABH compatibility had no significant effect on incidence of graft rejection or graft‐versus‐host disease. Fifteen recipients had pre‐transplant antibodies against donor ABH antigens. In 14, large volume plasma exchange and transfusion of donor‐type erythrocytes was successful in reducing the antibody titre to low or undetectable levels. In one patient, plasma exchange was unsuccessful and red cells were removed from the marrow inoculum by unit gravity sedimentation. This approach prevented transfusion reaction, and permitted engraftment of all haematopoietic cell lines despite persistently elevated antibody titres. Parallel in vitro studies revealed that antibodies to ABH antigens failed to inhibit the growth of progenitor cells committed to both granulocyte‐macrophage (CFU‐C) and erythroid (BFU‐E) development. These findings indicate that ABH‐antigens are not clinically important transplantation antigens and suggest that ABH antigens are not operationally present on hematopoietic stem cells.