bluuebunny/crossref_metadata_2025 huggingface dataset:
A Comparative Study of Acute Myeloblastic Leukaemia in Children and Adults
Authors: Ansari B. M. , Thompson E. N. , Whittaker J. A.
Published: March 2008

Summary.

The presenting features and clinical course of 89 adults and 15 children with acute myeloblastic leukaemia (AML) presenting to a Regional Leukaemia Centre has been analysed. Remission rate was related to age, being 40% for the total adult group and 60% for all children. Young adults and children had a particularly high remission rate, whilst elderly patients faired badly. Survival diminished with increasing age and patients who entered complete remission survived for a significantly longer time (P<0.001) than those who did not.

Adult AML differs from childhood AML, the adults having a lower remission rate, a significantly shorter survival (P< 0.005) and almost complete absence of second remissions. Adults showed no correlation between complete remission and initial WBC or initial blast cell count, but in children there was a significant correlation (P<0.05) between initial total WBC and complete remission. A significant correlation between initial platelet count and complete remission could not be demonstrated in either group. Although the numbers of children are small, preadolescent children may represent a favourable sub‐group, particularly those between 7 and 8 years of age.
The Intracellular Content of the Three Transcobalamins at Various Stages of Normal and Leukaemic Myeloid Cell Development
Authors: Zlttoun J. , Marquet J. , Zittoun R.
Published: March 2008

Summary.

A study of the three serum transcobalamins (TC) in myeloproliferative disorders and acute leukaemia has suggested a granulocyte origin for the three binders; the later stages of this cell line might be releasing ‘TC I’and ‘TC III’while ‘TC II’which is increased in sera from patients with acute myelogenous leukaemia, probably originates at least in part from myeloblasts.

The intracellular content of each TC has been studied in normal and leukaemic bone marrow cells after separation on a discontinuous bovine serum albumin gradient. Myeloblasts contain a TC II‐like protein almost exclusively but promyelocytes contain a TC I‐like protein which increases as the cells mature whilst ‘TC II’decreases. A TC III‐like binder parallels ‘TC I’but appears later and reaches a maximum in the polymorphonuclear cells. In various types of leukaemia, abnormal TC distributions are related either to marrow replacement by the blast cells or to a disorder of the maturation process in the leukaemic cell line.
Binding of Vitamin B12‐Rat Transcobalamin II and Free Vitamin B12 to Plasma Membranes Isolated from Rat Liver
Authors: Fiedler‐Nagy Christa , Rowley George R. , Coffey J. W. , Miller O. N.
Published: March 2008

Summary.

When dialysed rat serum which contains a single, low molecular weight binder for vitamin B12, rat transcobalamin II (rat TC‐II), was labelled in vitro with 57Co‐vitamin B12 and then incubated at 30°C (pH 7.5) with vesicles of highly purified plasma membranes separated from microsomal fractions of rat liver by density gradient centrifugation, the 57Co‐vitamin B12‐rat TC‐II complex bound to high affinity sites on the vesicles via a specific (binding after correction for ‘nonspecific’binding in the presence of a large excess of the non‐radioactive complex), saturable, and reversible interaction. The apparent affinity constant for the binding reaction was 5.5 × 109 M‐ 1. Using the same incubation conditions, free vitamin B12 also bound to the vesicles of plasma membranes via a specific, saturable, but apparently irreversible interaction. Preincubation of the membranes with free vitamin B12 did not interfere with the subsequent binding of the vitamin B12‐rat TC‐II complex to the membranes; however, preincubation with the vitamin B12‐rat TC‐II complex did interfere, to some extent, with the subsequent binding of free vitamin B12. Dialysed rat serum, perhaps the free rat TC‐II in the dialysed serum, also inhibited the binding of the vitamin B12‐rat TC‐II complex to the plasma membranes. The relationship of the binding sites identified in this report to the absorption of vitamin B12 by rat liver, and thus their physiological significance remains unknown until further work is done, perhaps using intact hepatocytes.
Plasma Heparin Levels after Low Dose Subcutaneous Heparin in Patients Undergoing Hip Replacement
Authors: Brozović Milica , Stirling Yvonne , Abbosh J.
Published: March 2008

Summary.

Plasma heparin levels were measured at 2 and 8 h after 5000 iu of heparin subcutaneously in 43 patients awaiting total hip replacement. At 2 h, 37 patients had plasma levels between 0.05 and 0.15 iu/ml, three patients had more than 0.2 iu/ml, and three had only trace levels. At 8 h, most patients had less than 0.1 iu/ml, and six patients had no detectable plasma heparin. There were no correlations between plasma heparin levels and age, sex, body weight or use of analgesics. Patients with high 2 h plasma heparin levels may bleed at operation, whereas those with very low plasma heparin concentrations may be at risk of post‐operative venous thrombosis; it may therefore sometimes be necessary to monitor plasma heparin levels in order to determine the optimal regime in individual cases.
Uptake of Transferrin‐Bound Iron by Rat Cells in Tissue Culture
Authors: Beamish M. R. , Keay L. , Okigaki T. , Brown E. B.
Published: March 2008

Summary.

The in vitro uptake of transferrin‐bound iron by rat liver cells and rat embryo cells in culture, and by rat reticulocytes, was studied using labelled rat serum.

The uptake of iron by rat liver cells was linear with time and showed a curvilinear relationship with the serum iron concentration. Over 40% of the cell radioactivity was found to be ferritin associated. Incubation of rat transferrin doubly labelled with 59Fe and 125l showed a difference between the uptakes of the two isotopes suggesting reflux of transferrin from the cell after iron uptake. In order to compare iron uptake from half saturated (Tf‐Fe 1) and fully saturated (Tf‐Fe2) transferrin, the three cell systems were each incubated in sera labelled with 55Fe at 10% saturation and with 59Fe at varying iron saturations ranging from 10% to 90%. Similar experiments were performed with sera having been randomly labelled with 55Fe and labelled with 59Fe but preincubated with reticulocytes in order to produce iron bound to non‐reticulocyte orientated transferrin binding sites. The data obtained confirms the presence of a functional heterogeneity with respect to the half and fully saturated transferrin. Thus iron uptake from fully saturated transferrin was almost twice that for the half saturated molecule with rat liver cells or reticulocytes, and over three times that of the half saturated molecule with rat embryo cells. In contrast no such differences could be detected when iron uptake from serum randomly labelled with iron and serum with labelled non‐reticulocyte orientated binding sites were compared.

Finally, the effect of pre‐incubating each of the three cell systems in unlabelled sera of varying percentage iron saturations on subsequent radio‐iron uptake was studied. An inverse relationship between percentage saturation of the preincubation serum and subsequent uptake was demonstrated indicating that cellular mechanisms also influence cell iron uptake.
Forms of Vitamin B12 in Blood and Bone Marrow in Patients with Pernicious Anaemia
Authors: Abe Tsukasa , Gibbs Bernard , Cooper Bernard A.
Published: March 2008

Summary.

The biologically‐active forms of vitamin B12 in blood and bone marrow and changes induced in these by injections of cyanocobalamin have been measured in patients with pernicious anaemia. Bone marrow methylcobalamin was low before therapy, and increased 24 h after therapy. The largest portion of bone marrow vitamin B12 was 5′deoxyadenosylcobalamin, and this increased more than did methylcobalamin during the 24 h after injection of cyanocobalamin. A single injection of roo μ of cyanocobalamin induced about 10 times the increase of the intracellular coenzyme forms of vitamin B12 in bone marrow than followed injection of 100 μg. Plasma methylcobalamin was extremely low before therapy, and increased only moderately 24 h after therapy; the majority of plasma vitamin B12 remaining as cyanocobalamin. In contrast, only a minority of intracellular bone marrow vitamin B12 was cyanocobalamin 24 h after injection of cyanocobalamin.

The degree of anaemia did not correlate with bone marrow methylcobalamin, nor did bone marrow cobalamin correlate significantly with cobalamin content of washed blood erythrocytes. Correlation was observed between intra‐erythrocyte vitamin B12 content and the degree of anaemia; the correlation being inverse with haemoglobin concentration in the peripheral blood. Inverse correlation also was observed between MCV and erythrocyte folate content.

These studies suggest that megaloblastic maturation appears at different concentrations of bone marrow vitamin B12 in different patients, presumably because in vitamin B12 deficiency, eventual limitation of normoblastic maturation may be determined by factors such as folate metabolism, vitamin B12 binders, and the affinity of vitamin B12 dependent enzymes. Although the clinical response to 1000 μg of cyanocobalamin does not differ from that to 100 μg, the concentration of vitamin B12 coenzymes in bone marrow cells was proportional to the cyanocobalamin injected.
Intracellular Localization of Hepatic Propionyl‐CoA Carboxylase and Methylmalonyl‐CoA Mutase in Humans and Normal and Vitamin B12 Deficient Rats
Authors: Frenkel Eugene P. , Kitchens Richard L.
Published: March 2008

Summary.

The intracellular localization of the enzymes in the vitamin B12 dependent pathway which involves the oxidation of propionate was studied in rat liver obtained from normal and vitamin B12 deficient rats as well as from man. The subcellular site of propionyl CoA carboxylase and the vitamin B12 dependent‐methylmalonyl CoA mutase were determined. All of the activity of these two enzymes was demonstrated to be in the mitochondria and those enzymes were shown to be loosely bound to the inner membrane‐matrix portion of the mitochondria. Vitamin B12 deficiency did not alter the subcellular localization. Finally, a rapid enzymatic assay for methylmalonyl‐CoA mutase was described.
Haem Synthesis in the Diamond‐Blackfan Syndrome
Authors: Freedman Melvin H. , Amato Dominick , Saunders E. Fred
Published: March 2008

Summary.

The response of bone marrow to erythropoietin (EPO) from five children with the Diamond‐Blackfan syndrome, also known as congenital hypoplastic anaemia (CHA), was tested in tissue culture by measurement of haem synthesis. Studies of 13 control marrows indicated that the maximum EPO effect occurred at approximately 70 h incubation using an EPO concentration of 0.2–0.3 units/ml and a nucleated cell concentration of 5 × 106 per culture. Under these conditions, haem synthesis was 121% greater in EPO‐stimulated than in unstimulated cultures. Patients with CHA with anaemia and diminished marrow erythroids had reduced or absent haem synthesis. In one patient, haem production became normal after a spontaneous remission, and was not inhibited by autologous plasma drawn at the time of diagnosis. Plasma from three patients did not show inhibitory activity when cultured with control marrow. In contrast, plasma from an adult with acquired pure red cell aplasia produced striking inhibition of haem synthesis when cultured with control marrow. We conclude that, in comparison to some cases of the adult acquired condition, CHA is not due to inhibitors or antibodies. When present, erythroid precursors in children with CHA are capable of responding normally to EPO with increased haem synthesis.
Iron Absorption: Present State of the Art
Authors: Jacobs A. , Worwood M.
Published: March 2008

Summary.

The availability of food iron is one of the major factors determining the total amount of iron absorbed. Iron is released from food in the stomach in an ionizable form or as haem. These forms are affected differently by both the composition of the diet and gastrointestinal secretions. Iron is taken up by the epithelial cells of the small intestine and is released from haem in the same cells. This uptake may be related to the iron status of the cells. Iron transport within these cells has been studied by both subcellular fractionation and electron microscopy with autoradiography. The biochemical studies have shown that much of the mucosal iron is associated with the mitochondria whereas absorbed radioiron is largely bound to ferritin in iron replete rats and to a transferrin‐like protein in iron deficient animals. It has been suggested that this latter protein is the intracellular ‘carrier’of iron. Serosal transport to the plasma is influenced by a number of factors and is not clearly understood. A review of the major problems in this area reveals more questions than answers.
Intestinal Absorption of Folic Acid Monoglutamates and Polyglutamates: a Brief Review of Some Recent Developments
Authors: Butterworth C. E. , Krumdieck C. L.
Published: March 2008

Summary.

Recent evidence indicates that there are important species differences in the digestion and intestinal absorption of pteroylpolyglutamates, the predominant form of the folate vitamin(s) in food. Birds appear to possess a pteroylpolyglutamyl hydrolase, or conjugase, in extracts of pancreas and intestinal mucosa, that acts as an endopeptidase and produces pteroyldiglutamate as a major end product. The precise anatomical site of pteroylpolyglutamate cleavage (whether intraluminal, intracellular, or at the cell surface) has not been established with certainty in the chick, although it has been assumed to occur in the lumen. Several lines of current investigation indicate that in mammalian systems conjugase is primarily an intracellular, lysosomal enzyme, and that it is not localized in brush‐border fractions of intestinal epithelium. There appears to be relatively little conjugase activity in the proximal small bowel lumen of humans or dogs. Human liver conjugase appears to act as an exopeptidase, cleaving terminal glutamate residues from the gamma‐glutamyl chain one at a time, with pteroylmonoglutamate as the end product. These and other observations are compatible with the view that (at least in some mammalian systems) pteroylpolyglutamates are absorbed intact by intestinal epithelial cells and cleaved to monoglutamates at an intracellular site.
Haemoglobin Tak: a β‐Chain Elongation
Authors: Lehmann H. , Casey R. , Lang A. , Stathopoulou R. , Imai K. , Tuchinda S. , Vinai P. , Flatz G.
Published: March 2008

Summary.

In Haemoglobin Tak two normal α‐chains are combined with two β‐chains elongated by 11 residues beyond the C‐terminus. Unlike in the α‐chain abnormal Hb Constant Spring, the elongation cannot result from a point mutation of a stop codon. It is probably due to an unequal crossing‐over near the 3′ end of the β‐chain structural gene. This could cause either a deletion of the normal stop codon or a shift in the reading frame. Oxygen dissociation of purified Hb Tak shows no cooperativity but in Hb A + Tak haemolysates there is no interaction between the two above 40% O2 saturation. Heterozygotes for Hbs A and Tak show an imbalance of globin chain synthesis (α/non α= 1.5), the synthesis of β Tak resembles that of the β‐chain in β+ thalassaemia.
Quantitation of Subclasses of Mononuclear Cells in Normal Human Blood by Membrane Receptor Studies
Authors: Habeshaw J. A. , Young Gillian A.
Published: March 2008

Summary. A technique for quantitating mononuclear cells expressing E, EA(IgG), EAC receptors, immunoglobulin carrying cells and phagocytic cells in normal human venous blood is described; normal values for each of these classes of mononuclear cell are established. The effects of different methods of cell preparation have been considered: these were found not to have a significant effect on the final values. In addition, we describe a second category of ‘fluorescent non‐capping’ cells and show that these are numerically equivalent to cells which ingest neutral red.
The Development of Radioimmunoassays for Fibrinogen Degradation Products: Fragments D and E
Authors: Gordon Y. B. , Martin M. J. , Landon J. , Chard T.
Published: March 2008

Summary. Fibrinogen degradation products, fragment D (FgD) and fragment E (FgE) have been measured in human serum by specific radioimmunoassays. In addition, the appearance of a neoantigenic determinant on FgD, revealed when fibrinogen is degraded by plasmin has been utilized to develop a specific radioimmunoassay for FgD in plasma (FgDneo). The reagents and conditions used in each assay are described in detail. The mean specific activity was 144 μCi/μg for 125I‐labelled FgE and 82 μCi/μg for 125I‐labelled FgD. Separation of antibody bound and free antigen was achieved using second antibody. The detection limits of the FgE, FgD and FgDneo assays were 0.8, 1.0 and 6.2 ng/ml respectively. The specificity of each assay with respect to fibrinogen and its degradation fragments has been assessed. Fibrinogen and fragment X cross‐reacted markedly in both the FgE and FgD assays, whereas the cross‐reaction of fibrinogen was abolished in the FgDneo assay, while the cross‐reaction of fragment X was 10%, indicating gradual emergence of the neoantigenic site during digestion of fibrinogen. The sensitivity, precision, and specificity of the radioimmunoassay systems described have major advantages over the existing procedures for the measurement of fibrinogen degradation products.
Globin Chain Synthesis in the Greek Type (Aγ) of Hereditary Persistence of Fetal Haemoglobin
Authors: Sofroniadou K. , Wood W. G. , Nute P. E. , Stamatoyannopoulos G.
Published: March 2008

Summary. Globin chain synthesis was studied in a family with both the Greek (Aγ) type of hereditary persistence of fetal haemoglobin and β thalassaemia. The ratio of α/(γ+β+δ) chain synthesis in the hereditary persistence of fetal haemoglobin (HPFH) heterozygotes was 0.97 while in the HPFHβ‐thalassaemia heterozygote it was 2.14. However, calculation of the amounts of haemoglobin synthesized per cell suggests that in the HPFHβ‐thalassaemia heterozygote, the β‐ and Aγ‐chain genes in cis to the HPFH determinant are unable to compensate for the deficiency of chains imposed by the β‐thalassaemia gene in trans and that the increased synthesis of Hb F is directed by the y‐chain genes located on the β‐thalassaemia chromosome. The data suggest that synthesis of β and Aγ chains in the Greek HPFH is fixed at a ‘preset’ level and indicate that the defect might be due to an abnormality in the rate of transcription of the closely linked β‐, δ‐ and γ‐chain genes.
Lymphocyte Transformation in Patients with Paraproteinaemia
Authors: Campbell A. E. , De Vine Julie , Azam L. , Hamid J. , Delamore I. W. , McFarlane H.
Published: March 2008

Summary. The transformation of lymphocytes in cultures with phytohaemagglutinin (PHA) or with pokeweed mitogen (PWM) was carried out during treatment on 28 patients with paraproteinaemia and on 20 control subjects. 78.5% of the peripheral blood lymphocyte short‐term cultures from the treated patients showed a subnormal PHA‐induced but high PWM‐induced lymphocyte transformation. The other 21.5% of the treated patients had a normal pattern of lymphocyte transformation. Eight of 12 untreated patients with multiple myelomatosis and four of five untreated patients with benign paraproteinaemia also had a subnormal lymphocyte transformation to PHA but a high response to PWM. Serial lymphocyte transformation studies in eight of 18 myeloma patients during chemotherapy revealed that the initial subnormal lymphocyte transformation to PHA subsequently became higher than that to PWM. 82% of the bone marrow cultures from the patients showed a higher PWM‐induced than PHA‐induced lymphocyte response, also suggesting a B cell type of disease. In both dose response and time dependent investigations peak transformation of peripheral blood lymphocytes did not always coincide.
Methaemoglobin in Man Living at High Altitude
Authors: Gourdin D. , Vergnes H. , Gutierez N.
Published: March 2008

Summary. Methaemoglobin has been found in the red cells of subjects living permanently at high altitudes (above 3500 m). The level of methaemoglobin appeared to be inversely related to the red cell count and was much increased in anaemic subjects. The methaemoglobinaemia disappeared when the subjects descended to low altitude. The mechanism of this methaemoglobinaemia is not clear. Its presence appears to be related to the hypoxia of altitude, for toxic factors, enzymopenia and the presence of haemoglobin M were eliminated. The regulation of the oxygen dissociation curve in this condition is discussed.
Changes in Folate Metabolism in Baboons Papio cynocephalus Treated with 2‐Methyl‐2‐aminopropanol‐B12
Authors: Siddons R. C.
Published: March 2008

Summary. After the intramuscular administration of the vitamin B12 analogue, 2‐methyl‐2‐aminopropanol‐B12, to vitamin B12‐deplctcd baboons (Papio cynocephalus) the effects on vitamin B12 and folate metabolism have been studied. The ‘apparent’ serum vitamin B12 and the scrum folate concentrations increased, the liver folate concentrations decreased, and the liver vitamin B12 concentrations were unchanged. The analogue caused a marked increase in the amount of formiminoglutamic acid excreted after a histidine load whereas the amount of methylmalonic acid excreted after a valine load decreased. The rate of red blood cell regeneration after venesection was normal in baboons treated with the analogue and there was no evidence of megaloblastic haemopoiesis. Two baboons given the analogue developed a partial loss of utilization and lack of co‐ordination in the hind limbs.
A Basophilic Micro‐organism Infecting Human Red Cells
Authors: Clark K. G. A.
Published: March 2008

Summary. Clusters and chains of basophilic micro‐organisms were found on the red cells of a woman who suffered from malignant melanoma. The infection was clinically silent for at least 7 years. During her last year of life the patient became febrile and anaemic. Her spleen enlarged and the basophilic bodies on her red cells became coarser. The infecting organism is referred to only as a ‘basophilic agent’, since its relationships to recognized genera are uncertain. Similar epicrythrocytic parasites occur in a wide variety of animals. Though these infections are often latent, they profoundly modify host immune responses.
Coagulation in an Agarose Gel and its Application to the Detection and Measurement of Factor VIII Antibodies
Authors: Bird Prudence
Published: March 2008

Summary. Evidence has been presented that fibrin formation may be detected directly in agarose gels containing citrated plasma when they are treated with thrombin or calcium chloride. A new assay is described for haemophilic factor‐VIII antibody based on inhibition of fibrin formation within an agarose gel containing normal plasma.
Platelet Kinetics in Idiopathic Thrombocytopenic Purpura (ITP) before and at Different Times after Splenectomy
Authors: Branehög Ingmar
Published: March 2008

Summary. Platelet kinetics were studied in 18 patients with idiopathic thrombocytopenic purpura (ITP) before splenectomy, 1 month and 8 months after splenectomy. A gamma function was used for the calculation of platelet mean life span (MLS).

Before splenectomy platelet MLS was greatly reduced and platelet production was 2.8 times normal. The peripheral platelet count and platelet recovery were significantly related to platelet MLS. One month after splenectomy platelet MLS increased appreciably while platelet production decreased but continued to be significantly above normal. These changes together with the removal of the splenic platelet pool resulted in a peripheral platelet count significantly higher than normal and a total platelet mass not differing from normal. Eight months after splenectomy platelet MLS increased slightly but was not significantly different from that found 1 month after splenectomy. Platelet production, however, decreased significantly and became normal. This resulted in a normal peripheral platelet count and a decreased total platelet mass which was significantly lower than in non‐splenectomized control subjects. The results suggest that platelet production is regulated by a feed‐back mechanism that is governed by the peripheral platelet count. The high platelet count encountered 1 month after splenectomy is due to a sluggishness of this regulatory mechanism.
Skin Window Studies in Polycythaemia Rubra Vera
Authors: Ghosh M. L. , Hudson G. , Blackburn E. K.
Published: March 2008

Summary. The inflammatory response was studied in patients with primary polycythaemia by means of a modified skin window technique. In untreated patients, the overall cellularity was a prominent feature and, as compared with the controls, the 48 h preparations showed a significantly greater percentage of granulocytes with a corresponding decreased percentage of macrophages. In the peripheral blood of these patients, both total white cells and granulocyte counts were significantly higher than in the control subjects. A highly significant relationship between the blood and skin window neutrophils was demonstrated. In polycythaemia patients in remission, the results were generally similar to the controls, except that there was still a highly significant increase in the percentage of skin window basophils.

The results provided no evidence of a defect in cell migration or in the phagocytic activity of macrophages in polycythaemia vera. Possible mechanisms of the cutaneous basophilia and its relevance to the manifestation of this disorder are discussed.
Observations on the Ultrastructure of Erythropoietic Cells and Reticulum Cells in the Bone Marrow of Patients with Homozygous β‐Thalassaemia
Authors: Wickramasinghe S. N. , Bush Valerie
Published: March 2008

Summary. An electron microscopic study of marrow fragments from patients with homozygous β‐thalassaemia has shown that 3% of early polychromatic erythroblast profiles and 20% of late polychromatic erythroblast profiles contain intracytoplasmic α‐chain precipitates. Various nuclear abnormalities were found including the loss of parts of the nuclear membrane and the presence of intranuclear α‐chain precipitates, and these abnormalities were virtually confined to the non‐dividing, late polychromatic erythroblasts. As most profiles of the proliferating early polychromatic erythroblasts did not contain intracytoplasmic or intranuclear α‐chain precipitates, it is suggested that the arrest of many of these cells in the G1 phase of the cell cycle may be related to the presence of an excess of free α‐chains rather than to the presence of α‐chain precipitates within them. The cytoplasm of the bone marrow reticulum cells contained early and late polychromatic erythroblasts at various stages of degradation, providing direct evidence of ineffective erythropoiesis.
A Congenital Haemolytic Anaemia with Thermal Sensitivity of the Erythrocyte Membrane
Authors: Zarkowsky H. S. , Mohandas Narla , Speaker Connie B. , Shohet S. B.
Published: March 2008

Summary Microspherocytes, measuring 2‐3 μm in diameter, and cells with blunted projections or triangular in shape characterized the erythrocyte morphology in three children with congenital haemolytic anaemia. Since the erythrocyte morphology resembled that associated with thermal injury, heat‐induced changes in erythrocyte morphology and membrane composition were studied. Erythrocytes developed filaments and spheroid bodies which fragmented, resulting in micro‐spherocyte transformation. Normal cells required exposure to 49° C, whereas, the patients’ cells fragmented at 45°incubation of patients’ cells at 37°C for 1711. The heat‐induced transformation of the patients’ cells was associated with an increase in the membrane cholesterol :phos‐ pholipid and cholesterol :protein ratios. The phospholipid :protein ratio was unchanged. This suggests that fragmentation produces a selective loss of membrane components. Splenectomy ameliorated the haemolytic process. We propose that the patients’ red‐cell morphology is the result of in vivo fragmentation, and that the spleen is the major site of microspherocyte and poikilocyte destruction.
Erythropoietin Responsiveness in Polycythaemia Vera
Authors: Golde David W. , Cline Martin J.
Published: March 2008

Summary A liquid bone marrow culture technique was used to study erythropoiesis in polycythaemia vera. Three patients with the diagnosis of polycythaemia vera treated only by phlebotomy were compared to three healthy volunteers. Erythropoiesis in vitro was assessed by counts of nucleated red cells and 3H‐thymidine incorporation into erythroid precursors. Human urinary erythropoietin was found to stimulate erythropoiesis in cultures from the polycythaemia patients as well as normals. The erythropoietin effect was blocked by a rabbit anti‐erythropoietin antibody. These results suggest that some erythroid precursor cells maintain an intact erythropoietin response mechanism in polycythaemia vera. They also support clinical studies of this disorder indicating that some regulation by erythropoietin may occur in vivo.
Effects of Vincristine on Normal and Stimulated Megakaryocytopoiesis in the Rat
Authors: Ebbe Shirley , Howard D. , Phalen Elizabeth , Stohlman F.
Published: March 2008

Summary Vincristine was given to rats in which thrombocytopoiesis was either normal or acutely or chronically stimulated by injections of heterologous antiplatelet serum. A single dose of 0.3 mg/kg was given intravenously. The drug produced an early and a delayed megakaryocytopenia suggesting that it was toxic to differentiated megakaryocytes as well as to proliferating stem cells. The results support the hypothesis that vincristine‐induced thrombocytosis may be due to homeostatic adjustments which, in turn, are activated as a result of drug‐induced cytotoxicity.
Identification of Haemoglobin S in Red Cells and Normoblasts, Using Fluorescent Anti‐Hb S Antibodies
Authors: Papayannopoulou TH. , Mcguire T. C. , Lim G. , Garzel E. , Nute P. E. , Stamatoyannopoulos G.
Published: March 2008

SUMMARY Anti‐haemoglobin S antibodies were raised in horses and purified by affinity chromatography. These antibodies recognize β6 valine, while they fail to bind to haemoglobins with a glutamyl (Hb A) or a lysyl (Hb C) residue in this position. The purified anti‐Hb S antibodies were composed of equine IgG(a, b) and IgG(T) subclasses and failed to cause precipitation with Hb S, evidently because of the bivalency of both antibodies and antigen. Identification of Hb S in single erythrocytes was achieved by reacting the fluorescein isothiocyanate‐conjugated anti‐Hb S antibodies with the haemoglobin antigen in fixed blood smears. All the red cells from persons with sickle cell trait and sickle cell anaemia are labelled with the anti‐Hb S‐FITC, while red cells from persons with normal haemoglobin or other mutant haemoglobin genotypes are not. Erythrocytes from new‐borns with sickle cell trait bind the anti‐Hb S antibody, although the intensity of labelling varies from cell to cell. This method of cellular identification of Hb S preserves the morphological characteristics of the reactive cells and permits observations of the pattern of haemoglobin synthesis among cells of various degrees of maturity. Fluorescent identification of Hb S in fixed blood smears may be applicable to prenatal diagnosis of haemoglobinopathies.
Quantitation of Ineffective Erythropoiesis from the Incorporation of [15N]Delta‐aminolaevulinic Acid and [15N]Glycine into Early Labelled Bilirubin: II. ANAEMIC PATIENTS
Authors: SAMSON Diana , HALLIDAY D. , Nicholson D. C. , Chanarin I.
Published: March 2008

SUMMARY The incorporation of [15N]δ‐aminolaevulinic acid and [15N]glycine into haemoglobin haem and early labelled bilirubin was measured in subjects with various haematological disorders. The clearance of [14C] bilirubin was used to measure bilirubin production rate, and the magnitude of the various sources of bilirubin production and the percentage ineffective erythropoiesis were calculated. Ineffective erythropoiesis was found to be a major factor in the production of the anaemia in patients with the following disorders: megaloblastic anaemia associated with the Lesch‐Nyhan syndrome, thalassaemia intermedia, sideroblastic anaemia, and the anaemia of chronic disorders. In three patients with iron‐deficiency anaemia ineffective erythropoiesis was increased, but was of minor importance in the production of the anaemia, while in two patients with aplastic anaemia and one with macrocytosis of alcoholism there was no increase in ineffective erythropoiesis.
The Cryopreservation of Leukaemia Cells: Morphological and Functional Changes
Authors: Lowenthal R. M. , Park Diana S. , Goldman J. M. , Th'ng K. H. , Hill R. S. , Whyte G.
Published: March 2008

SUMMARY. Stored autologous haemopoietic cells may be used to repopulate the bone marrow of patients in the advanced stages of different leukaemias who have received cytotoxic drugs. We have used a continuous flow blood cell separator to collect peripheral blood leucocytes from patients with chronic granulocytic leukaemia (CGL) before treatment. We also collected bone marrow cells from patients with CGL and from patients with acute myeloid leukaemia in complete remission. The collected cells were frozen at 1° per minute using dimethyl sulphoxide as cryoprotective agent and stored in liquid nitrogen. For reconstitution of frozen cells we found that the use of dextran 110 inhibited leucoagglutination. The viability and function of the reconstituted leucocytes were assessed by their morphological appearance, their capacity to phagocytose and kill Candida albicans organisms, their ability to reduce the dye nitroblue tetrazolium (NBT) in vitro and to incorporate tritiated thymidine into DNA and by the growth of colony forming units (CFUc) in agar culture. With this method of cryopreservation the phagocytic function of mature neutrophils is retained to some extent but their capacity to reduce NBT and their microbicidal activity are completely lost. In contrast CFUc may remain after storage for periods of at jeast 2 years.
Blast Crisis of Chronic Myeloid Leukaemia (CML): II. CELL SURFACE MARKER ANALYSIS OF ‘LYMPHOID’AND MYELOID CASES
Authors: Janossy G. , Greaves M. F. , Revesz T. , Lister T. A. , Roberts Marion , Durrant Jill , Kirk Barbara , Catovsky D. , Beard M. E. J.
Published: March 2008

SUMMARY. Fourteen cases of Philadelphia chromosome (Ph1) positive chronic myeloid leukaemia in blast transformation have been investigated using cell surface markers. Morphologically eight cases were lymphoid and the remainder myeloid in appearance. All cases were negative with surface markers for thymocytes and T and B lymphocytes. Five of the lymphoid cases reacted with an antiserum specific for acute lymphoid leukaemia (ALL) of non‐T non‐B type and were also weakly reactive with a lymphocyte reactive antiserum. A sixth patient, whose blast cells were anti‐ALL negative (ALL‐) at presentation, subsequently developed central nervous system leukaemia with anti‐ALL positive (ALL+) blast cells in the CSF. In all cases the leukaemic blast cells showed greatly diminished expression of cholera toxin receptors when compared to granulocytic cells from the chronic phase of CML. This parallels weak or negligible expression of the cholera toxin receptor in ALL and AML.

These results suggest that the blastic phase of CML may involve different cellular derivatives of a pluripotential stem cell in which the primary malignant/genetic changes reside. The blast crisis of CML can therefore be heterogeneous with respect to cellular expression and in a significant proportion of patients involves a cell which is by membrane markers and morphological criteria indistinguishable from that seen in the common form of ALL. In these cases the Philadelphia chromosome may be the only distinguishing cellular characteristic.
Studies on the Proportion and Synthesis of Haemoglobin G Philadelphia in Red Cells of Heterozygotes, a Homozygote, and a Heterozygote for both Haemoglobin G and α Thalassaemia
Authors: Milner P. F. , Huisman T. H. J.
Published: March 2008

SUMMARY. The proportion of Hb G Philadelphia (α68‐Asnalys) in heterozygotes has been found to have a well‐defined bimodal distribution around means of 33% and 46% Hb G. Microcytosis and hypochromia are consistently associated with the latter group, who also have a decreased ratio of α/β‐chain synthesis in the peripheral blood, but these characters are not linked to the Hb‐Gα gene, because a parent with microcytosis and 46% Hb Gα may have offspring with 33% Hb G without significant microcytosis. In one family a subject with Hb G and Hb G2 but no Hb A or Hb A2 is presumably a homozygote for αG. This subject has microcytosis and a decreased ratio of α/β chain synthesis. In another family a subject with Hbs H, G and G2 but without Hbs A or A2 is heterozygous for both Hb G and α thalassaemia 1. These findings are compatible with the hypothesis that the αG mutation occurs on a chromosome with only a single αchain locus and that the expression in heterozygotes as 46% or 33% Hb G is determined by the homologous chromosome in trans having either one or two normal αA genes respectively. The significance of this polymorphism for chromosomes carrying αchain genes is discussed.
Chelate Mediated Transfer of Iron from Transferrin to Desferrioxamine
Authors: Pollack Simeon , Aisen Philip , Lasky Fred D. , Vanderhoff Grace
Published: March 2008

SUMMARY. Desferrioxamine, widely used for the treatment of iron overload in Cooley's anaemia, binds iron so tightly that it should quantitatively remove iron from transferrin. Studies conducted in vivo and in vitro, however, have failed to demonstrate significant depletion of transferrin‐bound iron by a stoichiometric excess of desferrioxamine. However, low molecular weight chelating agents, capable of forming ternary complexes with transferrin and ferric iron, can promote a rapid transfer of iron from transferrin to desferrioxamine. A possible mechanism for this facilitated exchange is offered.
A Comparison of Platelet Size, Platelet Count, and Platelet 35S Incorporation as Assays for Thrombopoietin
Authors: Mcdonald T. P.
Published: March 2008

SUMMARY. Average platelet size, platelet count, and 35S‐incorporation into platelets were compared as methods for the measurement of thrombopoietin‐stimulated thrombopoiesis. In mice injected with rabbit anti‐mouse platelet serum (RAMPS) average platelet size was shown to be increased as mice were recovering from thrombocytopenia. Also, 35S‐measurements on platelets of these mice showed significant increases in cpm/average platelet 2–4 days after RAMPS treatment. Significant increases in 35S‐incorporation into the total circulating mass of platelets were found on days 3–4. In normal mice or mice in rebound‐thrombocytosis injected with thrombopoietin, platelet size remained unchanged, whereas the platelet count and 35S‐incorporation into platelets were shown to be significantly increased. Moreover, a dose‐response experiment in mice pretreated with RAMPS showed a slight increase in platelet count as the dose of TSF was increased, but platelet sizes were unaltered. The %35S‐incorporation into platelets showed a significant linear dose‐response, i.e. as the dose of thrombopoietin was increased, an increase in %35S‐incorporation into platelets was observed. These data indicated that of the three indirect measurements of thrombopoietin, the %35S‐incorporation into mouse platelets was the most sensitive, followed by platelet counting; the least sensitive measurement of thrombopoiesis was change in platelet size.
Binding of Thrombin to Human Platelets and its Possible Significance
Authors: Ganguly P. , Sonnichsen W.J.
Published: March 2008

SUMMARY Thrombin binds tightly to human platelets. The binding reaction is dependent on the thrombin concentration used. At a physiologically significant thrombin concentration, there are about 500 binding sites per platelet with an apparent dissociation constant of 0.02 u/ml. Autoradiography studies of platelets treated with labelled thrombin showed that the thrombin was located on the platelet surface. Separation of the subcellular fractions of platelets treated with labelled thrombin by density gradient centrifugation revealed that the membrane area contained over 80% of the radioactivity initially applied. Furthermore, isolated platelet membranes bind thrombin similar to intact platelets. These data suggest that the receptors for thrombin are located on the platelet membrane. Cytochalasin A, cytochalasin B or prostaglandin E1 did not have any effect on thrombin binding although these agents inhibited platelet aggregation. Thus, binding of thrombin is not sufficient for aggregation of platelets and other steps are involved. These agents do not affect the induction of stimulation but interfere at a later step in the thrombin‐platelet interaction. On the other hand, hirudin completely inhibited binding of thrombin to platelets. It appears that the platelet receptor recognizes that part of the thrombin molecule on its surface which is blocked by hirudin. Binding studies with serotonin loaded platelets showed a close correlation between thrombin binding and the release reaction.
The Significance of Plasma Lipoproteins on Erythrocyte Aggregation and Sedimentation
Authors: Scherer R. , Morarescu A. , Ruhenstroth‐Bauer G.
Published: March 2008

Increased erythrocyte aggregation can be induced by high concentrations of human lipoproteins. The dependence of aggregate formation on lipoprotein concentration was recorded by determination of erythrocyte sedimentation rate (ESR), by electrical measurement of the erythrocyte aggregation index (EAI) and by scanning electron microscopy. The lipoprotein concentrations necessary to induce a significantly increased ESR in an otherwise normal human plasma are much too high to be encountered in physiologic or even severe pathologic states. Therefore hyperlipoproteinaemia by itself cannot explain a raised ESR. In cases where the ESR is raised due to the presence of increased amounts of other erythrocyte aggregating plasma proteins (agglomerins), hyperlipoproteinaemia can contribute to a limited extent to the increase in ESR. The possible pathophysiological significance of the demonstrated erythrocyte aggregating capacity of human liproteins in a microvascular environment is noted.
Factor VIII and Factor‐VIII‐Related Antigen in Multiple Myelomatosis and Related Conditions
Authors: Gomperts E. D. , Shulman G. , Lynch S. R.
Published: March 2008

Factor VIII and factor‐VIII‐related antigen (FVIIIRA) were estimated in 19 patients with paraproteinaemia. The FVIIIRA levels were above the normal range in 12 cases with 11 of these showing an abnormal FVIII/FVIIIRA ratio. The patients could be subdivided into three groups according to their FVIIIRA levels: normal levels, levels greater than normal but less than 200%, greater than 200%. This subdivision correlated with the degree of bone involvement as assessed by radiological skeletal survey.
Effects of Sulphydryl Compounds on Abnormal Red Cell Pyruvate Kinase
Authors: Zanella A. , Rebulla P. , Giovanetti A. M. , Curzio M. , Pescarmona G. P. , Sirchia G.
Published: March 2008

The effectof some sulphydryl compounds on two new variants of red cell pyruvate kinase (ATP: pyruvate phosphotransferase, PK) is reported. In vitro a striking correction has been obtained of both the qualitative and, in one case, the quantitative defect of red cell PK. In vivo, a correction of the qualitative and quantitative abnormalities has been produced in both patients, with clinical improvement of one of them. These findings, together with the unexpected results in respect to the functional properties of PK found in the affected members of the two families studied, suggest that the PK abnormality is not the cause of the haemolytic anaemia, but an epiphenomenon of a primary unknown defect that apparently involves the red cell thiol groups.
Sodium Pertechnetate as a Red Cell Label: In Vitro and In Vivo Studies
Authors: Schmidt Peter , Lohrmann Hans‐Peter , Heimpel Hermann
Published: March 2008

In vitro investigations were performed to re‐examine the variables that influence the binding of sodium 99Tcm‐pertechnetate to human red blood cells. Irreversible binding of the radioisotope was observed only when red cells were incubated with tin chloride after their incubation with pertechnetate; binding of pertechnetate is completed within 10 min after its addition to red cells. Composition, temperature and possibly the oxygen saturation of the incubation medium affect the fraction of the isotope that becomes red cell‐bound.

Using a labelling method derived from these in vitro studies, the usefulness of pertechnetate‐labelled autologous red cells for the determination of red cell volumes was studied. Elution of pertechnetate occurred in vivo in an exponential fashion. Using the specific red cell activity, corrected for elution, of blood samples drawn 30 miu after injection of 99Tcm‐labelled red cells, calculated red cell volumes were almost identical to those determined simultaneously with radiochromate‐labelled cells. It is concluded that 99Tcm‐pertechnetate is a useful label for red cell volume determination; it may be less reliable when delayed mixing of the labelled red cell tracer in the patient's circulation is expected.
Distribution of Granulopoietic Activity in the Human Skeleton, Studied by Colony Growth in Agar Diffusion Chambers
Authors: Gordon M. Y. , Douglas I. D. C. , Clink H. M. , Pickering B. M. J.
Published: March 2008

The colony forming ability of bone marrow cells from 110 haemato‐logically normal patients was investigated by their growth in semi‐solid agar in intraperitoneal diffusion chambers. Marrow aspirated from the sternum showed a consistently higher colony yield than that from the iliac crest. Study of site variation in the individual was possible in donors for bone marrow transplantation. In each of four cases, sternal marrow produced more colonies than any other site. Examination of smears made from the aspirated samples suggested that the difference in yield was not due to varying dilution by peripheral blood on aspiration from different sites. It is concluded, therefore, that the incidence of colony precursor cells is not uniform throughout the active marrow.
Lysosomal Acid Hydrolases in Human Lymphocyte Subpopulations
Authors: Barr R. D. , Perry S.
Published: March 2008

Acid phosphatase and beta‐glucuronidase activities have been determined cytochemically in T, B and null lymphocytes as part of an effort to characterize the human haemopoietic stem cell. A combination of weak acid phosphatase activity and strong beta glucuronidase staining, in the form of a single large granule, has been shown to be specific for T cells. On the basis of this approach alone, non‐T cells could not be further subclassified. Further cytochemical evaluation is being explored.