bluuebunny/crossref_metadata_2025 huggingface dataset:
The Ultrastructure of Dyserythropoiesis in Aplastic Anaemia
Authors: Frisch B. , Lewis S. M. , Sherman D.
Published: March 2008

Summary Electron microscopy of erythropoiesis in aplastic anaemia has demonstrated some unique features of dyserythropoiesis at the ultrastructural level, which contribute to understanding the morphological features which are seen in the light microscope. The findings provide further evidence for the presence of qualitative defects in erythropoiesis in aplastic conditions. The alterations seen included bi‐and multinucleated cells, intercellular bridges containing microtubules, inter‐nuclear chromatin bridges, irregular nuclear shapes, juxta‐nuclear and peripheral cisternal structures, ringed sideroblasts, a diversity of intranuclear inclusions, nuclear clefts in both eu‐ and heterochromatin, and a variety of anomalies of the nuclear membrane. The significance of the occurrence of these anomalies in aplastic anaemia is considered and the role of the nuclear membrane and of endoplasmic reticulum in the pathogenesis of aplastic anaemia is discussed.
Isolation and Properties of a Low Molecular Weight Antiplasmin of Human Blood Platelets and Serum
Authors: Mui P. T. K. , James H. L. , Ganguly P.
Published: March 2008

Summary A low molecular weight antiplasmin has been detected in human blood platelets. The antiplasmin is dialysable and a similar material is present in normal plasma. A simple method for rapid isolation of the antiplasmin based on ultrafiltration is described. The inhibition of plasmin by these materials under different conditions has been studied. In the ultracentrifuge, both antiplasmins showed a single broad peak with an approximate sedimentation coefficient of 0.5S. Paper chromatography and paper electrophoresis indicated the isolated material to be heterogeneous. The individual components were isolated by paper chromatography and the antiplasmin activity was measured by the fibrin plate method. Preliminary studies on the fraction with maximal antiplasmin activity suggest that the inhibitory effect might be due to enzyme‐inhibitor complex formation. Based on the present data, it is concluded that the platelet antiplasmin and the serum antiplasmin are very similar. This antiplasmin material may be useful in fibrinolytic therapy.
Labile Aggregation Stimulating Substance (LASS): the Factor from Storage Pool Deficient Platelets Correcting Defective Aggregation and Release of Aspirin Treated Normal Platelets
Authors: Gerrard J. M. , White J. G. , Rao G. H. R. , Krivit W. , Witkop C. J.
Published: March 2008

Summary Mixtures of equal volumes of Hermansky‐Pudlak (HP) syndrome (adenine nucleotide storage pool deficient) and aspirin treated (AT) platelets (inhibited release of adenine nucleotides) undergo irreversible aggregation when exposed to collagen or adrenaline, whereas neither alone will do so. The present investigation has explored the basis for the mutual correction. Correction in the mixed system was accompanied by release of significant amounts of [14C]5‐hydroxy‐tryptamine from the AT platelets, whereas very little isotope was released when AT platelets alone were exposed to collagen. The correction of aggregation and the release of isotope could both be suppressed by pre‐treating HP platelets with aspirin. Labile aggregation stimulating substance (LASS) composed of two closely linked intermediates of prostaglandin (PG) E2 and PGF2a biosynthesis, produced using a microsomal fraction of HP platelets, could correct the aggregation and secretion defect of AT platelets exposed to collagen. These findings indicate that HP platelets when mixed with AT platelets and exposed to collagen secrete a substance which is responsible for the correction. LASS, identified as the factor involved, acted as an intercellular messenger which mediated the correction by overcoming the influence of aspirin on normal cells.
Anti‐Wrb, and Other Autoantibodies Responsible for Positive Direct Antiglobulin Tests in 150 Individuals
Authors: Issitt P. D. , Pavone B. G. , Goldfinger D. , Zwicker H. , Issitt C. H. , Tessel J. A. , Kroovand S. W. , Bell C. A.
Published: March 2008

SUMMARY Eluates from the red blood cells (and sera whenever free autoantibody was present) of 150 individuals with positive direct antiglobulin tests, have been studied for antibody specificity. Of 87 patients with AIHA, 64 had autoantibodies reacting with all red cell samples including Rhnull. Of these 64 anti‐dl autoantibodies, two were, and 32 contained, auto‐anti‐Wrb. Of 33 patients being treated with alphamethyldopa, who had developed positive direct antiglobulin tests, 23 had anti‐dl autoantibodies four of which contained auto‐anti‐Wrb. Of 30 haematologically normal donors with positive direct antiglobulin tests, 23 had anti‐dl autoantibodies, two of which were, and six of which contained, auto‐anti‐Wrb. The full specificities of autoantibodies, other than anti‐Wrb and anti‐dl, in the 150 patients are described, as are the natures of the protein red cell coatings that caused the positive direct antiglobulin tests. The presence of free serum autoantibody as a correlate of the three clinical conditions is reported.

Several observations on auto‐anti‐Wrb are documented. The antibody can cause gross red cell destruction in vivo, but can be benign on other occasions; it occurs with approximately the same frequency in AIHA patients and ‘normal’donors with positive direct antiglobulin tests, but in fewer patients with alpha‐methyldopa induced positive direct antiglobulin tests; it does not activate complement in vivo;, and finally it may eventually provide a clue to the aetiology of AIHA.
Simultaneous Inheritance of Mutant Isoenzymes of Erythrocyte Pyruvate Kinase Associated with Chronic Haemolytic Anaemia
Authors: Paglia D. E. , Gray G. R. , Growe G. H. , Valentine W. N.
Published: March 2008

SUMMARY The heterogeneity of pyruvate kinase (PK) deficiency associated with hereditary haemolytic anaemia is emphasized by studies of a kindred harbouring two distinct mutant forms of this enzyme, both of which were kinetically defective with markedly decreased affinities for the substrate, phosphoenolpyruvate. The two isoenzymes, designated PK‐Vancouver1 and PK‐Vancouver2, were primarily distinguishable from one another by differences in maximum in vitro activities and by variations in response to fructose‐1,6‐diphosphate activation. When combined in proband erythrocytes to the exclusion of any normal PK, the isoenzymes were associated with a severe chronic haemolytic process with many of the features of PK deficiency of the more common quantitative type. Clinical laboratory screening tests for detecting PK deficiency may be falsely negative or equivocal in such cases.
A Convenient and Reproducible Filtration Technique for the Determination of Erythrocyte Flexibility
Authors: Baar S.
Published: March 2008

SUMMARY. A micro‐cell filtration technique is described in which a silver membrane of 3 μm porosity is used. The apparatus consists of a membrane holder and disposable syringe only. A regeneration procedure which allows indefinite re‐use of one membrane is described. Filtration proceeds under the hydrostatic pressure of the fluid column only. A Locke‐albumin solution has been developed as a suspension medium and adapted for cell washing by the addition of fibrinogen and cholesterol. A plot of the volumes of cells filtered against log time shows a linear relationship for normal erythrocytes. The filtered red cells are virtually undamaged. The overall precision of the method is of the order of ± 2%.
Evidence that Collagen Releases Human Platelet Constituents by Two Different Mechanisms
Authors: AKBAR HUZOOR , Ardlie Neville G.
Published: March 2008

SUMMARY

Collagen is believed to be involved in the initial events in haemostasis and has been shown by others to cause platelet aggregation and release, and also to initiate the intrinsic pathway of coagulation. The present experiments provide evidence which suggests how these many effects of collagen may be involved in haemostasis.

It is shown here that collagen releases platelet constituents by two different pathways. Collagen causes platelets washed free of loosely adsorbed coagulation factors to release constituents. This activity is, therefore, independent of the intrinsic pathway of coagulation, and is not inhibited by heparin or hirudin. Collagen also releases platelet constituents by an alternative pathway which is inhibited by heparin and hirudin and is independent of factor XII, but is dependent on factor XI, subsequent factors in the intrinsic pathway of coagulation and calcium. These results suggest that collagen‐induced release of platelet constituents is in part due to a direct effect on the platelet, and, in part, to an indirect effect involving coagulation factors and mediated by thrombin. The present results suggest that irreversible aggregation by collagen is also mediated by thrombin.

The possible significance of this dual action of collagen in the haemostatic process is shown in Fig 7.
PROGRESSIVE MULTIFOCAL LEUCOENCEPHALOPATHY SUCCESSFULLY TREATED WITH CYTOSINE ARABINOSIDE
Authors: Buckman R. , Wiltshaw E.
Published: March 2008

Progressive multifocal leucoencephalopathy (PML) is a rare and fatal demyelinating disease of the central nervous system, that occurs in patients with reticuloendothelial neoplasms and in other diseases associated with defects of the immunological system (Åstrom et al, 1958). To date, two cases have been reported in which the patients have recovered from the PML after treatment with cytosine arabinoside (Bauer et al, 1973; Marriott et al, 1975). We would like to add a third case, that of a woman with Hodgkin's disease in whom a diagnosis of PML was made by exclusion.
Blast Crisis of Chronic Myeloid Leukaemia (CML): I. PRESENTATION SIMULATING ACUTE LYMPHOID LEUKAEMIA (ALL)
Authors: Beard M. E. J. , Durrant Jill , Catovsky D. , Wiltshaw E. , Amess J. L. , Brearley R. L. , Kirk Barbara , Wrigley P. F. M. , Janossy G. , Greaves M. F. , Galton D. A. G.
Published: March 2008

SUMMARY. Seven patients presenting as an acute leukaemia caused difficulty in diagnosis. The lymphoid appearance of the blast cells either initially or during treatment suggested acute lymphoid leukaemia (ALL). In each case the Philadelphia chromosome was shown to be present thus suggesting that these cases were examples of chronic myeloid leukaemia (CML) presenting in blast crisis without a detectable chronic phase.

The implications of these findings are discussed and the difficulty in achieving a precise diagnosis in the acute leukaemias is emphasized. Cytogenetic analysis should be carried out whenever the type of acute leukaemia present is of critical importance.
Alpha‐Thalassaemia Trait in Various Racial Groups in the United Kingdom: Characterization of a Variant of Alpha‐Thalassaemia in Indians*
Authors: Walford Diana M. , Deacon Rosemary
Published: March 2008

SUMMARY. Patients whose red‐cell indices are suggestive of thalassaemia trait, but who have a normal haemoglobin electrophoretic pattern, may be carriers of αthalas‐saemia. A diagnosis of αthalassaemia trait was made in 44 such patients, using the incorporation of [3H]leucine by reticulocytes to measure the relative rates of synthesis of the α and β‐chains of adult haemoglobin. Patients with αthalassaemia trait had a reduced rate of synthesis of the αchains, with a mean α/β specific activity ratio of 0.79 ± SD 0.07. The mean α/β specific activity ratio of 20 control subjects was 1.06 ± SD 0.08. The diagnostic value of the haemoglobin H (Hb H) preparation was assessed in proven αthalassaemia heterozygotes of various races. A high proportion of ‘false negative’results in Indian and Negro heterozygotes indicated that the Hb H preparation is a highly unreliable screening test for use in a multi‐racial population. There was no significant difference in the mean level of Hb A2 in αthalassaemia heterozygotes (2.0 ± SD 0.6) compared with that of the control group (2.1 ± SD 0.5). Comparison of data from patients with α and β‐thalassaemia traits showed that αthalassaemia trait is the milder disorder, in terms of its effects on red‐cell morphology, red‐cell indices and degree of globin chain imbalance. Amongst individual patients with αthalassaemia trait, there was no correlation between the α/β specific activity ratio and the red‐blood‐cell (RBC) count, mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH). This suggests that the α/β ratio cannot be used to distinguish between carriers of a mild gene (‘silent carriers’) and carriers of a more severe disease.

This is the first study to characterize αthalassaemia trait in Indians, in whom it appears to be a common disorder. Haematologically, αthalassaemia trait in Indians is milder than that seen in Chinese and the fact that haemoglobin Bart's hydrops fetalis does not occur in Indians makes it likely that the genetics of Indian αthalassaemia differ from those of the Chinese disease. A possible genetic model for Indian αthalassaemia is discussed and the identification of the homozygote is seen as the first step in the determination of the underlying molecular defect.
Iron Uptake and Ferritin Synthesis by Peripheral Blood Leucocytes from Normal Subjects and Patients with Iron Deficiency and the Anaemia of Chronic Disease
Authors: Summers M. R. , Jacobs A.
Published: March 2008

SUMMARY. The ferritin content of monocytes, lymphocytes and polymorphs is reduced in patients with iron deficiency anaemia. In patients with the anaemia of chronic disease a reduced serum iron concentration is associated with an increase in the ferritin content of all peripheral blood leucocytes. Iron uptake by all cell types is related to transferrin saturation. In iron deficiency anaemia lymphocyte iron uptake is greatly increased, possibly reflecting intracellular iron depletion. In patients with active rheumatoid arthritis and carcinomatosis there is no alteration in leucocyte iron uptake or ferritin synthesis.
Activation of Platelets by Platelet Activating Factor (PAF) Derived from IgE‐Sensitized Basophils
Authors: Henson Peter M. , Landes Richard R.
Published: March 2008

SUMMARY Platelet activating factor (PAF) derived from antigen‐stimulated, IgE‐sensitized rabbit basophils acts on platelets to induce aggregation and secretion of their content of granule‐bound vasoactive amines. Despite this, PAF did not activate platelet factor 3. In contrast, collagen induced aggregation, secretion and PF3 activation in the washed platelets. Other stimuli (ADP, C3b, thrombin) also initiated both secretion and PF3 activation. A wide dose range of PAF, including those giving maximal secretion and aggregation, were ineffective in making PF3 available and the possibility that PAF inhibited PF3, or its generation, was also excluded. It is concluded that PAF is a unique stimulus for platelets and that secretion and aggregation are not necessarily accompanied by PF3 generation.
Factor VIII and Human Platelet Aggregation
Authors: Bottecchia D. , Vermylen J.
Published: March 2008

SUMMARY Bovine factor VIII aggregates human platelets either in a strong single wave at high concentration (10 ug/ml platelet suspension) or in two waves at low wave at low concentration 2–1 ug/ml). The strong single wave of aggregation is not associated of [14C]serotonin or β‐glucuronidase; the high concentration retraction of reptilase‐clotted platelet‐rich plasma. With the low release of [14C]serotonin is observed just prior to the onset of the aggregation; release of β‐glucuronidase does not occur at any low concentration of bovine factor VIII induces moderate retraction of platelet‐rich plasma, which is inhibited by acetylsalicylic acid, in and apyrase, indicating that it is a consequence of release of platelet phosphate.

It has previusly been suggested that carbohydrate groups are involved in the bovine factor VIII interaction, since galactose oxidase and periodate the platelet aggregating activity of bovine factor VIII. The present these oxidizing substances also induce a degradation of bovine the exact role of carbohydrate groups in the aggregation process established.
Factor VIII and Human Platelet Aggregation
Authors: Bottecchia D. , Vermylen J.
Published: March 2008

SUMMARY During digestion of highly purified bovine factor VIII or neuraminidase‐treated human factor VIII by plasmin the procoagulant activity is destroyed more rapidly than the aggregating activity. At an intermediate stage, fragments are transiently formed, which inhibit platelet aggregation by the respective undigested materials, but without crossed inhibition. During proteolysis by plasmin, human factor VIII retains antigenic and ristocetin cofactor properties. Contrary to previous observations obtained with less purified preparations, plasmin digests of human or bovine factor VIII do not inhibit ADP‐induced platelet aggregation. Thrombin and reptilase do not modify the aggregating activities of bovine or neuraminidase‐treated human factor VIII.
Haemolytic Effect of Two Sulphonamides Evaluated by a New Method
Authors: Gaetani G. D. , Mareni C. , Ravazzolo R. , Salvidio E.
Published: March 2008

A technique to investigate drugs which could cause haemolysis in subjects deficient in glucosc‐6‐phosphate dehydrogenase (D‐glucose‐6‐phosphate: NADP oxidoreductase; G6PD) has been developed. The method is based on the technique of 14CO2 evolution during the incubation of normal erythrocytes in the presence of [I‐14C] glucose and their own serum, the latter containing the active metabolites of the drugs received by normal subjects. By this method agents causing a stimulation of the hexosemonophosphate pathway of normal erythrocytes should be regarded as potentially haemolytic for G6PD‐deficient subjects. Two sulphonamides, sulphormethoxine and sulphalene, of which until now no haemolytic effects have been reported, together with chloroquine, have been investigated. While chloroquine does not affect the hexosemonophosphate shunt of normal erythrocytes, the two sulphonamides stimulate this pathway. The results are confirmed by the reduction of the half‐life of 51Cr‐labelled G6PD‐deficient red cells (Mediterranean variant), after administration of the two sulphonamides.
Function and Ultrastructure of Platelets of Neonates: Enhanced Ristocetin Aggregation of Neonatal Platelets
Authors: Ts'ao Chung‐hsin , Green David , Schultz Klara
Published: March 2008

We have investigated platelet morphology and function in human maternal‐newborn pairs. Fibrinogen concentration and factor‐VIII activity in plasma were also determined. Our results showed that, compared to maternal platelets, neonatal platelets were poorly responsive to adenosine diphosphate, adrenaline and collagen. Uptake of labelled serotonin by neonatal and maternal platelets was the same, but release of the radioactivity was reduced in the former. Phagocytic activity of neonatal platelets, demonstrated with latex particles, was similar to that of maternal platelets. Although the ultrastructure of neonatal platelets approximated that of maternal platelets, immature appearing platelets were occasionally found in the neonatal samples. An unanticipated finding was that platelet aggregation induced by ristocetin was more vigorous in neonatal than in maternal platelet‐rich plasma samples. Furthermore, neonatal plasma, which had lower fibrinogen and factor‐VIII content than maternal plasma, facilitated maternal platelet aggregation by ristocetin and showed a greater ability than maternal plasma to promote ristocetin‐induced aggregation of platelets of a patient with von Willebrand's disease. These results indicate that the plasma of neonates contains large quantities of the ristocetin‐dependent platelet aggregation factor (RAF), probably more than is in maternal plasma, despite the higher levels of factor‐VIII procoagulant activity in the latter. Thus, in the newborn, there is a clear dissociation between factor‐VIII clotting activity and the RAF activity.
The Primary Rh0(D) Immune Response in Male Volunteers
Authors: Gunson H. H. , Stratton F. , Phillips P. K.
Published: March 2008

After a single antigenic stimulus, not greater than 5.0 ml R2 R2 red cells, anti‐D was detected in 79% of D‐negative volunteers increasing to 88% after subsequent spaced stimuli. The use of repeated antigenic stimuli at 2‐ or 4‐weekly intervals to induce the immune response did not appear to increase the frequency of responders. The results obtained are compared with those of other workers and evidence is presented to suggest that the immunogenicity of red cells from different donors may have a role in determining the frequency of responders with detectable anti‐D and is possibly associated with the R2 antigenic complex.
Inhibition of Membrane Peroxidation in Thalassaemic Erythrocytes by 2,3‐Dihydroxybenzoic Acid
Authors: Graziano J. H. , Miller D. R. , Grady R. W. , Cerami A.
Published: March 2008

Both iron and precipitated haemoglobin may catalyse the formation of free radicals, which in turn react with the polyunsaturated fatty acids of membranes leading to membrane failure and cell death. 2,3‐Dihydroxybenzoic acid (2,3‐DHB), a recently identified, orally effective iron‐chelating drug, inhibited membrane peroxidation in vitro in H2O2‐stressed crythrocytes from patients with beta‐thalassaemia major, beta‐thalassacmia intermedia, hacmoglobin Köln disease and sickle cell disease. We present evidence suggesting that the inhibition of peroxidation is due to the ability of 2,3‐DHB to scavenge free radicals via quinone formation, a mechanism analogous to that proposed for vitamin E.
The Patterns of Fetal Haemoglobin Production in Leukaemia
Authors: Sheridan B. L. , Weatherall D. J. , Clegg J. B. , Pritchard J. , Wood W. G. , Callender S. T. , Durrant I. J. , McWhirter W. R. , Ali M. , Partridge J. W. , Thompson E. N.
Published: March 2008

Elevated levels of haemoglobin F (Hb F) have been found in a wide range of haematological malignancies, but very high levels were found only in juvenile chronic myeloid leukaemia (JCML), and erythroleukaemia occurring in infancy. In both these disorders a reversion to a fetal form of erythropoiesis may occur, as judged by both the structure of the Hb F and by the disappearance of Hb A2 and the carbonic‐anhydrase isozymes during the course of the illness. The clinical picture of JCML is not always associated with a reversion to fetal erythropoiesis; there appears to be a heterogeneity of conditions with this clinical label. Thus the reversion to a completely fetal pattern of erythropoiesis seems to occur in a variety of leukaemias which start in early life. This change is associated with a uniformly bad prognosis. Of a group of 17 patients with acute myeloid leukaemia 15 developed an increase in the level of Hb F about 60 days after the commencement of treatment; significantly greater increases were observed in those achieving a clinical remission. The level of Hb F usually declined during remission but high levels persisted in a few cases. Increased levels of Hb F were found also in patients with other haematological malignancies who had undergone periods of marrow aplasia during treatment. In all cases the Hb F was heterogeneously distributed throughout the red cells. Analysis of γ15 or γCB3 peptides of Hb F from a variety of leukaemias gave glycine compositions ranging from 0.20 to 0.85 residues with many values in the fetal range; all cases with a reversion to fetal erythropoiesis had values in the fetal range. Attempts to confirm the ‘fetal’ origin of the cells containing Hb F by means of other markers was possible only in the cases of JCML and in one child with erythroleukaemia. These studies indicate that in some forms of leukaemia there may be a genuine reversion to fetal erythropoiesis while in others the emergence of cells containing Hb F appears to be part of a rapid regeneration process occurring after a period of marrow aplasia. The diagnostic and prognostic value of these observations is discussed.
Residual Marrow Damage: Possible Explanation for Idiosyncrasy to Chloramphenicol
Authors: Morley Alec , Trainor Kevin , Remes Judith
Published: March 2008

The effect of oral administration of chloramphenicol was studied in normal mice and mice having residual marrow damage. In the concentration used the drug had no detectable effect on the marrow of normal mice but caused a progressive fall in the numbers of pluripotential stem cells and granulocytic progenitor cells in mice bearing residual marrow damage. However, cells taken from such mice were no more sensitive than normal cells to the in vitro action of chloramphenicol. The results suggest that unrecognized residual marrow damage can be an aetiological factor in marrow failure due to drug idiosyncrasy.
Erythrocytosis Associated with Diffuse Parenchymal Lesions of the Kidney
Authors: Hoppin Edward C. , Depner Thomas , Yamuchi Hiroshi , Hopper James
Published: March 2008

Thirteen patients with diffuse parenchymal disease of the kidney and erythrocytosis are reported. In five the haematocrit was only minimally elevated because of an associated increase in plasma volume. The erythrocytosis in three patients was of particular interest: (1) transient appearance in a 23‐year‐old man with glomerulonephritis; (2) persistence despite the partial remission of nephrotic state in a 19‐year‐old man; (3) appearance following removal of a nephrosclerotic kidney from a 54‐year‐old woman. Because an associated expansion of plasma volume is not uncommon with parenchymal renal disease, it is suggested that the only reliable screening method for renal erythrocytosis is direct measurement of red cell volume.
The Role of Continuous Flow Centrifuge Leucapheresis in the Management of Chronic Lymphocytic Leukaemia
Authors: Fortuny I. E. , Hadlock D. C. , Kennedy B. J. , Theologides A. , McCullough J.
Published: March 2008

Continuous flow centrifuge leucapheresis (CFCL) provides a simple, mechanical technique for removing large numbers of circulating lymphocytes without cytotoxicity. Seven sequences of CFCL were performed on six patients with chronic lymphocytic leukaemia (CLL). In six, the median course consisted of six procedures in 10 days. A median total of 27.7 × 1011 leucocytes were removed and a 70% drop in leucocyte counts (from 433 × 109/l. to 130 × 109/l.) was observed with little fall in haemoglobin and platelet counts. Tissue masses decreased in two patients. A similar effect was seen in one patient managed less intensively with 21 procedures being performed over 73 days, removing a total of 87.6 × 1011 leucocytes. The peripheral leucocyte count decreased from 760 × 109/l. to 114 × 109/l. and hepatomegaly diminished markedly. One patient has survived for over 9 months since CFCL but stayed in control only briefly after each of two courses. Short courses of daily CFCL can produce short‐term benefit in the management of advanced CLL but at present its main value is as an investigational form of therapy which provides a unique opportunity to study the cytokinetics of advanced CLL without the complications of superimposed cytotoxic therapy.
Physiology and Ultrastructure of the Blood Platelet following Exposure to Hydrogen Peroxide
Authors: Rodvien Robert , Lindon Jack N. , Levine Peter H.
Published: March 2008

Summary.

Hydrogen peroxide in micromolar concentrations can induce shape change in human blood platelets, and can modify the aggregation and release reaction of these cells as induced by ADP or thrombin. In larger (millimolar) concentrations, H2O2 produces fusion of platelets with distortions in platelet morphology unlike those normally caused by aggregating agents. The production of H2O2in vivo by granulocytes or other cells could influence the processes of haemostasis or thrombosis.
Prophylaxis in Haemophilia: A Double‐blind Controlled Trial
Authors: Aronstam A. , Arblaster P. G. , Rainsford S. G. , Turk P. , Slattery M. , Alderson M. R. , Hall D. E. , Kirk P. J.
Published: March 2008

Summary.

A double‐blind controlled trial of prophylactic factor VIII therapy has been carried out on nine severe haemophiliacs at the Lord Mayor Treloar College. Infusions were given once weekly and calculated to give a post‐infusion plasma concentration of at least 0.25 I.U./ml of factor VIII. This regime reduced the overall bleeding frequency by 15%. The bleeding frequency in the first 3 days post‐infusion was reduced by 66%. A moderate overall reduction in morbidity was also achieved. It is calculated that to reduce the incidence of bleeding in severe haemophiliacs by 15% would require a 73% increased usage of therapeutic materials. More than twice this amount of material is likely to be needed to reduce the bleeding frequency of the same group by 66%.
Lymphocyte Response to Blood Transfusion in Man: a Comparison of Different Preparations of Blood
Authors: Hutchinson R. M. , Sejeny S. A. , Fraser I. D. , Tovey G. H.
Published: March 2008

Summary.

The appearance of atypical lymphocytes in post‐transfusion blood, their incorporation of tritiated thymidine in tissue culture and the elimination of cytotoxic antibody production, have been used as markers to show that frozen red cells are the least immunogenic when compared with dextran sedimented blood and whole blood donations. The absence of atypical lymphocytes and failure to produce lympho‐cytotoxic antibodies after transfusion of frozen cells is highly significant (P<0.001) when compared with whole blood donations.
Detection of Monocyte Specific Antigen on Human Acute Leukaemia Cells
Authors: Baker M. A. , Falk R. E. , Falk J. , Greaves M. F.
Published: March 2008

Summary.

Heteroantisera with specificity for human monocyte and macrophage antigens (HuMa) have been tested for activity against peripheral blood cells from patients with acute leukaemia. Using indirect immunofluorescence, blast cells from 25/27 patients with acute myelomonocytic leukaemia were reactive with antiHuMa antisera, whereas blasts from 19/20 patients with acute myeloblastic leukaemia and 13/13 patients with acute lymphoblastic leukaemia were not reactive. The F(ab1)2 fragment of the antibody retained specific activity. Activity of whole serum against myelomonocytic blast cells was specifically removed by absorption with macrophages. These observations indicate that circulating blast cells from patients with acute myelomonocytic leukaemia may carry differentiation antigens characteristic of monocytes and macrophages.
The Use of a Single Venous Blood Sample to Assess Oxygen Binding to Haemoglobin
Authors: Lichtman Marshall A. , Murphy Marion , Pogal Meredith
Published: March 2008

Summary.

The measurement of pH, Po2, Pco2 and So2 in a single venous blood sample can be used to determine the P50 at standard or at in vivo conditions. This technique makes it feasible for a physician, firstly, to make an assessment of the net adaptation of the red cell to reductions in blood oxygen content or flow and, secondly, to make an initial assessment of whether a haemoglobin with altered affinity for oxygen is present in subjects with polycythaemia or anaemia.
Induction of Haemolytic Anaemia by Substituted Phenylhydrazines
Authors: Itano H. A. , Hosokawa K. , Hirota K.
Published: March 2008

Summary.

The induction of anaemia and reticulocytosis by arylhydrazines was influenced by substituents on the benzene ring of phenylhydrazine. Arylhydrazines with ortho substituents, which would hinder the binding by haemoglobin of a ligand derived from the arylhydrazine, resulted in the least anaemia and reticulocytosis. These results are consistent with a previous finding that hydrazinobenzoic acid, which did not produce a ferrihaemochrome from ferrihaemoglobin, did not induce anaemia. The parallelism between the formation of a ferrihaemoglobin compound and the induction of anaemia supports the hypothesis that destabilization of haemoglobin by the binding of a ligand derived from the aryl portion of an arylhydrazine is an essential step in arylhydrazine‐induced haemolytic anaemia.
In Vitro Binding of Folates by Body Fluids
Authors: Retief F. P. , Heyns A. du P. , Oosthuizen Mariëtha , van Reenen O. R. , Badenhorst C. J.
Published: March 2008

Summary.

It is useful to differentiate between saturated folate binders in serum (carrying endogenous folate) and unsaturated binders (investigated in the present study). These two groups of binders need not necessarily be chemically identical and the unsaturated binder may even be an in vitro artifact, especially when measured with non‐physiological folates. Macromolecular binding of radio‐active N5‐methyl tetrahydrofolic acid (CH3H4PteGlu) and/or folic acid (PteGlu) by human serum and urine was assessed by means of exhaustive saline dialysis, haemoglobin‐coated charcoal adsorption, column chromatography with DEAE‐Sephadex A‐50, and sucrose gradient analysis. Binding was found to be minimal or absent. Charcoal adsorption showed a mean serum binding capacity of oμ/l. for PteGlu and 0.58 μ/1. for CH3H4PteGlu. In pregnancy the mean serum values were 0.23 μ/1. for PteGlu, 0.66 μ/1. for CH3H4PteGlu, and with folate deficiency 0.30 μ/1. for PteGlu, 0.49 μ/1. for CH3H4PteGlu. Mean urinary folate binding was minimal (<0.5 μ/1.), and red cell haemolysate similarly revealed very low binding on exhaustive dialysis. Column chromatography showed that tracer doses of [14C]PteGlu added to serum migrated distally to the protein zone; [14C]CH3H4‐PteGlu similarly showed no evidence of protein binding. On a sucrose gradient [14C]PteGlu also separated clear of the protein zone.
Inhibitors of Plasminogen Activation in Human Blood
Authors: Beattie A. G. , Ogston D. , Bennett B. , Douglas A. S.
Published: March 2008

Summary.

Plasma contained two inhibitors of plasminogen activation by urokinase when fractioned by gel filtration on Sephadex G‐200. The inhibitor in the lower molecular weight fractions was separate from the principal protease inhibitors of plasma: α1‐antitrypsin, α2‐macroglobulin, C1 inactivator and antithrombin III, and from factor XIII. This activation inhibitor was present in both plasma and serum and its recovery was not reduced by preincubating the serum for 6 h at 37°C. Its inhibitory activity was stable for several days at 4°C, and was enhanced in the presence of an increased saline concentration. Preparations of the inhibitor, active in a clot lysis system, failed to inhibit the esterase activity of urokinase on N‐α‐acetylglycyl‐L‐lysine methyl ester.
Nuclear DNA‐Polymerase Estimation in Human Leukaemic Myeloblasts
Authors: Wantzin Gunhild Lange , Karle Hans , Killmann Sven‐Aage
Published: March 2008

Summary. A technique which detects the presence of DNA‐polymerase in cell nuclei by measuring the incorporation of 3H‐thymidine‐5‐triphosphate (3H‐TTP) has been used to estimate the proportion of leukaemic myeloblasts which contains DNA‐polymerase and DNA capable of acting as primer‐template. In six cases of previously untreated acute myeloid leukaemia the 3H‐TTP labelling index (3H‐TTP LI) was much larger than the fraction in DNA synthesis. After a single ‘flash’ injection of cytosine arabinoside a pronounced decline was observed in the 3H‐TTP LI, which can be explained by a direct inhibition of DNA‐polymerase. No change was observed in the fraction of cells labelled with 3H‐thymidine. A decrease in 3H‐TTP LI was also observed after a single i.v. dose of methotrexate.
The Role of Disulphide Bonds in Heinz Body Attachment to Membranes
Authors: Chan E. , Desforges J. F.
Published: March 2008

Summary. The mechanism of binding of oxidant‐induced Heinz bodies to red cell membrane was studied. Heinz bodies induced by acetylphenylhydrazine in intact cells or in the presence of membrane remained attached to membrane when separated by a sucrose gradient. Incubation with sulphydryl reagents failed to free intact Heinz bodies from or prevent attachment to membranes, although the amount of haem was reduced. Thus disulphide bonds do not appear to be a major mechanism of attachment of Heinz bodies to red cell membrane.
Chelation Studies with 2,3‐Dihydroxybenzoic Acid in Patients with β‐Thalassaemia Major
Authors: Peterson C. M. , Graziano J. H. , Grady R. W. , Jones R. L. , Vlassara H. V. , Canale V. C. , Miller D. R. , Cerami A.
Published: March 2008

Summary. 2,3‐Dihydroxybenzoic acid was evaluated as a potentially useful, orally effective iron‐chelating drug by performing iron balance studies in patients with β‐thalassaemia major. The administration of this substance at 25 mg/kg/d to five patients for 8 d caused an average increase in iron excretion of 4.5 mg/d. When the drug was administered at 25 mg/kg q.i.d. to eight patients for 21 d, iron excretion increased to 6.5 mg/d. Chelation was highly specific for iron with changes in magnesium and calcium excretion being insignificant. The drug was well tolerated with side effects limited to gastrointestinal complaints which ameliorated when the drug was taken with food. These studies provide a rationale for further evaluation of 2,3‐dihydroxybenzoic acid in patients with iron overload.
Factor V in an Industrial Population
Authors: Brozovi Milica , Chakrabarti R. , Stirling Yvonne , Fenton Susan , North W. R. S. , Meade T. W.
Published: March 2008

Summary. Factor‐V levels have been measured in a random sample of 626 men and 307 women working in a variety of occupations in North West London. The method is an automated one‐stage assay using the same batch of freeze‐dried thromboplastin, all results being expressed in terms of the same freeze‐dried standard plasma; it has been shown that only one dilution of test plasma is necessary. Factor‐V levels are significantly higher when venepuncture is difficult than when it is satisfactory, the mean levels being about 130% and 117% respectively. Factor‐V levels are approximately normally distributed; they are similar in men and women and in blacks and whites, and increase significantly with age at the rate of about 0.6% per annum. Factor‐V levels are not affected by oral contraceptives or the menopause, and there are no differences according to blood group or secretor status.
Banked Blood Microfiltration I. MICROFILTER COMPOSED OF FIVE POLYURETHANE FOAM LAYERS WITH GRADED PORE SIZE
Authors: Guidoin R. G. , Gaylor J. D. S. , Borsanyi J.‐P.
Published: March 2008

Summary. Aggregates of amorphous material which develop with storage of banked blood may be a source of pulmonary microembolism in patients having massive transfusions. In order to remove such debris, blood microfilters have been developed and are in routine clinical use. This paper describes the evaluation of one such filter, the Bentley PF 127 model B, which is composed of five layers of polyurethane foam. The time of filtration as a function of the pressure applied, the number of particles removed, the observed changes in blood cells, and the results of scanning electron microscopy of the filter after filtration of human banked blood, are reported.
Banked Blood Microfiltration II. DACRON‐WOOL MICROFILTER
Authors: Guidoin R. G. , Gilchrist T.
Published: March 2008

Summary. It is well established that debris tends to accumulate in banked blood, the amount of debris increasing with storage time. If such a blood is transfused to a patient, it will be filtered by the capillary network of the lungs and could cause various intensities of microemboli. These can be prevented by microfiltration—the Swank IL200 transfusion filter made of Dacron wool is perfectly suitable. It removes more than 70% of the debris 29‐100 μm size from four blood units and its efficiency for particle removal is greatly increased by lactate priming.
The Utilization of a New Strength Citrate Anticoagulant during Centrifugal Plateletpheresis: I. ASSESSMENT OF DONOR EFFECTS
Authors: MISHLER JOHN MILTON , JANES A. WILLIAM , LOWES BERNARD , FARFAN CAROL , EMERSON PAULINE M.
Published: March 2008

Summary. A reduction of donor effects during centrifugal plateletpheresis with the Haemonetics Blood Processor was achieved by reducing the concentration of the citrate anticoagulant. Serum citrate and ionized calcium levels, immediately and I h post‐pheresis, were affected to a lesser extent by using 5.0 g total ionized citrate (TIC) than with either 8.0 g or 11.0 g. Total calcium, bicarbonate, prothrombin time, partial thromboplastin time, ECG, and platelet counts were affected to a similar degree by all three TIC formulations. The total number of platelets collected per litre of blood processed was not significantly different among the three TIC formulations. In vitro studies employing the screen filtration pressure (SFP) technique showed no evidence of platelet aggregates in whole blood collected into either 0.01 M or 0.005 M citrate and agitated or left stationary at room temperature for 5 h. The use of different citrate concentrations in plateletpheresis is discussed.
The Negro Variety of Hereditary Persistence of Fetal Haemoglobin is a Mild Form of Thalassaemia
Authors: CHARACHE S. , CLEGG J. B. , WEATHERALL D. J.
Published: March 2008

Summary. Further studies have been carried out on blood of the 15‐year‐old Negro male from Baltimore who was the first reported case of the homozygous state for hereditary persistence of fetal haemoglobin. His red cells contain only Hb F; Hbs A and A2 have never been detected. Over a 15‐year period of follow up the red cells of this individual have shown persistent microcytosis with reduced MCH and MCV values. His whole‐blood p50 value is decreased, probably because of lack of interaction between Hb F and 2,3‐diphosphoglycerate. However, his haemoglobin level at the age of 15 years is lower than would be predicted from the degree of increased oxygen affinity. Globin‐chain synthesis studies suggest that this is because he has a mild thalassaemia disorder with an α/γ‐chain production ratio of about 1.5, similar to that found in β‐thalassaemia heterozygotes. Thus Negro HPFH appears to be a well‐compensated form of δβ thalassaemia.
Serum ‘Uracil+Uridine’Levels Before and After Vitamin B12 Therapy in Pernicious Anaemia
Authors: PARRY T. E. , BLACKMORE J. A.
Published: March 2008

Summary. The serum ‘uracil+uridine’levels, expressed as uracil, have been measured in 10 cases of pernicious anaemia both before and after treatment, and compared with the levels in 97 normal subjects. The mean pre‐treatment value (8.82 μmol/l., range 6.0–12.0 μmol/l.) differed significantly from that of the normal controls (15.7 μmol/l., range 5.7–40.5 μmol/1., t= 8.8, P<0.001). This confirms the low serum uracil level previously reported in pernicious anaemia in relapse. The level rose progressively after treatment, reaching a maximum on the fourth day (mean 17.85 μmol/1., range 9.3–23.4 μmol/l.). This was not significantly different from the mean of the normal control group. The difference between the pre‐ and post‐treatment levels was significant on days 3, 4 and 5 (P<0.005, P<0.001 and P<0.005 respectively) and the rise preceded the reticulocyte response by 24 h. A further case was treated with physiological doses of vitamin B12 (2 μg daily for 6 d) and a similar rise in the serum uracil level noted.

These results are not explained by any of the known functions of vitamin B12. They are, however, similar to the changes in the serum methionine levels previously reported in pernicious anaemia. The latter were readily explained by the known action of vitamin B12 on ‘de novo’methionine synthesis and it is suggested that the synthesis of uracil, like that of methionine, might be influenced by vitamin B12 in man.
Microcytosis, Anisocytosis and the Red Cell Indices in Iron Deficiency
Authors: ENGLAND J. M. , WARD SUSAN M. , DOWN M. C.
Published: March 2008

Summary. Red cell volume distribution curves have been used to measure micro‐cytosis and anisocytosis in normal subjects, blood donors and patients with iron deficiency anaemia. These measurements were more sensitive than the conventional red cell indices for detecting blood donors with a low transferrin saturation. Three stages are suggested as iron deficiency progressively interferes with haemopoietic function. Anisocytosis and an increased percentage of microcytic cells are the first haematological abnormalities to occur and at this stage haemoglobin concentration is usually normal and transferrin saturation less than 32%. At the second stage the MCV and MCH decline, haemoglobin concentration is generally sub‐normal, though not below 9 g/dl, and transferrin saturation is usually below 16%. The final stage of iron deficiency is associated with a low MCHC, a haemoglobin concentration below 9 g/dl and a transferrin saturation of less than 16%.
The Heterogeneity of Erythrocyte Antigen Distribution in Human Normal Phenotypes: an Immunoelectron Microscopy Study
Authors: REYES F. , GOURDIN M. F. , LEJONC J. L. , CARTRON J. P. , GORIUS J. BRETON , DREYFUS B.
Published: March 2008

Summary. A and A1 antigen were detected on human blood erythrocytes by immunoelectron microscopy using peroxidase‐conjugated antibodies. Cells were obtained from various normal A subgroups, including rare weak A phenotypes and infant (cord blood) samples. Erythrocytes were fixed prior to incubation with specific reagents. The detection of surface antigens was carried out by an indirect method involving anti‐A and anti‐A1 antibodies and conjugated anti‐immunoglobulin antibodies. The surface labelling was seen as a diffuse dense layer. Haem peroxidase‐like activity resulted in a faint background which did not interfere at the level of ultrathin sections, with surface staining due to exogeneous peroxidase.

The most significant finding was the existence, in a given sample, of several populations of cells as revealed by their antibody‐binding capacity. The distribution of the various populations varied from one sample to another according to its subgroup. The progressive weakening of phenotype expression which characterizes the various subgroups from A1 to A weak was paralleled by a decreasing number of ‘antigen rich’cells, which were still detectable in weak phenotypes as a minor population.

This study confirms that a given normal phenotype in fact represents a mixture of antigenically different populations of erythrocytes.
Monocyte Recruitment in Tuberculosis and Sarcoidosis
Authors: Schmitt E. , Meuret G. , Stix L.
Published: March 2008

Monocytopoiesis and blood monocytes were investigated in nine patients with active tuberculosis and in six patients with active sarcoidosis in order to obtain information on monocyte consumption in these two types of granuloma. All patients with tuberculosis demonstrated a marked increase in proliferation activity of monocytopoiesis and premature monocyte marrow release. These changes indicate a high monocyte consumption which probably is caused by a high macrophage death rate due to the high macrophage‐toxicity of tubercle bacilli. Thus, tuberculous lesions are an example of a ‘high turnover granuloma’. In sarcoidosis monocytopoiesis showed no significant deviations from the normal. This indicates a low macrophage turnover or ‘low turnover granuloma’. Thus, any hypothetical agent assumed to be involved in the pathogenesis of sarcoidosis would have to possess low macrophage‐toxicity.
Neutrophil Accumulation in Vivo following the Administration of Chemotactic Factors
Authors: Jones D. G. , Richardson D. L. , Kay A. B.
Published: March 2008

Intravenously administered 51Cr‐labelled homologous neutrophils accumulated at guinea‐pig skin sites prepared by intradermal injection of factors derived from complement‐activated serum, possessing in vitro chemotactic activity. There was a strong correlation between the in vitro potency of Sephadex G‐100 fractions of the activated serum and their ability to evoke neutrophil accumulation in vivo. These experiments suggest that agents which are chemotactic for neutrophils in vitro also induce the localization of this cell in vivo.