bluuebunny/crossref_metadata_2025 huggingface dataset:
Loss of the Y Chromosome in Myelodysplasia: A Report of Three Cases Studied with Quinacrine Fluorescence
Authors: Rowley Janet D.
Published: March 2008

Summary. Three males with a variable proportion of 45, X cells in their bone marrow were observed among 100 males examined. All three were found in a group of 57 males who had some haematologic disorder. No Y‐minus males were observed in a group of 37 males who had other, non‐haematologic disorders, or among six volunteers. Quinacrine fluorescence was used to precisely identify the missing small acrocentric chromosome as the Y. One of the three males with a Y minus cell line was 55 yr; the other two were over 60 yr of age. All three males had thrombocytopenia which had been treated with prednisone and two had undergone splenectomy. One of the cases has shown a disappearance of his aneuploid cells over a 14 mth period. One other case has shown an increase in the proportion of 45, X cells during a 2 yr interval.
Temperature Gradient Dissociation of Red Cell Antigen–Antibody Complexes in the Polybrene Technique*
Authors: Lalezari Parviz , Oberhardt Bruce
Published: March 2008

Summary. The automated Polybrene technique for the detection of red cell antibodies has been adapted for the study of the thermal properties of various red cell antigen‐antibody reactions. A continuous flow of antibody‐induced red cell aggregates was subjected to gradually increasing temperature. The results produced ‘temperature gradient dissociation curves’ (TGDC) which represented not the influence of temperature on antibody uptake but the dissociation of already established immunological bonds. Temperature of dissociation was influenced by the thermal properties of the antibodies and by their concentration. Antibodies could be classified into ‘cold’, ‘intermediate’ and ‘warm’ reacting groups. The technique was also applicable to the study of antibodies that occur on sensitized red cells and, particularly, to patients with acquired haemolytic anaemia. Determination of TGDC has provided a new approach to the study of autoimmune diseases, and may have significant clinical implications.
The Specific Assay of Prothrombin Using the Taipan Snake Venom
Authors: Denson K. W. E. , Borrett R. , Biggs Rosemary
Published: March 2008

Summary. A simple one‐stage assay for the measurement of prothrombin employing the Taipan snake venom is described. The venom converts prothrombin to thrombin in the absence of all other known clotting factors.
Heterogeneity of Human Platelets: IV. IDENTIFICATION OF A YOUNG PLATELET POPULATION WITH [75Se]SELENOMETHIONINE
Authors: Amorosi Edward , Garg Sudershan K. , Karpatkin Simon
Published: March 2008

Summary. Heavy‐large platelets have previously been shown to be more active metabolically and functionally compared to light‐small platelets and to be present in increased per cent during conditions of enhanced thrombopoiesis. In the present report, the kinetics of platelet synthesis has been followed by in vivo labelling of megakaryocytes with the amino acid analogue, [75Se]selenomethionine. Kinetic measurements in three volunteer subjects indicated that heavy‐large platelets were young platelets and suggested a progression with age of heavy‐large platelets to light‐small platelets.
The Eosinophil Leucocytes in the Blood and Bone Marrow of Patients with Down's Anomaly
Authors: Archer R. K. , Engisch H. J. C. , Gaha T. , Ruxton J.
Published: March 2008

Summary. A study has been made of blood and bone‐marrow films obtained from mentally subnormal patients with Down's anomaly and without this anomaly. It was found that the eosinophils in all stages of maturation subsequent to the promyelocyte were about half as frequent in the mongol patients as in the controls. An investigation of the nuclear lobulation of eosinophils in these patients showed that there were at least twice as many with three or four lobes in mongol patients than in control patients; this finding is in contrast with that well established for the neutrophil leucocytes in mongolism.
Amyloidosis Associated with the Nephrotic Syndrome and Transfusion Reactions in a Haemophiliac
Authors: Prentice C. R. M. , Izatt Marian M. , Adams J. F. , McNicol G. P. , Douglas A. S.
Published: March 2008

Summary. It is suggested that the immunological response in a severe haemophiliac to longstanding incompatible plasma infusions may have been responsible for the development of amyloidosis which culminated in the nephrotic syndrome. Serological studies showed that the patient had a circulating anti‐Gm(1) antibody, and also that possibly he was on occasions reacting violently towards the infusion of immune complexes formed between the IgG1 (Fc region) antigens, Gm(1) and ISf(1) and their antibodies. Despite the fact that there was a marked reduction in the number and severity of transfusion reactions after treatment solely with selected Gm(‐1) plasmas, there was a progressive deterioration in the patient's renal function.
The Role of Ascorbic Acid in the Metabolism of Storage Iron
Authors: Lipschitz D. A. , Bothwell T. H. , Seftel H. C. , Wapnick A. A. , Charlton R. W.
Published: March 2008

Summary. The role of ascorbic acid in the metabolism of storage iron was investigated in guinea‐pigs. Ascorbic acid deprivation increased the total non‐haem iron concentration in the spleen and reduced it in the liver, and in both organs ferritin was diminished and haemosiderin increased. Replacing the ascorbic acid restored the normal distribution of iron between the two storage compounds, and in the spleen the total storage iron concentration returned to control levels within 24 hr. Evidence was obtained in experiments with 59Fe that the accumulation of iron in the spleen was due to a diminished release from reticulo‐endothelial cells. When 59Fe‐labelled haemoglobin in denatured red cells was injected, release of the isotope was inhibited in scorbutic animals. In contrast, after injecting labelled transferrin, 59Fe in the liver parenchymal cells was released to a greater extent than in normals. These observations may explain certain ferrokinetic peculiarities in patients with scurvy, and possibly also the predominantly reticulo‐endothelial localization of the iron in Bantu subjects with siderosis.
Apparent Folate Deficiency in Iron‐Deficiency Anaemia
Authors: Roberts P. D. , StJohn D. J. B. , Sinha R. , Stewart J. S. , Baird I. M. , Coghill N. F. , Morgan J. O.
Published: March 2008

Summary. Examination of 50 patients with iron‐deficient hypochromic anaemia showed evidence suggesting a high incidence of folate depletion. The peripheral blood films of 44 patients (88%) showed more than 3% of neutrophils with five nuclear lobes, 35 patients (70%) had a high mean neutrophil lobe count, 20 patients (40%) had hypersegmented neutrophils and 19 patients (38%) had giant metamyelocytes in the bone marrow. The serum folate was below 3 ng/ml in 12 patients and 3–6 ng/ml in 18. Red‐cell folate was subnormal in 15%, and 45% had a positive Figlu test.

Correlation between various tests for folate deficiency was not found, apart from a correlation between red‐cell folate levels and morphological changes in the neutrophils. The haemoglobin rise following intravenous iron therapy was smaller when the serum folate level was low. There was probably a similar relationship to the red‐cell folate level, but the numbers tested were small. The presence of neutrophil multilobing in the peripheral blood film and giant metamyelocytes in the bone marrow did not influence the response to iron therapy, neither did an abnormal Figlu excretion.

Following intravenous iron therapy, both neutrophil multilobing and marrow giant metamyelocytes were significantly reduced in number. This therapy did not significantly alter Figlu excretion measured 6 weeks after treatment, but both serum and red‐cell folate levels fell. Intravenous iron therapy did not produce any significant changes in the serum vitamin B12 levels. The part that iron deficiency itself may play in causing apparent folate depletion is discussed.
Sideroblastic Anaemia and its Association with Leukaemia and Myelomatosis: A Report of Five Cases
Authors: Catovsky D. , Shaw M. T. , Hoffbrand A. V. , Dacie J. V.
Published: March 2008

Summary.Five cases of sideroblastic anaemia associated with malignant diseases of the bone marrow are reported. The first patient had a 20‐yr history of ‘refractory anaemia’ and developed acute myeloid leukaemia as a terminal event, the second had chronic monocytic leukaemia for 6 yr, and the third suffered from myeloblastic leukaemia. The two remaining patients presented with severe refractory hypochromic anaemia, and myelomatosis was an unexpected finding on examination of the bone marrow. The histories of these five patients and those of similar patients reported in the literature suggest that sideroblastic anaemia can develop as the consequence of bone‐marrow stem cell mutations, which can lead to several types of cellular dysfunction. In some patients sideroblastic anaemia may be the only demonstrable change; in others, subsequent mutations may lead to overt leukaemia, while in still others sideroblast formation may be a minor phenomenon over‐shadowed from the start by an associated malignant disorder.
Epsilon‐Aminocaproic Acid Therapy for Dental Extractions in Haemophilia and Christmas Disease: A Double Blind Controlled Trial
Authors: Walsh P. N. , Rizza C. R. , Matthews J. M. , Eipe J. , Kernoff P. B. A. , Coles M. D. , Bloom A. L. , Kaufman B. M. , Beck P. , Hanan C. M. , Biggs Rosemary
Published: March 2008

Summary. The results are reported of a pilot study and two separate double blind controlled trials of the effectiveness of epsilon‐aminocaproic acid (EACA) in dental extractions in patients with haemophilia and Christmas disease. In the major trial 31 patients were studied; 23 at Oxford and eight at Cardiff. All patients received either EACA (6 g four times daily for 10 days at Oxford or for 7 days at Cardiff) or a placebo, in conjunction with a single preoperative dose of therapeutic materials expected to raise the plasma factor‐VIII or factor‐IX level to 50%. Post‐operative therapeutic materials were withheld unless intraoral bleeding occurred. Despite the fact that plasma factor‐VIII or factor‐IX levels were, on average, lower, the number of teeth extracted larger, the amount of therapeutic concentrates less and the postinfusion plasma factor‐VIII or factor‐IX levels lower in the EACA group at Oxford, the incidence of postoperative intraoral bleeding was lower and the requirements for postoperative therapeutic materials less in the group treated with EACA. Side‐effects were not a major problem. The number of patients studied at Cardiff was too few for statistical analysis but the results were similar to those at Oxford. The total conservation of therapeutic materials at Oxford in comparison to the amount utilized before EACA was used is estimated on the basis of these results to be approximately 12 000 factor‐VIII or factor‐IX units/patient, or approximately 190 units/kg/patient, equivalent, for each patient, to the amount derived from approximately 120 blood donations.

These results show that EACA in conjunction with preoperative therapeutic concentrates sufficient to raise the plasma factor‐VIII or factor‐IX level to 50% can be useful for all patients with haemophilia and Christmas disease undergoing tooth extraction. In some patients EACA therapy is contraindicated and for these patients adequate cover with therapeutic materials must be provided during the postoperative period.
Uptake of Tritiated Folates by Human Bone Marrow Cells in Vitro
Authors: Corcino J. J. , Waxman S. , Herbert V.
Published: March 2008

Summary. Using incubation periods up to 4 hr, it was demonstrated that uptake of tritiated pteroylglutamic acid ([3H]PteGlu) by human bone marrow cells in vitro was in the range of six‐fold greater than uptake by reticulocytes. Uptake was temperature‐dependent, increasing during 4 hr at 37°C but not at 4°C; a similar temperature dependence was found for the uptake of [3H]methyltetrahydrofolate ([3H‐CH3]H4PteGlu). The pH optimum for [3H]PteGlu uptake was in the range of 7.4. Percentage uptake decreased as concentration of [3H]PteGlu increased. Preincubation with unlabelled PteGlu reduced uptake of subsequently added [3H]PteGlu by twice as much as did preincubation with methotrexate, suggesting that methotrexate may only partly share the uptake mechanism for [3H]PteGlu. [3H]PteGlu uptake was not affected by preincubation with diphenylhydantoin or a sulphydryl inhibitor. Uptake of [3H‐CH3]H4PteGlu by human bone marrow cells appeared to be approximately twice the uptake of [3H]PteGlu. The findings support the concept of two mechanisms for folate uptake by human reticulocytes and bone marrow cells: an energy‐dependent, active carrier mechanism probably of primary physiologic significance, and a seemingly passive diffusion‐like mechanism, probably primarily of pharmacologic significance.
Study of in Vitro Lysis of Paroxysmal Nocturnal Haemoglobinuria (PNH) and PNH‐like Red Cells
Authors: Ferrone S. , Marubini E. , Mercuriali F. , Sirchia G.
Published: March 2008

Summary. In paroxysmal nocturnal haemoglobinuria (PNH) the intensity of haemolysis obtained in the acidified‐serum and sucrose‐haemolysis tests—the two procedures most commonly used for the laboratory diagnosis of the disease—varies according to the red cells, serum and test used. The influence of these three variables has been investigated on both PNH and PNH‐like lysis, that is lysis of PNH cells and of normal red cells treated in vitro with the sulphydryl compound 2‐aminoethylisothiouronium bromide (AET). The conclusions, almost identical for PNH and AET lysis experiments, suggest that for the laboratory diagnosis of PNH: (1) it is advisable to use both tests in parallel; (2) sera to be used in the two tests should be selected; (3) for this selection AET cells can be employed when PNH cells are not available.
Selective in Vitro Cytotoxicity of a Glycolytic Enzyme Inhibitor for Immature Lymphoid Cells
Authors: Mann J. R. , Simpson J. S. , McEvoy M. W. , Stuart J.
Published: March 2008

Summary. A specific lactate dehydrogenase inhibitor, sodium oxamate, which prevents glycolytic resynthesis of ATP, has been shown to possess a significant, in vitro cytotoxic effect for EB2 cells and for the acute leukaemia lymphoblast. This result was obtained at an oxamate concentration of 0.08 M in TC 199 medium adjusted to physiological osmolality, in comparison with control cultures containing an equimolar concentration of sodium chloride. No significant oxamate cytotoxicity was found for normal marrow myeloid precursors or for blood lymphocytes and neutrophils, although a significant reduction in glycolysis‐dependent neutrophil phagocytosis was obtained.
Detection of Splenic Anti‐Platelet Antibody Synthesis in Idiopathic Autoimmune Thrombocytopenic Purpura (ATP)
Authors: Karpatkin S. , Strick N. , Siskind G. W.
Published: March 2008

Summary. A significant decline in the serum anti‐platelet antibody titre, as measured by the platelet factor 3 (PF‐3) immuno‐injury technique, followed splenectomy in seven out of eight patients with autoimmune thrombocytopenic purpura (ATP).

Anti‐platelet antibody could be eluted from the splenic extracts of 11 of 13 patients with ATP. The anti‐platelet antibody detected could be adsorbed with washed human platelets and could be removed with rabbit anti‐human IgG (RAHIgG). Extracts of spleens from six ‘control’ patients (five with hereditary spherocytosis and one with Milroy's disease) had no detectable anti‐platelet antibody.

Minced spleen from nine patients with ATP and five ‘controls’ were cultured for 17 hr in a medium containing 14C amino acids. Washed platelets adsorbed at least five times more Ladioactivity from globulin fractions of cultured ATP spleens than did ‘control’ spleens. In contrast, washed granulocytes failed to adsorb significant radioactivity. Supernatants obtained from these splenic incubates were also tested for anti‐platelet antibody (PF‐3 test). Anti‐platelet antibody was detectable in all seven ATP patients tested and was absent in the three control subjects tested. The anti‐platelet antibody could be adsorbed with washed human platelets but not with washed human granulocytes or red blood cells. These data suggest that the spleen might be a significant site for anti‐platelet antibody synthesis in ATP.
Normal Haematological Values in the Central American Population
Authors: Viteri F. E. , de Tuna V. , Guzmán M. A.
Published: March 2008

Summary. A total of 6787 subjects were studied, representative of the Central American populations but without clinical evidence of haematological disease. Out of this sample, 452 individuals fulfilled the following conditions: they were hook‐worm free, with values of serum iron > 50 μg/100 ml; saturation of transferrin > 20%; serum folates (L. casei) > 5 ng/ml; and serum vitamin B12 (E. gracilis) > 150 Pg/ml. These individuals were considered adequately nourished from the erythropoietic point of view, and they constituted the ‘normal’ sample from which values for haemoglobin, packed cell volume, red blood cell count, and red blood cell indices were calculated. These subjects were grouped by age and sex, and according to the altitude above sea level of their place of residence (0‐750 and 751–1500 m).

Results from this highly selected sample indicate that haematological values are equal or higher to those reported for ‘normal’ individuals from well‐nourished, non‐tropical populations. To our knowledge, this is the only study where norms have been derived from a population sample with well‐documented ‘normal’ serum values for iron, % saturation of transferrin, folate and vitamin B12.

From these data, haematological values below which the risk for an individual having a subnormal haematological state increase, are derived and discussed.
Active Extrusion of Heinz Bodies in Drug‐Induced Haemolytic Anaemia
Authors: Amare M. , Lawson B. , Larsen W. E.
Published: March 2008

Summary. A patient with repeated episodes of sulphonamide‐induced haemolytic anaemia is reported. The haemolytic episodes were associated with a high percentage of Heinz bodies which had the unique feature of being actively extruded from the erythrocytes. Detailed morphological and biochemical studies are outlined, but the basis for the tendency to form Heinz bodies remains unclear. It is suggested that the phenomenon of active extrusion may provide an alternative route, independent of the reticuloendothelial system, for the removal of Heinz bodies. The close resemblance between the active expulsion of the Heinz bodies and the physiological denucleation of the maturing erythroblast is noted and a similar mechanism postulated.
Severe Allergic Pulmonary Oedema after Plasma Transfusion
Authors: Kernoff P. B. A. , Durrant I. J. , Rizza C. R. , Wright F. W.
Published: March 2008

Summary. Three patients with haemophilia suffered severe febrile reactions after transfusions of fresh frozen plasma and developed pulmonary oedema which did not appear to be due to circulatory overload. The chest radiographs were characterized by widespread patchy rounded opacities resembling bronchopneumonia. In one of the patients the reaction proved fatal; in the other two, treatment was followed by the rapid resolution of symptoms, clinical signs and the abnormal chest radiograph. In two of the patients there was evidence that the reactions might have been caused by transfer of antibodies to white cell or Gm factors from the donor to the recipient. It is recommended that haemophiliacs with a history of transfusion reactions should only be treated with concentrated preparations of factor VIII and not with fresh frozen plasma.
The Function of the Phagocytic Leucocytes in Leukaemia
Authors: Goldman J. M. , Catovsky D.
Published: March 2008

Summary. Various aspects of the function of the phagocytic cells (neutrophils and monocytes) in the peripheral blood of patients with leukaemia have been reviewed. Mobilization of neutrophils and monocytes to sites of experimentally induced inflammatory lesions appears to be defective in acute myeloid leukaemia (AML). Phagocytosis (uptake of bacteria) appears to be defective in the neutrophils of patients with untreated chronic granulocytic leukaemia. The killing of Candida albicans by neutrophils from patients with AML also seems to be impaired. Preliminary evidence suggests that monocytes from patients with acute monocytic leukaemia have an enhanced phagocytic and candidacidal activity after 7 days in culture.

Neutrophils and monocytes from patients with AML do not differ from normal leucocytes in their capacity to reduce nitroblue tetrazolium which suggests that cellular metabolism after phagocytosis is not greatly disordered in leukaemic cells: however, a possible deficiency in the neutrophils in chronic granulocytic leukaemia is suggested. On the other hand, some leukaemic neutrophils lack the enzyme myeloperoxidase and this abnormality is particularly associated with the myeloblastic‐promyelocytic type of AML. The contribution of these leucocyte abnormalities to the pathogenesis of infection is discussed.
Transferrin—Reticulocyte Binding: Evidence for the Functional Importance of Transferrin Conformation
Authors: Lane R. S.
Published: March 2008

Summary

Iron‐free and 59Fe‐labelled human and rabbit 125I‐transferrins have been chromatographed on DEAE cellulose at pH 7.9 and have also been incubated with rabbit reticulocytes in order to determine transferrin and iron uptake.

Chromatography of human apo‐ and iron‐transferrin indicated clear differences in their surface properties which are considered to be conformation‐dependent. These differences were not evident in rabbit transferrin samples.

Human and rabbit iron‐transferrins and rabbit apo‐transferrin all associated rapidly with rabbit reticulocytes; human apo‐transferrin was only weakly bound. Correlation of data from both experiments suggested that reticulocyte binding of human apo‐transferrin is limited by its molecular confirmation. The incubation studies confirmed results of other workers and indicated that the data from these experiments with human and rabbit transferrin are not in conflict.

Compared with rabbit iron‐transferrin, only half as much human iron‐transferrin was bound to rabbit reticulocytes in a comparable experiment. Iron transfer from bound human and rabbit transferrin took place at equal rate. It is argued that fewer receptors on rabbit reticulocytes are available for human transferrin because, when bound, the size and shape of a molecule limits further binding of transferrin molecules at adjacent sites.
The Anaemia of Kala Azar
Authors: Woodruff A. W. , Topley Elizabeth , Knight R. , Downie Campbell G. B.
Published: March 2008

Summary

It has been confirmed that during the active phase of kala azar the erythrocyte life span is shortened. At this time patients’ immunoconglutinin titres are raised, and their erythrocytes have been shown to be agglutinated by anti‐complement and anti‐non‐y‐globulin sera. Erythrocyte destruction in kala azar takes place largely in the spleen, as demonstrated by rising counts over the spleen relative to the praecordium and liver following labelling of the patients’ erythrocytes with 51Cr. This point is supported by the absence of anaemia observed in a patient with a heavy kala azar infection but whose spleen had been removed a year earlier. We postulate that an autoimmune mechanism is likely to be the explanation of these observations and of the reduced erythrocyte survival in kala azar. Such a mechanism provides a basis which may help to explain the development of anaemia in a number of other protozoal and helminthic infections in man.
Plasma Volume and Splenomegaly in Polycythaemia Vera
Authors: Bowdler A. J.
Published: March 2008

Summary

Blood volume studies in 23 patients with polycythaemia vera showed the mean plasma volume to be normal when compared with control subjects, and variation in plasma volume for the group as a whole was no greater than in normal subjects. However, the polycythaemic subjects proved to be heterogeneous with respect to plasma volume: in one subgroup, showing splenomegaly with demonstrable intrasplenic red cell pooling, there was a moderate increase in mean plasma volume. In addition, the effect of intrasplenic red cell pooling on the whole body haematocrit/venous haematocrit ratio was found to differ in this condition from the effect in non‐polycythaemic subjects: the splenic pool failed to concentrate red cells significantly above the levels present in the extrasplenic circulation, and did not result in a rise in the Hb/Hv ratio.
Effect of Amniotic Fluid on Blood Coagulation
Authors: Courtney L. D. , Allington M.
Published: March 2008

Summary

Amniotic fluids from, normal subjects and from patients with intrauterine death (IUD) were studied for their ability to shorten the clotting‐time of blood in vitro. The IUD amniotic fluid was different in that it seemed to require the presence of factor VII for its action whereas normal amniotic fluid did not. In this it resembled tissue factor in its action, whereas normal amniotic fluid resembled Russell Viper venom.
The Inclusions of the May‐Hegglin Anomaly and Dohle Bodies of Infection: an Ultrastructural Comparison
Authors: Cawley J. C. , Hayhoe F. G. J.
Published: March 2008

Summary

Marked differences in the ultrastructures of the inclusion of the May‐Hegglin anomaly and the Dohle bodies of infection are described. Previously these two inclusions have been considered essentially similar, and the inclusions of the May‐Hegglin anomaly have been called‘Döhle bodies’ . It is suggested that in future the term‘Döhle body’ should be restricted to the basophilic inclusions seen in neutrophils in a variety of infectious states.

The distribution of the two inclusions in the different haematological cell types is described, and their functional significance is discussed.
The Clinical and Biosynthetic Characterization of αβ‐Thalassaemia
Authors: Knox‐Macaulay H. H. M. , Weatherall D. J. , Clegg J. B. , Bradley J. , Brown M. J.
Published: March 2008

Summary

A Cypriot family is described in which three thalassaemia genes, α‐thalassaemia 1, α‐thalassaemia 2, and β–thalassaemia, are segregating. Two siblings are heterozygous for both α‐thalassaemia 1 and β‐thalassaemia while a third child has typical haemoglobin H disease. The α‐thalassaemia β‐thalassaemia combination, which is associated with an α/β chain production ratio of unity, produces a moderate degree of anaemia with marked hypochromia and morphological changes of the red cells together with increased rates of flux of potassium across the membranes, but the red cell survival is normal. These changes m red cell morphology and metabolism are very similar to those found in the sibling with haemoglobin H disease in whom there is gross imbalance of globin chain synthesis and shortened red cell survival. These results suggest that imbalanced globin chain production is the primary cause of shortened red cell survival in thalassaemia and that changes in membrane permeability are probably of secondary importance and may, at least in part, result from factors other than globin chain imbalance.
Assay of the Immune Inhibitor in Classic Haemophilia: Application of Virus‐Antibody Reaction Kinetics
Authors: Pool Judith G. , Miller Rupert G.
Published: March 2008

Summary

The kinetics of virus neutralization by antibody are shown to apply to factor‐VIII neutralization by the immune antibody of haemophilic patients. The analysis established for virus neutralization has been used as a model for designing a general system for inhibitor assay based on mixtures in which antigen is present in excess, rather than with antibody in excess, as has been the case with systems previously proposed. The new system permits conversion of inhibitor potency results from other systems based on the use of different mixtures of factor VIII and inhibitor as long as factor VIII is in excess and as long as constant physical conditions of time and temperature of incubation are applied. It also takes account of the fact that some antibodies reveal a‘residual resistant fraction’ of antigen with which they cannot react. The procedure for converting assay data into‘Oxford units’ is described.
Experimental Enterogenous Cyanosis and Anaemia
Authors: Westphal Robert G. , Azen Edwin A.
Published: March 2008

Summary

Rats with jejunal pouches develop an anaemia which js partially haemo‐lytic in nature and accompanied by small amounts of methaemoglobm (MHb) and sulphaemoglobin (SHb). These pigments increase upon sterile incubation of the blood. The presence of these abnormal pigments indicates that oxidativc damage has occurred. Most of the abnormalities improve on feeding neomycin. Rats with blind pouches are more sensitive than normal rats to the oxidant effects of phenacetin on RBCs. RBCs incubated in urine from rats with pouches developed more MHb than RBCs incubated in urine from normal rats. We conclude that bacterial overgrowth in the intestine, by means yet undetermined, can cause oxidative damage to RBCs and that this effect can be produced by constituents of the urine of rats with pouches. This toxic effect of bacterial overgrowth may condition RBCs to oxidant: drug damage.
Studies with Ferritin‐Labelled Dolichos biflorus Lectin on the Numbers and Distribution of A Sites on A1 and A2 Erythrocytes, and on the Nature of its Specificity and Enhancement by Enzymes
Authors: Williams M. A. , Voak D.
Published: March 2008

Summary. The conjugation of Dolichos biflorus extract with ferritin produced an electron dense anti‐A reagent that was used to study the number and distribution of A sites on A1 and A2 red cells; there were five times as many ferritin‐labelled lectin molecules on A1 as on A2 cells (about 800,000: 150,000). The arrangement of sites on both A1 and A2 cells was diffuse but not random, as pattern analysis of the ferritin molecules revealed a contagious distribution of the sensitized A sites. It is suggested that the anti‐A1 saline test activity shown by the Dolichos reagent is due to the inefficiency of the small lectin molecules to bring about agglutination of the A2 cells. A2 cells have fewer sites and consequently a larger mean distance between their A sites than have A1 cells. Thus the probability of ‘bridging’ between the A sites of adjacent cells is reduced. Enzyme treatment increased lectin uptake and the A2 and B cells both bound over 8 million molecules per cell. The A2 cells gave agglutination reactions like untreated A1 cells but the B cells were not agglutinated. In areas of agglutination between the cells lectin molecules probably occurred mainly as a monolayer; however, in the other areas they occurred in multiple layers and only the first layer was in contact with cell membrane sites.
Studies on Blood Coagulation, Fibrinolysis and Platelet Function Following Exercise in Normal and Splenectomized People
Authors: Prentice C. R. M. , Hassanein A. A. , McNicol G. P. , Douglas A. S.
Published: March 2008

Summary

The effect of a short period of strenuous exercise on the haemostatic mechanism has been studied. Following exercise there was an increase of factor VIII which was not affected by prior splenectomy or by aminocaproic acid, an inhibitor of plasminogen activator. Although exercise produced increased plasminogen activator concentrations, as measured by the euglobulin lysis time, there was no alteration in the FR‐antigen (fibrinolytic degradation product) concentration. Platelet adhesiveness, and ADP‐induced aggregation were significantly increased after exercise and in the latter test the phase of platelet disaggregation was impaired.

An increase of platelet factor 3 availability was noted after exercise, and differential centrifugation studies indicated that this was due to the release of micro‐particulate material which may have represented membrane complexes released from platelets.
Comparison of Three Methods for Measuring Vitamin B12 in Serum: Radioisotopic, Euglena gracilis and Lactobacillus leichmannii
Authors: Raven J. L. , Robson M. B. , Morgan J. O. , Hoffbrand A. V.
Published: March 2008

Summary. The vitamin B12 content of 481 sera was estimated by a radioisotopic assay and 478 of the sera were also assayed by the Euglena gracilis method and 396 by the Lactobacillus leichmannii method. The serum vitamin B12 level in normal subjects ranged from 235 to 1470 pg/ml, mean 497 pg/ml (radioisotopic), 170–1144 pg/ml, mean. 460 pg/ml (E. gracilis) and 155–1075 pg/ml, mean 471 pg/ml (L. leichmannii). In general, the radioisotopic assay gave higher results than the other two methods and the E. gracilis assay gave the lowest results. The E. gracilis assay gave the clearest distinction between normal subjects and patients with untreated pernicious anaemia. A large number of post‐gastrectomy and folate deficient patients and occasional patients with other conditions gave subnormal results by one or both of the microbiological assays but normal values by the radioisotopic assay.

The serum vitamin B12 level measured by both the E. gracilis and radioisotopic assays rose with folic acid therapy in two folate deficient patients. In one of them, the values by the two assays were initially 100 pg/ml apart and remained this degree apart during the response, while in the other patient the two assays gave almost identical results both before and during the response to folic acid therapy.
Neuraminidase‐induced Thrombocytopenia in Rats
Authors: Choi Sun‐I , Simone J. V. , Journey L. J.
Published: March 2008

Summary. The purpose of this study was to elucidate the effect of neuraminidase on circulating platelets in rats and the pathogenesis of this effect. Injection of bacterial, purified bacterial and viral neuraminidase caused thrombocytopenia of various degrees. Thrombocytopenia was less pronounced in rats given viral neuraminidase and in splenectomized animals. A dose‐response effect was seen only with purified bacterial neuraminidase injected intravenously. Ultrastructural studies of blood platelets were performed after injection of neuraminidase. Fifteen minutes after injecting neuraminidase, before a definite fall in the count, platelets showed evidence of structural damage such as irregular shapes, centripetal migration and fusion of granules and bleb formation. The structural alterations were progressive, reaching a maximum at about 2–4 hr. At no time were large platelet aggregates observed in the circulating blood. Four hours after injection of neuraminidase, when the platelet count dropped significantly, many platelets had normal ultrastructurc. By 24 hr, the majority of circulating platelets appeared normal; however, the splenic sinusoids and macrophages contained platelets with the same structural abnormalities seen in the circulation 15 min to 2 hr after injection of neuraminidase. We conclude that neuraminidase alters the platelet membrane resulting in its rapid and permanent removal by the reticuloendothelial system.
Quantification of α‐Chain Excess in Erythrocytes in β‐Thalassaemia by Microinterferometry*
Authors: Yataganas X. , Fessas P. , Gahrton G.
Published: March 2008

Summary: Microinterferometry was used to determine the dry mass of α‐chain precipitates in erythrocytes from four patients with homozygous β‐thalassaemia.

α‐Chain precipitates represented on the average 40% of the total dry mass of cells with measurable amounts of precipitation. The maximum load of α‐chain inclusions in a single cell was 75% of the dry mass. A linear correlation was established between the amount of total haemoglobin and non‐α‐chains in individual cells, and the amount of α‐chain precipitation increased linearly with decreasing amounts of soluble haemoglobin, thus indicating an association between impaired haemoglobin production and lack of non‐α‐chains at undisturbed α‐chain production.

The mean ratio of the quantity of α/(β+γ+δ) chain in individual patients varied between 2.3 and 3.0. These ratios are lower than expected from synthetic rate studies, thus indicating the possibility of some precipitate degradation in vivo.
An Unstable Haemoglobin with Reduced Oxygen Affinity: Haemoglobin Peterborough, (βiii (G13) Valine → Phenylalanine, its Interaction with Normal Haemoglobin and with Haemoglobin Lepore
Authors: King Marguerite A. R. , Wiltshire Barbara G. , Lehmann H. , Morimoto H.
Published: March 2008

Summary: A new unstable haemoglobin, Hb Peterborough, is described in father and son. In the former it was found with Hb A but in the latter with Hb Lepore, the mother being a heterozygote for Hb A and Hb Lepore Boston (or Washington). In this respect it differs from other unstable haemoglobins so far described which have been observed in heterozygotes together with normal adult haemoglobin. It was therefore possible to compare the clinical and biochemical interactions of Hbs A and Peterborough, A and Lepore, and Peterborough and Lepore.

Hb Peterborough has been identified as βiii (G‐13) Val → Phe. It has a low oxygen affinity, but the Bohr effect and haem‐haem interaction appear to be normal.
The Response of W/Wv and Sl/Sld Anaemic Mice to Haemopoietic Stimuli
Authors: Harrison D. E. , Russell E. S.
Published: March 2008

Summary: The percentage of haem‐containing nucleated RBC precursor cells in marrows and spleens of W/Wv and Sl/Sld anaemic mice increased in response to hypoxia produced by reduced pressure, red‐cell loss, or red‐cell destruction by phenylhydrazine. The response in the spleens of W/Wv mice occurred several days later than the marrow response, in contrast to normal or Sl/Sld mice in which the marrow and spleen responses occurred simultaneously. These responses were also simultaneous in W/Wv mice whose anaemia had been cured by an injection of normal stem cells. After RBC destruction by phenylhydrazine the duration and degree of tissue hypoxia necessary to stimulate the Sl/Sld response varied widely among individual Sl/Sld mice. In Sl/Sld mice with anaemias alleviated by implants of intact normal spleens, most erythropoiesis was contained within these normal spleen grafts.

Polycythaemic W/Wv and Sl/Sld anaemic mice gave defective responses to erythropoietin; the defect was not caused by delayed erythroid maturation. Serum samples from anaemic mice responding to RBC destruction by phenylhydrazine contained high concentrations of erythropoietin in quantities inversely proportional to the donor's PCV (packed cell volume), but did not contain any factors, other than erythropoietin, to stimulate erythropoiesis in polycythaemic Sl/Sld anaemic mice.
The Effects of Repeated. Doses of Bacterial Endotoxin on Erythropoiesis in the Normal and Splenectomized Mouse
Authors: Twentyman P. R.
Published: March 2008

Summary: A study has been made of the effects of repeated doses of bacterial endotoxin on erythropoiesis in the mouse. It was found that, in the intact mouse, 10 μg/day of Lipopolysaccharide B from S. abortus equi caused a fall in haemoglobin level, reticulocytosis, a severe reduction in bone‐marrow erythropoiesis, and an increase in splenic erythropoiesis. In the splenectomized mouse a fall in haemoglobin occurred but it was not accompanied by reticulocytosis, and the reduction in bone‐marrow erythropoiesis was much less marked.

There was no great increase in red‐cell production over a. period of 20 days in the intact mouse and a raised percentage of 59Fe into circulating blood at 10 days was due mainly to a raised percentage of iron‐incorporating cells in peripheral blood rather than a change in production rate. Random destruction of circulating red cells was not significantly increased in endotoxin‐treated animals.

Bone‐marrow erythropoiesis remained depressed throughout 20 days of endotoxin administration. Possible mechanisms for such a shift of erythropoiesis, maintained over a considerable period, are discussed.
Platelet and Fibrinogen Kinetics with [75Se]‐Selenomethionine in Patients with Myeloproliferative Disorders
Authors: Brodsky Isadore , Ross Evelyn M. , Petkov Goldie , Kahn S. Benham
Published: March 2008

Summary: Simultaneous platelet and fibrinogen kinetics were determined with a cohort label, [75Se]selenomethionine, in three patients with chronic myelocytic leukaemia (CML), three with subacute myelogenous leukaemia (SML) and three with myelofibrosis with myeloid metaplasia (MF). In the CML group platelet survival was normal to increased (10.5–14.0 days), platelet turnover was essentially within the control range (22 000–51 000 platelets/μ/day), fibrinogen survival was prolonged (11.0–19.0 days) and fibrinogen turnover decreased (0.12–0.32 mg/ml/day). In two patients with MF platelet survival was decreased (5.0–7.5 days) and platelet turnover sharply increased (215 000–385 000 platelet/μl/day): one of these patients had multiple thrombotic episodes with an associated decrease in fibrinogen survival (4.0 days) and increased fibrinogen turnover (0.85 mg/ml/day): the other patient had no haemorrhagic or thrombotic episodes and his fibrinogen survival and turnover were within normal limits. Four patients, three with SML and one with MF, presented with thrombocytopenia. Kinetic studies were performed to aid in selecting those patients that might benefit from splenectomy. Splenectomy was performed in two with excellent results. Platelet and fibrinogen kinetics are important adjuncts to the study of the pathogenesis, differential diagnosis, and therapy of the myeloproliferative disorders.
Albumin Density Gradient Separation and Washing of Platelets and the Study of Platelet Coagulant Activities
Authors: Walsh P. N.
Published: March 2008

Summary: Platelets were centrifuged into a specific gravity gradient made by mixing the interface between platelet rich plasma (PRP) and a 40–45% aqueous solution of bovine albumin. By this method of albumin density gradient separation (ADGS) it was possible to centrifuge platelets without disruptive squashing upon a hard surface. The separated platelets were washed in calcium‐free Tyrode's solution without releasing coagulant activity. Platelet yields after multiple washings were high. Morphology and ADP‐induced aggregation of platelets washed by ADGS (in contrast to platelets washed by conventional methods) were indistinguishable from those of unwashed platelets tested as PRP.

The activities of factors V and XI remained in close association with platelets after four successive washes by ADGS, whereas factors VIII, IX and XII were entirely removed.
The Effect of Dilution of Plasma on Coagulation: The Significance of the Dilution‐Activation Phenomenon for the Study of Platelet Coagulant Activities
Authors: Walsh P. N.
Published: March 2008

Summary: The coagulation of non‐contacted plasma is enhanced by dilution with citrate‐saline solution, an effect termed the ‘dilution‐activation phenomenon (DAP)’. When platelets rather than phospholipid were added to plasma, the dilution effect was greatly attenuated. A large clotting time difference between platelet and phospholipid additions was therefore evident when undiluted plasma was tested in plastic tubes. This difference disappeared when the plasma was diluted and tested in glass tubes. Thereby the‘DAP’masked a platelet coagulant activity which appeared to be distinct from platelet factor 3 activity.

In order to make meaningful use of a test system for platelets which eliminated the ‘DAP’, it was desirable to ascertain what factors accounted for the clot‐promoting effect of dilution. The experimental evidence suggests that a physicochemical alteration results in factor‐XII activation when plasma is diluted. Physicochemical factors shown to be important in enhancing the‘DAP’and presumably contact activation are alkaline pH, low ionic strength and negative electrochemical charge, whereas positive charge, high ionic strength and acid pH inhibit clotting in the system employed.
The Kinetic Behaviour of Transfused Donor Bone Marrow Neutrophils and their Precursors in Neutropenic Patients
Authors: Chikkappa G. , Greenberg M. L. , Rubin A. , Taub R. N. , Fotino M.
Published: March 2008

Summary. Six neutropenic patients (eight studies) with hypo‐ or aplastic bone marrow (BM) received 3H‐thymidine (3H‐TdR) and/or 32P‐diisopropylfluorophosphate (DF32P) labelled donor BM cells intravenously. The donor‐recipient leucocyte histocompatibility (HL‐A) match was A in three studies, B in one study and C in another four studies. Leucocyte specific activity (LSA) and autoradiography of serial blood and BM specimens were used to follow the behaviour of the labelled donor neutrophilic cells in the recipient. In two patients (three studies), who received HL‐A A and B matched cells, the donor neutrophil precursors were shown to divide and feed mature cells into the circulation for 10–11 days. The pattern of the entry of labelled segmented neutrophils into the blood was similar to the patterns noted after the i.v. injection of these isotopes in normal individuals except for the rapid emergence of the 3H‐TdR labelled cells into the blood and shortening in all the three phases of the DF32P LSA curves. Studies of this type can be used as an aid in the rapid evaluation of the fate of the donor's transplanted mitotable granulocytic cells in the recipient. Labelled donor cells did not survive in the other patients.

Repopulation of the BM did not ensue in the two patients (three studies) with ideal donor‐recipient pairing (identical twins, negative serum cytotoxicity tests), perhaps due to persisting infectious hepatitis virus or effects therefrom in one patient whose BM aplasia followed infectious hepatitis.
Surface Microfragmentation of Red Cells as a Mechanism for Complement‐Mediated Immune Spherocytosis
Authors: Brown D. L. , Nelson D. A.
Published: March 2008

Summary. The adherence site between red cells and liver macrophages (Kupffer cells) was studied by light‐ and electron‐microscopy at intervals after the intravenous injection of complement‐coated red cells (EC43) into rabbits. The injected red cells adhered to the cytoplasm of the Kupffer cells and became grossly distorted at the point of contact. At an early stage (2 min post injection) protrusions from the red cells were surrounded by macrophage cytoplasm, while later (7–25 min post injection) round fragments of red cells were seen lying apparently separated in the Kupffer cell cytoplasm.

The appearances seen suggest that the adherent red cells lose fragments or ‘microspheres’ to the Kupffer cells without lysis, and, as a result of a proportionally greater surface‐loss than volume‐loss, the adherent red cells become more spherical. The time course of such a microfragmentation process is consistent with the observation that spherocytes appear in the peripheral blood shortly afterwards.
Ultrastructural Studies in β‐Thalassaemia Major
Authors: Polliack A. , Rachmilewitz E. A.
Published: March 2008

Summary. The peripheral red blood cells of 11 patients with β‐thalassaemia major were studied by electronmicroscopy, and a wide spectrum of intracellular changes was observed. Findings were striking in the six splenectomized individuals and most prominent in reticulocytes and late normoblasts. There was marked accumulation of iron in different forms, either as free particles or as aggregates of ferritin and haemosiderin within membrane‐bound particles or mitochondria. Glycogen accumulation was often found in normoblasts. Cells were grossly distorted and deformed, showing indentations and infolding of the plasma membrane with marked vacuole formation.

The most striking finding was the presence of Heinz bodies in various stages of development. These were unlike those encountered in phenylhydrazine induced haemolytic anaemia; no large marginated Heinz bodies were found attached to the cell membrane. The implications of this finding in relation to the present concept of sequestration of Heinz bodies in the spleen are discussed. Another prominent feature was the presence of many bizarre membrane forms and myclin figures which may represent attempts at autodigestion of excess intracellular inclusions, particularly in the absence of the spleen.

The wide spectrum of ultrastructural changes found may be an expression of multiple intracellular defects involving the biosynthesis of globin, haem, glycoprotein, and membranes in β‐thalassaemia.