bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood Coagulation and Fibrinolysis in Acute Leukaemia
Authors: Brakman P. , Snyder J. , Henderson E. S. , Astrup T.
Published: March 2008

Summary. Selected parameters of blood coagulation and fibrinolysis were determined in blood plasma both from 26 patients with untreated acute leukaemia and from patients in remission. Normal, as well as unusually low or high, levels of fibrinogen were observed in the untreated group. Factor VIII was consistently increased in the active state of the disease and remained elevated in patients in long‐term remission. Plasminogen was low in six untreated patients, five of whom died soon after admission. The plasma euglobulin fibrinolytic activity showed wide variations with an increased average value, though four untreated patients (and three patients in long‐term remission) had unusually low activities. In the untreated group fibrinolytic activity of the ‘peptone’ precipitable euglobulins was usually lower than in regular euglobulin precipitates, a reversal of the pattern observed in normal persons. Inhibition of fibrinolysis was decreased in a few patients. There was no consistent relation between occurrence of bleeding episodes and defects in coagulation or fibrinolysis. This, taken together with the variability of the data, would suggest that haemorrhage in acute leukaemia is the result of a fortuitous combination of haemostatic defects. Determinations of factor V and plasminogen may be of prognostic value.
Tissue Typing Using a Routine One‐Step Lymphocyte Separation Procedure
Authors: Harris R. , Ukaejiofo E. O.
Published: March 2008

Summary. A reliable rapid one‐step method is described for the preparation of lymphocytes from 0.25 ml aliquots of defibrinated whole venous blood for tissue typing. An average yield of 70% lymphocytes is obtained relatively free from platelets, erythrocytes and granulocytes even in the presence of large numbers of granulocytes in blood obtained from cadaver donors. Using a high‐speed micro‐centrifuge lymphocytes can be prepared from defibrinated blood within 10 min and it is thus possible to complete a tissue typing test within 75 min. Using this method sufficient lymphocytes for typing purposes can be obtained from 0.5 ml of capillary blood thus greatly facilitating family studies where infants are involved.
Ontogeny of the Haemopoietic System: Yolk Sac Origin of In Vivo and In Vitro Colony Forming Cells in the Developing Mouse Embryo*
Authors: Moore Malcolm A. S. , Metcalf Donald
Published: March 2008

Summary. The mouse yolk sac has been shown to contain in‐vivo colony forming cells capable of producing granulocytic, megakaryocytic and erythroid spleen colonies; in‐vitro colony forming cells producing granulocytic and mononuclear‐macrophage colonies in agar; and cells capable of repopuiating the lymphoid and myeloid tissue of lethally irradiated hosts. Similar haemopoietic precursor cells were also demonstrated in the blood at the time of initiation of the circulation and in the early embryonic liver.

Organ cultures of 7 day embryos with intact yolk sacs, and embryos or yolk sacs after separation have shown the autonomous nature of the development of haemopoiesis in the yolk sac and the dependence of intra‐embryonic haemopoiesis, particularly in embryonic liver, on colonization by yolk sac haemopoietic cells.

Both in‐vivo and in‐vitro colony forming cells have been involved in the first migration stream, between yolk sac and embryonic liver, and evidence has been presented for the role of local environmental factors in controlling the differentiation of these cell types.

These results support the view that development of haemopoietic organs in both embryo and adult is dependent on colonization by circulating cells and that these circulating stem cells originate initially in the yolk sac. This indicates that the yolk sac is the only site of genuine de novo formation of haemopoietic stem cells.
An Example of Polyagglutinability Due to the Tn Antigen
Authors: Gunson H. H. , Stratton F. , Mullard G. W.
Published: March 2008

Summary. A case of atypical polyagglutinability due to the Tn antigen is described. The patient, although apparently healthy, has leucopenia and thrombocytopenia. Her cells presented the usual difficulties of polyagglutinable cells due to the presence of anti‐Tn in typing reagents whether of human or animal origin. The practical advantages, particularly in ABO typing, of using papain to destroy the T and Tn receptors has been demonstrated. Anti‐Tn, like anti‐T, is a 19S and probably a γM globulin.

It has been clearly shown that Tn cells differ from T‐activated polyagglutinable cells and Cad positive cells, but it has not been possible to convert normal cells to Tn cells. Certain aspects of both typical and atypical polyagglutinability have been discussed.
Factor XIII Assay by an Isotope Method
Authors: Dvilansky Alexander , Britten Anthony F. H. , Loewy Ariel G.
Published: March 2008

Summary. The urea and acetic acid solubility tests to detect deficiency of factor XIII were compared. The urea test is more specific, but the acetic acid test is more sensitive, quicker, and therefore more convenient.

A radioisotope assay method for factor XIII (transamidase) is described. Incorporation of 14C putrescine into casein provides a substitute for the linkage by transamidation of fibrin monomers. Fresh plasma, fresh frozen plasma and out‐dated bank plasma have similar transamidase activity. Slight instability is apparent after storage for 3 days at room temperature. The normal range of plasma factor XIII is wide. Aged serum contained 12.5% of normal plasma transamidase. Platelets are a rich source of transamidase activity, particularly after disruption. Approximately half the total whole blood transamidase resides in the platelets and the remainder in the plasma. Leucocytes and red cells contain no transamidase. After lysis, however, red cells do reveal activity in this assay system. This cross reaction requires further elucidation.

The venom (‘Reptilase’) of the Brazilian viper, Bothrops jararaca, can activate factor XIII in similar fashion to thrombin.
The Content of Vitamin B12 in Adult and Foetal Tissue: A Comparative Study
Authors: Rappazzo Mary E. , Salmi Heikki A. , Hall Charles A.
Published: March 2008

Summary. The vitamin B12 content of 17 tissues in 28 adults and 17 foetuses was measured by a Euglena gracilis bioassay. The former were taken after sudden deaths, many accidental, and the latter after therapeutic abortions for mild psychiatric reasons. In the adult, the liver had the highest level of vitamin B12 followed by the kidney, adrenal, pancreas and heart. The tissues of greatest vitamin B12 concentration in the foetus were the liver, adrenal, kidney and lung. Foetal spleen, lung and colon were significantly higher than their adult counterparts, while foetal liver, kidney, heart and brain were significantly lower in comparison with the adult. Differences in vitamin B12 levels may reflect adult and foetal differences in circulation, metabolic maturity or plasma vitamin B12 transport.
Lyophilized Plasma as an Assay Standard for Factor VIII: Calibration and Control with Fresh Plasma Samples
Authors: Hills M. , Ingram G. I. C. , Matchett Myrtle O.
Published: March 2008

Samples of fresh human citrated plasma were lyophilized, mixed and stored in batches at 4°C under vacuum. This material, reconstituted with water, was examined as a standard for factor‐VIII assays. Its initial potency was about 30% of average normal plasma and it lost potency at the rate of 2.6% per month at 4°C and 16.3% per month at room temperature. It is therefore not suitable as a factor‐VIII standard.

The experimental error of the assay was estimated as 83‐120% of the mean. This interval includes approximately 90% of estimated potency ratios whose true values are 100%. The results confirmed that the factor‐VIII concentration in a single individual is reasonably constant and that the range of concentrations in a group of normals is approximately 50‐200% of the mean. The distribution of the values was approximately log‐normal. It is suggested that, in the absence of a more satisfactory stored standard, a single subject who has been calibrated against a population of normals could serve as a factor‐VIII standard.
Continuous Intravenous Infusion of Factor VIII in Classic Haemophilia
Authors: McMillan Campbell W. , Webster William P. , Roberts Harold R. , Blythe William B.
Published: March 2008

The effects of continous intravenous infusion of factor VIII were studied under varied conditions in five children with classic haemophilia. Factor‐VIII activity was stable at room temperature (73–76°F) up to 12 hr in cryoprecipitates from fresh plasma and up to 27 hr in glycine‐precipitated fractions (Fraction AA and Method 4, Hyland). Constant infusion (17 ml/hr) of different concentrations of these fractions produced levels of plasma factor‐VIII activity which were proportional to the dose‐rate and became relatively steady after 12‐18 hr. In separate studies of two of the subjects, dental extraction and laminectomies were uneventfully supported by an initial dose of factor VIII followed by continuous infusions. It is suggested that continuous intravenous infusion of factor VIII is useful for studying the regulation of factor‐VIII levels in plasma and for maintaining steady plasma factor‐VIII activity during replacement therapy.
Thrombokinetics in Idiopathic Thrombocytopenic Purpura*
Authors: Harker Laurence A.
Published: March 2008

Summary Measurements of platelet production in 16 patients with idiopathic thrombocytopenic purpura (ITP) demonstrate that megakaryocytopoiesis (total thrombopoiesis) and platelet turnover (effective thrombopoiesis) are increased in parallel to as much as 8 times normal. The marrow megakaryocytes show changes characteristic of stimulated thrombopoiesis.
Heterogeneity of Human Platelets III. GLYCOGEN METABOLISM IN PLATELETS OF DIFFERENT SIZES
Authors: Karpatkin Simon , Charmatz Arthur
Published: March 2008

Summary Glycogenolysis, glycogenesis, glyconeogenesis and the enzymes phosphorylase, glycogen synthetase, and fructose‐1–6‐diphosphatase were investigated in heavy and light platelet populations in order to determine the presence or extent of metabolic heterogeneity. The mean volume of the heavy platelets was approximately 2.4‐fold greater than that of the light platelets. Heavy platelets contained 3.4‐fold more glycogen and had a 5.9‐fold greater rate of glycogenolysis per g wet weight, than light platelets. Heavy platelets incorporated 5.9‐fold more [14C]glucose into platelet glycogen when data were expressed per g wet weight, and 1.7‐fold more [14C]glucose when data were expressed per μmole of glycogen. This difference in glycogen synthesis was paralleled by a 1.8‐fold greater glycogen synthetase activity. Heavy platelets incorporated 3.0‐fold more [14C]citrate into platelet glycogen when data were expressed per g wet weight, and 1.4‐fold more [14C]citrate when data were expressed per μmol of glycogen. Similar results were obtained utilizing [14C]pyruvate as precursor. This difference in glyconeogenesis was paralleled by a 2.5‐fold greater fructose‐1–6‐diphosphatase activity for heavy platelets.
Studies of Folate Uptake by Phytohaemagglutinin‐Stimulated Lymphocytes
Authors: Das K. C. , Hoffbrand A. V.
Published: March 2008

Summary Cultures of lymphocytes with and without phytohaemagglutinin (PHA) were used as an in vitro model system to study the cellular uptake of tritiated folic acid, [3H]PteGlu and of14C labelled 5 methyltetrahydrofolic acid, 5 [methyl‐14C]‐H4PteGlu. Total folate uptake by PHA‐stimulated transformed lymphocytes, measured both by liquid scintillation counting and by autoradiography, was much greater than by non‐stimulated mature lymphocytes. This suggests that growing and dividing cells take up folate more avidly than mature non‐dividing cells. Uptake of both folate compounds, measured over a 4 hr period, was (a) approximately 80% less at 4°C than at 37°C, (b) exhibited saturation kinetics, and (c) was inhibited by methotrexate in concentrations of 10−3 M, 10−4 M, but was not affected by methotrexate at a concentration of 10−6 M.

The results suggest that an active transport mechanism may be at least partly involved in cellular folate uptake. The two folate compounds, PteGlu and 5 methyl H4PteGlu, however, do not appear to share the same uptake pathway, since PteGlu did not inhibit the uptake of 5 [methyl‐14C]H4PteGlu by PHA‐stimulated lymphocytes. Further, the uptake of 5 [methyl‐14C]H4PteGlu by PHA‐stimulated lymphocytes from six patients with untreated pernicious anaemia was found to be significantly impaired, whereas the uptake of [3H]PteGlu by those cells was normal. Diphenylhydantoin did not show any consistent effect on folate uptake by lymphocytes. A sequential relation was found between the peak rates of RNA synthesis, folate uptake and DNA synthesis by PHA‐stimulated lymphocytes. The peak rate of folate uptake occurred at 44–48 hr of culture, and preceded peak DNA synthesis by 24 hr and followed peak RNA synthesis by 24 hr. Folate uptake, however, did not appear to be directly coupled to either DNA or RNA synthesis, since actinomycin D inhibited both DNA and RNA synthesis by transformed lymphocytes over a 4 hr period without significantly affecting folate uptake.
Effect of Oxidized Glutathione and Acetylphenylhydrazine on Haemoglobin from Normal and Glucose‐6‐Phosphate Dehydrogenase Deficient Individuals
Authors: Zaidman Jacob
Published: March 2008

Summary After incubation of haemolysates of normal and G6PD‐deficient red cells with oxidized glutathione, the GSSG is differently bound to the various HbA3 minor components (as demonstrated by Amberlite CG‐50 column chromatography) of the two kinds of red cells. In normal haemolysates the interaction is with the HbA3e component, and in G6PD‐deficient haemolysates with the HbA3d and HbA3e components.

After incubation of normal and G6PD‐deficient red cells with acetylphenylhydrazine, in the presence of glucose, where GSH is oxidized to GSSG, interaction is observed only in the enzyme‐deficient red cells with the same components as observed in the incubation with GSSG.
In Vivo Effects on Maturation Time of Rabbit Reticulocytes in the Circulation
Authors: Cividalli Lia , Danon David
Published: March 2008

Summary Reticulocyte rich blood with [32P]RNA was transfused into control and experimentally treated host rabbits, to find out whether the maturation time of reticulocytes is predetermined before they enter the circulation or is influenced by extracellular factors. It was found that in controls as well as in rabbits made anaemic by bleeding, splenectomized or subjected to hypoxia or hyperoxia, the maturation time was unaffected by treatment of the host rabbit. In contradistinction maturation was enhanced in iron‐deficient host rabbits. In irradiated host rabbits, maturation was delayed.

It is suggested that the maturation time of reticulocytes in the circulation is not influenced by conditions affecting the cell function as carrier of oxygen, but rather by changes in the protein synthesis apparatus of the reticulocytes, at least as far as iron deficiency is concerned.
Identification of the First Example of Anti‐Cob
Authors: Giles Carolyn M. , Darnborough J. , Aspinall P. , Fletton M. W.
Published: March 2008

Summary A new blood group antibody has been detected in the serum of a male patient who has been transfused on several occasions. In addition to anti‐E + Cw, the patient's serum contains anti‐Cob.
Absorption of Vitamin B12 by the Guinea‐Pig I. SUBCELLULAR LOCALIZATION OF VITAMIN B12 IN THE ILEAL ENTEROCYTE DURING ABSORPTION*
Authors: Peters T. J. , Hoffbrand A. V.
Published: March 2008

Summary.

Guinea‐pig intestinal mucosa was fractionated into brush borders, nuclei, mitochondria, lysosomes, microsomes and a soluble fraction; the mitochondria were subfractionated into inner and outer membranes. Vitamin B12, assayed micro‐biologically using Euglena gracilis, was found to be predominantly localized to the inner mitochondrial membranes. In the fasted animal, the concentration of vitamin B12 was similar in the jejunum and ileum.

Approximately 25% of 10 ng of [58Co]cyanocobalamin given orally was retained after 7 days, when measured by a whole‐body counting technique.

Fasted guinea‐pigs were fed 8–11 ng of [57Co]cyanocobalamin and the distribution of radioactivity throughout the gastro‐intestinal tract was studied. Although radioactivity was present in the ileal mucosa within 1 hr of feeding, no significant hepatic activity was present until 3 hr, confirming the presence of a mucosal delay in vitamin B12 absorption, a characteristic finding of intrinsic factor mediated vitamin B12 absorption. Seventy per cent of the vitamin B12 was found in the colon at 3 and 4 hr indicating that approximately 30% of the dose was absorbed.

The ileal mucosa showed a highly significant rise in mitochondrial specific activity of labelled vitamin B12 during the first 2 hr post‐feeding. This subsequently decreased coincidentally with the rise in hepatic activity. It is suggested that the mitochondria play a hitherto unsuspected role in vitamin B12 absorption. No significant lysosomal radioactivity occurred during the absorptive period; this observation does not support the suggestion that vitamin B12 is absorbed by pinocytosis.
Immunoglobulins in Chronic Cold Haemagglutinin Disease
Authors: Cooper A. G. , Hobbs J. R.
Published: March 2008

Summary.

Total serum IgM levels in 32 patients with chronic cold haemagglutinin disease ranged from 100 to 2400 mg/100 ml; follow‐up in nine patients showed slowly progressive increases.

The excess IgM was mostly cold agglutinin and although there was considerable variation in serologic activity on a molecular basis, specific lytic activity against enzyme‐treated red cells was very similar. The cold agglutinin IgM seemed well produced and closely parallel to normal IgM; few 7S units and little Bence‐Jones proteinuria were detected. Non cold‐agglutinin IgM levels were mostly normal, but 31% of patients showed subnormal levels of IgA or IgG especially when the IgM level was greater than 1000 mg/100 ml.
Red Cell Acetylcholinesterase Activity in Autoimmune Haemolytic Anaemias
Authors: Sirchia G. , Ferrone S. , Mercuriali F. , Zanella A.
Published: March 2008

Summary.

Red cell acetylcholinesterase activity was determined in 59 patients suffering from autoimmune haemolytic anaemia, classified according to the types of erythrocyte‐coating globulins. The enzyme activity was found significantly subnormal in the γG and the γG+C' types of AIHA, while it was not significantly lower than normal in the group of patients where complement (C') alone coated the red cells. These results suggest that different types of autoantibodies are able to cause different biochemical alterations of the red cells.
Intracellular Hydrogen Transport Systems in Acute Leukaemia
Authors: Stuart J. , Simpson J. S. , Mann J. R.
Published: March 2008

Summary. A cytochemical study of intra‐ and extra‐mitochondrial dehydrogenase enzymes in human leukaemic lymphoblasts has shown an increase in activity, relative to the small lymphocyte, of those enzymes which are extramitochondrial in location. In contrast, there was no significant increase in the activity of intramitochondrial respiratory dehydrogenases or of an α‐glycerolphosphate dehydrogenase which is thought to regulate hydrogen transport across the mitochondrial membrane. The findings are in keeping with a hypothesis that leukaemic blast cells show diminished hydrogen transport to the intramitochondrial respiratory pathway. A compensatory increase in lactate dehydrogenase activity in these cells may signify a degree of metabolic dependence on this enzyme, and its selective inhibition by oxamate analogues may therefore result in a cytotoxic effect.
An Explanation of the Failure of the Direct Antiglobulin Test to Detect Erythrocyte Sensitization in ABO Haemolytic Disease of the Newborn and Observations on Pinocytosis of IgG Anti‐A Antibodies by Infant (Cord) Red Cells
Authors: Voak D. , Williams M. A.
Published: March 2008

Summary.

Group A cord cells sensitized with large amounts of IgG anti‐A/B antibodies will give strong antiglobulin test reactions. The failure of the direct antiglobulin test in cases of ABO haemolytic disease is due to the relatively lower level of antibodies sensitizing the cells.

The conjugation of ferritin with IgG anti‐A/B globulin gave an electron dense reagent which was used to study the distribution of A sites on adultand infant group A red cells. Similar contagious patterning of A sites was found on both the adult and infant cells’ surface. The levels of conjugated antibodies on the cells of the different bloods were in agreement with the fact that A1 adult cells have more A sites than A2 adult or infant A cells and that A2 adult cells have similar numbers of A sites to A1 infant cells. The distribution of A sites on infant and adult cells shows similar degrees of dispersion. Thus the average distance between A sites must be greater on infant A1 cells than on adult A1 cells.

A combination of the small molecular size of IgG antibodies and the relatively greater distances between the A sites of A1 infant cells compared to those of A1 adult cells is proposed to be the cause of the failure of the direct antiglobulin test to give strong reactions with the sensitized cells from ABO HD cases.

Pinocytotic activity shown by group A infant cells exposed to IgG anti‐A antibody levels similar to those found in ABO haemolytic disease cases was mainly confined to the more primitive cells containing ribosomes.
Immunocytological Demonstration of Lysozyme (Muramidase) in Human Leukaemic Cells
Authors: Asamer H. , Schmaizl F. , Braunsteiner H.
Published: March 2008

Summary.

Using the immunofluorescent method the intracytoplasmic lysozyme content of leukaemic cells in the various forms of leukaemia was compared with the biochemically assayed serum and urinary lysozyme levels. This enzyme was demonstrated in mature leukaemic monocytes, in promyelocytes and in myelo‐monocytic cells in concentrations varying according to the particular case under study, in the myelogeneous forms of patients with chronic granulocytic leukaemia, but not in immature leukaemic monocytes, myeloblasts or lymphoblasts. Thus the lysozyme content of a single leukaemic cell depends not only upon the type of cell involved but also upon the degree of cellular maturation as evaluated by morphological and histochemical methods. There was an excellent correlation between the amount of cellular lysozyme demonstrated immunocytologically, and the lysozyme level in serum and urine.
The Oxygen Affinity of Haemoglobin in Chronic Renal Failure
Authors: Mitchell T. R. , Pegrum G. D.
Published: March 2008

Summary. The position of the oxygen dissociation curve of haemoglobin was established in 42 healthy subjects and 30 anaemic patients with chronic renal failure, 20 of whom were being treated by maintenance haemodialysis.

Twenty‐four patients showed right‐shifted dissociation curves with a different distribution pattern from that in normal subjects.

This study has shown a negative correlation between the degree of anaemia and the half saturation value of the dissociation curve (r = ‐ 0.59) in chronic renal failure. In patients with undialysed chronic renal failure the patient's own hydrogen ion concentration was found to be important in determining the position of the curve.

The mean advantage in the calculated haemoglobin concentration due to the right shift in the curve was 0.5 g/100 ml more than that measured photometrically. These changes could result in as much as 25% saving in cardiac output in a resting subject. We therefore consider that these compensatory changes are helpful in ameliorating tissue hypoxia in anaemic subjects.
Haematological Effects in Mice of Partially Purified Colony Stimulating Factor (CSF) Prepared from Human Urine
Authors: D. Metcalf  , Stanley E. R.
Published: March 2008

Summary. Single injections of partially purified human urine preparations containing 10 000–20 000 units of colony stimulating factor (CSF) produced a monocytosis and polymorph leucocytosis in neonatal C57BL mice which was maximal at 48 hr. Adult C57BL mice developed a monocytosis, and although no polymorph leucocytosis developed, labelling studies suggested an increased rate of granulopoiesis. In adult mice the injected CSF preparations did not alter total bone‐marrow cellularity but increased the percentage of primitive granulocytes and caused an increase in the number of in vitro colony‐forming cells and granulocytic cluster‐forming cells. The preparations also increased the number of bone‐marrow cells able to initiate colony and cluster formation immediately after culture of the bone‐marrow cells in agar. The results suggest that CSF in vivo may stimulate both granulopoiesis and monocyte formation in an action similar to its action in agar cultures of bone‐marrow cells.
The Significance of Lysozyme Estimations in Acute Myeloid and Chronic Monocytic Leukaemia
Authors: Catovsky D. , Galton D. A. G. , Griffin C.
Published: March 2008

Summary. Serum and urine concentrations of lysozyme (muramidase) were estimated by a turbidimetric method in 17 adults suffering from acute myeloid leukaemia (AML) and six from chronic monocytic leukaemia (CMoL). AML case were classified on cytological and cytochemical criteria as myeloblastic (AMbL), myelomonocytic (AMML) or monoblastic‐monocytic (AMoL) leukaemia.

Serum lysozyme was within normal limits in AMbL, was increased from moderate to high levels in AMML and greatly increaed in all but one AMoL; CMoL had also raised concentrations. The concentrations of lysozyme in 24 hr collections of urine corresponded in general with the serum levels. When serum levels were very high (above 400 μg/ml) the ratio urine/serum was raised 15–40 times.

Repeated lysozyme estimations made for periods of 1–12 mth during the course of treatment in seven patients with AMML or AMoL demonstrated three patterns of changes: (a) in four cases the serum concentrations fell during remission and rose again during relapse; when the remission was not complete they remained above the upper limit of the normal range; (b) in one case the changes were as (a), but when the relapse occurred the serum and urine concentrations remained normal; (c) in two cases of AMML with only moderately increased lysozyme levels, the changes in the serum concentration followed only the changes in the monocyte counts, thus reflecting only partially the activity of the disease.

Lysozyme estimations are useful in classifying AML and in assessing the degree of remission achieved as a result of treatment. Additional observations regarding the presence of abnormalities in the neutrophils, lymphadenopathy, and gum hypertrophy were also found to be useful in the classification of AML, the former being more common in AMbL and the latter in AMoL.
Clinical Investigation of Intermediate‐ and High‐Purity Antihaemophilic Factor (Factor VIII) Concentrates
Authors: Johnson A. J. , Karpatkin Margaret H. , Newman J.
Published: March 2008

Summary. Intermediate‐ and high‐purity antibaemophilic factor (factor‐VIII) concentrates developed by the American National Red Cross (ANRC) have undergone extensive clinical investigation. When administered to severely affected haemophiliacs, the recovery in vivo and the metabolic half‐disappearance time were similar to those achieved with plasma.
Reptilase®‐R—A New Reagent in Blood Coagulation
Authors: Funk C. , Gmür J. , Herold R. , Straub P. W.
Published: March 2008

Summary. Reptilase®‐R, a purified thrombin‐like snake‐venom enzyme from Bothrops atrox, has been tested for its usefulness as a substitute for thrombin in two coagulation assay systems. Reptilase was found to be more stable than thrombin and not to be inhibited by heparin and hirudin.

Performance of the Reptilase time allows the rapid exclusion of heparin contamination in plasma samples with prolonged thrombin times. In the presence of fibrinogen/fibrin split products, the Reptilase time is less prolonged than the thrombin time, whereas in patients with prolonged thrombin times because of congenital dysfibrinogenaemia the inverse is found. Thus, performance of both Reptilase time and thrombin time allows the rapid identification of three major causes for a prolongation of the thrombin time. In patients on heparin therapy because of a defibrination syndrome, the Reptilase time may replace the thrombin time as a guide for the degree of coagulation inhibition by FDP, although our results show that it is somewhat less sensitive.

Reptilase can also be substituted for thrombin in the rapid chronometric fibrinogen assay. Unfortunately the advantages of reagent stability and insensitivity to heparin are balanced by a lesser reproducibility, especially in the presence of FDP.
The Sequestration Site of Platelets in Idiopathic Thrombocytopenic Purpura: its Correlation with the Results of Splenectomy
Authors: Najean Y. , Ardaillou N.
Published: March 2008

Summary. The clinical usefulness of external scanning data after infusion of 51Crlabelled platelets into patients with idiopathic thrombocytopenic purpura (ITP) is a matter of controversy. Observations have been made in 575 patients with ITP. Short‐term (6 mth) results of splenectomy were assessed in 206 subjects, and longterm (1‐3 yr) in 153. It appears that the site of sequestration is neither a direct function of the severity of the disease nor of the duration of the disease from the clinical onset. Diffuse sequestration, which cannot be taken as an indication for or against splenectomy, is frequently seen in recent and severe cases. Splenic sequestration is more often observed in young patients (72.5% under 30 yr of age) than in older subjects (36% over 30 yr of age).

A good correlation was found between the site of sequestration and the shortand long‐term results of splenectomy: success in more than 90% of cases with splenic sequestration but complete failure in 70% with hepatic sequestration. In any patient with ITP splenectomy should be undertaken only after a careful study of the platelet sequestration site.
Some Factors Affecting the Release of Iron from Reticuloendothelial Cells
Authors: Lipschitz D. A. , Simon M. O. , Lynch S. R. , Dugard J. , Bothwell T. H. , Charlton R. W.
Published: March 2008

Summary. Factors modifying the release of iron from reticuloendothelial cells were studied in rats by injecting heat‐denatured erythrocytes containing [59Fe]haemoglobin. The cells were rapidly taken up by the liver and spleen, and a proportion of the 59Fe was released into the plasma, the maximum rate being between 1 and 4 hr after injection. The remaining 59Fe was incorporated into storage compounds. A 10‐fold variation in the load of denatured erythrocytes produced a proportional change in the amount of iron released, the percentage remaining constant. Percentage release of 59Fe was enhanced in venesected rats and diminished in hypertransfused rats. Release was inhibited by injecting either unlabelled denatured erythrocytes or iron bound with nitrilotriacetic acid (NTA‐iron) before the 59Fe‐labelled cells, the maximum effect being obtained if the interval between the two injections was 3‐9 hr. Release was also inhibited by injecting NTA‐iron 30 min after the denatured labelled erythrocytes. Inhibition was always preceded by a rise in the serum‐iron concentration, and was associated with an increase in the percentage of 59Fe incorporated into ferritin. It is postulated that the shortage of free transferrin binding sites for iron delays the entry of liberated haemoglobin iron into the plasma, and consequently there is enlargement of a ‘pre‐release’ iron pool. Other workers have shown that iron induces the synthesis of ferritin; the presence of a stimulated mechanism for ferritin synthesis within the reticuloendothelial cells would result in the diversion of an increased percentage of erythrocyte iron into storage compounds.
Eosinophilia, Splenomegaly and Cardiac Disease
Authors: Shepherd A. J. N. , Walsh C. H. , Archer R. K. , Wetherley‐Mein G.
Published: March 2008

Summary. Three cases of eosinophilia, splenomegaly and cardiac disease are presented. Cases of a similar nature have been reported in the literature and the question as to whether this syndrome is an eosinophil leukaemia or a primary connective tissue disorder is discussed.
Studies of the Effect of Sulphydryl and Other Inhibitors on Reticulocyte Uptake of Doubly‐labelled Transferrin
Authors: Edwards Shirley A. , Fielding J.
Published: March 2008

Summary. Inhibition of iron uptake from transferrin by reticulocytes has been further studied using doubly‐labelled 59Fe‐125I‐transferrin. The inhibition by p‐hydroxy mercuribenzoate, previously reported, is shown to occur also with the sulphydryl inhibitors p‐hydroxymercuriphenylsulphonate and N‐ethylmaleimide. The pattern of ‐SH inhibition is compared with and found to differ from that for two other inhibitors, trypsin, which alters membrane binding, and sodium fluoride, a metabolic inhibitor. When assessed by three variables, namely the effect of inhibitor concentration, the time of inhibitor exposure, and the effect on the rate of uptake, sulphydryl inhibitors are shown to act differentially on transferrin binding and on iron uptake. The latter is far more sensitive and is affected at lower concentrations. It is concluded that ‐SH groups are active in the dissociation of iron from its transferrin bond after surface binding has occurred. The effect of‐SH inhibitors on transferrin binding may be secondary to changes in membrane function induced by higher concentrations of these inhibitors.
Immunological Homogeneity of Membrane Proteins from Hereditary Spherocytic, Hereditary Elliptocytic, and Normal Red Cells
Authors: Gomperts E. D. , Metz J. , Zail S. S.
Published: March 2008

Summary. Haemagglutination inhibition and immunodiffusion techniques were applied to the study of red‐cell membrane protein obtained from normal human subjects, patients with hereditary spherocytosis, patients with hereditary elliptocytosis, and also from calves and monkeys.

These techniques failed to detect differences between the normal and abnormal human red cells but did show differences between the red cells of the human and the two other mammalian species.
Coexistence of Pernicious Anaemia and Chronic Myeloid Leukaemia: An Experiment of Nature Involving Vitamin B12 Metabolism
Authors: Corcino J. J. , Zalusky R. , Greenberg M. , Herbert V.
Published: March 2008

Summary. Chronic myeloid leukaemia (CML) and pernicious anaemia (PA) coexisted in a 51‐yr‐old man (who also had myasthenia gravis). His serum vitamin B12 level was normal instead of the high value expected in CML, or the low value expected in PA. Short‐term culture of his peripheral blood cells showed deranged DNA synthesis of the type observed in vitamin B12 deficiency, indicating that his ‘normal’ serum vitamin B12 level was not associated with normal amounts of vitamin B12 available to his CML cells. A tracer dose of 0.45 μg of[57 Co]vitamin B12 disappeared abnormally slowly from his serum over 24 hr (as previously observed by others in patients with either CML or PA). A therapeutic injection of vitamin B12 appeared to disappear abnormally slowly from his serum over a period of 2 yr. His serum showed a markedly elevated unsaturated vitamin B12 binding capacity, with a less sharp increase in percentage of vitamin B12 binding α as compared to β globulin than usually expected in CML.

Withholding therapeutic vitamin B12 for 2 yr has been associated with stabilization of his WBC at the 40 000‐50 ooo/μl range. Although this association could be chance rather than cause and effect, the patient does represent an ‘experiment of nature’ involving vitamin B12 metabolism and does raise the question as to whether his leukaemia is retarded by his deficiency of vitamin B12, which results from inadequate absorption of vitamin B12 and possibly from elevated serum vitamin B12 binding a globulin rendering his circulating vitamin B12 metabolically inert.
Studies on the Adsorption of Factor VIII: Application to the Purification of Bovine Factor VIII
Authors: Simonetti C. , Casillas G. , Pavlovsky A.
Published: March 2008

Summary. The adsorption on tricalcium phosphate and on barium sulphate of factor VIII from bovine and human plasma and from human factor VIII concentrates was studied. A technique for the preparation of a concentrate of bovine factor VIII by direct adsorption of plasma with tricalcium phosphate and then filtration on agarose gel has been developed. The use of EACA, glucose and glycine markedly increase the yield and make possible the lyophilization of highly purified factor VIII. The technique can be used on a preparative scale using 10–30 litres of bovine plasma.
Clinical and Ultrastructural Aspects of Congenital Dyserythropoietic Anaemia Type I
Authors: Lewis S. M. , Nelson D. A. , Pitcher C. S.
Published: March 2008

Summary. A case of congenital dyserythropoietic anaemia (CDA) type I is described This is characterized by erythroblastic multinuclearity and ineffective erythropoiesis. By contrast to ‘HEMPAS’ or CDA type II, the acidified‐serum test is negative. The morphological features of the bone marrow have been studied by electron microscopy. These include loss of nuclear envelope, uneven condensation of nuclear chromatin with spongy appearance, chromatin bridges connecting nuclei of divided cells and also cytoplasmic connection between cells by bundles of microtubules. The persistence of microtubules in the cytoplasm and presence of cytoplasmic organelles within the nuclear area are other unusual features. The significance of these findings is discussed in relation to speculation concerning the pathogenesis of the disease.
The Mode of Action of Antibodies which Destroy Factor VIII
Authors: Biggs Rosemary , Austen D. E. G. , Denson K. W. E. , Rizza C. R. , Borrett R.
Published: March 2008

Summary. In 1959 Biggs & Bidwell proposed the hypothesis that the reaction between antibody and factor VIII could be interpreted as a second‐order reaction and that in antibody excess the amount of antibody present could be measured using a linear calibration graph relating the logarithm of residual factor VIII to antibody concentration after incubation for a fixed period of time. This hypothesis has been re‐examined because antibody is seldom present in excess in most routine test systems and in view of a new and attractive hypothesis put forward by Pool & Miller (1972). All the evidence seems to suggest that the original hypothesis of Biggs & Bidwell is adequate for assessing the potency of antibodies of the type occurring in many haemophilic patients.
The Mode of Action of Antibodies which Destroy Factor VIII
Authors: Biggs Rosemary , Austen D. E. G. , Denson K. W. E. , Borrett R. , Rizza C. R.
Published: March 2008

Summary. Antibodies which destroy factor VIII may follow second‐order kinetics (Biggs et al, 1972) or a more complex type of reaction may occur. The present communication concerns the complex reactions and evidence is presented which supports the hypothesis that these antibodies may react with factor VIII to form a complex with some factor‐VIII activity.
A Comparison of Erythrocyte Characteristics in Sickle Cell Syndromes in Jamaica
Authors: Serjeant G. R. , Serjeant B. E.
Published: March 2008

Summary. Red‐cell characteristics were studied in the steady state in three sickle‐cell syndromes, homozygous sickle‐cell disease (SS), sickle‐cell/haemoglobin‐C disease (SC), and sickle‐cell/β‐thalassaemia (S/thal). Hb‐SC disease had the highest haemoglobin levels, red cell counts, and mean corpuscular haemoglobin concentrations, all of which may contribute to the high thrombotic tendency noted in this disease. The two types of S/thal (with and without Hb A) generally had different haematological features. The non‐Hb‐A type of S/thal, which may resemble SS disease on electrophoretic techniques and present a diagnostic problem, was distinguishable on many red cell characteristics reported here.
In Vitro Studies of Bone Marrow in Refractory Anaemia
Authors: Sultan C.
Published: March 2008

Summary. Bone marrows of six patients with refractory anaemia have been studied for their capacity to form colonies in vitro on agar gel medium. They are not capable of forming colonies, and in this they behave as do bone marrows from patients with acute myeloblastic leukaemia in relapse.
The Subcellular Distribution of 59Fe in Small Intestinal Mucosa: Studies with Normal, Iron Deficient and Iron Overloaded Rats
Authors: Worwood M. , Jacobs A.
Published: March 2008

Summary

Iron deficient and iron loaded rats were given 59Fe by stomach tube and the subcellular distribution of 59Fe in the intestinal mucosa was investigated. There was considerable concentration of 59Fe in the soluble fraction shortly after administration of the dose but concentration in the mitochondria at 12 and 18 hr. Concentration in the mitochondria was particularly marked in the iron deficient rats. After intravenous injection of 59Fe normal, iron deficient and iron loaded rats all showed the greatest concentration of the isotope in the mitochondrial fraction. The incorporation of 59Fe into ferritin was not greater in the iron loaded rats than in normal rats, although the absorption of 59Fe was only one third of the normal level. In iron deficiency very little radioiron was incorporated into ferritin. These results support the suggestion that ferritin formation may not be a major controlling factor in iron absorption and demonstrate the need to consider the role of mitochondria in iron transport within the intestinal epithelial cell.
Studies of the Antibody on the Platelet Surface in Patients with Factor‐VIII Inhibitors
Authors: Ali M. A. M. , Blajchman M. A.
Published: March 2008

Summary

An IgG $kP immunoglobulin was present on the surface of the platelets in four of five patients with factor‐VIII inhibitor. A similar antibody was found in the plasma of two out of three patients. Serum IgM was subnormal in two out of three patients with‘idiopathic’ acquired factor‐VIII inhibitor, while the two patients with haemophilia A (with inhibitors) had normal serum immunoglobuhns.
Storage of Human Platelets: Effects on Metabolically Active ATP and on the Release Reaction
Authors: Kotelba‐Witkowska Barbara , Holmsen Holm , Murer Erik H.
Published: March 2008

Summary

During storage platelets rapidly lose their viability and their functions of aggregation, adhesion and secretion. This loss of function does not correlate with the small decrease in total ATP occurring. Platelet adenine nucleotides probably exist in two pools, one metabolically active and one inactive, so that only 40% of total ATP actually participates in the energy reactions. The metabolic pool was labelled by incubation of platelet‐rich plasma (PRP) in acidified citric acid‐sodium citrate‐dextrose (ACD) with trace amounts of radioactive adenine. Platelets thus labelled in acidified ACD‐PRP were stored in the same medium for 24 hr at 4°C, and the effect of storage on radioactive ATP and on the release reaction induced by collagen and thrombin was studied. The total amount of ATP decreased by only 10% during storage, whereas the metabolic ATP was reduced by 40–50% with a corresponding formation of hypoxanthine. The reactivity in the release reaction also dropped by 50%. When the acidified ACD‐PRP was stored with 9 him inosine a 20% reduction of metabolic ATP and 25% reduction in release ability occurred. Storage with 8 mM adenosine also counteracted the loss in release ability, but 50% of the ATP radioactivity was lost. Adenosine increased the total ATP level during storage. Thus, the metabolic ATP pool might have been preserved intact under these circumstances, while exchange of radioactive ATP with nonradioactive ATP synthesized from adenosine took place. These results indicate strongly that during storage of platelets the metabolic ATP pool is lost by degradation to hypoxanthine in quantities that might well account for the loss in function.
Platelet and Fibrinogen Kinetics with [75Se]Selenomethionine in Thrombocytopenic States. II
Authors: Brodsky Isadore , Ross Evelyn M. , Kahn S. Benham , Petkov Goldie
Published: March 2008

Summary

Simultaneous platelet and fibrinogen kinetics were determined with a cohort label, [75Se]selenomethionine, in 19 thrombocytopenic patients divided into three groups. Group 1 consisted of six patients with idiopathic thrombocytopenic purpura, five were splenectomized and one treated with corticosteroids after the isotope study; Group 2 consisted of four patients with hypersplenism who were splenectomized after the isotope study; and Group 3—nonsplenectomized patients—consisted of two with hepatic cirrhosis, six with either leukaemia or lymphoma and one with paroxysmal nocturnal haemoglobinuria (PNH). A control group, consisting of patients with non‐haernatologic disease and normal coagulation profiles, had a mean platelet survival of 10.5 days and a mean fibrinogen survival of 7.8 days. In both Groups 1 and 2 the mean platelet survivals of 2.4 and 5.2 days respectively were sharply decreased as compared to the controls and also to the mean platelet survival of 11.0 days in Group 3. Mean platelet turnovers in all three groups were either decreased or normal as compared to the control range. Platelet survival curves in Groups 1 and 2 suggested random destruction and in Group 3 a combination of random plus senescent destruction. In Groups 1 and 2 splenectomy or corti‐costeroid therapy resulted in excellent haematologic remissions. In all three groups the mean fibrinogen survivals were within the control range of 6.5–9.5 days. However, one patient, in Group 2 had a substantial decrease in both platelet and fibrinogen survival, probably indicative of low‐grade disseminated intravascular coagulation. A hypercoagulable state was not demonstrated in the patient with PNH. Simultaneous platelet and fibrinogen kinetics are important adjuncts to the study of the pathogenesis, differential diagnosis, therapy, and criteria for splenectomy in thrombocytopenic patients.
Production of Spherocytosis and Increased Osmotic Fragility in Normal Erythrocytes by Sodium Stress
Authors: Ikoku N. B. , Fielding J.
Published: March 2008

Summary

Red cells from normal human subjects were exposed to hypertonic sodium chloride solutions for varying periods of time. The cell changes commonly observed in hereditary spherocytosis were induced, including microspherocyte formation with corresponding increase in osmotic fragility. Solutions of magnesium sulphate of equal osmolarity did not produce these changes. These observations lend support to the theory that excessive sodium permeability is concerned in the pathogenesis of hereditary spherocytosis. Increased influx of sodium and water into erythrocytes leads to spherocytosis, but only increased efflux of sodium leads to irreversible microspherocytosis.