bluuebunny/crossref_metadata_2025 huggingface dataset:
The in Vitro Action of Dapsone and its Derivatives on Normal and G6PD‐deficient Red Cells
Authors: Scott G. L. , Rasbridge Marian R.
Published: March 2008

Summary. In an attempt to define the mechanism of dapsone‐induced haemolysis the in vitro action of dapsone (4,4′, diaminodiphenylsulphone) and some of its derivatives on normal and G6PD‐deficient red cells has been investigated. Dapsone, its N′ acetyl‐ and hydroxy‐derivatives have no measurable effect on metabolism, haemoglobin or the membrane in either normal or G6PD‐deficient red cells. 4, hydroxylamino‐4′ aminodiphenylsulphone (HADS) and 4, hydroxylamino‐4′ acetaminodiphenylsulphone (HMDS) have oxidant effects on red cells. Both of these compounds readily form methaemoglobin but in normal red cells higher levels of methaemoglobin are achieved with HMDS than with HADS. With both compounds less methaemoglobin is formed in glucose‐deprived or in G6PD‐deficient red cells. In normal red cells hexose monophosphate pathway (HMP) activity is increased, HADS being a more potent stimulus than HMDS. It is suggested that methaemoglobin formation is dependent upon the recycling of these compounds via the HMP.

Normal red cells incubated with the two hydroxylamine compounds did not show any change in reduced glutathione (GSH), globin or the membrane, apart from increased sensitivity to peroxide lysis. G6PD‐deficient red cells, on the other hand, did lose GSH rapidly and underwent increased autohaemolysis. The mechanism of these effects is discussed.

Some, but not all, of the in vivo effects of dapsone can be reproduced by the hydroxylamine derivatives in vitro. It is concluded that either some other derivatives are involved or that the effect of long exposure to these oxidant compounds in vivo is different.
Effect of Warfarin Anticoagulation on Vitamin‐K1 Metabolism in Man
Authors: Shearer M.J. , Mc Burney A. , Barkhan P.
Published: March 2008

Summary. Three normal men were injected intravenously with 1 mg [1′,2′‐3H2]‐vitamin K1 before and after treatment with therapeutic doses of warfarin for 8–10 days. Warfarin treatment caused a delayed clearance of lipid‐soluble radioactivity from the plasma. This was found to be due to the accumulation in the plasma of a metabolite with the properties of vitamin K1 oxide as shown by thin‐layer chromatography: the rate of clearance of vitamin K1 from the plasma was not affected. Under warfarin treatment there was about a two‐fold increase in the urinary excretion of radioactivity.
Folated Deficiency in Pernicious Anaemia Measured by Determination of Decreased Serine Synthesis in Lymphocytes
Authors: Ellegaard Jøgen , Esmann Viggo
Published: March 2008

Summary. A functional folate deficiency was determined in four of five patients with untreated pernicious anaemia by a new method (Ellegaard & Esmann, 1972a) for determination of intracellular folate activity utilizing the folate‐dependent incorporation of 14C‐formate into serine of lymphocytes under standardized conditions. The decreased folate activity of lymphocytes was improved within 3 days of treatment with vitamin B12. These observations support the theory of an intracellular, functionally inert pool of 5‐methyl‐tetrahydrofolate in vitamin B12 deficiency due to impaired methionine synthesis.
Effect of ADP‐Induced Aggregation on 32PO4 Incorporation into Phosphatidic Acid and the Phosphoinoistides of Rabbit Platelets
Authors: Lloyd J. V. , Nishizawa E. E. , Joist J. H. , Mustard J. F.
Published: March 2008

Summary. The time course of the ADP‐induced changes in 32P‐labelling of phosphatidic acid (PA), monophosphatidylinositol (MPI), diphosphatidylinositol (DPI) and triphosphatidylinositol (TPI) in suspensions of washed rabbit platelets was investigated. Washed rabbit platelets were labelled with 32PO4, washed, and resuspended in a medium containing unlabelled phosphate. When this suspension was kept at 37°C for 40 min the specific activity of PA slowly decreased. Seven seconds after the addition of ADP, an increase in the labelling of PA was detected. In suspensions that contained no apyrase, deaggregation was slow or absent and the ADP‐induced increase in labelling of PA was still present up to 450 s after the addition of ADP. In suspensions that contained apyrase, ADP‐induced platelet aggregation was followed by deaggregation; the subsequent return of the platelets to a disc shape was associated with the virtual disappearance of the ADP‐induced increase in labelling of PA. In a similar experiment in which 32PO4 was added to the suspending medium, ADP also caused an increase in labelling of PA which was followed by a decrease. An increase in labelling of MPI could be detected 180–450 s after the addition of ADP and an increase in labelling of DPI at 30, 120 and 450 s after the addition of ADP. The changes induced by ADP in the labelling of phospholipids were not due to changes in the amounts of these phospholipids. The results for PA indicate that ADP may induce the synthesis of a small highly labelled pool of PA. The changes found are consistent with a possible role for these phospholipids in the mechanism of ADP‐induced platelet aggregation.
Proliferative Activity and Glycogen Accumulation of Erythroblasts in β‐Thalassaemia
Authors: Yataganas X. , Gahrton G. , Fessas Ph. , Kesse‐Elias M. , Thorell B.
Published: March 2008

Summary. A combined microfluorometric, microspectrophotometric and autoradiographic method was used to determine the glycogen content, measured as the fluorescence intensity after a fluorescent periodic acid‐Schiff reaction (F‐PAS), of erythroblasts in homozygous β‐thalassaemia. F‐PAS‐positive erythroblasts were found most frequently in the G1‐phase of early polychromatic erythroblasts and seem to account for the increased number of cells in this phase of DNA‐synthesis in β‐thalassaemia as compared to normal. The fraction of F‐PAS‐positive G1‐early polychromatic erythroblasts was higher and the glycogen content of F‐PAS‐positive cells was greater in severe as compared to less severe cases, which seems to be of prognostic importance. It is postulated that the accumulation of glycogen represents a storage of unutilized energy in erythroblasts that are blocked in the G1‐phase of the cell cycle.
A Solid‐Phase Radioassay for Factor‐XIII Activity (Fibrin Stabilizing Factor) in Human Plasma
Authors: Schmer G.
Published: March 2008

Summary. Casein, insolubilized to scpharosc, was used as a substrate for measuring factor‐XIII activity. Activated factor XIII incorporated [14C]putrescine into the insolubilized substratc. After the removal of non‐incorporated putrescine, the insolubilized 14C‐tagged cascin was readily digested off the gel by pronase and counted in a beta counter. The rate of [14C]putrescinc incorporation was linear for up to 4 hr. Standard deviations of the assay were ±4% for 35 single determinations. The assay is easy to perform, highly reproducible and inexpensive, which makes the method suitable for routine use in a coagulation laboratory.
Direct Determination of Red Cell Bound Antibody Specificity
Authors: Lalezari Parviz
Published: March 2008

Summary. A new method is described for direct determination of specificity of red cell‐bound antibodies without the need for antibody elution. This method employs Polybrene in an Auto Analyzer continuous flow system. Washed antibody‐coated cells are mixed, in different proportions, with uncoated indicator cells of known blood type. If the indicator cells possess appropriate receptor sites for the cell‐bound antibody, ‘coagglutination’ occurs between the antibody‐coated and indicator cells. In contrast, in the absence of specific receptor sites, indicator cells not only fail to coagglutinate but physically interfere with the expected agglutination of antibodycarrying cells. Quantitative data are provided which appear to be related to the amount of cell‐bound antibody. The technique has been used to identify antibodies with Rh and penicillin specificities and antibodies associated with acquired haemolytic anaemia.
Perturbation of Generation Cycle of Human Leukaemic Lymphoblasts in Vivo by l‐Asparaginase
Authors: Ernst Peter
Published: March 2008

Summary. In four patients with acute lymphoblastic leukaemia the cytokinetic effect on leukaemic lymphoblasts of l‐asparaginase has been studied in vivo, by a method of combined autoradiography and microspectrophotometry. During an observation period of 30 hr the labelling index, the mitotic index and the mean grain count decreased, and in one case studied a rapid decline in the peripheral blast cell count occurred. The interpretation of the observations is that l‐asparaginase arrests cells in S phase, whereas no preferential effect is exerted on cells in G1 and mitosis. Besides this action l‐asparaginase has a generalized lytic effect on lymphoblasts independent of proliferative activity. Differentiated myeloid and erythroid cells seem to be resistant to the action of l‐asparaginase.

In the study a microspectrophotometric method by which it is possible to measure in the autoradiograms the DNA content of single cells labelled with 3H‐nucleoside without interference from the autoradiographic silver grains is described.
Studies on the Mechanism of Erythrocytosis Associated with a Uterine Fibromyoma
Authors: Ossias A. Lawrence , Zanjani Esmail D. , Zalusky Ralph , Estren Solomon , Wasserman Louis R.
Published: March 2008

Summary. Erythrocytosis has occasionally been associated with uterine fibromyoma and attributed to an inappropriate elaboration of erythropoietin (Ep) by the tumour. Whether Ep per se, its precursor substrate, or activator analagous to the renal erythropoietic factor (REF) is produced, has not been defined. In a 54‐yr‐old woman with erythrocytosis and uterine fibromyoma, plasma, urine and tumour extracts were examined to clarify these possible mechanisms. Preoperatively, elevated serum and urinary Ep were present with constant levels over a range of haematocrits induced by serial phlebotomy. Extracts of the tumour did not generate Ep when incubated with normal serum, nor did they contain substrate activity for known REP. Tumour extracts did, however, possess Ep activity which was abolished with anti‐Ep. The patient has remained haematologically normal more than 1 yr following surgery. The data show that the erythrocytosis associated with uterine fibromyoma is mediated through the direct production of Ep by the tumour and not by a substrate‐REF‐like mechanism as found in the normal kidney.
Chronic Lymphocytic Leukaemia Associated with Acute Myelomonocytic Leukaemia
Authors: Roberts P. D. , Forster P. M.
Published: March 2008

Summary. An elderly male, with no previous medical history of note, presented with haematological changes of chronic lymphocytic leukaemia combined with acute myelomonocytic leukaemia. This patient had never received cytotoxic drugs or radiotherapy.
Quantitative Haemagglutination
Authors: Greenwalt T. J. , Steane E. A.
Published: March 2008

Summary. NANA is known to be part of the M and N antigenic receptors. Despite this, the oldest population of red cells in a blood sample agglutinates more strongly with anti‐M and ‐N reagents even though the total NANA content of the cells is lower than that of the youngest population. Removal of small amounts of NANA by the action of neuraminidase results in increased agglutinability with human anti‐M and ‐N and with rabbit anti‐N but not with rabbit anti‐M. Each reagent (human and rabbit anti‐M; human, rabbit and lectin anti‐N) produces a different pattern of agglutination with progressively neuraminidase treated cells, probably indicating that antisera from human, rabbit and seed sources react with different portions of the receptors. The variations in behaviour of human and rabbit anti‐M and anti‐N reagents can be more easily explained by these observations.
Glucose‐6‐Phosphate Dehydrogenase Deficiency in Maltese Newborn Infants
Authors: Grech J. L. , Vicatou Maria
Published: March 2008

Summary. A survey was carried out to establish the incidence of erythrocyte glucose‐6‐phosphate dehydrogenase deficiency in newborn infants in the Maltese islands. Of the infants born during the period of the survey 28.7% were tested: 1147 samples were obtained from male and 1016 samples from female infants. Some degree of enzyme deficiency was detected in 5.1% of the samples, and 1.0% were completely enzyme deficient; 5.8% of the male infants and 4.1% of the female infants were enzyme deficient, but only 1.5% of the males and 0.4% of the females were completely deficient. The results are compared with those of a previous survey carried out on the child and adult populations of the islands. G‐6‐PD deficiency was not found in association with the foetal haemoglobin variant Hb F(Malta).
Phenobarbitone Sensitivity of Jaundice in Haemolytic Patients
Authors: Perona G. , Corrocher R. , Frezza M. , Falezza G. C. , Cellerino R. , Tiribelli C. , Fusaro A. , De Sandre G.
Published: March 2008

Summary. Eleven cases of chronic haemolytic jaundice treated with phenobarbitone are reported. In all patients, except one, the drug caused a significant reduction of hyperbilirubinaemia in spite of the persistence of an increased haemolysis. Radioactive bilirubin (3H) kinetic studies demonstrated a bilirubin transfer defect from plasma to liver in seven patients which improved after phenobarbitone therapy along with the reduction of bilirubinaemia. It is concluded that a phenobarbitone‐sensitive hepatic mechanism frequently operates in producing jaundice in chronic haemolytic syndromes.
Early Onset of Pernicious Anaemia in Two Siblings: Genetic and Autoimmune Aspects
Authors: Hall Charles A. , Beebe Richard T.
Published: March 2008

Summary. Two sisters were found to have classical pernicious anaemia developing prior to the age of 20. Their mother and maternal grandmother both had spontaneous hypothyroidism. The father had classical pernicious anaemia at the age of 47. The two sisters had antibodies against thyroid tissue, parietal cells and intrinsic factor. One sister had anti‐thyroglobulin as well. The father and mother had parietal cell antibodies in insignificant titres; all other antibodies in the mother were negative. The maternal grandmother was likewise free of antibodies; probably the long duration of hypothyroidism in the mother and grandmother permitted the decline of antibodies. It is postulated that the presence of autoimmune disease in both sides of the family potentiated a genetic predisposition to autoimmune disease. As a consequence, the offspring developed pernicious anaemia at an unusually early age and had antibodies against several tissues and/or their products including thyroid tissue but no overt thyroid disease.
Isoprecipitins: Precipitating Antibodies Reacting with Blood Group Specific Substances
Authors: Pintera J. , Vacl J.
Published: March 2008

Summary.

A precipitating iso‐anti‐A was discovered in the serum of a female blood donor when routine tests were being performed for anti‐Au HAA antibody by means of counter‐immunoelectrophoresis. The isoprecipitin reacted with the sera of blood group A and AB donors and with dilute solutions of blood group A substance. Using the same technique, a significantly higher instance of anti‐A and anti‐B isoprecipitins was found in mothers of children suffering from haemolytic disease of the newborn due to ABO incompatibility. Regular screening of antenatal sera for precipitating anti‐A and anti‐B is recommended for possible antenatal prediction of affected infants. False positive reactions might be obtained when known Au HAA antigen‐positive sera used for screening for the presence of human anti‐Au HAA also contain A or B specific substances which could react with corresponding precipitating antibodies.
125I‐Transferrin Turnover in Rabbits with Haemolytic Anaemia
Authors: Shepp Margaret , Yamada Hideo , Berenfeld Marion , Gabuzda Thomas G.
Published: March 2008

Summary.

The metabolism of purified 125I‐labelled homologous transferrin in rabbits with haemolytic anaemia was compared to that in control rabbits and no significant differences were found. During 3 weeks of haemolysis plasma transferrin concentrations did not change appreciably. The fractional turnover rate was 43% of the intravascular pool per day. The intravascular pool was 45% of the total exchangeable transferrin.
Metabolic Studies on Red Cells from Patients with Chronic Renal Disease on Haemodialysis
Authors: Wallas Charles H.
Published: March 2008

Summary. Studies were performed which suggest intracorpuscular metabolic alterations in red cells from patients with chronic renal disease. In a buffered salt solution, the red cell metabolic rate of uraemic subjects was significantly greater that that of normal subjects. Furthermore, the metabolic rate of uraemic red cells was not diminished when incubated in normal plasma. Finally, incubation of control and uraemic red cells in a buffered salt medium with 10 mM phosphate did not obliterate the differences in metabolic rate between control and uraemic cells. When the activities of several rate‐limiting glycolytic enzymes were measured, red cells from the uraemic subjects had a normal level of phosphoglycerate kinase and a significant increase in the activities of hexokinase, glucose‐6‐phosphate dehydrogenase, pyruvate kinase and glutamic‐oxaloacetic transaminase compared to control red cells. These results indicate that red cells from patients with chronic renal disease are intrinsically hypermetabolic, and the pattern of increased enzyme activities suggests the presence of a red cell population with a younger mean cell age due to preferential loss of older cells.
Changes in Megakaryocyte Development Following Thrombocytopenia
Authors: MacPherson G. G.
Published: March 2008

Summary. Rats were made thrombocytopenic by total‐body X‐irradiation and changes in megakaryocyte development studied in shielded tibial bone marrow. The platelet count fell from the fourth day after irradiation to reach a minimum value of 19.5% of the initial count 7 days after irradiation. No changes in megakaryocyte development were observed 5 days after irradiation. Megakaryocyte size was increased at day 7, and the increment maintained until day 12. Megakaryocyte numbers were only significantly increased 12 days after irradiation, but differential counts and electron‐microscope autoradiographs suggested increased influx at day 9. Changes in the labelling index of megakaryocytes suggested that some cells already present were undergoing one or more extra endoreduplications. No evidence was obtained for an accelerated megakaryocyte maturation rate or for a sudden release of platelets from mature megakaryocytes. Morphological changes, signifying increased synthesis of protein and demarcation membranes were apparent only in young cells 7 days after irradiation but were present in mature cells by day 12, demonstrating that the response to thrombocytopenia is confined to immature cells.
The Separation of Free and Bound Vitamin B12
Authors: Adams J. F. , Mcewan Fiona C.
Published: March 2008

Summary. The separation of free and bound vitamin B12 was studied using four binders and two forms of vitamin B12 and effecting separation by charcoal adsorption, bag dialysis, ultrafiltration and gel filtration. In charcoal separation systems the results could be significantly affected by a change in the binder, the form of vitamin B12, the type of charcoal, the mass of charcoal or the presence of albumin. In systems with gastric juice and cyanocobalamin one charcoal separation procedure yielded results which were comparable to those obtained by all other procedures but this did not occur with gastric juice and hydroxocobalamin or with other binders and both forms of vitamin B12.

Results obtained using dialysis for separation varied with the membrane employed. Results obtained by gel filtration were significantly different from those obtained by ultrafiltration and those obtained by one ultrafiltration procedure were always significantly different from those yielded by another. Separation methods which give insignificantly different results in one system do not necessarily yield insignificantly different results in a system with a different binder or different form of vitamin B12.
The Role of Complement in Endotoxin Shock and Disseminated Intravascular Coagulation: Experimental Observations in the Dog
Authors: Garner R. , Chater B. V. , Brown D. L.
Published: March 2008

Summary.

The endotoxin shock response of Beagle dogs, characterized by profound hypotension and progressive fatal disseminated intravascular coagulation (DIC) was compared in anaesthetized normal dogs and in dogs previously decomplemented with purified cobra venom factor. The hypotension and the early thrombocytopenia were abolished in the decomplemented group, while the subsequent DIC was greatly ameliorated. Factor VII was apparently activated in decomplemented dogs but consumed in normal dogs. An observation as yet to be defined in relation to the shock syndrome was that dog platelets lost their collagen aggregation throughout the period of total decomplementation. It is suggested that complement activation is a major requirement for the full expression of fatally progressive DIC in endotoxin shock and that the activation of the coagulation system occurs, at least in part, via the extrinsic pathway.
Effect of Various Factors on Iron Absorption in Mice with X‐linked Anaemia
Authors: Sorbie Janet , Hamilton Donald L. , Valberg Leslie S.
Published: March 2008

Summary. The sla mouse has a hereditary iron deficiency anaemia and inappropriately low absorption of dietary iron. The response of the iron transport system in sla to a variety of factors that stimulate iron absorption in the normal mouse was investigated. The administration of an iron deficient diet for 5 days significantly increased the absorption of iron from graded oral doses of 59Fe ascorbate in both sla and normal mice compared with control groups fed an iron supplemented diet. Curvilinear dose‐absorption relationships indicated a saturable component in the iron transport system of both sla and normal mice fed the iron deficient diet. This suggests that the actual transport process is intact and can be activated by dietary iron deprivation. Phenobarbital enhanced the response to an iron deficient diet in both normal and sla but had no effect in animals given an iron containing semi‐synthetic diet. Blood loss, erythropoietin and hypoxia did not affect iron absorption in sla mice fed an iron supplemented cube diet, in contrast to a marked increase in normal mice. It appears that the iron transport pathway in sla mice has a limited capacity when saturated with dietary iron, but normal regulatory mechanisms operate when the pathway is not saturated. Whether the primary defect lies in a mucosal intracellular component or in an extracellular stimulus to activate iron transport is not clear.
The High Frequency of Anti‐Bga
Authors: Eska Patricka L. , Grindon Alfred J.
Published: March 2008

Summary. Anti‐Bga is detectable in 1.5% of donor and patient samples, when tested with a strongly reacting Bg(a+) cell. If, as suggested, this antibody is of little clinical significance, red cells used for antibody detection should lack this strong Bga antigen.
Simultaneous Measurement of Bactericidal Activity and Glucose Oxidation in the Human Granulocyte
Authors: Territo Mary C. , Davidson Warren D. , Tanaka Kouichi R.
Published: March 2008

Summary. A new technique for simultaneously measuring phagocytosis‐stimulated glucose oxidation and bactericidal activity in granulocytes is presented. This procedure provides a simple means for rapidly determining phagocytosis‐stimulated glucose oxidation and allows for correlation of that variable with bactericidal activity on a single blood sample.
Globin Synthesis in the Jamaican Negro with Beta‐Thalassaemia
Authors: Friedman Sh. , Schwartz E. , Ahern V. , Ahern E.
Published: March 2008

Summary. Globin synthesis was studied in three Jamaican Negro families with 18 heterozygotes and five homozygotes for β‐thalassaemia. Synthesis of the β‐chain of Hb A in the peripheral blood of heterozygotes was equal to that of α‐chain in 10 patients and was decreased in the remainder. In one patient with Hb C β‐thalassaemia, the β/α ratio was normal. These findings were similar to those in American Negroes, but differed from those in Caucasians with β‐thalassaemia trait, in each of whom the β/α ratio was decreased. Globin synthesis was balanced in the bone marrows of Negro and Caucasian heterozygotes. Despite the milder clinical disease in Negro homozygotes as compared to Caucasian patients, the β/α synthesis ratios were similar in both groups.

The presence of α‐thalassaemia combined with β‐thalassaemia in Negro heterezygotes is not a likely explanation for the high incidence of balanced globin synthesis ratios. The expression of relative β‐ to α‐chain synthesis in Negro heterozygotes appears to be modified by a factor which is not linked to the δ‐chain locus. The nature of this factor is not known at present.
Further Evidence for the Lymphocytic Nature of Leukaemic Reticuloendotheliosis (Hairy‐Cell Leukaemia)
Authors: Haak H. L. , De Man J. C. H. , Hijmans W. , Knappp W. , Speck B.
Published: March 2008

Summary. A case of hairy‐cell leukaemia was studied by conventional cyto‐chemical, electronmicroscopic and immunofluorescence techniques. This condition is often known as leukaemic reticuloendotheliosis (LRE) but the nature of the proliferating cells has been in doubt. The accumulation of immunoglobulin‐synthesizing cells in the peripheral blood suggests that the involved cells are lymphocytes and no evidence for a reticular or endothelial origin of the LRE cell could be found.
Mechanisms of Resistance of Human Acute Leukaemia Cells to Cytosine Arabinoside
Authors: Tattersall M. H. N. , Ganeshguru K. , Hoffbrand A. V.
Published: March 2008

Summary. Biochemical differences between human acute myeloblastic leukaemia cells sensitive and resistant to cytosine arabinoside have been studied. Bone marrow cells from seven patients with leukaemia resistant to cytosine arabinoside showed deoxycytidine kinase activity either below normal or in the low normal range and the mean level of this enzyme was significantly lower in cells from resistant than sensitive cases. On the other hand, deoxycytidine deaminase activity tended to be high in resistant cells but the difference from sensitive cells was not statistically significant. Deoxycytidylate deaminase activity also did not vary significantly with degree of sensitivity to cytosine arabinoside. The free concentrations of the four deoxyribonucleoside triphosphates were, in general, lower in resistant than sensitive marrow cells. The deoxycytidine triphosphate level, however, was relatively increased in the resistant cells.

We conclude that a number of biochemical changes may be present in cytosine arabinoside resistant leukaemia cells. It is likely that kinetic differences are the basis for some of these changes, and that low proliferative rate may be the major cause of cytosine arabinoside resistance. Selection of cells with raised deoxycytidine deaminase levels and relatively increased deoxycytidine triphosphte concentrations may also contribute to resistance.
Characterization of Fibrin Degradation Products in Patients on Ancrod Therapy: Comparison with Fibrinogen Derivatives Produced by Plasmin
Authors: Prentice C. R. M. , Edgar W. , McNicol G. P.
Published: March 2008

Summary. The fibrin digestion products which appear in serum of patients having treatment with ancrod have been compared with derivatives of human fibrinogen produced by plasmin by means of immunoelectrophoresis, immunodiffusion and gel filtration techniques. In ancrod‐treated patients the large molecular weight derivatives X and Y, as well as fragments D and E were seen 4 hr after the start of therapy. By 24 hr the main serum component was fragment D with low concentrations of large molecular weight derivatives still present, but fragment E had been cleared from the circulation. The identity of these fragments to the known products of fibrinogen proteolysis by plasmin has been confirmed. It is suggested that the initial degradation of fibrin micro‐clots produced in the circulation during ancrod infusion is mediated by the fibrinolytic enzyme system.
A Serum Haemolytic Factor Demonstrable at Low Ionic Strength
Authors: Riedler G. F. , Straub P. W.
Published: March 2008

Summary. In the sucrose haemolysis test incubation of patient serum with normal compatible erythrocytes led to the discovery of a new serum haemolytic factor (SHF). Normal erythrocytes in the presence of sucrose 1% haemolysis when incubated with sera of 55 normal individuals, but more than 4% haemolysis with sera of 17 of 37 patients with acute leukaemia, four of 17 with chronic leukaemia and three of eight with aplastic anaemia. Three additional patients with aplastic anaemia and a positive SHF test subsequently developed acute myeloid leukaemia. In three cases of acute myeloblastic ledtaemia the test became negative during remission. In all eight patients with Coombs‐positive haemolytic anaemia the test was negative.

The SHF is active only in the presence of sucrose. Its activity is increased with rising temperature; the pH optimum is between 7.4 and 7.8. The factor is labile upon storage except at ‐ 70°C and it is inactivated at 56°C. For incubation times below go min, haemolysis is a linear function of the serum concentration.
Functional Immaturity of Bone Marrow Bands and Polymorphonuclear Leucocytes
Authors: Altman Arnold J. , Stossel Thomas P.
Published: March 2008

Summary. Neutrophilic granulocytes from peripheral blood and from bone marrow were compared with respect to their ability to ingest paraffin oil droplets coated with bovine serum albumin or E. coli lipopolysaccharide. Granulocytes were incubated with unopsonized droplets or with droplets opsonized with fresh human serum. While myeloblasts and promyelocytes did not ingest either type of droplet, some myelocytes and metamyelocytes engulfed droplets; bands and polymorphonuclear leukocytes were the most active phagocytes. Morphologically mature granulocytes of bone marrow, in contrast to those of peripheral blood, had an ingestion defect for unopsonized droplets that was only partially corrected by heat‐labile opsonin. A dissociation between functional and morphologic maturity can therefore be demonstrated in bone‐marrow granulocytes.
In Vitro Stimulation of Chronic Lymphocytic Leukaemic Lymphocytes
Authors: Perera D. J. B. , Pegrum G. D.
Published: March 2008

Summary. The response of chronic lymphocytic leukaemic (CLL) lymphocytes to various mitogens in 5‐day cultures has been studied. Concanavalin A (Con.A) and pokeweed mitogen (PWM) were not superior to phytohaemagglutinin (PHA) in their stimulatory activity. Suboptimal levels of PHA and PWM together caused greater RNA synthesis than either mitogen at their optimal levels. The response to PHA could be chanced by the addition of a specific number of red blood cells and by glutathione. Using transformed nuclear material a reaction approaching the level found in normal lymphocytes was attained clearly indicating marked stimulation of the leukaemic cell population.
Cytotoxic Ability of Peripheral Blood Lymphocytes from Patients with Chronic Lymphocytic Leukaemia
Authors: Perera D. J. B. , Pegrum G. D.
Published: March 2008

Summary. Lymphocytes from patients with chronic lymphocytic leukaemia (CLL) could be induced to exhibit cytotoxicity against Chang target cells. This reactivity was provoked by transformed nuclear material, whereas stimulation with phytohaemagglutinin (PHA), pokeweed mitogen (PWM) or Concanavalin A (Con.A) had no significant effect.
Studies on the Mechanism of Cyanate Inhibition of Reticulocyte Protein Synthesis
Authors: Freedman Michael L. , Schiffman Fred J. , Geraghty Michael
Published: March 2008

Summary. Sodium cyanate promptly inhibited protein synthesis in both human sickle‐cell reticulocytes and rabbit reticulocytes but the human cells were sensitive to lower concentrations of the drug. Investigation of the ribosomal aggregation pattern in rabbit reticulocytes sliowed that cyanate induced a conversion of the polyribosomes to monoribosomes, a portion of which remained attached to mRNA. This conversion required ribosomal movement along mRNA. Cyanate's major effect, therefore, appears to be on initiation and/or an early clongation step of translation. While the drug also decreased reduced glutathione (GSH) concentration in these cells, this alone cannot explain the inhibition of protein synthesis. A generalized inhibition of translation of this type might result in a diminished crythropoetic response as well as severe toxicity in other tissues. Many more studies are necessary, therefore, before cyanate may be recommended for widespread use in patients.
Australia Antigen (HB‐Ag) Subtyping by a Sensitive Tanned Cell Haemagglutination‐Inhibition Technique
Authors: Hopkins R. , Das P. C.
Published: March 2008

Summary. The tanned cell haemagglutination‐inhibition technique was used to subtype Hepatitis B antigen (HB‐Ag) sera according to their D (ad +, y−) and Y (ay +, d−) specificity. The results were in agreement with those obtained by workers in the United States and Sweden using immunodiffusion. The majority (35 out of 39 tested) of HB‐Ag blood donors detected during the last 2 yr in South‐East Scotland were found to possess the D subtype. The Hepatitis B antibody subtype content of two preparations of immunoglobulin prepared for clinical use has also been studied.
A Comparison between Radioimmunoassay and Other Immunological Techniques for the Measurement of Fibrinogen/Fibrin Degradation Products in Serum
Authors: Ratky Susan M. , Martin Marion J. , Gordon Y. B. , Baker L. R. I. , Leslie J. , Chard T.
Published: March 2008

Summary. Fibrinogen/fibrin degradation products (FDP) have been measured in serum by radioimmunoassay for fragment E (FgE), one of the terminal fragments in plasmic digestion of fibrinogen, and the results compared with those determined by both a tanned red cell haemagglutination inhibition immunoassay (TRCHII) and a latex particle agglutination inhibition immunoassay. The detection limit of the FgE assay was 0.8 ng/ml, that of TRCHII was 625 ng/ml and the latex particle agglutination inhibition immunoassay was 10 μg/ml. All the samples measured had detectable levels of FDP with the FgE assay, whereas only 88% were measurable with the TRCHII and 2% by the latex particle agglutination inhibition immunoassay. A comparison of those samples giving a positive result with both the TRCHII and FgE assay showed overall agreement between the results of the two types of assay, but there was considerable scatter of FgE levels at each point of the TRCHII. The major advantages of the radioimmunoassay system are greater sensitivity, specificity and reproducibility.
The Procoagulant Factor of Leukaemic Promyelocytes: Demonstration of Immunologic Cross Reactivity with Human Brain Tissue Factor
Authors: Gouault‐Heilmann M. , Chardon E. , Sultan C. , Josso F.
Published: March 2008

Summary. A procoagulant activity was found in the immature cells from patients with acute promyelocytic leukaemia. It was demonstrated that this activity was related to tissue factor. The protein component of tissue factor from brain extract was purified by Nemerson's technique and injected into rabbits to obtain anti‐TF antibodies. Similar antibodies were produced against the promyelocyte extract. The anti‐brain tissue factor antibodies neutralized the tissue factor activity of promyelocyte extract, and antibodies against immature cells were able to neutralize the tissue factor activity of human brain extract. In immunoprecipitation studies a reaction of partial identity appeared between one component of promyelocyte extract and one component of brain tissue factor. The data demonstrated that the promyelocyte procoagulant is antigenically related to brain tissue factor.
The Oxygen Affinity of Haemoglobin Hammersmith
Authors: May A. , Huehns E. R.
Published: March 2008

Summary. Oxygen dissociation studies were carried out on red cells and lysates from a patient heterozygous for Hb Hammersmith. The oxygen affinity of the cells at pH 7.1 was decreased, partly by an increased cellular concentration of 2, 3‐DPG and more importantly by an intrinsic low affinity of Hb Hammersmith. Haemhaem interactions were reduced and an abnormal Bohr effect (pH 7.1–7.4) was found in the cells which would bring about an additional decrease in oxygen affinity at physiological pH. Oxygen dissociation studies on the lysates showed a low oxygen affinity, a normal Bohr effect at 50% saturation, slightly decreased haem‐haem interactions and a normal interaction with 2, 3‐DPG. Estimation of the percentage of Hb Hammersmith in the lysates of the patient's cells by selective precipitation of the abnormal β chain with p‐chloromercuribenzoate (PCMB) showed the presence of 30–33% abnormal haemoglobin. Assuming no interaction between the Hb A and the Hb Hammersmith, oxygen dissociation curves were calculated for Hb Hammersmith alone and these indicated that it has greatly decreased oxygen affinity, a normal Bohr effect, normal 2, 3‐DPG interaction and (somewhat) decreased haem‐haem interactions.
Haemoglobin C/α Thalassaemia: Haematological and Biosynthetic Studies
Authors: Steinberg Martin H.
Published: March 2008

Summary. A family with genes for haemoglobin C (Hb C) and a thalassaemia was studied. The mother had Hb‐C trait. The father also had Hb‐C trait but in addition displayed microcytosis, elevated Hb‐F levels and a concentration of Hb‐C less than usual for heterozygotes. The proband was homozygous for Hb‐C but had Hb‐F levels far exceeding those present in Hb‐C disease. Biosynthetic studies of globin synthesis in both father and daughter showed a deficit of α chains relative to non‐α chains, confirming the presence of α thalassaemia. The coexistence of α thalassaemia influences the level of mutant haemoglobin in haemoglobinopathies in which Hb C is present, in a fashion similar to that observed in sickle‐cell trait.
In Vitro Spontaneous Thrombin Generation in Human Factor‐IX Concentrates
Authors: Sas Geza , Owens Rosemary E. , Smith James K. , Middleton Sarah , Cash John D.
Published: March 2008

Summary.

A series of in vitro studies designed to ascertain the potential in vivo thrombogenicity of human factor IX‐containing concentrates is described. Using concentrates obtained from several different Centres the fibrinogen clotting time with some preparations was less than 6 h and/or the recalcification time of normal plasma was shortened. In some preparations, however, the plasma recalcification time was lengthened.

Further studies revealed that all diluted factor‐IX concentrates generated thrombin after recalcification, and that the rate of thrombin generation appeared to be characteristic of a particular preparation. This characteristic has been designated the TGt50, which is the incubation period in minutes, after recalcification, required to obtain a 50 s clotting time of a fibrinogen substrate. The TGt50 was found to correlate most strongly with recalcification time of celite exhausted plasma (P < 0.001), but no correlation was observed between it and the immunological antithrombin III or factor‐VIII antigen levels. Evidence is presented which suggests that the thrombin generation test and recalcification time of celite exhausted plasma may represent suitable in vitro quality control assays for factor‐IX concentrates.
Fibrin‐Blood Platelet Interaction in a Contracting Clot
Authors: Cohen Isaac , Gabbay Jacob , Glaser Tova , Oplatka Avraham
Published: March 2008

Summary.

The interaction of fibrin and blood platelets was studied by measuring tension development in clots containing different ratios of these two components. Isometric tension attained a saturating value (MT) when plotted as a function of the concentration of either component at a constant concentration of the other. The value of MT increases linearly with the minimal platelet concentration (Ps) required in order to reach saturation at various different fibrin concentrations. A linear relationship also holds in a plot of Ps versus the logarithm of the corresponding fibrin concentration. When the cylindrical clots contracted isotonically, the velocity, normalized with respect to the momentary length, was found to be constant. This velocity decreased in an hyperbolic manner with increasing fibrin concentration at a constant concentration of blood platelets. The results are interpreted in terms of a three‐dimensional network in which the blood platelets serve as contractile cross‐bridges connecting fibrin fibres. Tension development is assumed to be accompanied by the reduction of the angle between fibrin fibres. An attempt was made to calculate the isometric tension developed by a platelet.
Abnormal Membrane Surface Properties during Maturation of Rat Reticulocytes Elicited by Bleeding as Measured by Partition in Two‐Polymer Aqueous Phases
Authors: Walter Harry , Krob Eugene J. , Ascher Galli S.
Published: March 2008

Summary.

Partition of cells in two‐polymer aqueous phases is an extremely sensitive method for the separation of cells and for tracing subtle changes in the cells' membrane surface properties (primarily surface charge) as a function of in vivo processes (e.g. differentiation, maturation, ageing). Combining isotope labelling and cell countercurrent distribution techniques we have examined the membrane surface properties of rat reticulocytes produced in response to erythroid stress by bleeding. It was found that the rapid increase in the partition coefficient of normal reticulocytes subsequent to release into the peripheral blood (which reflects a rapid increase in membrane surface charge) is absent in reticulocytes produced in response to bleeding. In this way the reticulocytes behave like those produced in response to repeated phenylhydrazine injections. ‘Stress reticulocytes’ never mature to erythrocytes having normal membrane surface properties as judged by partition. The experiments show that our previous results are not due to phenylhydrazine per se but rather to the production of ‘stress reticulocytes’. Further, if remodelling of the phlebotomy‐induced reticulocyte cell membrane occurs in the circulation as has been suggested it does not lead to a cell with normal membrane surface properties. Whether the abnormal membrane surface properties of ‘stress reticulocytes’ and of the erythrocytes to which they give rise affect the cells' life‐span is not clear since phenylhydrazine‐induced ‘stress reticulocytes’ have been reported to have at least a component of short‐lived cells while the phlebotomy‐induced cells are said to have a virtually normal life‐span. It is likely that reticulocytes produced in severe anaemias, in general, mature to erythrocytes having abnormal surface properties (i.e. lower membrane surface charge).
The Effect of Diet and Alcohol on the Development of Folate Deficiency in the Rat
Authors: McGuffin Robert , Goff Paul , Hillman R. S.
Published: March 2008

Summary.

Studies of the rate of depletion of serum and tissue methylated and nonmethylated folates were carried out in rats maintained for long periods on either a folate deficient (sucrose‐water/sulphathiazole) diet or a deficient diet plus high alcohol intake. By means of implantation of a feeding gastrostomy tube, it was possible to sustain constant blood ethanol levels of between 50 and 300 mg/dl for 3–4 weeks with relatively normal calorie intake and without death of the animal. Using this animal model, which closely resembles severe alcoholism in man, a very rapid depression in serum 5‐methyl tetrahydrofolate was observed similar to that reported in alcoholic man. At the same time, release of folates from liver stores was unimpaired by alcohol ingestion. Liver folate store depletion rates were identical for alcoholic and folate starved animals. The explanation for the sudden alcohol suppression of serum folate levels must, therefore, be sought at a point in the internal metabolic sequences of folate other than the delivery of folate stores to plasma.
Stimulation by Propylthiouracil of the Hexose Monophosphate Shunt in Human Polymorphonuclear Leucocytes during Phagocytosis
Authors: Tsan Min‐Fu , McIntyre Patricia A.
Published: March 2008

Summary.

The effect of propylthiouracil on glucose metabolism in human polymorphonuclear leucocytes was studied. At a therapeutically achievable concentration (0.1 mm), propylthiouracil stimulated hexose monophosphate shunt activity in normal leucocytes during phagocytosis but not in resting cells. However, in the presence of hydrogen peroxide it stimulated hexose monophosphate shunt activity in resting cells, and in the soluble fraction when reduced glutathione and reduced nictotinamide adenine dinucleotide phosphate (NADPH) were also present. Propylthiouracil had no effect on glucose‐1‐C oxidation in either phagocytosing or resting leucocytes obtained from two male patients with chronic granulomatous disease. Stimulation of the hexose monophosphate shunt activity in normal leucocytes during phagocytosis also was demonstrated with methimazole, thiouracil and thiourea, but not with adenine, uracil or urea. There was an apparent minimal common structure requirement in thiourea. Propylthiouracil had no effect on phagocytosis, formate oxidation, glucose‐6‐phosphate dehydrogenase, 6‐phosphogluconate dehydrogenase or catalase activities. Thus, the stimulation of the hexose monophosphate shunt activity by propylthiouracil is dependent on hydrogen peroxide and is best explained by its stimulation or participation in the glutathione cycle.
Absorption and Transport of Vitamin B12
Authors: Gräsbeck Ralph
Published: March 2008

Summary.

A review of recent advances is presented, mostly dealing with physiological mechanisms and the chemical and functional properties of the transport proteins. A new nomenclature is proposed for the R‐proteins and transcobalamin II. Scepticism is expressed as to whether we are warranted in accepting the different R‐proteins as separate entities.
The Immunological Localization of Factor V in Human Tissue
Authors: Giddings J. C. , Shearn S. A. M. , Bloom A. L.
Published: March 2008

Summary. Antibody to factor V was produced by immunizing rabbits with purified factor V from human plasma. Various tissues were examined for the presence of factor‐V antigen using this antiserum. It was consistently demonstrated in homogenates of liver and spleen by means of an antibody (coagulation inhibitor) neutralization technique. The antigen was further localized histologically by the indirect fluorescent antiglobulin technique. It was present on the endothelium of normal blood vessels in all organs examined. In the liver it was detected in hepatic parenchymal cells and a distinctive pattern of fluorescence in the spleen suggested that it was being detected on platelets. Results were negative in all other tissues examined. The findings confirm the presence of factor V in hepatic parenchymal cells and support the suggestion that endothelial coagulation factors may play a role in haemostasis and thrombosis.
Haemoglobin Bart's in Saudi Arabia
Authors: Pembrey M. E. , Weatherall D. J. , Clegg J. B. , Bunch C. , Perrine R. P.
Published: March 2008

Summary. The haemoglobin (Hb) patterns of 345 Shiite Saudi Arab cord bloods were examined by alkaline starch‐gel electrophoresis. A fast‐moving component, identified by structural analysis as Hb Bart's, was found in 52% of cases, the highest incidence of this variant yet recorded. The levels of Hb Bart's ranged from 0.5 to 16% of the total haemoglobin. The relative rates of synthesis of the α, β and γ‐chains, measured by [3H]lcucine incorporation, were estimated in 12 newborn Arab infants. There was an excellent correlation between the amount of Hb Bart's and the α/non‐α‐globin‐chain production ratio. Furthermore there was a significant correlation between the level of Hb Bart's and morphological abnormalities of the red cells and the mean cell haemoglobin (MCH). These findings indicate that elevated levels of Hb Bart's in this population are due to the presence of a thalassaemia. The absence of hydrops fetalis and the rarity of Hb‐H disease despite the intense inbreeding in this population, points to an a‐thalassacmia genotype that is, in terms of phenotypic expression, intermediate between the heterozygous state for α‐thalassacmia 1 and Hb‐H disease. A possible molecular basis for this genotype is suggested.