bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement of Polymorphonuclear Leucocyte Oxidative and Bactericidal Functions in Chronic Granulomatous Disease with 4‐Amino‐4′‐4 Hyndroxylaminodiphenyl Sulphone
Authors: Ismail Ghazally , Boxer Laurence A. , Allen John M. , Baehner Robert L.
Published: March 2008

Summary. In an effort to restore oxidant‐dependent capabilities to chronic granulomatous disease (CGD) polymorphonuclear leucocytes (PMN), we studied a dapsome derivative, 4‐amino‐4′‐hydroxylaminodiphenyl sulphone (DDS‐NOH), known to generate H2O2. After incubation of CGD PMN with 0.2 and 1.0 mm DDS‐NOH for 30 min, the rate of glucose‐I‐14C oxidation via hexose monophosphate (HMP) shunt increased 2‐4‐fold and ‐ that of iodination of ingested zymosan particles 1.5‐2.7‐fold. Both effects could be further enhanced by superoxide dismutase (SOD) but inhibited by catalase. In three patients, 0.2 mM DDS‐NOH improved in vitro killing of Staph. aureus. DDS‐NOH 0.02 mM induced capping of Concanavalin A (Con A) receptor complexs suggesting interference by the drug with microtubule‐associated function. Thus, optimal concentrations of DDS‐NOH my be employed as an oxidant to improve metabolic and bactericidal functions of PMN from patients with CGD.
Organelle Pathology in Primary and Secondary Haemochromatosis with Special Reference to Lysosomal Changes
Authors: Seymour Carol A. , Peters T. J.
Published: March 2008

Summary. The organelle pathology of liver biopsy specimens from patients with either primary or secondary haemochromatosis was investigated by analytical subcellular fractionation in combination with enzymic microanalysis. The most striking changes were found in the lysosomes. Increased total activities but decreased latent activities of enzymes selectively localized to the high density population of lysosomes was demonstrated in the iron overloaded biopsies. Depletion of the iron, where possible, by venesection was accompained by a return to normal of these changes.

The other subcellular organelles, plasma membrane, endoplasmic reticulum, biliary canaliculi, mitochondria, peroxisomes and the low density population of lysosomes appear to be relatively unaffected. The minor changes demonstrated are similar to those seen in other forms of chronic liver disease. It is suggested that iron mediated lysosomal disruption is implicated in the pathogenesis of the tissue damage in haemochromatosis.
Iron Deficiency Anaemia in Newborn sla Mice: a Genetic Defect of Placental Iron Transport
Authors: Kingston P.J. , Bannerman Catherine E. M. , Bannerman R. M.
Published: March 2008

Summary. . Newborn mice with X‐linked anaemia (gene symbol sla) have lower haemoglobin levels at birth than normal and carrier mice but there is considerable overlap. Serial observations showed that the haemoglobin values of segregating male mice separate into a bimodal distribution by 42 d of age, and 50 d values were used to assign genotypes retrospectively. The anaemia in newborn sla mice is attributable to iron deficiency, since their total body iron is lower than in normal newborn mice, while their birth weights are almost identical.

Haemoglobin levels at birth in normal, anaemic and carrier mice are also influenced by the mother's genotype and phenotype, and the haemoglobin value was progressively lower according to the sla gene dose of the mother.

Materno‐fetal iron transfer was examined by labelling pregnant carrier females with radioiron in various ways. When given as single or intermittent doses by injection no clearcut differences emerged in apparent iron transfer to anaemic as compared to non‐anaemic fetuses. However, when radioiron was administered continuously in food a significant reduction in iron transfer to anaemic fetuses was demonstrated.

The sla gene is already known to have a major effect in reducing iron transport in the small intestine. The present studies provide evidence of an analogous defect in placental iron transport.
Trimethoprim and Sulphamethoxazole Inhibition of Haematopoiesis in Vitro
Authors: Golde David W. , Bersch Noelle , Quan Shirley G.
Published: March 2008

Summary. The effect of trimethoprim and sulphamethoxazole on haematopoiesis was studied in vitro using cloning techniques for human and murine erythroid and granulocytic precursor cells. Trimethoprim was found to inhibit granulopoiesis and erythropoiesis in vitro in a dose‐dependent fashion with approximately 50% inhibition of human erythroid and granulocytic colonies at a therapeutically achievable concentration of 7 μg/ml. Sulphamethoxazole was also shown to impair haematopoiesis in vitro. The inhibition caused by both these constituents of co‐trimoxazole was completely reversed by folinic acid. The data suggest that co‐trimoxazole can impair human haematopoiesis by inhibition of tetrahydrofolate synthesis. These observations suggest that the clinical haematopoietic toxicity of trimethoprim‐sulphamethoxzole can be abrogated by simultaneous administration of folinic acid.
A Case of Atypical Polyagglutinability due to Tk‐Transformation
Authors: Mullard G. W. , Haworth Clare , Lee D.
Published: March 2008

Summary. A case of acquired transient polyagglutinability in a group O, Rh(D) positive female patient, Wa, is described. An unusual finding in the preliminary serological investigation was the non‐reaction of the cells with peanut (Arachis hypogaea) extract. The patient's red cells were strongly agglutinated by normal human and rabbit sera from which anti‐T had been removed. Electrophoretic mobility studies and tests with polybrene indicated that their sialic acid content was normal. The use of a specific antiserum raised in a rabbit to Wa cells and cross‐absorption tests showed that the patient's erythrocytes shared similar antigenic determinants to red cells treated with supernatants from pure cultures of Bacteroides fragilis. The in vitro‐modified red cells reacted strongly with BS II lectin prepared from Bandeiraea simplicifolia seeds thus providing confirmatory evidence for Tk transformation on Wa cells.

It is concluded that the Tk receptor is probably a more frequent feature of acquired transient polyagglutinability than has hitherto been recognized.
Sedimentation Cytology in Central Nervous System Leukaemia with a New Simple Apparatus
Authors: Nishimura Kozo , Hosoya Ryohta , Nakajima Kiyotaka
Published: March 2008

Summary. The clinical usefulness of a newly devised, simple apparatus, the Nishimura fluid sedimentation chamber, is described. The latter consists of an open‐ended plastic tube, an absorbent paper with a central hole, and a glass slide which are stuck to each other with double‐sided plastic adhesive tape with a central hole of equal size. The chamber is set to allow 0.5 ml of cerebrospinal fluid (CSF) to be absorbed in 15–20 min. The principle of the sedimentation relies on the absorption of the fluid portion into the absorbent paper with the cellular elements sedi mented onto the glass slide. Thin stationery paper is used as the absorbent material which controls sedimentation speed and minimizes cellular loss. 190 CSF specimens obtained from 28 patients with acute leukaemia were processed by this method. The results obtained showed that sedimentation using this simple chamber is quite reliable for the recognition of small number of abnormal cells in the CSF producing little or no noticeable cellular damage. This technique may be applied to other clinical situations in which fluid cytology is useful in establishing a diagnosis or in evaluating clinical progress.
Quantitative Abnormality of an Aα Chain Molecular Weight Form in the Fibrinogen of Cirrhotic Patients
Authors: Weinstein Mark J. , Deykin Daniel
Published: March 2008

Summary. The molecular weight heterogeneity of fibrinogen from the whole plasma of 12 normal and seven cirrhotic individuals was examined by means of a novel two‐dimensional sodium dodecyl sulphate (SDS) gel electrophoretic technique. Fibrinogen was first separated from other plasma proteins on a large pore gel, cut out of the gel, reduced, and separated into its component Aα, Bβ and γ chains on a second gel. Fibrinogen was resolved into two major bands, I and II, on the first gel. The ratio of fibrinogen II to fibrinogens I plus II was approximately 0.3 (range 0.2‐0.35) for both normals and cirrhotic patients. Two major molecular weight (mol wt) forms of Aα chain were present in normal fibrinogen I: Aα/I and/or Aα/2, mol wt 7 × 104 and 6.7 × 104 respectively. Normal fibrinogen II contained either one of these Aα chains plus one of the smaller Aα chains, Aα/6‐10, accounting for the 3–4 × 104 mol wt difference between bands I and II. Aα/2 comprised 33% of the total Aα chains (range 27–41%) in normal fibrinogen I and approximately 25% of the Aα chains in normal fibrinogen II. In contrast, fibrinogen I from six out of the seven patients contained a lower percentage of Aα/2 (range 10–25%). Similarly fibrinogen II from these patients was deficient in Aα/2, although the protein contained normal levels of lower mol wt Aα derivatives. No correlation was found between per cent fibrinogen II and per cent Aα/2 in either normal or cirrhotic subjects. These results suggest that at least two independent processes are responsible for the observed levels of Aα heterogeneity in normals and cirrhotics and that the process controlling Aα/2 production is abnormal in cirrhotic individuals. This decrease in Aα/2 does not affect the coagulability of fibrinogen. Fibrin monomer aggregation studies indicate that a serum component is, in part, responsible for the abnormally transparent clot formed from the plasma of cirrhotics.
Isolation of Human Antibodies to Factor VIII
Authors: Lavergne J. M. , Meyer Dominique , Koutts J. , Larrieu Marie‐José
Published: March 2008

Summary. It has been claimed that human anti‐VIII:C antibodies do not form stable complexes with factor VIII and this fact has hampered in the past the isolation of such antibodies. In this study the purification of human anti‐VIII:C antibodies appearing in haemophiliac patients following replacement therapy has been achieved using two different systems. In a liquid phase system, purified human factor VIII was mixed with IgG from a haemophilic patient with a high titre antibody. Specific anti‐VIII:C antibodies were recovered following filtration of the antigen‐antibody complexes on Biogel A‐5m, dissociation of complexes at pH 3.5 and final isolation by filtration on Sephadex G‐200. In a solid phase system, the same IgG fraction was specifically bound to insolubilized human factor VIII. Purified anti‐VIII:C antibodies were subsequently recovered by elution of antigen‐antibody complexes with magnesium chloride. The results demonstrated that stable complexes form between anti‐VIII:C antibodies and either the whole factor VIII molecule, or VIII:C dissociated by previous interaction with the antibodies. It is postulated that, in vivo, similar antigen‐antibody complexes may form following replacement therapy in haemophilic patients with antibody.
Erythroblast Kinetics in Homozygous and Heterozygous Beta‐Thalassaemia
Authors: And P. Dörmer , Betke K.
Published: March 2008

Summary

The kinetics of erythroblast proliferation have been investigated in four patients with homozygous and two patients with heterozygous β‐thalassaemia. One case was studied prior to and following splenectomy. Another patient was included in the study in whom an unstable haemoglobin was assumed according to the clinical data. The cell‐kinetic technique is based on short‐term in vitro incubation of suspended bone marrow cells with [14C]thymidine and 5‐fluorodeoxyuridine. By means of quantitative autoradiography using 14C‐referencc sources, the rates and time of DNA synthesis were determined in individual cell compartments classified according to morphological criteria. Cell loss was estimated by comparing the ratios of the relative rates of cell production in successive cytological compartments with the corresponding ratios during normal erythropoiesis.

In both homozygous and heterozygous β‐thalassaemia the erythroblasts proceed through all morphological compartments at a reduced rate. This kinetic impairment is considered to be independent of intracellular precipitation of excess α‐chains. A normal or increased rate of proliferation in the early compartments in the case of haemoglobinopathy underlines the significance of the findings in β‐thalassaemia, which become the more prominent the more the erythroblasts approach the non‐proliferating stage. In the homozygous disease only 15–30% of these cells escape destruction during their proliferative life. In heterozygous β‐thalassaemia intramedullary cell death seems to be confined to the non‐proliferating stages. On the basis of the kinetic parameters accessible through the method applied, the heterozygous disorder is largely comparable to iron deficiency anaemia.
Variations in Globin Synthesis in Delta‐Beta‐Thalassaemia
Authors: Kinney T. R. , Friedman Sh. , Cifuentes E. , Kim H. C. , Schwartz E.
Published: March 2008

Summary

Peripheral blood globin synthesis studies were done in 11 patients with δβ‐thalassaemia trait, Hb S‐δβ‐thalassaemia or δβ/βº‐thalassaemia from two black and two Caucasian families. All patients had elevated Hb F and normal or decreased Hb A2 levels and 10 had family studies confirming the diagnosis. In addition, four unrelated non‐thalassaemic patients with elevated Hb F levels also had peripheral blood globin synthesis studies. The β/α specific activity globin synthesis ratios in the three blacks with δβ‐thalassaemia trait were 0.60–1.04. In the four Caucasians with δβ‐thalassaemia trait, the β/α ratios were 0.58–1.02. These results demonstrate a wide range of ratios overlapping those of normal controls (0.99 ± 0.06). The βs/α ratios in three blacks with Hb S‐δβ‐thalassaemia ranged from 0.66 to 1.00, similar to those of patients with δβ‐thalassaemia trait. In the black patient with δβ/βº‐thalassaemia, the γ/α ratio was 0.67. The β/α peripheral blood ratios in the four non‐thalassaemic patients with elevated Hb F ranged from 1.00 to 1.1, similar to those of normal controls. These studies indicate that a decreased β/α ratio is not an invariable finding in δβ‐thalassaemia in blacks or Caucasian patients and that globin synthesis data alone is insufficient to diagnose definitively heterozygotes for δβ‐thalassaemia or to distinguish this trait from non‐thalassaemic haematological disorders associated with a normal percentage of Hb A2 and an elevated level of Hb F.
Surface Properties of Rat Reticulocytes during Maturation as Measured by Partition: Membrane Charge‐Associated Changes Discerned prior to Membrane Lipid‐Related Changes
Authors: And Harry Walter , Krob Eugene J.
Published: March 2008

Summary

Partition of cells in dextran‐polyethylene glycol aqueous phases is an extremely sensitive method for separating cells and for tracing subtle changes in the cells’membrane surface as a function of differentiation, maturation and ageing. By appropriate manipulation of polymer concentration and ionic composition and concentration one can select, to a great extent, the membrane properties (e.g. charge‐associated, lipid‐related) that will determine the cells’partition coefficient. Combining radioiosotopic labelling and cell countercurrent distribution techniques we have previously reported rapid increases in charge‐associated partition coefficients during rat reticulocyte maturation followed by slower decreases in the partition coefficients during the entire life span of the mature erythrocyte. In the present experiments red blood cell populations containing radioisotopically labelled cells of different and distinct ages have been subjected to countercurrent distribution in phase systems reflecting either membrane charge‐associated or lipid‐related characteristics. It was found that: (a) membrane lipid‐related surface properties change rapidly during reticulocyte maturation; (b) membrane charge‐associated changes during reticulocyte maturation are discerned prior to those that are lipid‐related; and (c) lipid‐related membrane changes continue slowly throughout the entire life‐span of the erythrocyte.

The sensitivity with which red cell membrane lipid‐related differences or changes can be detected and measured by cell partition suggests great usefulness of this simple method in the study of abnormalities in red cell lipids.
Identification of the Hairy Cells of Leukaemic Reticuloendotheliosis by an Esterase Method
Authors: Higgy K. E. , Burns G. F. , Hayhoe F. G. J.
Published: March 2008

Summary

A distinctive pattern of alpha‐naphthyl butyrate esterase positivity was observed in all of a series of 14 patients with HCL. This pattern was not observed in a range of other haematological malignancies including nine cases of CLL, two cases of Sézary's syndrome, six cases of null cell ALL, two cases of Schilling‐type monocytic leukaemia and one case of lymphosarcoma cell leukaemia. The potential diagnostic value of this simple cytochemical test is discussed in relation to existing methods.
Efficacy of Platelets Collected by Semi‐continuous Flow Centrifugation (Haemonetics Model 30)
Authors: Slichter Sherrili J.
Published: March 2008

Summary

The viability of both autologous and donor platelets collected by means of a semi‐continuous flow centrifuge (Haemonetics Model 30) was measured. Radiochromium labelled pooled platelets harvested from six passes and reinfused into their normal donors showed recoveries and survivals of 58%± 8 and 9.6 d ± 0.8 compared with normal values of 65%± 5 and 9.5 d ± 0.6, respectively. When the viability in aplastic thrombocytopenic recipients of platelets collected from normal donors by the Haemonetics Model 30 was compared with that of platelets collected from the same donors by multiple bag plateletpheresis the results were very similar: recoveries of 82%± 18 and 80%± 25 and survivals of 3.9 d ± 2.0 and 3.8 d ± 1.3, respectively. The in vitro function of the collected platelets in response to aggregating agents was normal when measured within 60 min of collection at pH 7.4. The relationship between bleeding time and post‐transfusion platelet count in aplastic, thrombocytopenic patients showed that in vivo function was maintained.
Prolymphocyte Leukaemia of T‐Cell Type: Immunological, Enzymatic and Ultrastructural Morphometric Characteristics
Authors: Woessner Soledad , Lafuente Rosario , Sans‐Sabrafen J. , Vives J. , Rozman C.
Published: March 2008

Summary. A case of prolymphocytic leukaemia with immunological characteristics of T‐cell type is reported. Three noteworthy findings can be emphasized: the presence of C3 receptors on the T‐prolymphocytes, the study of the acid‐phospha‐tase isoenzymatic pattern, which showed an increased band 3 with absence of band 3b, and the morphometric ultrastructural investigation. Cytochemistry and ultra‐structural morphometry may be useful for a more precise characterization of prolymphocytic leukaemia and help to distinguish it from other lymphoprolifera‐tive disorders.
Erythrocyte Populations in Pyruvate Kinase Deficiency Anaemia following Splenectomy
Authors: Leblond Pierre F. , Lyonnais Jean , Delage Jean‐Marie
Published: March 2008

Summary. Light and scanning electron microscopic examination of fresh erythrocytes from the blood of four patients splenectomized for severe pyruvate kinase (PK) deficiency haemolytic anaemia has revealed three morphologically different cell populations. Most striking in these patients was the presence of crenated red cells (30–50% of total) which fit current criteria for definition as cchinocytes, but also with unusual surface retraction compatible with the existence of intracellular shrinkage, a phenomenon known to occur after incubation of PK deficient red cells in vitro. Additional cell types include immature reticulocytes (7–8% of total) and thin macrocytic discocytes (45–65% of total) none of which are considered specific for the disease. These morphological findings are in sharp contrast with the paucity of red cell shape abnormalities usually described in PK deficient patients before splenectomy and may provide insight into the mechanisms leading to accelerated erythrocyte destruction in this disease.
Inhibition of Plasmin by Antithrombin‐Heparin Complex
Authors: Collen D. , Semeraro N. , Telesforo P. , Verstraete M.
Published: March 2008

Summary. The role of antithrombin as an inhibitor of plasmin in the presence and absence of heparin was studied during thrombolytic therapy in 12 patients with vascular occlusive disease. The extent of plasmin‐antithrombin–(heparin) complex formation was studied by intravenous injection of iodine‐labelled antithrombin (5–20 /μCi) and quantitation of the amount of antithrombin bound to plasmin. In the absence of heparin, less than 0.8% of the labelled antithrombin was recovered in the plasmin‐antithrombin complex but between 1.2 and 4.8% following injection of 5000 iu of heparin. Thus only between 3 and 11% of the in vivo formed plasmin is neutralized by antithrombin–heparin complex. Repeated activation of the fibrinolytic system resulted in a shortening of the plasma radioactivity half‐life of labelled antithrombin from 2.45 to 2.03 d in the absence of heparin (three patients), and from 3.13 to 2.35 d following heparin administration (two patients). This increased turnover does not result in a decrease of the plasma antithrombin level.
Radioimmunoassay of Platelet Factor 4 and β‐Thromboglobulin: Development and Application to Studies of Platelet Release in Relation to Fibrinopeptide A Generation
Authors: Kaplan Karen L. , Nossel Hymie L. , Drillings Michael , Lesznik George
Published: March 2008

Summary.Platelet and fibrinogen survival and turnover studies have shown that platelet activation and fibrin formation may occur to different degrees in different thrombotic disorders. More direct evidence of differential involvement of platelet activation and fibrin formation should be provided by specifically measuring the products of these reactions, i.e. released platelet proteins and fibrinopeptide A. Two platelet proteins, platelet factor 4 (PF4) and β‐thromboglobulin (βTG), were isolated and characterized, and sensitive and specific radioimmunoassays were developed to measure them. These assays were employed, along with the radioimmunoassay for fibrinopeptide A (FPA), to study the release of PF4 and βTG in relation to FPA cleavage. PF4 and βTTG were released by ADP and collagen with time course and concentration dependence similar to that of [14C]serotonin release. FPA was not cleaved from fibrinogen during ADP or collagen‐induced platelet release. Thrombin caused release of PF4 and βTTG as well as cleavage of FPA. Cleavage of FPA occurred with concentrations of thrombin about 100 times less than did release of PF4 and βTG, and release of [14C]serotinin required still higher thrombin concentrations. Release of [14C]serotonin and platelet proteins was similar as a function of time. Sodium citrate was found to inhibit platelet release induced by thrombin.
Hairy Cell Leukaemia: Surface Markers and Functional Capacities of the Leukaemic Cells Analysed in Eight Patients
Authors: Rieber Ernst Peter , Hadam Martin R. , Linke Reinhold P. , Saal Johannes G. , Riethmüller Gert , von Heyden Hans W. , Waller Hans D.
Published: March 2008

Summary. In eight patients with hairy cell leukaemia (HCL) peripheral blood cells and in two patients also spleen cells were analysed for surface markers and functional capacities. Only cells containing the tartrate resistant isoenzyme 5 of the acid phosphatase were considered. Hairy cells (HC) of all patients were found to adhere spontaneously to glass and plastic surfaces and to spread after adherence like monocytes. They ingested latex particles of more than 1 μm diameter, but, in contrast to monocytes, did not phagocytose erythrocytes sensitized either by IgM or by IgG antibodies. HC of all patients bore Fc‐receptors with a high binding affinity for aggregated IgG. Using 125I‐labelled F(ab′)2‐fragments of monospecific antibodies in autoradiography, only one light chain type was detected on HC of individual patients. In four patients μ‐ and δ‐chains were simultaneously expressed on HC, whereas in two patients only γ‐chains and in one case only μ‐chains were observed on HC. One patient showed a combination of γ‐ and δ‐chains on his HC. A great variation in density of surface immunoglobulins of HC was observed within individual patients. After removal by capping, surface immunoglobulin reappeared on HC during cell culture, but more slowly than on normal B‐lymphocytes. As shown in two patients by internal labelling, HC secreted immunoglobulin light chains, but no heavy chains. On the basis of these findings the classification of HC as belonging to the B‐cell lineage, rather than to the monocytic lineage, seems to be justified.
Kinetics of Red Cell Membrane Phosphorylation: Altered Affinity of HS Membrane Protein Acceptors
Authors: Ali S. A. , Gordon‐Smith E. C. , Selhi H. S.
Published: March 2008

Summary. Membrane ghosts were prepared from red blood cells of normal and hereditary spherocytosis (HS) subjects. Time dependent phosphorylation studies of the membrane (in the presence of γ labelled AT32P) showed that the HS membrane appeared to incorporate less phosphate than the normal membrane but the results were not significantly different. Using the initial linear rate of phosphorylation and varying the ATP concentration, it was found that the Michaelis constant (Km) for the normal membrane was 24.8 μM ATP and Vmax was 0.120 nmol phosphate bound/mg membrane protein/min. These values for the HS membrane were found to be 39.0 and 0.118 respectively. In one family the affected mother and son each showed a positive cooperativity effect for similar reciprocal plots. This indicates that phosphorylation of various HS membrane proteins may be an ordered rather than random process and suggests biochemical heterogeneity of the HS condition. The constants, Km and Vmax, however, were found to be similar to the rest of HS subjects studied.

Crude extracts of the enzyme protein kinase, which catalyses the membrane phosphorylation, showed no significant difference in the Km and Vmax value in HS compared with that from normal red cells. The kinetic difference in phosphorylation is probably due to abnormalities in the membrane protein(s) which accept the phosphate from protein kinase in HS red cells.
Vitamin B6Metabolism in Idiopathic Sideroblastic Anaemia and Related Disorders
Authors: Solomon Lawrence R. , Hillman Robert S.
Published: March 2008

Summary.Patients with idiopathic anaemias associated with abnormal sideroblasts were defined according to morphologic and ferrokinetic criteria and the haematologic and biochemical effects of vitamin B6therapy were evaluated. While all patients presented similar clinical pictures, peripheral blood changes and bone marrow abnormalities, two distinct groups were identified by sideroblast morphology and ferrokinetics. Patients with more than 5% true ring sideroblasts in the marrow (IRSA) uniformly had marked ineffective erythropoiesis, while those with abnormal sideroblasts but few true ring forms were hypoproliferative. Measurements of red cell pyridoxine kinase (PnK) and intracellular pyridoxal 5‐phosphate availability (PLP) as assessed by the activity of the PLP‐dependent enzyme aspartate aminotransferase (EGOT), revealed slightly decreased PnK levels in IRSA subjects but normal intracellular PLP activities in both groups. Furthermore, when treated with pyridoxine, all patients showed increases in both red cell PnK and EGOT activities which were similar to those seen in normal subjects. Treatment with PLP also effectively increased erythrocyte vitamin B6activity. Even so, neither pyridoxine nor intramuscular PLP improved erythropoiesis as determined by serial haematocrits, reticulocyte counts and erythron iron turnover measurements. However, since both therapies increased red cell protoporphyrin levels and the excretion of urinary coproporphyrin in a number of subjects, the possibility that impaired haem synthesis in the sideroblastic anaemias is associated with abnormal vitamin B6metabolism at the level of the mitochondrion must still be considered.
Detection of Carriers of Haemophilia B
Authors: ØRstavik K. H. , Veltkamp J. J. , Bertina R. M. , Hermans J.
Published: March 2008

Summary. Plasma levels of factor IX activity and factor IX antigen were determined in 18 definite carriers of haemophilia B‐, 10 definite carriers of haemophilia B+ and 40 control subjects. Factor IX antigen was determined by the electroimmunoassay technique of Laurell using a rabbit antiserum against factor IX. In haemophilia B‐, statistical tolerance regions were constructed containing 90% of the factor IX activity and factor IX antigen values for the carriers and the control subjects. Of the 18 carriers, 10 were outside the tolerance region for the control subjects, and of the 40 control subjects, 17 were outside the tolerance region for the carriers. A better separation of the probabilites of being a carrier was obtained for the B‐ carriers and the control subjects when factor IX antigen was determined in addition to factor IX activity. In haemophilia B+, factor IX antigen was present in excess of factor IX activity in nine of the 10 carriers. The values for factor IX activity and factor IX antigen for these nine carriers fell outside the 90% tolerance region for the control subjects. It is concluded that the quantitative determination of factor IX antigen may be of value in the detection of carriers of both haemophilia B+ and haemophilia B‐.
ADCC (K‐Cell) Lysis of Human Erythrocytes Sensitized with Rhesus Alloantibodies
Authors: Urbaniak S. J.
Published: March 2008

Summary. An ADCC system has been developed using anti‐D and papainized group O rhesus (D) positive red cells as the targets. Monocyte depleted mononuclear cell suspensions were effective in lysing appropriately sensitized red cells and papainization considerably enhanced the degree of specific lysis. Variation in culture volume and incubation in tubes or microplates was not critical to the degree of specific lysis obtained provided that the number of effector cells and target cells per culture was constant and the anti‐D not diluted below the optimal concentration. Cytolytic activity was seen down to levels of 3 ng anti‐D per culture. Specificity for lysis resided with the anti‐D and not the effector cells. Several sources of anti‐D were effective in inducing lysis of D positive red cells although individual variation was noted. Anti‐c and anti‐E were also shown to be effective in inducing specific lysis of red cells with the appropriate antigens.
ADCC (K‐Cell) Lysis of Human Erythrocytes Sensitized with Rhesus Alloantibodies
Authors: Urbaniak S. J.
Published: March 2008

Summary. The mechanism of lysis of anti‐D coated human erythrocytes by human mononuclear K‐cells was investigated. Red cell lysis was measureable after 30 min incubation and reached a maximum by 18–20 h. Cell‐to‐cell contact was necessary for lysis, phagocytosis was not a prerequisite, and intact microfilament function was required. The divalent cations Ca2+ and Mg2+ were both required for lysis to occur. Studies with metabolic inhibitors indicate that some RNA and protein synthesis is required for maximum expression of ADCC and intact microtubule function is essential. In the present system lysis was mediated by IgG1 anti‐D antibodies and was signficantly inhibited by IgG1 and IgG3 subclasses with some inhibition by IgG2 but not by IgG4, IgA or IgM. This suggests that the K‐cell Fc receptor is specific for IgG but that there is major cross‐reactivity between IgG1 and IgG3. The inhibiting effect of hydrocortisone suggests that ADCC inhibition may be one mode of action of corticosteroids in ameliorating autoimmune haemolytic anaemia.
Variations in Granulocyte Colony Forming Cell Numbers in Adult Blood
Authors: Barrett A. J. , Faille A. , Ketels F.
Published: March 2008

Summary. Blood granulocyte colony forming units (CFU‐C) were studied in normal adults to establish: (i) a normal range; (ii) variability due to the culture technique; (iii) variability of blood CFU‐C within individuals. Thirty men studied had 98 (range 8–300) CFU‐C × 103/1, and 28 women studied had 44 (range 0–260) CFU‐C × 103/1. This difference was significant (P<0.001). There was also a significant sex difference in the total number of cells forming colonies and clusters per litre; and in the incidence of colony formers and of cluster formers in buffy coat and mononuclear cell blood fractions. CFU‐C were assayed in four subjects over a 10 week period. When buffy coat cells were used as a source of colony stimulation the week to week variation in the combined growth of the four subjects was wide (±36%) but with conditioned medium the variation was smaller (±14%).

In all subjects colony and cluster growth varied in the same way (r=0.77, P=0.001) but there was no correlation with the total leucocyte count. A 3–4 week cyclical change in CFU‐C/1 was found which was independent of the variation inherent in the technique. The physiological significance of the sex difference and the apparent cyclical changes in blood CFU‐C are not explained, but the results emphasize the wide fluctuations in CFU‐C that may occur in normal individuals.
Elevation of Pyrimidine Enzyme Activities in the RBC of Patients with Congenital Hypoplastic Anaemia and their Parents
Authors: Zielke H. Ronald , Ozand Pinar T. , Luddy Ruth E. , Zinkham William H. , Schwartz Allen D. , Sevdalian David A.
Published: March 2008

Summary. The activities of orotate phosphoribosyl transferase (OPRT) and orotidine monophosphate decarboxylase (ODC) were significantly elevated (P<0.001) in erythrocytes (RBC) from five patients with prednisone‐responsive congenital hypoplastic anaemia (CHA). (OPRT: patients=10.1–64.2 nmol/h/109 RBC; controls=2.8·0.3 (mean×SEM, n=37); ODC: patients=30–124 nmol/h/109 RBC; controls=10.2×0.7 (mean×SEM, n=37).) Two patients had a less pronounced, but significant, increase of aspartate transcarbamylase activity and three patients had marginal increases of dihydroorotase activity. Dihydroorotate dehydrogenase activity was not detected in any CHA patient or control. In one patient prior to prednisone therapy, the OPRT and ODT activities were elevated 10‐fold and remained elevated 3‐fold after 16 months of therapy. An elevated enzyme pattern similar to that of RBC from CHA patients was observed in three parents of three CHA patients, but not in three parents of two other CHA patients. The activities of all five pyrimidine enzymes were normal for one patient with transient erythroblastopenia of childhood. In contrast, the activities of all the pyrimidine biosynthetic enzymes were elevated in blood from patients with a young RBC population: sickle cell anaemia, sickle‐β‐thalassaemia, hereditary spherocytosis, and DiGuglielmo syndrome and from the newborn. It is postulated that factors which affect the activities of pyrimidine enzymes in CHA may also result in diminished erythropoiesis.
The Effect of Nitrous Oxide on the Cell Cycle in Human Bone Marrow
Authors: Cullen M. H. , Rees G. M. , Nancekievill D. G. , Amess J. A. L.
Published: March 2008

Summary. The effect of 24 h exposure to nitrous oxide on the cell division cycle in human bone marrow has been studied in vivo using the technique of DNA flow microfluorimetry. All patients who received nitrous oxide showed a significant increase in the proportion of early S‐phase cells with a decrease in late S, G2 and mitotic cells. These changes resemble those seen following the use of S‐phase‐specific cytotoxic drugs. Control patients showed no such effect. Parallel studies have suggested that interference with the function of vitamin B12 underlies this response. Nitrous oxide may provide a convenient method for studying the cell kinetic aspects of acute B12 deficiency and the possibility of using it to increase the therapeutic index of antitumour drugs is discussed.
Megakaryoblastic Leukaemia (Acute Myelofibrosis): a Report of Three Cases
Authors: Den Ottolander G. J. , Velde J. te , Brederoo P. , Geraedts J. P. M. , Slee P. H. T , Willemze R. , Zwaan F. E. , Haak H. L. , Muller H. P. , Bieger R.
Published: March 2008

Three patients with megakaryoblastic leukaemia are described. All three presented with pancytopenia, a few blast cells in the peripheral blood and absence of overt hepatosplenomegaly. In two of them bone marrow aspiration yielded a dry tap. Histological investigation of the bone marrow indicated that the megakaryocytic cell line was the dominant proliferating lineage. Cytochemical and EM investigation supported these findings. The isomorphic isoenzyme pattern of the elevated serum lactic dehydrogenase might be of diagnostic importance. Despite chemotherapy, there was a rapidly fatal terminal leukaemic phase with high blast cell counts. The differentiation from other haematological malignancies, especially acute (aleukaemic) leukaemias and the accelerated phase of primary (chronic) myelofibrosis, is discussed. The picture appears to be identical with acute (malignant) myelofibrosis.
Blood Eosinophils and Eosinophil Cationic Protein after Acute Myocardial Infarction or Corticosteroid Administration
Authors: Hällgren R. , Venge P. , Cullhed I. , Olsson I.
Published: March 2008

Measurements of eosinophil cationic protein in serum (S‐ECP) have been made in patients with acute myocardial infarction. In spite of low numbers of blood eosinophils in the acute phase of the disease S‐ECP levels fluctuated widely with often markedly raised levels. Peak levels of S‐ECP were significantly correlated (P < 0.001) to peak numbers of blood eosinophils, although the former usually occurred within the first 2–3 d of illness and the latter usually after the sixth day. Intravenous injection of 0.5 g methylprednisolone in healthy individuals reduced blood eosinophil counts for at least 24 h, but S‐ECP levels remained within normal limits. These results suggest that ECP is released from eosinophils in vivo. The raised levels of S‐ECP in patients with acute myocardial infarction are probably due to the active participation of eosinophils in the inflammatory process.
Determinants of Haemoglobin Level in Sickle Cell‐Haemoglobin C Disease
Authors: Bannerman R. M. , Serjeant Beryl , Seakins M. , England J. M. , Serjeant G. R.
Published: March 2008

Summary. The determinants of steady state haemoglobin levels in sickle cell‐haemoglobin C (SC) disease were investigated by measuring routine haematological and biochemical indices, red cell survival, oxygen affinity, pitted erythrocytes, and red cell and plasma volumes in 31 adult patients (15 male; 16 female).

Red cell survival was shortened in all subjects, and was positively correlated with haemoglobin level. However, many haemoglobin values were within the normal range, especially in male subjects. Palpable splenomegaly, which occurred in 53% of patients, did not appear to affect haemoglobin level, red cell survival, plasma volume, or red cell volume, but was associated with lower platelet counts and decreased pitted red cells. Sex related differences were found in total haemoglobin, packed cell volume, conductivity cell volume, red cell count, and in the blood volume measurements.

Red cell, plasma and total blood volumes in patients varied with weight and cube of height in manner observed in normal subjects, although red cell volumes were lower and plasma volumes were greater than in normal subjects of given height and weight. Anaemia in SC disease is related to the haemolytic rate but the major determinant appears to be an inappropriate increase in plasma volume.
Haemophilia ‘A’in a 46,X,i(Xq) Female
Authors: Mori P. G. , Pasino M. , Vadalà C. Rosanda , Bisogni M. C. , Tonini G. P. , Scarabicchi S.
Published: March 2008

Summary. A phenotypically normal female, with negative family history for bleeding disorders, was found to be affected by severe haemophilia A. All laboratory tests performed confirmed the diagnosis. Chromosome analysis showed the presence of an X isochromosome of the long arm in every cell. It is hypothesized that the propositus inherited the affected X from her mother, a probable carrier, and the isochromosome, of paternal origin, was not able to mask the abnormal gene.
Studies of Ineffective Erythropoiesis and Peripheral Haemolysis in Congenital Dyserythropoietic Anaemia Type II
Authors: Barosi G. , Cazzola M. , Stefanelli M. , Ascari E.
Published: March 2008

Summary. Erythrokinetic parameters were estimated in six patients suffering from congenital dyserythropoietic anaemia type II (CDA II) by means of a mathematical model of iron kinetics. A wide variation in effectiveness of erythroid activity was observed, and a significant negative correlation was found between ineffective erythropoiesis and peripheral haemolysis. In four patients with prominent peripheral haemolysis, splenectomy was carried out. Marked improvement in their clinical condition and in haemoglobin level resulted.
Phagocytosis of Nucleated and Mature β Thalassaemic Red Blood Cells by Mouse Macro‐phages in Vitro
Authors: Knyszynski Ahuva , Danon David , Kahane Itzhak , Rachmilewitz Eliezer A.
Published: March 2008

Summary. Physiological or experimental decrease in sialic acid (SA) content on the red blood cell (RBC) membrane is believed to play an important role in the recognition of these cells by macrophages. Since there is a 20–30% decrease in the SA content on the membrane of thalassaemic RBC, the interaction between macrophages and these RBC was studied in vitro. Using mouse peritoneal macrophages, it was found that these macrophages ‘recognize’and phagocytize thalassaemic RBC while RBC from normal donors are hardly phagocytized. The average level of phagocytosis of thalassaemic RBC from splenectomized patients was found to be 22‐fold higher than that of RBC from normal donors. The phagocytized cells consisted of both mature and nucleated RBC. Mouse peritoneal macrophages seem to be a useful in vitro system for the study of the accelerated sequestration and shortened life span of thalassaemic RBC.
Variables Involved in Cryoprecipitate Production and their Effect on Factor VIII Activity
Authors: Gunson H. H.
Published: March 2008

Summary. The Regional Transfusion Centres in England and Wales have studied the following variables in the preparation of cryoprecipitate: blood group of the donor, choice of anticoagulant (ACD or CPD), conditions of freezing, thawing and centrifugation of the plasma and age of the starting plasma.

Mean factor VIII activity in cryoprecipitates derived from group A plasma was significantly higher than from group O plasma. CPD may lead to increased potency of factor VIII when freshly collected plasma is used to prepare cryoprecipitates; rapidity of thawing may also be advantageous.

None of the other variables studied produced significant effects on the factor VIII activity of the cryoprecipitates and particular attention has been drawn to the results of experiments concerned with the age of starting plasma.
The Significance of Ph1 Mosaicism: a Report of Six Cases of Chronic Granulocytic Leukaemia and Two Cases of Acute Myeloid Leukaemia
Authors: Wayne A. W. , Sharp J. C. , Joyner M. V. , Sterndale H. , Pulford K. A. F.
Published: March 2008

Summary. Six cases of chronic granulocytic leukaemia (CGL) and two cases of acute myeloid leukaemia (AML) with dual populations of karyotypically normal and Philadelphia (Ph1) chromosome‐positive cells are described. GTG and QF‐banding characterized the Ph1 as resulting from a 9/22 translocation in all eight cases. Four of the patients suffering from CGL presented with 100% Ph1‐positive bone marrows, and after receiving intensive chemotherapy, karyotypically normal cells were demonstrated. The other two patients with CGL showed Ph1 mosaicism at presentation. The two patients with AML exhibited Ph1 mosaicism at presentation and throughout the course of the disease. In both of these patients, a marker No. 10 chromosome was found in some of the Ph1‐positive cells and in one hyperdiploidy was observed to have developed only in the clone with the additional chromosome anomaly.
37°C E Rosettes in Various Malignant and Non‐malignant Disorders
Authors: Palutke Margarita , Tabaczka Pamela , Varadachari Chandra , Carrillo Gilda
Published: March 2008

Summary. T lymphocytes, forming sheep erythrocyte rosettes at 37°C, have been described in thymus glands, mitogen and allogeneic cell stimulated lymphocyte cultures, and acute T cell lymphoblastic leukaemia. This paper describes the finding of such lymphocytes in lymph nodes of a variety of disorders including Hodgkin's disease, Lennert's lymphoma, malignant lymphoma of large transformed T lymphocytes, immunoblastic lymphadenopathy, metastatic carcinoma, and other disorders. A large number of tonsils, but not reactive lymph nodes, also contained a significant number of those T lymphocytes. The significance of these findings in relation to interpretation of immune red cell rosettes and to the subclasses of T lymphocytes is discussed.
Cross‐Linking of Membrane Proteins of Metabolically‐Depleted and Calcium‐Loaded Erythrocytes
Authors: Coetzer Theresa L. , Zail Solam S.
Published: March 2008

Summary. The membranes of erythrocytes undergoing metabolic depletion or an influx of calcium undergo several changes in structure and function. In erythrocytes incubated without substrate we find extensive cross‐linking of membrane proteins by disulphide bonding occurring after 24–48 h, involving all major membrane proteins as well as haemoglobin. Aggregates of mol wt 40 × 106 or greater are formed. These changes are partially reversible by repletion with adenosine. Rapid introduction of calcium (intracellular concentrations approximately 0.6 mm) into metabolically replete erythrocytes with the ionophore A23187 results in transgluta‐minase‐dependent cross‐linking of membrane proteins. Cellular calcium concentrations of approximately 0.3 mm have no cross‐linking effect. Cells undergoing metabolic depletion show a progressive loss of transglutaminase activity to undetec‐table levels at 12 h, so that influx of calcium into such cells cannot cause cross‐linking by a transglutaminase‐mediated reaction. These studies suggest that the metabolic state of the cell and the rate and degree of calcium influx into erythrocytes are critical factors in determining the type of membrane protein cross‐linkage.
Electron Microscopy of Metal‐shadowed Fibrinogen Molecules Deposited at Different Concentrations
Authors: Mosesson Michael W. , Escaig Jacques , Feldmann Gérard
Published: March 2008

Summary. The hypothesis that fibrinogen molecules are elongated (47.5 nm) trinodular structures is founded upon electron microscopic analyses of air‐dried metal shadowed preparations in which collections of three discrete linearly aligned particles were commonly observed and were assumed to represent an intact molecule. In our study, human fibrinogen at concentrations ranging from 0.1 to 150 μg/ml in 0.1% ammonium acetate buffer, pH 9.3 ± 0.1, was deposited on carbon films, air‐dried, shadowed with platinum, and then examined by electron microscopy. When deposited at a concentration of 10 μg/ml or higher, most images showed considerable polydispersity, evidently due to overloading, that was characterized by multinodular shapes varying in length, width, and height; discrete particulate forms were also commonly observed. At a lower concentration in the range of 1 μg/ml, the degree of polydispersity was much less and the predominant form was a discrete particle with a mean diameter and height of 13.7 ± 1.4 nm and 5.4 ± 0.8 nm, respectively. Only rarely were three such particles aligned linearly so as to suggest a trinodular molecular shape. At still lower fibrinogen concentrations in the range of 0.1 μg/ml, the number of particles per unit of surface was considerably diminished, but virtually all forms observed were individual particles that were well separated (> 20 nm) from one another. Approximation of the volume of each particle indicates that an anhydrous molecule having the size of fibrinogen can be accommodated. These observations plus quantitative considerations of particle distribution do not support the conclusion that molecules of fibrinogen are elongated trinodular structures but suggest instead that the particulate forms reflect single molecules of fibrinogen and that multinodular forms represent collections of two or more molecules.