bluuebunny/crossref_metadata_2025 huggingface dataset:
Circulating pluripotent haemopoietic cells in patients with myeloproliferative disorders
Authors: Douer Dan , Fabian Ina , Cline Martin J.
Published: March 2008

Summary. Peripheral blood cells of 17 patients with myeloproliferative disorders were grown in vitro to form mixed colonies (CFU‐mix). CFU‐mix were found in four of nine patients with chronic phase chronic myelocytic leukaemia (CML) at levels 28‐ to 132‐fold higher than the upper limits of normal subjects. These patients also had higher than normal levels of circulating granulocyte‐macrophage (CFU‐GM) and erythroid (BFU‐E) progenitors. Development of blastic crisis was associated with disappearance of CFU‐mix from the blood.

CFU‐mix was detected in the blood of three of five patients with polycythaemia vera (PV) and two patients with myelofibrosis with myeloid metaplasia (MMM) at levels slightly higher than those of normal subjects. Our results are consistent with a clonal expansion of multipotent haematopoietic precursor cells in CML and abnormal kinetics of this cell in CML, PV and MMM.
Transition from granulocyte to macrophage predominance in bone marrow cultures
Authors: Shulman Lawrence N. , Robinson Stephen H.
Published: March 2008

Summary. These studies examine the mechanisms underlying the transition from granulocytes to macrophages that characterizes cultures of bone marrow cells. Normal mouse marrow was cultured for several weeks in diffusion chambers (DCs) implanted in previously irradiated host mice. Previous studies with this system have shown that there is a period of logarithmic cell growth for 7 d followed by a plateau in both total cell number and the numbers of CFU‐S and CFU‐C. The present studies show that this plateau in cell growth is associated with high cycling rates of both CFU‐S and CFU‐C. The plateau phase ends with a rapid decrease in the numbers of these progenitor cells. This is associated with a decline in the number of granulocytes, whereas macrophages persist at approximately the plateau level for the entire 48 d of culture. Transplantation of plateau phase cells into secondary DC cultures leads to a more rapid decline in the numbers of CFU‐S and CFU‐C followed by the total disappearance of granulocytes; in contrast, macrophage number undergoes a small increase which is maintained thereafter. When regular injections of 3H‐thymidine are given to host mice bearing late phase secondary cultures, macrophage labelling rises to a plateau of only 30–40%, indicating that most macrophages at this stage of culture are long‐lived cells. The kinetics of the stem cell changes implicate the CFU‐C as the immediate precursor of granulocyte development in DC cultures but suggest that the CFU‐S is the major determinant of overall cell renewal in this system. Macrophages doubtless also originate from CFU‐C but, unlike granulocytes, these are often long‐lived cells probably with some capacity for continuing cell division. These properties account for the often observed propensity of macrophages to repopulate bone marrow cultures from which stem cells and granulocytes have been depleted.
Inhibition of the pentose phosphate shunt by 2,3‐diphosphoglycerate in erythrocyte pyruvate kinase deficiency
Authors: Tomoda Akio , Lachant Neil A. , Noble Nancy A. , Tanaka Kouichi R.
Published: March 2008

Summary. Pentose phosphate shunt activity was studied by the release of 14CO2 from 14C‐1‐glucose and 14C‐2‐glucose in the red cells of five patients with pyruvate kinase deficiency and found to be significantly decreased after new methylene blue stimulation when compared to high reticulocyte controls. Incubated Heinz body formation was increased and the ascorbate cyanide test was positive in blood from these patients. The activity of glucose‐6‐phosphate dehydrogenase (G6PD) as well as that of 6‐phosphogluconate dehydrogenase (6PGD) was inhibited to 20% of baseline in normal red cell haemolysate by 4 mM 2,3‐diphosphoglycerate at pH 7·1. 2,3‐Diphosphoglycerate was a competitive inhibitor with 6‐phosphogluconate (Ki= 1·05 mM) and a noncompetitive inhibitor with NADP (Ki= 3·3 mM) for 6PGD. Since the intracellular concentrations of glucose‐6‐phosphate, 6‐phosphogluconate and NADP are below their Kms for G6PD and 6PGD, the kinetic data suggest that increased concentrations of 2,3‐diphosphoglycerate in pyruvate kinase deficient red cells are sufficiently high to suppress pentose phosphate shunt activity. This suppression may be an additional factor contributing to the haemolytic anaemia of pyruvate kinase deficiency, particularly during periods of infection or metabolic stress.
Cellular abnormalities of folate deficiency
Authors: Steinberg Stephen E. , Fonda Susan , Campbell Caryl L. , Hillman Robert S.
Published: March 2008

Summary. To trace the development of folate‐deficient abnormalities of morphology and DNA synthesis, Friend erythroleukaemia cells were grown in media containing 102, 103 and 104 ng of [3H]PteGlu1/ml and then transferred to folate‐free media. Parameters examined were: intracellular folate levels; growth potential; morphology; dU suppression; and DNA content by flow microfluorimetry. The most sensitive indicators of folate‐deficient cell growth were those related to DNA synthesis (dU and flow microfluorimetry). These became abnormal at intracellular folate levels of 0.2–0.5 ng/106 cells and markedly so below 0.1 ng/106 cells. Morphological criteria were less sensitive. Cells became megaloblastic at intracellular folate levels below 0.06 ng/106. The capacity of the cells to replicate in folate‐free media was a function of the intracellular folate (ICF): duplications = 4.01+ln(ICF)/0.67 (r = 0.993, P<0.001).

These studies demonstrate that regardless of initial intracellular folate levels, cellular stigmata of folate deficiency appear when cellular folate falls below 3 × 105 molecules per cell (dU and flow microfluorimetry) and cells lose the capacity for further replication below 7–10 × 105 molecules. The intracellular folate level not only predicts early defects, but also determines the replicative capacity.
Ultrastructural localization of the transferrin receptor and transferrin on marrow cell surfaces
Authors: Parmley Richard T. , Hajdu Ivan , Denys Francis R.
Published: March 2008

Summary. The monoclonal antibody OKT9 (a known transferrin receptor antibody) and a monoclonal antibody to transferrin (ATfn) were used to localize the transferrin receptor and transferrin on marrow cells. After incubation of cell suspensions with the antibody, the cells were reacted with an affinity purified Fab fragment of goat anti‐mouse IgG conjugated to horseradish peroxidase (GAM‐HRP), which was in turn visualized by reaction with 3,3′‐diaminobenzidine (DAB). Erythroblast cell surfaces stained intensely with OKT9‐GAM‐HRP‐DAB, weaker staining was observed on reticulocyte surfaces, whereas erythrocyte surfaces lacked staining. Staining was present on surface caveolae, which often contained endogenous ferritin particles. Moderate to strong OKT9 staining was observed on less than 10% of presumed lymphoid cells. Monocytes, macrophages, promyelocytes, granulocytes, megakaryocytes and platelets consistently lacked OKT9 staining. ATm‐GAM‐HRP‐DAB staining of erythroid cells was similar to that observed with OKT9 staining; however, in contrast to the latter staining, ATfn stained the surfaces of megakaryocytes, platelets, monocytes and most lymphocytes. Promyelocytes stained weakly, whereas late granulocytes lacked staining. These results indicate that the T9 transferrin receptor (1) is largely confined to erythroid cells in marrow, (2) is diffusely distributed on the surface of early erythroid cells, (3) decreases with cell maturation, and (4) is lost when haemoglobin synthesis is completed. Transferrin appears in a similar distribution on erythroid cell surfaces but also appears to bind to some cell surfaces lacking the T9 receptor.
Analysis of the Hb M Milwaukee mutation at the DNA level
Authors: Horst J. , Schäfer R. , Kleihauer E. , Kohne E.
Published: March 2008

Summary. Restriction endonuclease mapping of cellular DNA with the enzyme Sst I has been used to detect the haemoglobin (Hb) Milwaukee mutation directly. Instead of a normal 15.5 kilobase pairs (kb) fragment which contains the normal β‐globin structural genes, in heterozygous Hb M Milwaukee DNA two additional fragments of 9.0 kb and 6.5 kb were obtained that are diagnostic for this anomaly. The position of Sst I sites within the β‐globin gene region could be established.
Unusual combination of genetic defects in a Sicilian family: β‐thalassaemia, haemoglobin Lepore Boston‐Washington and heterocellular hereditary persistence of fetal haemoglobin
Authors: Schilirò G. , Musumeci S. , Romeo M. A. , Gregorio F. Di , Russo A. , Testa R. , Russo G.
Published: March 2008

Summary. This paper reports a Sicilian family in which β‐thalassaemia, haemoglobin Lepore Boston‐Washington and heterotocellular hereditary persistence of fetal haemoglobin (HPFH) were present in various combinations. The most interesting combination was that of Hb Lepore and heterocellular HPFH, which has not been previously reported. This subject was clinically normal, with the haematological picture of Hb Lepore trait and an unusually high level of Hb F due to an increased number of circulating F cells.
The effects of alpha‐thalassaemia in HbSC disease
Authors: Steinberg M. H. , Coleman M. B. , Adams J. G. , Platica O. , Gillette P. , Rieder R. F.
Published: March 2008

Summary. In HbSC disease, as in sickle cell anaemia, there is a spectrum of clinical severity. A reduced mean corpuscular volume and haemoglobin concentration, traits typical of thalassaemia, might retard sickling. We therefore ascertained the prevalence of α‐thalassaemia in 53 adults with HbSC disease and related α‐globin gene deletion to the haematologic and clinical findings. Alpha‐globin genotype was identified by restriction endonuclease gene mapping. Indirect ophthalmoscopy and fluorescein angiography were used to document the presence of proliferative retinopathy. Bone necrosis and infarction were determined roentgenographically or by radionuclide scanning. Either heterozygous or homozygous alpha‐thalassaemia‐2 was present in 26% of patients. There was no relationship between α‐globin genotype and haematocrit, pain crises, bone lesions, proliferation retinopathy or clinical severity score.
Interference of IgG, IgG aggregates and immune complexes in tests for platelet autoantibodies
Authors: Helmerhorst F. M. , Smeenk R. J. T. , Hack C. E. , Engelfriet C. P. , Borne A. E. G. Kr. von dem
Published: March 2008

Summary. Three techniques, based on the antiglobulin principle, used for the detection of autoantibodies against platelets, were compared; the antiglobulin consumption assay (QACA), the platelet radioactive antiglobulin test (PRAT) and the platelet suspension immunofluorescence test (PSIFT).

Upon incubation of normal donor platelets with purified IgG, in concentrations higher than that in serum, an increased amount of platelet‐associated IgG was demonstrated only in the QACA. Upon incubation with aggregated IgG, all three tests became positive, but the PSIFT only with high concentrations of aggregates. Binding of soluble C1q‐binding immune complexes (IC), which consisted of tetanus toxoid and IgG antitetanus antibodies (TaT) to normal donor platelets, was only detectable in the QACA. However, a positive result was obtained in all three tests with platelets incubated with soluble DNA‐IgG‐antiDNA antibodies (DaD) IC. Fixation of the platelets with paraformaldehyde prevented the binding and the detection of the DaD‐IC, but not of IgG, aggregated IgG or TaT‐IC.

Eluates from platelets incubated with aggregated IgG, TaT‐ or DaD‐IC did not react with normal donor platelets in the three techniques, in contrast to eluates from platelets sensitized with platelet antibodies.
Steroid modulation of naturally occurring diurnal variation in circulating pluripotential haematopoietic cells (CFU‐GEMM)
Authors: Lasky Larry C. , Ascensao Joao , McCullough Jeffrey , Zanjani Esmail D.
Published: March 2008

Summary. The number of committed granulocyte‐monocyte precursors (CFU‐GM) in circulation has been shown to increase following steroid administration in humans. To see if steroids could be used to improve the collection of peripheral blood stem cells for haematopoietic reconstitution, their effect on circulating CFU‐GEMM was investigated. Six healthy young adults, three men and three women, were given 60 mg of prednisone orally at 8 a.m. on day 1. Blood CFU‐GEMM and committed precursors (CFU‐GM and BFU‐E) were cultured in methylcellulose. Samples were taken at 4 p.m. the day before and at 8 a.m. on the day of steroid administration, and at 4, 8, 24 and, in a few cases, 48 h after steroid. Pre‐treatment CFU‐GEMM levels (per unit volume of blood) at 4 p.m. was 180 ± 23% (mean ± SEM) of that at 8 a.m., showing a significant (P < 0·025) diurnal variation. 8 h after steroids there was a fall in CFU‐GEMM to 28 ± 4·5% of the 8 a.m. presteroid level. 24 h following steroid administration, CFU‐GEMM rose significantly (P < 0·05) to 188 ± 33% of 8 a.m. baseline values; CFU‐GM and BFU‐E changes generally paralleled those noted for CFU‐GEMM. These results suggest that blood levels of CFU‐GEMM exhibit a significant diurnal variation. Oral prednisone given 24 h in advance of collection increases the 8 a.m. value to that found at 4 p.m. The steroid effect may be due to a resetting of the diurnal control mechanism. Use of this information may be important in collection of circulating haematopoietic stem cells for use in bone marrow reconstitution in man.
Effect of coumadin‐induced coagulopoietin plasma on vitamin K‐dependent carboxylation of liver microsomes
Authors: Karpatkin S. , Chang R.‐J. , Pierce W. , Karpatkin M.
Published: March 2008

Summary. Coumadin‐treated rabbits have a humoral substance(s) (coagulopoietin) which is capable of elevating vitamin K‐dependent coagulation factors when injected into recipient rabbits (Karpatkin & Karpatkin, 1973). Biologic levels of coagulation factors II, V, VII and X; immunologic levels of factors II and X; and vitamin K‐dependent liver microsomal carboxylase activity were measured in recipient rabbits receiving coumadin‐induced coagulopoietin plasma. Factor II biologic activity increased 3·5‐fold compared to the increase in immunologic activity. Factor X biologic activity increased 1·7‐fold compared to the increase in immunologic activity. This indicates an increase in specific activity of factors II and X. Coumadin‐induced coagulopoietin plasma had no effect on vitamin K‐dependent liver microsomal carboxylase activity in vitro. However, livers obtained from recipient animals treated with coumadin‐induced coagulopoietin plasma enhanced their carboxylase activity (compared to control animals) 2·4‐fold employing endogenous microsomal precursor for carboxylation, and 6·2‐fold employing synthetic substrate, phe‐leu‐glu‐glu‐val. Thus, coumadin‐induced coagulopoietin plasma enhances the biologic activity of vitamin K‐dependent coagulation factors II, VII, and X as well as the ex vivo vitamin K‐dependent carboxylase activity of liver microsomes.
Chronic myelodysplastic syndrome: short survival with or without evolution to acute leukaemia
Authors: Weisdorf Daniel J. , Oken Martin M. , Johnson Gerhard J. , Rydell Robert E.
Published: March 2008

Summary. The myelodysplastic syndromes represent a prognostically diverse group of disorders. Their study has recently been facilitated by the classification proposed by the French‐American‐British (FAB) Cooperative Group. Using this scheme it is now possible to define more precisely their natural history and clinical relationship to acute leukaemia. Using the longitudinal case control technique, we reviewed the clinical data and morphology of 69 patients (all elderly males) with chronic irreversible haematological cytopenia and dysplasia. Applying FAB criteria we found: refractory anaemia (RA) in 43%; sideroblastic anaemia (RA‐S) in 33%; refractory anaemia with excess blasts (RAEB) in 13%; RAEB in transformation (RAEB‐T) in 9% and chronic myelomonocytic leukaemia (CMML) in 1%. The median survival for the entire group was 27 months (RA, 52; RA‐S, 29; RAEB, 12; RAEB‐T 11; and CMML, 2 months). Short survival was predicted by transfusion requirement and other manifestations of severe cytopenia, as well as by myeloid immaturity. The presence or absence of sideroblastosis did not correlate with survival. Acute leukaemia developed in only eight patients (12%), six of whom initially had RA. Leukaemic transformation was not predicted by progressive cytological immaturity. This study demonstrates that even in the absence of leukaemic transformation, chronic myelodysplasia is a lethal haematological disorder.
A new type of congenital dysfibrinogenaemia with defective fibrin lysis—Dusard syndrome: possible relation to thrombosis
Authors: Soria Jeannette , Soria Claudine , Caen Jacques P.
Published: March 2008

Summary. Congenital dysfibrinogenaemia is described in three members of a family presenting with recurrent thrombosis and in two other young members not yet affected. An abnormality in the polymerization of fibrin monomers was noted. In addition, the pathological fibrin clots were found to be less sensitive to degradation by a post venous occlusion euglobulin solution than normal fibrin. After fibrin clot incubation with lys‐plasminogen at different concentrations, the biological activity of plasminogen in patient fibrin clot on S 2251 after SK‐addition, was less than that observed with normal fibrin. It is speculated that defective in vivo thrombolysis might explain the recurrent thrombosis observed in this family. This finding represents a new concept in understanding thromboembolic diseases.
Granulocytic cryopreservation: further studies on the pathogenesis of impaired cellular function
Authors: McCarthy D. M. , Skacel P. , Raja K. , Martin F. , Peters T. , Goldman J. M.
Published: March 2008

Summary The effects of different methods of purifying peripheral blood granulocytes on their functional capacity after cryopreservation have been studied. The standard method of isolating granulocytes by centrifugation through Lymphoprep sensitizes cells to damage induced by subsequent cryopreservation. Some of this damage occurs during addition of cryoprotectant and we describe how to add and remove dimethyl sulphoxide without impairing cell function. The granulocytic function that suffers most during cryopreservation is granulocyte migration. This impairment may be partially due to uncontrolled entry into the cell of calcium. In contrast the bactericidal capacity in vitro is not so greatly reduced and ability to reduce nitroblue tetrazolium redox dye even less so.
Defect in glycosylation of erythrocyte membrane proteins in congenital dyserythropoietic anaemia type II (HEMPAS)
Authors: Fukuda Michiko N. , Papayannopoulou Thalia , Gordon‐Smith E. C. , Rochant Henri , Testa Ugo
Published: March 2008

Summary Congenital dyserythropoietic anaemia type II (HEMPAS) is a hereditary disease believed to be caused by a membrane abnormality of erythroid cells. Since the molecular basis of this membrane abnormality has not yet been defined, membrane glycoproteins of HEMPAS erythrocytes were analysed by cell surface labelling and endo‐β‐galactosidase digestion in this study. HEMPAS erythrocytes showed an abnormal glycoprotein profile when cells were labelled by the galactose oxidase/NaB[3H]4 method; Band 3 and Band 4·5 glycoproteins in HEMPAS are labelled but with less intensity although normally these proteins are the major components revealed by the same method. Instead, in HEMPAS, labelled lactosaminoglycans were found as a lower molecular weight glycoconjugate (HEMPAS glycan). HEMPAS glycan was characterized by micelle formation, a monomer molecular weight of 4000, susceptibility to endo‐β‐galactosidase and resistance to protease. These characteristics suggest that HEMPAS glycan has the nature of macroglycolipid. Proteins of Band 3 and the glucose transport protein (a component of Band 4·5), which were detected by antibodies showed a slightly decreased molecular weight in HEMPAS erythrocytes compared to those from normal erythrocytes, which was consistent with the decreased glycosylation of these proteins. The results indicate that anomalies in glycosylation occurred specifically in lactosaminoglycan glycoproteins of HEMPAS erythrocytes.
The haemoglobin H disease mental retardation syndrome: molecular studies on the South African case
Authors: Bowcock Anne M. , Van Tonder Sue , Jenkins Trefor
Published: March 2008

Summary The child with haemoglobin H (HbH) disease and multiple congenital abnormalities reported by Borochowitz et al (1970) has been more fully investigated in view of the recent report by Weatherall et al (1981) of a ‘new’syndrome of HbH disease and mental retardation. Restriction enzyme analysis indicates that the child's HbH disease results from the inheritance of an α‐thalassaemia 2 chromosome (‐ α/) from his mother and, from his father, a chromosome which has undergone a deletion including the ζ‐ and α‐globin genes as well as an undefined Ijsngth of DNA. Striking similarities between our patient and Weatherall et al's patients at the clinical level and between our patient and their patient number 1 at the molecular level, confirm that a new syndrome has been defined.
Molecular heterogeneity of beta thalassaemia in the Italian population
Authors: Giampaolo A. , Mavilio F. , Massa A. , Gabbianelli M. , Guerriero R. , Sposi N. M. , Caré A. , Cianciulli P. , Tentori L. , Marinucci M.
Published: March 2008

Summary Fifty‐one subjects originating from Southern Italy and affected by Cooley's anaemia have been studied in order to define the degree of heterogeneity of β thalassaemia mutations in this high incidence area. Restriction endonuclease mapping has been carried out on genomic DNA by the Southern blot technique both to exclude the existence of gross deletions or rearrangements and to establish the relative frequency of four polymorphic restriction sites (i.e. Gγ and Aγ Hind III, β Ava II and β Bam HI) within the γδβ gene region. In 28 subjects unequivocal linkage of the four polymorphic sites has been determined leading to the identification of seven different chromosome haplotypes, six of which had previously been reported associated with specific β0 and β+ thalassaemia mutations. Globin chain synthesis studies on peripheral blood reticulocytes indicated that subjects carrying the same genotype may behave differently as far as the β chain production is concerned relative to both the a and the non‐α chains. Thus, β thalassaemia turns out to be quite heterogeneous even in this limited geographical area. β+ mutations appear to be predominant, particularly those affecting nuclear precursor RNA splicing to mature β globin mRNA.
Fibrinogen Adelaide: a familial hypodysfibrinogenaemia associated with abnormal alpha chains
Authors: Exner Thomas , Barber Sylvia , Sage Robert E. , Kronenberg Harry
Published: March 2008

Summary A familial hypodysfibrinogenaemia occurring in four females with occasional haemorrhagic problems in an Adelaide family was investigated. Affected family members had slightly prolonged thrombin time, prothrombin time and Reptilase time tests, and apparently elevated levels of fibrin degradation products (FDPs). Fibrinogen assessed by reactivity with thrombin (Clauss method) was significantly less than fibrinogen determined by various other methods, though even by immunoquantitation fibrinogen levels were only slightly above half normal in affected family members. Isoelectric focussing (IEF) of the reduced patient's fibrinogen in urea/polyacrylamide gel revealed minor components with higher isoelectric points (pI) than present in normal fibrinogen or fibrin. These were shown to have a molecular weight similar to the alpha chain of fibrin by subsequent sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE). The abnormal species tended to remain in plasma after clotting with Reptilase or thrombin although polymerization of the clotted fibrin was normal. The magnitude of the pI shift relative to normal fibrin alpha chain indicated an increased positive charge on the abnormal species.
Acute leukaemia with mixed lymphoid and myeloid phenotype
Authors: Pui Ching‐Hon , Dahl Gary V. , Melvin Susan , Williams Dorothy L. , Peiper Stephen , Mirro Joseph , Murphy Sharon B. , Stass Sanford
Published: March 2008

Summary Three children with acute leukaemia had blasts that expressed both lymphoid and myeloid markers. The blasts met immunological criteria for acute lymphoblastic leukaemia (ALL)—common ALL antigen+, HLA‐DR+, terminal deoxynucleotidyl transferase+–but their cytochemical features, including positive myeloperoxidase and Sudan black B, were those of acute nonlymphoblastic leukaemia (ANLL) as defined by the French‐American‐British Group. 30% of the blasts from one of two patients tested reacted with a monoclonal antibody specific for nonlymphoid cells (MCS‐2). The wide overlap in the percentages of blasts expressing lymphoid or myeloid markers indicates that some leukaemic cells in each child had a mixed phenotype. There were no consistent cytogenetic findings, and the Philadelphia chromosome was not present. Complete remission was induced by treatment effective for either ALL (two patients) or ANLL. These three cases appear to represent a rare leukaemia subtype that we have designated acute leukaemia with mixed lymphoid and myeloid phenotype. Its recognition may be important in treatment, since two patients achieved remission with standard therapy for ALL. These cases demonstrate further the phenotypic heterogeneity that may be seen in leukaemic cell differentiation.
Platelet antigens in chronic idiopathic thrombocytopenic purpura
Authors: Donna Mason  , McMillan Robert
Published: March 2008

Summary. IgG produced by splenic cells from five patients with chronic immune thrombocytopenic purpura (ITP) and from four control subjects was radiolabelled and incubated with target platelets. Eluates from these platelets were incubated with solubilized proteins from the same platelets immobilized on nitrocellulose paper. After washing, putative antigens were localized by autoradiography. Positive results were obtained in three of the five ITP patients; in two patients three radiolabelled bands of high molecular weight were noted (greater than 200 000) and in the third patient two different bands (molecular weight 140000 and 105000) were seen. Control studies were negative. These studies show that patients with chronic ITP produce antibody reactive with antigens associated with human platelet proteins and that the antigenic pattern differs between patients.
Availability, distribution and kinetics of the transferrin receptors of rabbit reticulocytes
Authors: Egyed A.
Published: March 2008

Summary. The binding of transferrin by reticulocytes was studied at 3°C and 37°C. It was demonstrated that 50% of the total number of receptors are in an exposed position on the outer side of the plasma membrane and 50% of them are in a masked position. All of the receptors can take either of the positions. The transition between these positions is temperature dependent and can be observed as the progressive phase of the transferrin binding. It is suggested that the transferrin receptors are in dimers, one of the subunits being in the exposed position the other in the masked position. The subunits change position simultaneously. This movement of the receptors is independent of the presence of transferrin. In the light of these results a new model for transferrin‐cell cycle is proposed.
Simultaneous demonstration of membrane antigens and cytochemistry at ultrastructural level: a study with the immunogold method, acid phosphatase and myeloperoxidase
Authors: Robinson D. , de Castro J. Tavares , Polli N. , O'Brien M. , Catovsky D.
Published: March 2008

Summary. We describe a method for electron microscopy that combines myeloperoxidase or acid phosphatase cytochemistry with the labelling of cell surface antigens with monoclonal antibodies and colloidal gold. This technique was tested in samples of normal mononuclear cells, leukaemic T cells with a mature or immature phenotype and an acute myeloid leukaemia. This method allows the demonstration at a single cell level of ultrastructural morphology, cytochemical reaction and the presence of a membrane antigen. It will improve further the analytic power of electron microscopy in the characterization of leukaemic cells particularly in cases of mixed leukaemias and in the study of normal haemopoietic differentiation with monoclonal antibodies.
Hyperleucocytic retinopathy in chronic granulocytic leukaemia: the role of intensive leucapheresis
Authors: Mehta A. B. , Goldman J. M. , Kohner E.
Published: March 2008

Summary. We describe three patients with chronic granulocytic leukaemia who developed impaired visual acuity in association with high leucocyte counts. Two of the patients were in the chronic phase of their disease and the third developed visual symptoms at the time of blastic transformation. Fundoscopy showed retinal haemorrhages and exudates and a vascular picture consistent with hyperviscosity. We propose the term ‘hyperleucocytic retinopathy’ for these appearances. Two of the patients derived rapid benefit following leucapheresis and the visual acuity of the third patient improved following treatment with cytotoxic drugs.
The agglutination of human platelets by botrocetin: evidence that botrocetin and ristocetin act at different sites on the factor VIII molecule and platelet membrane
Authors: Howard Margaret A. , Perkin Jean , Salem Hatem H. , Firkin B. G.
Published: March 2008

Summary. Botrocetin caused a factor VIII (FVIII) dependent platelet agglutination which was associated with a reduction in the plasma levels of all FVIII parameters as a result of specific binding of FVIII to the platelets. The site of binding of FVIII to the platelet in response to ristocetin or botrocetin involves the glycoprotein 1 complex.

This is suggested by the inability of chymotrypsin treated platelets or platelets from patients with the Bernard‐Soulier syndrome to agglutinate in response to ristocetin.

These platelets responded to botrocetin, but this was greatly reduced compared to normal. Crossed immunoelectrophoretic analysis indicated that in the presence of botrocetin most multimetric forms of FVIII bound to the platelet, whereas ristocetin caused binding of high and intermediate molecular weight forms. The antibiotic vancomycin inhibited platelet agglutination by ristocetin but had no effect on that caused by botrocetin. Assays of FVIII von Willebrand factor (VIII:vWf) using botrocetin compared well with those obtained using ristocetin in plasmas from normal individuals and from patients with classical von Willebrand disease (vWd).

However, a patient with variant vWd demonstrated 100% botrocetin cofactor activity and 0% ristocetin cofactor activity. This suggested that the site of interaction on the FVIII molecule for botrocetin and ristocetin are different. Therefore the diagnosis of some von Willebrand variants cannot be excluded on the basis of a normal botrocetin cofactor assay.
Characterization of leucocyte phagocytic stimulatory material released by activated human platelets
Authors: Sakamoto H. , Firkin F.
Published: March 2008

Summary. The action of material released by activated human platelets on the phagocytic activity of human neutrophils and monocytes has been examined under conditions where there was a low background level of exposure of the leucocytes to platelet release products (PRPr). Incubation with PRPr produced an approximate three‐fold stimulation of particle ingestion by both cell types. Material with stimulatory activity passed through ultraflltration membranes capable of retaining molecules greater than 500 Daltons. This low molecular weight stimulator was inactivated by ATP diphosphopyrolase to suggest it could be one or more of the nucleotide di‐ or triphosphates in PRPr. Comparison with the action of specific nucleotides revealed only ADP and ATP exerted equivalent stimulatory effects at concentrations consistent with those of nucleotides normally present in PRPr. This effect was mediated by induction of a change in leucocyte behaviour which was sustained in the absence of continued exposure to the stimulatory agent. Stimulation of phagocytosis was also produced by high molecular weight material in PRPr, but this appeared to be mediated by dissociation of low molecular weight stimulator from a high molecular weight complex. These observations extend the range of biological functions known to respond to nucleotides in PRPr, and have implications for the interpretation of leucocyte phagocytic activity assessed in vitro in view of the potential modification of this process by exposure of leucocytes to PRPr during the isolation of leucocytes from peripheral blood.
Detection of haemophilia carriers: multivariate analysis compared with discriminant analysis using up to five factor VIII variates
Authors: Duncan B. M. , Tunbridge L. J. , Duncan E. M. , Lloyd J. V.
Published: March 2008

Summary. Multivariate analysis is potentially superior to the linear discriminant analysis which is commonly used to identify carriers of haemophilia. Our aim was to compare these two statistical methods, and to find which factor VIII variates most effectively partitioned carrier and normal subjects. In this study we assayed one‐ and two‐stage factor VIII coagulant activity, factor VIII related antigen by electroimmunoassay and by fluoroimmunoassay, and ristocetin co‐factor in 50 normal females and 50 carriers of haemophilia. From the results we calculated multivariate ellipses which circumscribed the normal and the carrier populations, and we displayed these on the monitor of a microcomputer. These ellipses separated the two populations better than linear discriminants calculated on the same data. Multivariate analysis correctly identified 94% of the carriers whereas discriminant analysis correctly identified only 84%. Discriminant analysis gave poorer results because the statistical assumption of equal variance was breached, whereas the assumption of multivariate normality was upheld. Of the five factor VIII variates, two‐stage factor VIII coagulant activity and factor VIII related antigen by electroimmunoassay correctly identified the most subjects. Ristocetin co‐factor did not improve the diagnostic ability, either when in lieu of or when added to factor VIII related antigen.
Radioimmunoassay of fragment E‐related neoantigen: validation studies and clinical application
Authors: Chen James P. , Hanna Wahid T. , Williams Teresa K. , Krauss Stephen
Published: March 2008

Summary. Measurement of fibrinogen‐fibrin degradation products (FDP) levels in plasma may provide a direct index of plasmin action, and increased levels of FDP would indicate coagulopathy. We have established an E‐neoantigen radioimmunoassay (Eneo RIA) that can determine normal and pathological plasma levels of E‐related FDP. The assay employs rabbit antiserum produced against fragment E derived from a plasmin digest of fibrinogen and subsequently absorbed with fibrinogen. The absorbed antiserum contains antibodies which are equally reactive with fibrinogen derived E (Fg‐E) and fibrin derived E (Fb‐E) but not with fibrinogen at 1 mg/ml. The Eneo RIA was validated by assay parallelism and by recovery experiments. Plasma Eneo immunoreactivities in 14 normals were 4–22 ng/ml (mean 12.7 ng/ml). Plasma Eneo levels in 23 of 24 patients with neoplastic and haematological diseases were elevated above normal (range 27–2027 ng/ml). Unusually high Eneo values were observed with three patients whose diseases were complicated by either disseminated intravascular coagulation (DIC) or deep vein thrombosis. After heparin therapy, the Eneo level of a patient with chronic DIC declined. A pathological plasma was eluted from a Sephadex G‐200 column and Eneo immunoreactivity was determined on the eluates. The gel filtration pattern of Eneo indicates that E‐related FDP is a family of plasmic fragments derived from crosslinked fibrin.
Atherosclerosis and von Willebrand factor
Authors: Mannucci P. M. , Bloom A. L. , Larriel M. J. , Nilsson I. M. , West R. R.
Published: March 2008

Summary. The prevalence of von Willebrand's disease (VWD) in Western European countries and Israel was assessed. Possible patients were identified initially by questionnaire and those with plasma levels of von Willebrand factor antigen (WFAg; factor VIII related antigen) less than 1 u/dl (<1%) were confirmed by immunoradiometric assay performed at four reference centres. The prevalences of severe VWD in Scandinavian countries were similar to each other and about 10 times greater than those in other Western European countries. The prevalence in Israel was intermediate. The Registry will form the basis for future non‐invasive epidemiological studies of atherosclerosis in these individuals.
Book Reviews
Authors: Catovsky Daniel , Turner M. W. , Gunson H. H. , Hoffbrand A. V.
Published: March 2008

Functional T Cell Subsets Defined by Monoclonal Antibodies. Immunological Reviews, Vol. 74 (Edited by G. Möller).

Primary and Secondary Immunodeficiency Disorders. Edited by R. K. Chandra.

Progress in Clinical and Biological Research. Volume 108. Massive Transfusion in Surgery and Trauma. Edited by J. Collins, K. Murawski and A.W. Shafer.

The Cobalamins. Edited by Charles A. Hall.
(Aγδβ)° thalassaemia: similarity of phenotype in four different molecular defects, including one newly described
Authors: Trent R. J. , Jones R. W. , Clegg J. B. , Weatherall D. J. , Davidson R. , Wood W. G.
Published: March 2008

Summary. Globin gene mapping of DNA from families with (Aγδβ)° thalassaemia has revealed a previously unreported gene deletion responsible for this condition. The deletion removes the Aγ, δ and β genes and while its 5’ end is in a similar position to that described in a previous deletion of this type, the 3’ ends of the two deletions are quite different. In addition we have observed further examples of two other previously described deletions which result in this disorder. Phenotypic comparisons of families with (Aγδβ)° thalassaemia, in which the molecular basis has been defined, show a remarkable similarity among the four different deletion defects, with important implications with regard to the mechanism by which deletions allow the continued expression of γ genes.
Clinical and immunological study of non‐Hodgkin T‐cell lymphomas (cutaneous and lymphoblastic lymphomas excluded)
Authors: Brouet Jean‐Claude , Rabian Claire , Gisselbrecht Christian , Flandrin Georges
Published: March 2008

Summary. We present here the immunologic, morphologic and clinical features of 16 T‐derived adult non‐Hodgkin lymphomas (NHL) (lymphoblastic and cutaneous lymphomas being excluded) observed in an unselected series of 260 NHL. Malignant cells bore T cell antigens (16 cases) but formed E rosettes in 14 cases only. In nine cases studied with monoclonal antibodies to T cell antigenic subsets, the phenotype of malignant cells was homogeneous; in seven cases the cells had a clear‐cut helper or suppressor/cytotoxic phenotype; in one case cells had a cortical thymocyte phenotype. No T‐cell subset antigens were detected on malignant cells from the last patient. Prominent morphologic features were a striking variation in tumour cell sizes, vascular proliferation and admixture of a large number of macrophages; most often, those lymphomas with a diffuse growth pattern could not be easily assigned to a given NHL subtype. The course of the disease was aggressive in most patients, only four having experienced a sustained complete remission. Waldeyer's ring involvement, waxing and waning nodes, polyclonal hypergammaglobulinaemia and skin infiltrates may be distinctive clinical features in some patients.
Reduced helper (OKT4 +): suppressor (OKT8 +) T ratios in aplastic anaemia: relation to immunosuppressive therapy
Authors: Kuriyama K. , Tomonaga M. , Jinnai I. , Matsuo T. , Yoshida Y. , Amenomori T. , And Y. Yamada , Ichimaru M.
Published: March 2008

Summary. T cell subset composition of peripheral blood and bone marrow from 22 patients with aplastic anaemia (AA) was studied by monoclonal antibodies (OKT3, OKT4 and OKT8). In the peripheral blood, OKT3 (pan‐T). OKT4 (helper/inducer‐T) and OKT8 (suppressor‐T) cells varied widely in number. The ratios of OKT4:OKT8 exhibited the same tendency. However, a subgroup of AA with a reduced ratio of lower than 1.0 was present both in cases with or without prior prednisolone therapy. Of the eight patients treated with immunosuppressants, four with reduced ratios responded, whereas the other four with normal or higher ratios did not. The ratios of two of the four responders gradually reached the normal level. These results suggest that the reduced OKT4: OKT8 ratio may be related to the cell‐mediated immunosuppressive mechanism postulated as a cause of stem cell inhibition in a subgroup of AA, and indicate prospectively the effectiveness of immunosuppressive therapy.
Probable in vivo induction of differentiation by retinoic acid of promyelocytes in acute promyelocytic leukaemia
Authors: Nilsson Bo
Published: March 2008

Summary. A 30‐year‐old woman was diagnosed as having an acute promyelocytic leukaemia in September 1981. Chemotherapeutic courses of daunomycin, ara‐C, thioguanine and prednisolone were administered, resulting in a complete remission, A relapse occurred in January 1982, and chemotherapy did not lead to a second complete remission; neutropenia persisted with a marked left shift of the marrow granulopoiesis. Courses of chemotherapy were given throughout the study. In September 1982, marrow promyelocytes markedly increased, making up 51% of the nucleated cells. One month later the situation was unchanged, and 13‐cis‐retinoic acid (1 mg/kg) was administered by mouth. Gradually the marrow proportion of promyelocytes decreased to normal levels. The peripheral blood and marrow were still normal after 20 weeks of treatment with retinoic acid. Thus, retinoic acid seemed to have been inducing differentiation in an abnormally increased, maturation‐deficient population of promyelocytes in a patient with acute promyelocytic leukaemia.
Leu‐10 (HLA‐DC/DS) antigen distribution in human leukaemic disorders as detected by a monoclonal antibody: correlation with HLA‐DR expression
Authors: Al‐Katib Ayad , Koziner Benjamin
Published: March 2008

Summary. Reactivity of the monoclonal antibody anti‐Leu‐10 that detects the human equivalent of the murine 1‐A subregion antigen(s) was studied and correlated with anti‐HLA‐DR expression on 83 cases of acute and chronic leukaemias, leukaemic non‐Hodgkin's lymphomas (NHL) and seven human cell lines. Peripheral blood and/or bone marrow leukaemic cell suspensions were stained by indirect immunofluorescence for both monoclonal antibodies and analysed by flow cytometry. Leu‐10, like HLA‐DR, was absent from T‐cell acute lymphoblastic leukaemia (ALL) (two cases). It was expressed on: 33% of TdT +, CALLA + ALL cases (8/24); 27% (4/15) of acute non‐lymphocytic leukaemia (ANLL); 85% (24/28) of HLA‐DR+B‐cell chronic lymphocytic leukaemia (CLL); and on 9/14 (64%) B‐cell NHL cases. There were no differences in clinical characteristics between Leu‐10+and Leu‐10–patient subgroups. We were able to induce Leu‐10 expression on ‘Josh’ cell line by culturing it with 12‐O‐tetra‐decanoylphorbol‐13‐acetate (TPA). Our data indicate that Leu‐10 expression on leukaemic cells is more restricted than HLA‐DR and is likely to be differentiation related, since it can be induced to be expressed at a later stage than HLA‐DR.
Platelet factor‐4 excretion in myeloproliferative disease: implications for the aetiology of myelofibrosis
Authors: Burstein Samuel A. , Malpass Thomas W. , Yee Esther , Kadin Marshall , Brigden Malcolm , Adamson John W. , Harker Laurence A.
Published: March 2008

Summary. Experimental evidence suggests that the fibroblastic proliferation often associated with the myeloproliferative disorders is not part of the neoplastic process, but is secondary to an unknown stimulus. This stimulus may be a factor derived from platelets which promotes the proliferation of fibroblasts in vitro (PDGF). Plateletderived growth factor is localized to platelet α‐granules together with PF4 and β‐TG. As an indicator of α‐granule release, we have measured PF4 levels in plasma. platelets and urine in 46 normal subjects and 49 patients with myeloproliferative disorders, secondary thrombocytosis and miscellaneous malignancies. All 11 patients with elevated urinary PF4 excretion exhibited myelofibrosis, whereas 11 of 22 patients with documented myelofibrosis had urinary PF4 excretion in the normal range. No correlation was seen between marrow fibrosis and plasma levels or the platelet content of PF4. The data are consistent with the possibility that release of mitogen(s) from platelet or megakaryocyte α‐granules in some patients with myeloproliferative disorders is pathogenetically related to the development of marrow fibrosis.
Sudan black B positive acute lymphoblastic leukaemia
Authors: Stass S. A. , Pui C.‐H. , Melvin S. , Rovigatti U. , Williams D. , Motroni T. , Kalwinsky D. , Dahl G. V.
Published: March 2008

Summary. The presence of greater than 3% Sudan black B (SBB) positivity in leukaemic blasts has been considered diagnostic of acute non‐lymphocytic leukaemia (ANLL). A rare report has indicated that this finding may not be specific for ANLL. In order to determine whether SBB could be found in acute lymphoblastic leukaemia (ALL) the data on 350 patients with newly diagnosed ALL were reviewed. Six patients (1.6%) were found to have 5% or greater SBB positive blasts. The diagnosis of ALL was supported by morphology, cytochemistries, immunologic markers, therapeutic response, and in one case immunoglobulin gene rearrangement. It is important to recognize the fact that SBB is not specific for AML and may be found in the blasts of patients with ALL.
Bone marrow histology in myelodysplastic syndromes
Authors: Tricot G. , Wolf‐Peeters C. De , Hendrickx B. , Verwilghen R. L.
Published: March 2008

Summary. Forty bone‐marrow trephine biopsies of patients with myelodysplastic syndromes (MDS) were studied and compared with bone marrow smears. It proved possible to make the diagnosis of MDS on the basis of bone‐marrow biopsies. Features of abnormal erythropoiesis and myelopoiesis, although different from those observed in smears, were easily detectable. Moreover, obvious advantages of bone‐marrow biopsy over bone‐marrow smears were found. Firstly, the biopsy in MDS gives an exact assessment of the cellularity; secondly, the presence of reticulin fibres, increased in the majority of the cases, can be evaluated; thirdly, dysmegakaryopoiesis is more easily detected in trephine biopsies than in smears; finally, abnormal behaviour of myeloblasts, clustering centrally in the bone marrow, is frequently observed before the smears show an excess of myeloblasts. The bone‐marrow biopsy also has a role in difficult cases, where the diagnosis of MDS can not be established on just bone‐marrow smears. The five types of MDS, as described by the FAB group, do not represent recognizable histological entities.
‘Pitted’ erythrocytes: impaired formation in splenectomized subjects with congenital spherocytosis
Authors: O'Grady J. G. , Harding B. , Egan E. L. , Murphy B. , O'Gorman T. A. , McCarthy C. F.
Published: March 2008

Summary. ‘Pitted’ erythrocyte counts are widely used in the quantitative assessment of functional hyposplenism and in the prediction of recurrence of splenic function in splenectomized subjects. Both of these functions require counts from electively splenectomized controls for comparison. The ‘pitted’ erythrocyte counts in patients splenectomized for congenital spherocytosis are lower than those found in subjects whose spleens were removed electively for other reasons. This appears to be due to the specific membrane defect which impairs the formation of the vacuoles which are responsible for the appearance of ‘pitted’ erythrocytes under the interference‐phase microscope.