bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphocytes from patients receiving lithium do not inhibit CFU‐C growth
Authors: Irvine A. E. , Crockard A. D. , Desai Z. R. , Ennis K. T. , Fay A. C. , Morris T. C. M. , Bridges J. M.
Published: March 2008

Summary. Lymphocyte subset levels and function were examined in 12 patients on lithium therapy and in 11 healthy hospital personnel. Co‐culture of allogeneic human bone marrow cells with monocyte‐depleted lymphocyte preparations revealed that CFU‐C formation was significantly reduced (mean 43% inhibition) in the presence of normal lymphocytes but not with the patients’ lymphocytes (< 5% inhibition). This did not reflect numerical changes in lymphocyte subsets, since these were similar for control and lithium subjects. T colony formation was significantly depressed in the patient group (P <0.05), whereas B colony numbers were similar in both groups (P > 0.1). The possible role of HLA‐incompatability affecting CFU‐C growth was investigated in co‐culture experiments, using lymphocytes from HLA‐identical twins, one of whom was receiving lithium. In four separate co‐culture experiments, the inhibitory effect was shown with lymphocytes from the non‐lithium twin but was not demonstrated by the lithium subject. Addition of lithium in vitro to co‐cultures of normal marrow and lymphocytes was found to negate the inhibitory phenomenon in a dose‐related manner. It is postulated that granulocytosis induced by the administration of lithium may be a manifestation of changes in a lymphocytic control system.
Intracellular forms of iron during transferrin iron uptake by mitogen‐stimulated human lymphocytes
Authors: Bomford Adrian , Young Stephen , Williams Roger
Published: March 2008

Summary. Transferrin receptors expressed by mitogen stimulated human lymphocytes mediate the uptake of transferrin iron into haem, ferritin and a non‐haem, non‐ferritin component. In spite of different rates of iron uptake by cells from different individuals, the proportional incorporation of 59Fe into these components was similar, suggesting that there was an obligatory relationship between the different forms of iron in the cells. By 3 h over 60% of the iron taken up was incorporated into ferritin while less than 10% was found in haem. Initially (10 min) non‐haem, non‐ferritin iron comprised 70% of total iron and this diminished to 30% by 3 h. At 10 min 80% of iron in the non‐haem, nonferritin component was retained by anti‐transferrin affinity columns indicating it was transferrin‐bound. The proportion retained fell to reach a steady state level of 50% by 60 min. These results indicate that 10–20% of the iron in the cells was not recognized as transferrin, ferritin or haem iron. The finding that iron incorporation into this unidentified pool reached equilibrium while that into haem and ferritin increased suggests the iron may act as a precursor for functional and storage compounds.
Studies on immunological assay of vitamin‐K dependent factors.
Authors: Mikami S. , O'Brien D. P. , Mellars G. , Goodall A. H. , Tuddenham E. G. D.
Published: March 2008

Summary. Two‐site immunoradiometric assays (IRMAs) for factor IX antigen (IX:Ag) were developed using a monoclonal antibody (RFF‐IX/1) on the solidphase and either another monoclonal antibody (RFF‐IX/4) or a human polyclonal inhibitor antiserum as tracer (M‐M and M‐I IRMA respectively). The lower sensitivity limits of these two assays for IX: Ag in normal reference plasma were 4 × 10−4 (M‐M IRMA) and 2 × 10−4 (M‐I IRMA) units/ml.

In 20 samples of normal plasma, levels of factor IX coagulation activity (IX:C) and of factor IX antigen measured by both IRMAs were highly correlated. Mean values of approximately 1·0 units/ml were obtained in all three assays. In normal serum, IX: Ag levels were lower with means of 0·84 (M‐M IRMA) and 0.83 (M‐I IRMA) units/ml. 4/25 patients with haemophilia B were CRM neg., two were CRM+ and the remaining 19 patients were CRMR variants. In two of these, IX: Ag was detectable by M‐I IRMA whilst IX:C and IX:Ag measured by M‐M IRMA were undetectable. In plasma from a fetus subsequently terminated on eugenic grounds, IX:C and IX:Ag by both M‐M and M‐I IRMA were undetectable.

In warfarin‐treated plasma (n= 12), the level of IX:C was low (mean 0·39 units/ml). The levels of IX: Ag measured by M‐M IRMA (mean of 0·80 units/ml) and by M‐I IRMA (0.70 units/ml) showed a discrepancy. M‐M IRMA reflects thereal amount of IX:Ag in warfarinized plasma because both monoclonal antibodies bind to epitopes distant from the light chain carboxylated region.

Western blotting of denatured factor IX demonstrated that RFF‐IX/1 binds an epitope that is lost after XIa activation. RFF‐IX/4 binds the heavy chain.
Complete remission in plasma cell leukaemia
Authors: Montecucco Carlomaurizio , Riccardi Alberto , Merlini Giampaolo , Ascari Edoardo
Published: March 2008

Summary. Two patients with primary plasma cell leukaemia who achieved complete remission are reported. They were treated with induction therapy consisting of a multipeptide derivative of sarcolysin, Peptichemio, given intravenously, combined with vincristine and/or prednisone, followed by conventional melphalan‐prednisone therapy. 5–7 months following the beginning of therapy, both patients attained a complete remission which lasted 23 and 6 months; second remission was not achieved. Survival from starting therapy was 57 and 16 months respectively. These cases indicate that intravenous alkylating agents can induce a complete remission in plasma cell leukaemia similar to that achieved in other acute leukaemias.
Prenatal diagnosis of hereditary red cell membrane defect
Authors: Morris S. A. , Ohanian V. , Lewis M. L. , Chahwala S. B. , Rodeck C. H. , Mibashan R. S. , Gratzer W. B.
Published: March 2008

Summary The red cells of a severely anaemic 2‐year‐old child of a white British family showed high haemolytic fragility with poikilocytosis. The cells showed markedly impaired thermal stability. The mother was phenotypically normal, but the father's red cells showed mild elliptocytosis. The spectrin from the latter, extracted at low temperature, was 30% dimeric (cf. 5–10% in normals). Tryptic digests of the spectrin from both father and daughter showed a reduction in the fragment of 80 000 molecular weight, derived from the terminus of the α‐chain, and the elevation of a fragment of molecular weight 46 000, as well as one of 53 000. These characteristics and the autosomal recessive inheritance lead to a diagnosis of type II hereditary pyropoikilocytosis, so far reported only in two black American families (Lawler et al, 1983). The spectrin from the father was examined with respect to thermal conformational stability, and was found to be normal. The spectrin from the cells of the daughter gave evidence of the presence of oxidative (disulphide) cross‐links, as well as of extensive noncovalent aggregation. Blood was obtained from the umbilical cord vein of the 19‐week fetus of the pregnant mother: 250 μl of blood was used for preparation of red cell membranes for SDS‐gel electrophoresis and for extraction of spectrin. Analysis of the spectrin by gel electrophoresis in the native state revealed that the proportion of dimer was within the normal range, and the fetus therefore did not possess the hereditary pyropoikilocytosis phenotype. It is suggested that the procedures described could be generally applied to the prenatal identification of phenotypes associated with severe haemolytic anaemias.
Fibronectin‐mediated inhibition of human platelet activation by crosslinked immunoglobulin G
Authors: Sypcher M. O. , Lerch P. G. , Nydegger U. E.
Published: March 2008

Summary. The effect of purified human plasma fibronectin on stimulation of human platelets with crosslinked immunoglobulin G (IgG) was tested by means of aggregometry, 14C‐serotonin release and fibronectin‐to‐platelet binding experiments. Incremental additions of fibronectin to gel‐filtered platelets (8x108/ml) followed by 25 μg/ml bisdiazoniumbenzidine (BDB)‐crosslinked IgG produced a dose‐related inhibition of platelet aggregation and suppression of 14C‐serotonin release from the platelets. Evidence was obtained that upon stimulation of the platelets with BDB‐IgG, fibronectin becomes bound to its receptor and that further platelet activation is inhibited when a sufficient number of receptors is occupied.
The haemorrhagic and antithrombotic effects of dermatan sulphate
Authors: Fernandez F. , Ryn J. Van , Ofosu F. A. , Hirsh J. , Buchanan M. R.
Published: March 2008

Summary. Heparin and dermatan sulphate are effective antithrombotic agents but the clinical use of heparin is complicated by haemorrhage. The haemorrhagic effect of dermatan sulphate is unknown. In this study we compared the antithrombotic, haemorrhagic and anticoagulant effects of heparin and dermatan sulphate in rabbits. The antithrombotic effect was measured as prevention of venous thrombus formation. The haemorrhagic effect was measured as 51Cr‐blood loss from standardized cuts in rabbit ears. The anticoagulant effect was measured as changes in the APTT, TCT and circulating anti‐factor Xa level, and the formation of 125I‐thrombin/inhibitor complexes ex vivo. The effect of heparin and dermatan sulphate on collagen‐induced platelet aggregation was measured ex vivo. Maximal antithrombotic effects of heparin and dermatan sulphate were achieved with 70 and 500 μg/kg respectively. A 20‐fold increase in heparin dose caused an 8‐fold increase in blood loss and higher doses (40‐ and 80‐fold increases) caused further dose‐related increases in blood loss (13‐ and 35‐fold increases respectively). In contrast, a 20‐ to 40‐fold increase in the antithrombotic dose of dermatan sulphate did not increase blood loss and an 80‐fold dose increase caused only a 7‐fold increase in blood loss. There was no relationship between the antithrombotic and haemorrhagic effects of either heparin or dermatan sulphate and their anticoagulant activities. In contrast, there was a relationship between the dose‐related enhancement of blood loss by these glycosaminoglycans and the inhibition of collagen‐induced platelet aggregation ex vivo. These results suggest that dermatan sulphate is less haemorrhagic than heparin at equivalent antithrombotic doses, and that the haemorrhagic effect is associated with a glycosaminoglycan‐induced platelet defect.
Increasing haemoglobin oxygen affinity to prevent sickling: abnormal haemoglobin variants as models
Authors: Franklin I. M. , Huehns E. R. , Rosemeyer M. A.
Published: March 2008

Summary. TWO abnormal high oxygen affinity haemoglobins have been studied for their ability to inhibit the polymerization of deoxy‐Hb S. They were used as models to predict the effect of chemically modifying haemoglobin to increase oxygen affinity since the oxy(R)‐conformation is the most potent inhibitor known of cell sickling. The participation of these variants in deoxy‐Hb S polymers has been described quantitatively in terms of an exclusion coefficient, f, that relates their behaviour to that of deoxy‐Hb S, and qualitatively in terms of an exponent of hybridization.

Hb Bethesda was a potent inhibitor of polymerization, its behaviour being similar to that of Hb F studied previously. Hb Radcliffe demonstrated atypical behaviour, with hybrid molecules of the formula α2βsβRad participating in the polymerization as effectively as Hb S, as has been shown for Hb C.

These data have implications for the development of anti‐sickling agents designed to increase oxygen affinity by covalent binding to Hb S.
Monocytic differentiation in acute myeloid leukaemia: a cause of Fc binding of monoclonal antibodies
Authors: Lawlor E. , Finn T. , Blaney C , Temperley I. J. , McCann S. R.
Published: March 2008

Summary. Cells from 17 patients with acute myeloid leukaemia (AML) were investigated using the following IgG2a monoclonal antibodies cALL, B1 and Leu 1. Cells from six patients showed binding of cALL, five showed B1 and three showed Leu 1 positivity. Binding of the monoclonal antibodies was shown to be due to Fc binding as it was blocked by prior incubation with aggregated human IgG. Measures such as prior incubation with AB serum or incubation at 37°C were insufficient to block Fc binding in all cases. Fc binding correlated with evidence of monocytic differentiation. Our findings suggest that unless Fc binding is outruled false positive results may be obtained in the investigation of lineage infidelity in AML particularly in monocytic leukaemias.
Acquired aplastic anaemia: a PNH‐like disease?
Authors: Nissen Catherine , Gratwohl Alois , Speck Bruno , Würsch Andreas , Moser Yolanda , Weis Johanna
Published: March 2008

Summary. Bone marrow from 20 patients with aplastic anaemia at different stages of disease and from three patients with paroxysmal nocturnal haemoglo‐binuria (PNH) was incubated in isosmolar sucrose with 5% autologous serum prior to culture in methylcellulose. If fresh serum was used, colony formation by granulocyte‐macrophage colony forming cells (GM‐CFC) and immature eryth‐roid precursors (BFU‐E) was reduced to approximately 50% in all patients tested, at any stage of disease, including complete autologous bone marrow recovery. Heat inactivation and complement inactivation with EDTA completely abrogated this inhibitory serum effect. Selective inactivation of the classical, antibody dependent complement pathway with Mg2+ EGTA reduced the inhibitory effect by 50%. Complement sensitivity of haemopoietic precursors is a known feature of PNH. Since the majority of our patients did not have PNH as judged by a negative sucrose‐test on mature erythrocytes, we conclude that, in aplastic anaemia, haemopoietic cells express a PNH‐like defect at a primitive level.
Neutrophils are involved in the increased vascular permeability produced by activated complement in man
Authors: Williamson Lorna M. , Sheppard Kelvyn , Davies John M. , Fletcher John
Published: March 2008

Summary. TO investigate the role of neutrophils in complement‐induced changes in vascular permeability, skin wheal and flare responses to intradermal injection of autologous activated serum complement were measured in normal and neutropenic subjects. In normal subjects, responses were dose‐dependent and were abolished by removal of C5 from serum. Biopsy of a wheal revealed neutrophils adherent to vascular endothelium. In neutropenic subjects (neutrophil count < 0.5 × 109/1), responses to complement‐activated serum or a low molecular weight fraction from it were significantly reduced. This could not be accounted for by a reduction in concentration of C5 conversion products. In one subject with chronic granulomatous disease a normal response was produced. Local injection of the anti‐histamine (Hi) drug clemastine produced only partial inhibition of responses, while almost totally abolishing histamine‐induced wheals. Systemic anti‐inflammatory drugs had no effect. The data suggest that the microvascular response to activated complement in man is at least partly due to an interaction between C5 fragments and neutrophils.
Autologous bone marrow transplantation for acute leukaemia in remission
Authors: Gorin N. C. , Herve P. , Aegerter P. , Goldstone A. , Linch D. , Maraninchi D. , Burnett A. , Helbig W. , Meloni G. , Verdonck L. F. , Witte T. de , Rizzoli V. , Carella A. , Parlier Y. , Auvert B. , Goldman J.
Published: March 2008

Summary. Between 1980 and 1985, 175 patients with acute leukaemia in first or subsequent complete remission (CR) were treated by chemotherapy or chemoradiotherapy followed by transfusion of autologous bone marrow cells that had been collected days or months previously. In 85 cases, autologous marrow cells were treated ex vivo with cytotoxic drugs or monoclonal antibodies with the intention of removing residual leukaemic cells. The actuarial relapse‐free rate was 52% at 2 years. Of 89 patients autografted for acute non‐lymphocytic (myeloid) leukaemia (ANLL), 60 were treated in first remission and 18 in second CR; their relapse‐free rates at 2 years were 67% and 41% respectively (P<0‐001). In contrast, of 77 patients autografted for acute lymphoblastic leukaemia (ALL), 32 were treated in first CR and 28 in second CR and their actuarial relapse free rates at 2 years were 56% and 55% respectively (P= NS). There was no significant difference in leukaemia relapse rates between patients autografted with purged and those autografted with non‐purged marrow cells. These preliminary results suggest that autologous bone marrow transplantation may be valuable if offered to patients with ANLL in first CR or to patients with ALL in first or second CR but the need for marrow purging remains uncertain.
An evaluation of the acidified glycerol lysis test
Authors: Rutherford C. J. , Postlewaight B. F. , Hallowes M.
Published: March 2008

Summary

The acidified glycerol lysis test (AGLT), as a screening procedure for spherocytosis, has been assessed for sensitivity and specificity. AGLT identified 24 of 26 subjects with hereditary spherocytosis and produced 19 abnormal AGLTs out of a total of 52 normal blood donors. The claims for the test as being both 100% sensitive and specific could not be confirmed.
Studies on a circulating anticoagulant inhibiting factor XI in a patient with congenital deficiency and carcinoma of the prostate
Authors: Chediak Juan , Madej‐Zevin Patricia , Ratnoff Oscar D. , Goldsmith George H. , Eversox Barbara
Published: March 2008

An inhibitor of plasma thromboplastin antecedent (PTA, factor XI), measured in coagulant and radioimmunoassays, was detected in a 60‐year‐old man with carcinoma of the prostate who had no evidence of a bleeding tendency. Family studies indicated that the patient was either a homozygote or a heterozygote for hereditary factor XI deficiency. In contrast to earlier described patients with factor XI deficiency in whom inhibitors were detected, the patient was unaware of having been transfused with blood or blood products at any time before the discovery of the inhibitor. The inhibitor of factor XI in the patient's plasma appeared to be predominantly in the IgG4 fraction and to be directed at a locus on the factor XI molecule other than the active site; it did not block the amidolytic properties of activated factor XI (XIa). Rather, it appeared to block adsorption of factor XI to negatively charged surfaces. The inhibitor interfered with measurement of other components of the intrinsic pathway of thrombin formation, perhaps explaining the low titres of other coagulation factors of the intrinsic system reported in patients with strong inhibitors directed against factor XL.
Immunological abnormalities in myelodysplastic syndromes II. COEXISTENT LYMPHOID OR PLASMA CELL NEOPLASMS: A REPORT OF 20 CASES UNRELATED TO CHEMOTHERAPY
Authors: Copplestone J. A. , Mufti G. J. , Hamblin T. J. , Oscier D. G.
Published: March 2008

Summary

We report a series of 20 patients with the myelodysplastic syndrome (MDS) each with a coexistent lymphoid or plasmacytic neoplasm. In none of the patients did chemotherapy play a part in the pathogenesis. The possible reasons for the coincidence of these conditions are discussed. The frequency and variety of associated neoplasms reinforces the idea that in MDS there are genetically unstable progenitor cells liable to clonal differentiation along a variety of pathways given appropriate stimuli.
Factor VIII concentrates contain factor VIII procoagulant antigen bound to phospholipid
Authors: Kemball‐Cook G. , Barrowcliffe T. W.
Published: March 2008

Summary

Fractionation of a human antibody to factor VIII :Ag by immuno‐absorption with factor VIII/phospholipid (PL) complex has produced two pools of labelled Fab’fragments which can be used in fluid‐phase immunoradiometric assays (IRMAs). One pool binds only to the PL‐binding sites on factor VIII :Ag (and thus measures only that factor VIII: Ag in a sample not bound to PL), while the second binds to other antigenic sites. Parallel assays of factor VIII‐containing materials using these two pools provide estimates of the proportion of factor VIII :Ag bound to PL in those materials. Five batches each of nine brands of factor VIII concentrates from eight different manufacturers were tested for PL‐bound factor VIII :Ag by this method: all contained substantial amounts, ranging from 28% to 54% of the total factor VIII :Ag present. In addition, varying amounts of the factor VIII: Ag present in both non‐activated and activated prothrombin complex concentrates (PCCs) from four manufacturers were PL‐bound.
Bone marrow transplantation in two multiply transfused patients with thalassaemia major
Authors: Brochstein Joel A. , Kirkpatrick Dahlia , Giardina Patricia J. , Weinberg Rona S. , Alter Blanche P. , Driscoll Catherine , Wolfe Lorraine , Shank Brenda , O'Reilly Richard j.
Published: March 2008

Summary

Bone marrow transplantation has generally been unsuccessful when applied to patients with thalassaemia major over the age of 6 years. We report here two successful transplants for this disorder in a 7‐year‐old boy and an 11‐year‐old girl following a pre‐transplant cytoreductive/immunosuppressive regimen of total body irradiation and cyclophosphamide. Complete durable engraftment of donor haematopoietic and lymphoid populations was documented through several approaches, including cytogenetic analysis, haemoglobin electrophoresis, globin chain synthetic ratios, red cell typing and DNA restriction enzyme analysis. Both patients are surviving in good health, 28 and 9 months from transplantation. The successful outcome in these patients demonstrates the feasibility of marrow transplantation for the treatment of thalassaemia in multiply transfused and, presumably, highly sensitized patients.
Normal levels of ATP, total nucleotides and activities of three enzymes related to nucleotide metabolism in fetal erythrocytes
Authors: Lestas A. N. , Nicolaides K. H. , Rodeck C. H. , Bellingham A. J.
Published: March 2008

Summary

Pure fetal blood was obtained by direct‐vision fetoscopy from 30 fetuses at 17–23 weeks gestation. The erythrocyte concentrations of ATP and total nucleotides and the activities of the enzymes pyrimidine‐5′‐nucleotide nucleosidase (Fyr5N), phosphoribosylpyrophosphate (PRPP) synthetase and adenylate kinase (AK) were analysed by established techniques to find the normal ranges for this gestational age. The ranges were relatively narrow and could serve as reference values for the prenatal diagnosis of defects in nucleotide metabolism.
The role of aggressive chemotherapy in the treatment of the myelodysplastic syndromes
Authors: Tricot G. , Boogaerts M. A.
Published: March 2008

Summary

Fifteen patients with the initial diagnosis of myelodysplastic syndrome (MDS) received aggressive chemotherapy with high dose cytarabine or with a standard acute myeloid leukaemia (AML) regime. Cases treated with aggressive chemotherapy were either younger individuals with refractory anaemia with excess of blasts (RAEB) or patients, irrespective of age in advanced stages of MDS (RAEB in transformation or after evolution to frank AML), who did not have a major infection at the time of presentation. Age seemed to be the most important factor in determining the outcome of aggressive remission induction chemotherapy in MDS: 86% of the patients <50 years entered complete remission, compared to only 25% in the older age group. In spite of intensive consolidation therapy the duration of complete remission was short. We conclude that young patients (<50 years) with excess of bone marrow blasts should be treated with aggressive chemotherapy even in the early stages of the disease. Elderly patients in advanced stages of MDS should be treated with less aggressive chemotherapy.
Alpha thalassaemia in an Italian population
Authors: Velati C. , Sampietro M. , Biassoni M. , Cappellini. M. D. , Wainscoat J. S. , Higgs D. R. , Fiorelli G.
Published: March 2008

Summary

The incidence of α‐thalassaemia in an Italian population has been determined by a survey of random cord bloods for the presence of Hb Bart's. 144 out of 4730 (3%) had detectable amounts of Hb Bart's. Furthermore, α‐globin gene analysis of 100 random cord bloods showed that five out of 100 had the common type of α‐thalassaemia caused by a single α‐globin gene deletion (—α). The molecular basis of α‐thalassaemia was also determined in a selected group of 34 newborns with detectable levels of Hb Bart's. 25 of these cases had the —α3.7 deletion type of α‐thalassaemia and nine had nondeletion types of α‐thalassaemia in four of which the molecular defect was detectable directly by restriction enzyme analysis.
The myelosuppressive effect of recombinant interferon γ in short‐term and long‐term marrow cultures
Authors: Coutinho L. H. , Testa N. G. , Dexter T. M.
Published: March 2008

Summary

The effect of a highly purified human gamma IFN (r‐γ‐IFN) on the growth of haemopoietic progenitors was examined in soft agar assays and in long‐term bone marrow cultures. r‐γ‐IFN reduced colony formation by progenitor cells of the granulocyte/macrophage lineage (GM‐CFC). the eryth‐roid lineage (BFU‐E) and multipotent cells (GEMM‐CFC), and suppressed haemopoiesis in long‐term culture in a dose‐related fashion. At high doses (1000 units/ml) r‐γ‐IFN appeared to be toxic to the stromal cells of the bone marrow.
Immunohistological localization of factor VIII in placental endothelial cells
Authors: Stel Herbert V. , Oudejans Cornelis B. M. , Veerman Enno C. I. , Meijer Chris J. L. M.
Published: March 2008

Summary

DNA‐recombinant studies have established that factor VIII‐mRNA is present in liver and placental tissue. In a previous immunohistological study using monoclonal antibodies we have localized factor VIII in liver sinusoidal endothelial cells. In this paper we demonstrate that also the endothelial cells lining the fetal vessels in human placenta contain factor VIII. Based on the combined results of the present study and a previous study we favour‐the concept that factor VIII is synthesized in placental endothelial cells.
Immunohistochemical study of basement membrane proteins and type III procollagen in myelofibrosis
Authors: Apaja‐Sarkkinen Meeri , Autio‐Harmainen Helena , Alavaikko Martti , Risteli Juha , Risteli Leila
Published: March 2008

Summary

In this study the distribution of type IV collagen in the marrow is compared with that of laminin, another basement membrane protein. In addition, incompletely processed type III procollagen is identified with specific antibodies.
Failure of transferrin to enhance iron absorption in achlorhydric human subjects
Authors: Bezwoda W. R. , Macphail A. P. , Bothwell T. H. , Baynes R. D. , Derman D. P. , Torrance J. D.
Published: March 2008

Summary

There is evidence in experimental animals that transferrin, produced either by gastrointestinal cells or derived from bile, mediates the luminal absorption of iron. The applicability of these findings to human subjects was tested by administering diferric transferrin labelled with 3 mg 59Fe to seven patients with pernicious anaemia. Achlorhydric subjects were chosen to ensure that the iron transferrin complex did not dissociate in the stomach. The geometric mean absorption of 1.4% was similar to that of 3 mg iron given as ferric chloride (1.9%) and much less than that of ferrous ascorbate (18.9%). These findings suggest that transferrin does not play a physiological role in the absorption of iron in human subjects.
Congenital plasminogen deficiency associated with venous thromboembolism: therapeutic trial with stanozolol
Authors: Mannucci P. M. , Kluft C. , Traas D. W. , Seveso P. , D'Angelo A.
Published: March 2008

Summary

We describe the case of a 34‐year‐old man with a history of venous thrombotic disease and demonstrate that his plasma plasminogen was reduced to about half‐normal levels but was functionally intact. The deficiency was inherited as an autosomal trait and the propositus and his sister are heterozygotes. The propositus, but no other family member, had an associated deficiency of plasminogen activator activity, probably due to heightened plasminogen activator inhibition in his plasma. A therapeutic trial with the anabolic steroid stanozolol transiently normalized plasminogen and plasminogen activator levels.
Alteration of red cell membrane organization in sickle cell anaemia
Authors: Choe Hye‐Ryun , Schlegel Robert A. , Rubin Elizabeth , Williamson Patrick , Westerman Maxwell P.
Published: March 2008

Summary

Bee venom phospholipase A2 and the fluorescent probe merocyanine 540 were used to examine plasma membrane phospholipid organization in the spicules released by deoxygenation and reoxygenation of sickle red cells, as well as in reversibly and irreversibly sickled erythrocytes. Digestion of phosphatidyl ethanolamine in spicules was comparable to that of phosphatidyl choline, and these structures were stained by the fluorescent probe. Both assays suggest that membrane lipid asymmetry is disrupted in spicules. The residual cells, from which the spicules were derived, retain the normal asymmetry in phospholipid distribution between the outer and inner leaflets of the plasma membrane bilayer. Comparable experiments with cell fractions enriched in irreversibly sickled cells revealed a partial enhancement of phosphatidyl ethanolamine digestion, confirming the similar experiments of Lubin et al (1981). Staining of these cells with merocyanine 540, however, did not reveal a subfraction of stainable cells, indicating that this increase in phosphatidyl ethanolamine digestion is not due to the presence of a small fraction of cells which have completely lost their membrane asymmetry.
A role for glycoprotein IIb‐IIIa complex in the binding of IgE to human platelets and platelet IgE‐dependent cytotoxic functions
Authors: Ameisen Jean Claude , Joseph Michel , Caen Jacques P. , Kusnierz Jean‐Pierre , Capron Monique , Boizard Bernadette , Wautier Jean‐luc , Levy‐Toledano Sylviane , Vorng Han , Capron André
Published: March 2008

Summary. A possible relationship between binding sites for Immunoglobulin E (IgE) on human platelets, involved in IgE‐dependent cytotoxic functions of platelets against helminth parasites, and well‐characterized platelet constituents involved in haemostasis, was investigated. We first explored the interaction with IgE of platelets from patients with rare inherited deficiencies of defined platelet constituents and functions: Glanzmann's thrombasthenia, Bernard‐Soulier and grey platelet syndromes. We report that only type I and II thrombasthenic platelets, which lack the membrane glycoproteins (GP) lIb and IIIa, failed to bind IgE and to exhibit IgE‐dependent effector functions. Since thrombasthenic monocytes, however, showed normal interaction with IgE, this defect appeared restricted to platelets. Polyclonal and monoclonal antibodies directed against GP Ilb‐IIIa complex, but not monoclonal antibody directed against GP Ib, inhibited the binding of IgE to normal platelets, and their IgE‐dependent cytotoxicity. Taken together, these findings indicate a relation between the GP Ilb‐IIIa complex and the expression of IgE binding sites and IgE‐dependent effector functions in human platelets.
Immune thrombocytopenic purpura: autoimmune or immune complex disease?
Authors: Klefel Volker , Spaeth Peter , Mueller‐Eckhardt Christian
Published: March 2008

Summary. To assess the pathogenic role of circulating immune complexes (CIC) in idiopathic thrombocytopenic purpura (ITP), 39 patients with ITP were compared to 17 patients with other forms of thrombocytopenia (hypersplenism (N= 12), impaired thrombopoiesis (3), thrombocytopenia of unknown origin (2)) and six nonthrombocytopenic subjects. In all patients, platelet mean life span (MLS), platelet associated IgG (PAIgG), as well as circulating anti‐platelet antibodies and Clq binding activities were determined. In most cases, immune complex solubilization capacity (ICSC) and immune complex precipitation inhibition capacity (ICPIC) of sera were also assessed. All patients with ITP had a reduced platelet MLS, but PAIgG was elevated in only 16 out of 24 patients with chronic ITP, in six out of 10 patients with acute ITP and in four out of five patients with secondary ITP. In the group of patients with thrombocytopenia due to splenomegaly, seven out of 12 patients had elevated PAIgG while the platelet MLS was only slightly reduced. Of the 39 patients with ITP only one with secondary ITP had Clq binding material in his serum, as opposed to six out of 12 thrombocytopenic patients with splenomegaly. Whereas only three patients with ITP had abnormal immune‐complex modulating capacities, such deviations were found in seven out of 12 patients with thrombocytopenia due to splenomegaly. We conclude that our data render the role of CIC in the pathogenesis of ITP very questionable.
Standardization of T‐cell depletion in HLA matched bone marrow transplantation
Authors: Gilmore M. J. M. L. , Patterson J. , Ivory K. , Brenner M. K. , Graphakos S. , Janossy G. , Hoffbrand A. V. , Prentice H. G.
Published: March 2008

Summary. The IBM 2991 Blood Cell Processor has been used to isolate a mononuclear cell (MNC) fraction from the marrow of 31 allogeneic donors. The MNC fraction was then incubated with a combination of two murine monoclonal antibodies MBG6 (CD6) and RFT8 (CD8) followed by two rounds of treatment with rabbit complement resulting in a marrow inoculum significantly reduced in the number of T‐lymphocytes. We report here new specifications for the use of Ficoll‐Metrizoate, the method used to calculate T‐lymphocyte depletion and the details of our attempts to improve T‐depletion. Following marrow transplantation with this T‐depleted fraction, 29 patients are evaluable for engraftment, one patient failed to engraft and one died too early for evaluation. Twenty‐two had no acute graft‐versus‐host disease (aGvHD), at a minimum of 60 d, six had grade I acute GvHD and one grade III. No correlation was found between the absolute number of MNC infused and time to engraftment, nor any relationship between the number of residual viable T‐lymphocytes in the infused marrow and the incidence of GvHD, but the patient with the most severe aGvHD also had the highest number of T‐lymphocytes infused.
Chronic myelogenous leukaemia: haematological remissions with alpha interferon
Authors: Talpaz Moshe , McCredie Kenneth , Jian Hagop Kantab , Trujillo Jose , Keating Michael , Gutterman Jordan
Published: March 2008

Summary. Twenty‐seven consecutive patients with previously untreated, or minimally treated benign phase Philadelphia‐chromosome‐positive, chronic myelogenous leukaemia (CML) were treated with partially purified human leucocyte (alpha) interferon; 24 of the 27 patients responded to therapy achieving either haematological remission (20 patients) or partial haematological remission (four patients). In the responding patients the peripheral white blood cells declined from a median of 89.6 × 10 × 109/1 to 4.5 × 10 × 109/1. The serum lactate dehydrogenase declined from a mean of 8.36 Katal/1 (492 mu/ ml) to 2.8 Katal/1 (165 mu/ml), and the vitamin B12 levels declined from 1492 pg/ml to 838 pg/ml. Fifteen patients had splenomegaly. The spleen size normalized in four and decreased by a median of 30% in 10 additional patients. The bone marrow cellularity fell from a median of 100% to a median of 62%. In seven of the 24 responding patients, followed for ≤ 6 months, the percentage of Ph1‐positive cells in the bone marrow declined to a median of 70% (range 5–75%). Alpha interferon was found to be an effective therapeutic agent for controlling the myeloid proliferation in CML, and in partially restoring the nonclonal haematopoietic cells in some of the patients.
The morphological spectrum of T‐prolymphocytic leukaemia
Authors: Matutes E. , Talavera J. Garcia , O'Brien M. , Catovsky D.
Published: March 2008

Summary. The morphology of the cells from 29 cases of T‐prolymphocytic leukaemia (T‐PLL) was studied by light (LM) and transmission electron microscopy (TEM) and was compared with that of 3 3 B‐cell PLL. The membrane phenotype of T‐PLL cells was T4 +, T8 ‐ in two‐thirds of the cases, others being T4‐ T8+ or T4+ T8+. Two morphological types of T‐PLL were defined according to the nuclear features: regular (55% of cases) and irregular (45% of cases). T‐PLL cells with a regular, round or oval, nuclear outline resembled B‐PLL cells but had less abundant cytoplasm and a higher nucleo‐cytoplasmic ratio. Irregular T‐prolympbocytes displayed a distinct convoluted nucleus. A 'small‐cell’variant of T‐PLL was recognized by TEM in six cases in which the diagnosis was uncertain by LM. A characteristic of all types of T‐prolymphocytes by LM was the presence of a deep basophilic cytoplasm which by TEM corresponded to clusters of ribosomes and endoplasmic reticulum. No differences in clinico‐haematological features or membrane markers were apparent between the morphological types of T‐PLL, although it was noted that the three T4‐ T8+ cases had irregular cells and four of the small cell variant were T3‐ T4+. TEM permits a more precise assessment of the cytoplasmic organelles and nucleolus than LM analysis and facilitates the distinction between T‐PLL and other leukaemias with a mature T‐cell phenotype, namely adult T‐cell leukaemia/lymphoma, Sezary syndrome and T‐chronic lymphocytic leukaemia.
Antigenic determinants on myeloid leukaemia colony‐forming cells resemble those of normal myeloid progenitor cells and differ from those of circulating blast cells
Authors: Robak Tadeusz , Dowding Charles , Garewal Gurjivan , Hibbin Jill A. , Th'ng Kang Hock , Goldman John M.
Published: March 2008

Summary. We studied the antigenic characteristics of leukaemic colony‐forming cells (CFU‐L) from the blood of patients with chronic granulocytic leukaemia (CGL) in blastic transformation (BT) and acute myeloid leukaemia (AML) by in vitro culture techniques after complement‐mediated lysis with one anti‐DR and 10 selected myeloid monoclonal antibodies (McAbs), all of which were cytotoxic in the presence of complement. At the same time we studied the antigenic characteristics of the circulating blast cells from the same patients using in addition one non‐complement fixing antibody (BI.3C5) with standard immunofluorescence and immunoalkaline phosphatase techniques. We also used myeloid progenitor cell assays in conjunction with cytotoxic McAbs to investigate the antigenic determinants on Day 7 CFU‐GM, Day 14 CFU‐GM and BFU‐E from the blood of patients with CGL in chronic phase (CP) and from normal bone marrow. We found that two of the McAbs, S4–7 and WGHS29.1, recognized a higher proportion of CFU‐L from the blood of AML patients than from patients with CGL‐BT. However, the patterns of reactivity for CFU‐L from CGL‐BT and AML patients with the other McAbs quite closely resembled those observed in CFU‐GM and BFU‐E from normal individuals and patients with CGL in CP. A McAb with DR specificity and one of the myeloid McAbs, 54/39, recognized both CFU‐L from CGL‐BT and AML and reacted also with circulating blast cells from the same patients. In contrast, six of the other myeloid McAbs that recognized CFU‐L failed to label the corresponding blast cells. We conclude
Bone marrow purging procedure in Burkitt lymphoma with monoclonal antibodies and complement: quantification by a liquid cell culture monitoring system
Authors: Favrot M.‐C. , Philip I. , Philip T. , Pinkerton R. , Lebacq A. M. , Forster K. , Adeline P. , Doré J. F.
Published: March 2008

Summary. Using B1, Y29/55 and AL2 monoclonal antibodies (MoAbs) to target Burkitt lymphoma (BL) cell lines, we defined optimal conditions to lyse, in the presence of baby rabbit complement, BL cells in excess bone marrow (BM). After the purging procedure, down to one residual BL cell in 106 normal ones was detectable with a liquid cell culture assay. Using a cocktail of three MoAbs, on five different cell lines were observed more than 4 log BL cell depletion in samples contaminated with 1% BL cells and only one failure of the procedure on 17 experiments. However, a sixth line was constantly resistant to the procedure.
Variable differentiation patterns of acute myelogenous leukaemia blasts in liquid suspension cultures
Authors: Baer Maria R. , Dessypris Emmanuel N. , Stein Richard S. , Cousar John B. , Krantz Sanford B.
Published: March 2008

To study the ability of acute myelogenous leukaemia blasts to spontaneously differentiate in vitro, bone marrow and/or blood mononuclear cells from 63 patients with acute myelogenous leukaemia were incubated in liquid suspension cultures containing human serum, without addition of chemical inducers of differentiation. Cultures were examined weekly for disappearance or persistence of blasts, and for appearance of morphologically recognizable granulocytes and macrophages. Culture outcomes were extremely variable, ranging from lack of appearance of differentiated cells to complete disappearance of blasts with replacement by mature cells. In 50 cases an increase (25–185%) in the absolute number of differentiated cells in culture was noted during the culture period. Full differentiation was seen exclusively in cultures from 13/48 (27·1%) patients studied at diagnosis, as compared to 0/20 patients studied at relapse (P<0·01). The ability to fully or partially differentiate in culture was lost to a significant degree at relapse (13/20 patients) as compared to diagnosis (48/48 patients, P= 0·0001). At diagnosis full differentiation in culture was associated with a significantly higher remission rate than partial differentiation (89% versus 40%. P<0·02). Origination of mature cells from leukaemic rather than normal precursors was suggested by the appearance of Auer rods in mature cells in seven cases, by the correlation of types of differentiated cells seen in culture with the FAB class of leukaemia and by cytogenetic data in one case.
Heterogeneity of large granular lymphocyte proliferations: morphological, immunological and molecular analysis in seven patients
Authors: Lauria F. , Foa R. , Migone N. , Giubellino M. C. , Raspadori D. , Buzzi M. , Casorati G. , Gobbi M. , Tazzari P. L. , Tura S.
Published: March 2008

The clinical, morphological, immunological and molecular features of seven patients with a stable picture of chronic granular lymphocytosis, observed over a period of up to 4 years, were studied. Mild splenomegaly was detected in one patient, while lymphoadenopathy and hepatomegaly were absent. Surface marker analysis showed in five patients the common membrane phenotype of granular T‐cell lymphocytosis (T3 +, T4‐, T8 +, Leu‐7 +); of the remaining two, one presented an unusual phenotype (T3 +, T4 +, T8 +) and the other showed a marked positivity with the Leu‐11 and M1 monoclonal antibodies, but lacked the T3, T4, T8 antigens. Three cases had a low (<30%) expression of the T1 antigen. Functional studies showed that the proliferative response to PHA and the NK function were reduced in four of the seven cases. Molecular analysis, performed in six cases, revealed a monoclonal rearrangement of the T‐cell receptor β‐chain gene in three, a polyclonal T‐cell configuration in two and a germ‐line arrangement in the last. All three monoclonal cases showed a depressed NK activity and two a reduced PHA response. The results of this study document the heterogeneity of granular lymphocyte expansions and suggest that the clonal or reactive nature of these often indolent proliferations, suspected on the basis of immunologic functional studies, may be recognized at the DNA level.
Comparative effect of heparin and heparan sulphate on two abnormal antithrombin III type 3 variants
Authors: Fischer A. M. , Beguin S. , Sternberg C. , Dautzenberg M. D.
Published: March 2008

Two families were found with an antithrombin III that was unresponsive towards heparin (type 3 AT III variants). The abnormal species were purified using affinity chromatography on Sepharose bound anti‐AT III antibodies. This yielded active proteins, as judged by their progressive antithrombin activities. In an attempt to explain the throm‐botic tendency observed in this abnormality we compared the effect of heparin and heparan sulphate on these abnormal AT III, since, unlike heparin, heparan sulphate is a naturally occurring anticoagulant in the human. In normal plasma the heparan sulphate used in this study had a heparin‐like activity of 50 U/mg by anti‐F.XA and anti‐F.IIa amidolytic assays. Full expression of the heparin cofactor activity in normal plasma could be obtained at a final concentration of 0·024 mg/ml of heparan sulphate (equivalent to 0·007 mg/ml of heparin). At this concentration of heparan sulphate the two abnormal AT III still exhibit a heparin cofactor activity below 10%. This absence of binding of heparan sulphate to abnormal AT III of type 3 could explain why some patients with this abnormality suffer from thrombo‐embolic episodes while their AT III acts normally in the absence of heparin.
Immunohistological diagnosis of a case of composite lymphoma
Authors: Salter D. M. , Sheehan T. , Krajewski A. S. , Ludlam C. A.
Published: March 2008

A case of composite lymphoma consisting of Hodgkin's disease and B cell lymphocytic lymphoma is described. A diagnosis of lymphocyte predominant Hodgkin's disease was made initially by histology. The presence of a composite lymphoma was only appreciated once immunohis tochemistry was available. The findings illustrate the importance of the use of immunological markers for the accurate diagnosis of lymphoma.
Inhibition of platelet aggregation by myeloid leukaemic cells demonstrated in vitro
Authors: Fauldt R. , Ankerst J. , Zoucas E.
Published: March 2008

The effect of myeloid leukaemic cells, cells of the promyelocytic cell line HL‐60 and normal polymorphonuclear granulocytes (PMN), enclosed in dialysis tubes, on the aggregation of allogeneic normal platelets after induction with arachidonic acid (AA) and with adenosine diphosphate (ADP) was investigated in vitro. Inhibition of aggregation was seen after preincubation of the platelets with leukaemic blood or bone marrow cells from 7/14 patients belonging to various FAB groups and with HL‐60 cells, but not with normal PMN (14/14 cases). A dose‐dependent inhibition was seen after lysis of some leukaemic cells with anti‐human rabbit antiserum and active complement. Lysis of normal PMN inhibited platelet aggregation slightly and inconstantly and only at higher cell concentrations. Platelet inhibitory activity was not related to a higher rate of metabolism of the leukaemic cells which inhibited platelet aggregation since they did not differ from the cells not inhibiting aggregation with respect to heat production. Neither was a non‐specifically increased cell membrane permeability the cause of the release of platelet inhibitory factor(s) since the release of 51Cr‐labelled leukaemic cells was not related to the inhibition of platelet aggregation.
Blunted erythropoietin response to anaemia in rheumatoid arthritis
Authors: Baer Alan N. , Dessypris Emmanuel N. , Goldwasser Eugene , Krantz Sanford B.
Published: March 2008

Summary. The relationship of serum immunoreactive erythropoietin to haemoglobin concentration was defined for 54 patients with rheumatoid arthritis (RA) and 41 patients with anaemia of varying aetiology (excluding pregnancy and renal insufficiency), not associated with RA. Significant inverse correlations between the logarithm of serum immunoreactive erythropoietin and the haemoglobin concentration were noted for the anaemic patients in both groups. The regression line for the RA patients had a similar slope, but a significantly lower y‐intercept as compared to that for the non‐RA patients. Erythropoietin levels were also significantly lower for the group of RA patients than for the group of non‐RA patients when matched for comparable haemoglobin concentrations. These studies suggest that the erythropoietin response to anaemia in RA is intact but blunted relative to that for anaemia of other aetiologies. Lower levels of serum erythropoietin in anaemic RA patients may contribute to the pathogenesis of their anaemia.