bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum crosslinked fibrin (XDP) and fibrinogen/fibrin degradation products (FDP) in disorders associated with activation of the coagulation or fibrinolytic systems
Authors: Hunt Frederick A. , Rylatt Dennis B. , H art Rosemary‐Anne , Bundesen Peter G.
Published: March 2008

Summary

Soluble crosslinked fibrin derivatives (XDP) in serum were determined by enzyme immunoassay utilizing monoclonal antibodies and compared with serum fibrinogen/fibrin degradation products (FDP) assayed by conventional techniques.

In healthy subjects and patients with miscellaneous disorders not usually associated with activation of the haemostasis mechanism, mean XDP levels were 45 and 70 ng/ml respectively. However, elevated levels of XDP occurred in conditions commonly associated with intravascular and possibly extravascular activation of the coagulation system. Markedly raised mean XDP values (677–6900 ng/ml) occurred in treated pulmonary embolism, disseminated neoplasia, severe inflammatory disorders and complicated postoperative states, and lesser but significant elevation (mean 150–400 ng/ml) in treated venous thrombosis, uneventful postsurgical states, localized neoplasia, liver disease and symptomatic arterial disease. Levels during initial streptokinase therapy (mean 24 000 ng/ml) fell tenfold as treatment was continued.

The degree of XDP elevation over normal values was significantly higher than that of FDP in conditions with a propensity for venous thrombosis (post‐operative states, disseminated neoplasia and inflammatory diseases) than in liver disease, localized neoplasia or patients receiving heparin therapy for venous thromboembolism.
Agarose gel method: its usefulness in assaying factor VIII inhibitors, evaluating treatment and suggesting a mechanism of action for factor IX concentrates
Authors: Coots Macie C. , Glueck Helen I. , Miller Mary Ann
Published: March 2008

Summary

Plasmas from 14 patients with factor VIII inhibitors, 10 haemophiliacs and four non‐haemophiliacs, were assayed by both the agarose gel and Bethesda methods. Good correlation was observed in 34 samples from 13 patients, but there was poor correlation in three samples from a single haemophilic patient. The sensitivity of the method was increased by diluting normal platelet rich plasma (PRP) with congenital factor VIII deficient plasma. With this modification, as little as 0.4 of a Bethesda unit (Bu) could be measured accurately. The agarose method is easier to perform and requires much less technician time than the Bethesda assay (Ba). Inhibitory activity can be measured even in the presence of large amounts of transfused factor VIII or factor IX concentrates. To study the effect of factor IX concentrates on the inhibitor, the method was modified by incorporating into the gels plasmas specifically deficient in either factor VII, IX or X. Our data suggest that factor VII is probably responsible for the ‘bypassing’activity of factor IX concentrates.
Heterogeneity of erythropoietin‐dependent erythrocytosis: case report in a child and synopsis of primary erythrocytosis syndromes
Authors: Kulkarni Vivekanand , Ritchey Kim , Howard Donald , Dainiak Nicholas
Published: March 2008

Summary

To investigate the pathogenesis of polycythaemia in a child with isolated, primary erythrocytosis, we measured serum erythropoietin activity and in vitro erythroid progenitor cell responsiveness to erythropoietin. Unstimulated erythropoietin activity was markedly elevated (1.8 IU/ml), and isovolaemic phlebotomy induced a four‐fold increment above this level. In contrast to findings in our index case with this syndrome, normal erythroid colony growth patterns were present in patient marrow cultures. The primary mechanism of polycythaemia in this individual is similar to that reported in the index case: an inappropriately elevated regulatory set point for erythropoietin production. Since an additional defect of progenitor cell hypersensitivity to erythropoietin is not always present, we conclude that abnormalities at single or multiple sites of the erythropoietic regulatory axis may occur in primary erythropoietin‐dependent erythrocytosis.
Features affecting outcome during remission induction of acute myeloid leukaemia in 619 adult patients
Authors: Swirsky D. M. , Bastos M. de , Parish S. E. , Rees J. K. H. , Hayhoe F. G. J.
Published: March 2008

Summary. Six hundred and nineteen patients with de novo acute myeloid leukaemia, entered into the Medical Research Council's eighth trial of therapy have been studied. All patients were treated with the same remission induction regimen. Pretreatment variables comprising age, clinical status, haematologi‐cal status and a detailed marrow cytology and cytochemistry score have been analysed. Poorer remission rates have been found in older patients, in those with lower Karnofsky scores and in patients with a platelet count of less than 25 × 109/1. Leukaemias showing evidence of cytoplasmic maturation along the granulocyte and monocyte lines, as evidenced by granules, Auer rods, a high percentage of Sudan black positive blast cells and morphologïcal and cytoche‐mical abnormalities of neutrophils were associated with a higher remission rate. Marrow eosinophilia was a good prognostic feature. Nuclear features of immaturity, i.e. increasing numbers and prominence of nucleoli were associated with a low remission rate. Abnormalities of the erythroid series, notably Periodic acid‐Schiff positivity which was present in 13 3 cases (22% of the total), was associated with a low remission rate. Patient age and pretreatment Karnofsky score were the most useful predictors of treatment outcome.
Treatment of acute lymphoblastic leukaemia in adults
Authors: Barnett M. J. , Greaves M. F. , Amess J. A. L. , Gregory W. M. , Rohatiner A. Z. S. , Dhaliwal H. S. , Slevin M. L. , Biruls R. , Malpas J. S. , Lister T. A.
Published: March 2008

Summary.Between 1972 and 1982, 112 consecutive previously untreated adults (aged 15–69 years, median 26) commenced therapy for acute lymphoblastic leukaemia (ALL) at St Bartholomew's Hospital. The first 63 patients entered into the study received initial treatment which comprised four cycles of adriamycin and vincristine, prednisolone and L‐asparaginase with the first cycle (OPAL). In 1978, six cycles were given, with escalating doses of adriamycin and cyclophosphamide from cycle 3 (HEAV'D). Central nervous system (CNS) prophylaxis incorporated intrathecal methotrexate and cytosine arabinoside with cranial irradiation. Maintenance chemotherapy consisted of 6‐mercaptopurine, cyclophosphamide and methotrexate for 3 years.

Results obtained with the OPAL and HEAV'D regimens were not significantly different. The overall complete remission (CR) rate was 66% (73/111), factors correlating unfavourably with achievement of CR being advanced age (P<0.001) and L3 morphology/B‐ALL immunophenotype (P<0.01). Fifty‐three patients have relapsed, the bone marrow being the primary site in 43. Extramedullary relapse alone occurred in 10 (seven CNS, two testicular and one skin). Only three of the 64 patients who had complete CNS prophylaxis subsequently relapsed in the CNS as an isolated site. One patient died in CR, 19 remain in continuous CR between 2.5 and 10.5 years. The median duration of remission of the 73 patients who achieved CR was 18.5 months, factors correlating favourably with duration of CR being low blast cell count at presentation (P<0.002) and common ALL immunophenotype (P<0.04). Twenty‐four patients remain alive, with a median survival of all patients of 18 months.
Progression of a myelodysplastic syndrome to pre‐B acute lymphoblastic leukaemia with unusual phenotype
Authors: Bonati Antonio , Delia Domenico , Starcich Riccardo
Published: March 2008

Summary. This study shows the progression of a myelodysplastic syndrome (MDS) to pre‐B acute lymphoblastic leukaemia (ALL) with an unusual phenotype. On diagnosis of leukaemia bone‐marrow mononuclear cells were labelled with murine monoclonal antibodies HLA‐DR, VIL‐A1 (CALLA), 3813, VIM‐D5 and with a rabbit antiserum to TdT using a double colour indirect immunofluorescence technique. In addition simultaneous detection of cytoplas‐mic μ chains (Cyμ) and of TdT was carried out and a direct immunofluorescence analysis for surface membrane immunoglobulins (Smlg) was performed. Two main populations were present: the major one being HLA‐DR+, Cyμ+, VIM‐D5+, TdT−, CALLA−, SmIg−; the minor one HLA‐DR+, Cyμ+, VIM‐D5−, TdT+, CALLA−, SmIg−. The progression of our case to acute leukaemia with a population of leukaemic ceils each of which demonstrated features of lymphoid and myeloid cells suggests that MDS would originate at the pluripotential stem cell level.
Danazol fails to increase factor VIII or IX levels in a double‐blind crossover study of patients with haemophilia A and B
Authors: Nugent Diane J. , Bray Gordon L. , Counts Richard B. , Clements Maribel J. , Thompson Arthur R.
Published: March 2008

Summary. Twenty‐one patients, 14 with haemophilia A and seven with haemophilia B, completed a double‐blind crossover study to evaluate the effects of danazol on factor VIII and factor IX levels. Clotting and immunoradiometric assays were used to measure factor levels at baseline, 2 weeks and 8 weeks on both danazol and placebo. Fibrinogen, plasminogen and activated partial thromboplastin time were measured on all patients during placebo and danazol treatment. Although plasminogen levels rose significantly (P<0.01) and fibrinogen decreased (P<0.01), factor VIII and IX levels did not change. While on danazol, three patients had increased bleeding and shortened euglobulin lysis times compared to their baseline levels. We conclude that danazol does not raise factor VIII or IX levels and increases bleeding in some patients.
T‐cell origin of Lennert's lymphoma
Authors: O'Connor N. T. J. , Feller A. C. , Wainscoat J. S. , Gatter K. C. , Pallesen G. , Stein H. , Lennert K. , Mason D. Y.
Published: March 2008

Summary. The arrangement of the T‐cell receptor and immunoglobulin genes has been analysed in five cases of Lennert's lymphoma. All cases showed rearrangement of the gene coding for the β chain of the T‐cell receptor and a germline configuration of the immunoglobulin genes. This provides strong evidence that Lennert's lymphoma is a T‐cell lymphoma.
Autologous bone marrow transplantation following high dose chemotherapy for the treatment of adult patients with acute myeloid leukaemia
Authors: Goldstone A. H. , Anderson C. C. , Linch D. C. , Franklin I. M. , Boughton B. J. , Cawley J. C. , Richards J. D. M.
Published: March 2008

Summary. 24 adult patients with acute myeloid leukaemia (AML) were treated with intensive chemotherapy followed by autologous marrow rescue. The procedure was repeated twice in eight patients. 11 of 16 patients treated in first remission continue in first unmaintained remission (9–54 months, median 17 months). Eight patients treated at relapse or second remission have relapsed again and died within 14 months of their first autologous bone marrow transplant (ABMT). This form of intensification therapy would appear valuable for adult AML patients in first remission.
Cytogenetic evidence for partially committed myeloid progenitor cell origin of chronic myelomonocytic leukaemia and juvenile chronic myeloid leukaemia: both granuloctye‐macrophage precursors and erythroid precursors carry identical marker chromosome
Authors: Amenomori T. , Tomonaga M. , Yoshida Y. , Kuriyama K. , Matsuo T. , Jinnai I. , Ichimaru M. , Omiya A. , Tsuji Y.
Published: March 2008

Summary. We used a micromethod for cytogenetic analysis from single haematopoietic colonies to study two adults with chronic myelomonocytic leukaemia (CMML) and a boy with juvenile chronic myeloid leukaemia (JCML) carrying distinct chromosome abnormalities, 7q—, der(21), and trisomy 21. We wanted to know if both granulocyte‐macrophage (GM) and erythroid precursors are involved in the abnormal clone. In all three patients, their chromosome abnormality was seen in almost all metaphases obtained from GM‐colonies and erythroid bursts. Peripheral blood leucocytes stimulated with phytohaemagglutinin exhibited only normal karyotypes. Clones of B‐cell produced by Epstein‐Barr virus had only normal karyotypes in the CMML patients. These findings indicate that CMML and JCML are clonal haemopathies that originate in a partially committed myeloid progenitor cell.
Granulopoiesis‐supporting effects of marrow stromal cells in continuous culture from patients with primary myelofibrosis
Authors: Hotta Tomomitsu , Murate Takashi , Yamao Hisami , Yamada Hideo , Saito Hidehiko
Published: March 2008

Summary. The granulopoiesis‐supporting effects of marrow adherent cells from seven patients with primary myelofibrosis (PMF) were studied by a continuous allogeneic co‐culture system in which the survival of donor granulocyte‐macrophage colony‐forming cells (CFU‐GM) depends upon the supporting ability of adherent cells. Marrow adherent cells from all these seven patients were able to sustain the same number of CFU‐GM as those from control subjects. Cytochemical studies showed that colonies grown from cells sustained on fibrous marrow adherent cell layers were predominantly neutrophilic, as were those in control cultures, although many eosinophil colonies grew from patients' bone marrow. These results indicate that marrow stromal cells from PMF patients function normally in their ability to support granulopoiesis.
Multiple chromosomally distinct cell populations in myelodysplastic syndromes and their possible significance in the evolution of the disease
Authors: Mecucci Cristina , Rege‐Cambrin Giovanna , Michaux Jean‐Louis , Tricot Guido , Van den Berghe Herman
Published: March 2008

Summary. Serial chromosome investigations performed in untreated myelodysplastic syndromes (MDS) revealed that: (a) multiple chromosomaily unrelated cell populations may emerge during the course of the disease; (b) chromosome changes of the clonal type may disappear or become undetectable, and may or may not reappear during further evolution. The former of these phenomena was found in four cases and was consistently associated with clinical‐haematological progression of the disease to a more malignant form, including acute leukaemia. The second was apparently unrelated to clinical and haematological changes in the evolution pattern.

These observations are in favour of the hypothesis that MDS evolve as a multistep process in which several events, including the acquisition of sometimes unrelated chromsome changes determine the progression of the disease.
Acute promyelocytic transformation of chronic myeloid leukaemia with an isochromosome 17q
Authors: van der Merwe Tessa , Bernstein Renee , Derman Denis , Stanley Anthony , Dukes Ivan , Murray Jill , Weaving Ann
Published: March 2008

Summary. Transformation to an acute promyelocytic leukaemia occurred in a patient approximately 2 years after having been diagnosed as suffering from chronic myeloid leukaemia (CML). At this time, in addition to the Ph1 chromosomal aberration, an isochromosome 17q [i(17q)] was noted. The t(15;17) was absent. The implications of this are discussed.
Studies on immunological assay of vitamin K dependent factors
Authors: Mikami S. , Tuddenham E. G. D.
Published: March 2008

Summary. We describe and compare five assay systems for Protein C (PC) in human plasma; a functional assay for PC activity, Laurell electroimmunoassay with EDTA or calcium (EDTA‐Laurell or Ca‐Laurell), radioimmunoassay (RIA) and immunoradiometric assay (IRMA). The lower limit of sensitivity of PC in normal reference plasma was 2 × 10−3 units/ml by RIA and 1 × 10−4 units/ml by IRMA, but the ratio of maximum binding to blank binding was superior in RIA.

In normal plasma (= 20), there was no significant discrepancy of PC levels between RIA, Laurell (EDTA− and Ca−) and functional assay. In normal serum, PC antigen (PC:AG) measured by Ca‐Laurell showed a high value (1.37 0.24 units/ml) compared to the other assays. PC:AG of a purified sample increased its level by Ca‐Laurell method during activation by thrombin. Activated PC (PCa) has decreased affinity for anti‐PC rabbit IgG, similarly to decarboxy (warfarinized) PC.

In warfarin‐treated individuals (= 12) who were anticoagulated orally for more than 3 weeks, functional activity was lower (0.27 0.12 units/ml) than that measured by RIA (0.64 0.12 units/ml) or EDTA‐Laurell (0.62 0.06 units/ml), whereas PC:AG measured by CA‐Laurell had a normal value of 0.96 0.40 units/ml. After stopping administration, the activity showed a gradual increase to the normal at 2 weeks, while PC:AG increased more rapidly to normalize in 1 week.
Isolation of a new specific plasminogen activator in hibitor from pregnancy plasma
Authors: Lecander Ingegerd , Åstedt B.
Published: March 2008

Summary. A specific plasminogen activator inhibitor was isolated from the plasma of pregnant women by matrix‐bound, cross reacting monoclonal antibodies against placental plasminogen activator inhibitor. The pregnancy plasma plasminogen activator inhibitor (PP‐PA‐I) was found to be immunologically different from the inhibitor produced by endothelial cells. Its molecular weight was 70 000 daltons. It formed complexes with urokinase (u‐PA) and with plasminogen activator of the tissue type (t‐PA), similar to those formed by the placental plasminogen activator inhibitor (Pl‐PA‐I). It did not inhibit plasmin. For measuring PP‐PA‐I, an enzyme‐linked immunosorbent assay (ELISA) was designed using monoclonal and polyclonal antibodies against the placental inhibitor. Concentrations of PP‐PA‐I increased successively during pregnancy, and fell sharply after delivery. This inhibitor could not be detected in normal non‐pregnancy plasma. The results indicate that the inhibitor isolated from pregnancy plasma is responsible for the depressed fibrinolytic activity during pregnancy, and that the placenta is the source of the inhibitor.
Growth fraction of tumour cells and infiltrationdensity with natural killer‐like (HNK1+) cells in non‐Hodgkin lymphomas
Authors: Greil R. , Gattringer C. , Knapp W. , Huber H.
Published: March 2008

Summary. Two markers for cells in the growth fraction, the T9 antigen (i.e. the transferrin receptor) and the T10 antigen were investigated in frozen tissue sections of 105 non‐Hodgkin lymphomas (NHL). The results were correlated with the histological subtype and the pattern of tumour infiltration by reactive cells. Special attention was directed to the density of natural killer (NK)‐like cells using the anti‐HNK1 (Leu7) antibody since the transferrin receptor (tfr) or other growth‐associated membrane structures may serve as target for NK cells. Our study confirms a relationship between number of tumour cells with the T9 marker and tissue infiltration by HNK1 cells in NHL of low (chroniclymphocytic leukaemia, hairy‐cell leukaemia, immunocytic lymphoma, centroblastic‐centrocytic lymphoma) and intermediate (centrocytic and centroblastic lymphoma) but not in NHL of high malignant grade (immunoblastic and lymphoblastic lymphoma). Comparable results were obtained with the T10 antigen although the correlation was less close. The percentage of cells in the growth fraction, defined by the expression of the T9 and T10 marker, corresponded with prognostically unfavourable subgroups with the remarkable exception of follicular NHL of centroblastic‐centrocytic type. This lymphoma showed high numbers of cells with the T9 and the T10 marker in a microenvironment resembling normal germinal centres in many aspects.
Status of major red cell blood group antigens on neutrophils, lymphocytes and monocytes
Authors: Dunstan Robert A.
Published: March 2008

Summary. Previous investigators have reported that a number of red cell (RBC) antigens are found on neutrophils (PMN) and lymphocytes. However, there is alack of consensus in the literature. Furthermore, few data are available concerning the occurrence of RBC antigens on monocytes. To address this problem, leucocyte fractions prepared from donors of known RBC antigen phenotype were analysed by fluorescence flow cytometry. Based on recent reports that PMN do not express ABH antigens and on the data presented here, the occurrence of major RBC antigens on leucocytes may be defined as follows: (1) lymphocytes express A, B. Lea and Leb antigens, depending on the ABH secretor (Se) status of the donor; (2) ABH and Lewis antigens cannot be detected on monocytes and PMN regardless of Se status; (3) D, E, e, C, c, Fya, Fyb, Fy5, Jka, Jkb, Jk, K, k, M, N, S, s, U, Vel, Coa, Lan, Jk3 Yta, Dib, Ge, Sc:1 or Lub antigens were not detected on lymphocytes, monocytes and PMN; (4) lymphocytes, monocytes and PMN all express I, i, P and P1 antigens. The absence of selected Rhesus, Duffy, Kell, Kidd and other antigens on PMN is at variance with some previous reports. Furthermore, the distribution of RBC antigens on lymphocytes and monocytes has not been previously characterized using immunofluorescence flow cytometry.
Effect of ethanol on thrombopoiesis
Authors: Levine Richard F. , Spivak Jerry L. , Meagher Richard C. , Sieber Fritz
Published: March 2008

Summary. Chronic ethanol abuse causes thrombocytopenia but the underlying mechanism is unknown. To determine the target cells involved, we examined the effects of the drug in vitro on both megakaryocyte progenitor cells (CFU‐Meg) and isolated, maturing megakaryocytes. In the presence of ethanol concentrations of 0.05–2.0 g/dl, megakaryocyte colony formation by mouse CFU‐Meg in soft agar was normal. At 5 g/dl ethanol, colony formation was reduced by 50%; with 7 g/dl ethanol, no megakaryocyte colonies were formed. Acetaldehyde did not inhibit colony formation unless very high concentrations (100 mg/dl) were employed. Isolated guinea‐pig megakaryocytes can maintain their viability and incorporate 3H‐leucine into TCA‐precipitable protein for at least 24 h. Incubation of these maturing megakaryocytes with ethanol did not affect their viability, but at concentrations greater than 120 mg/dl ethanol progressively inhibited protein synthesis. At 0.5 g/dl ethanol, protein synthesis was decreased by 23% while viability was still 93% of control. Like CFU‐Meg, maturing megakaryocytes were resistant to the toxic effects of acetaldehyde. To determine the in vivo correlates of these results, guinea‐pigs were fed 5 g/dl ethanol in a liquid diet. by 11 d, when blood ethanol levels were 20–150 mg/dl, platelet counts in the animals were reduced by 17–29%, while the number of marrow megakaryocytes was unaltered. These data indicate that the site of action of ethanol in suppressing thrombopoiesis is at the level of the maturing megakaryocyte.
Changes in the granulocyte membrane following chemotherapy for chronic myelogenous leukaemia
Authors: Baker Michael A. , Kanani Amita , Hindenburg Alexander , Taub Robert N.
Published: March 2008

Summary. Granulocytes from patients with chronic myelogenous leukaemia (CML) have been previously shown to have aberrant sialylation of membrane glyoproteins. We have examined the granulocytes from CML patients receiving intermittent chemotherapy to determine the relationship of the oligosaccharide changes to treatment. Compared to cells from non‐leukaemic patients, granulocytes from untreated CML patients showed less adherence to nylon wool, decreased reactivity with peanut lectin, and decreased binding of the synthetic chemotactic peptide formyl‐methionine‐leucine‐phenylalanine (FMLP). Granulocytes from CML patients treated with chemotherapy showed nylon wool adherence, peanut lectin reactivity and FMLP binding comparable to non‐leukaemic cells. Chemotherapeutic agents may interfere with oligosaccharide synthesis with a resulting change in the composition of cell surface glycoconjugates.
An identical translocation between chromosome 1 and 15 in two patients with myelodysplastic syndromes
Authors: Mecucci Cristina , Tricot Guido , Boogaerts Marc , Van den Berghe Herman
Published: March 2008

Summary. An identical translocation between the long arm of chromosome no. 1 and the short arm of chromosome no. 15 was found in two unrelated patients with refractory anaemia type I, according to the FAB classification of myelodysplastic syndromes. In the first patient the typical translocation was associated with anomalies commonly found in preleukaemic states, i.e. a 5q — and a 20q— chromosome. Furthermore, in both patients the long arm of chromosome no. 1 was trisomic. Cytogenetic follow‐up in the second patient demonstrated a proliferative advantage of the cells bearing a t(1;15) translocation over the cells with trisomy 8 as well as over normal cells. This karyotypic evolution, however, was not accompanied by a transformation of the haematological disorder into acute leukaemia.
Lithium stimulation of in vitro granulopoiesis: evidence for mediation via sodium transport pathways
Authors: Gallicchio Vincent S.
Published: March 2008

Summary. Lithium (Li) stimulates granulopoiesis both in vitro and in vivo by increasing the number of committed granulocyte‐macrophage colony forming stem cells (CFU‐GM) either through direct or indirect mechanisms. In this report are described further studies designed to investigate if this Li stimulation could be altered by modulating Li transport using agents known to influence monovalent cation transport. Ouabain, a Na/K ATPase inhibitor, added to nonadherent bone marrow cells either before or immediately after the addition of ultra‐pure Li (1 mEq) produced an irreversible reduction in CFU‐GM indicatingthe importance of the Na/K ATPase in these processes. Further studies using the sodium and potassium ionophores gramicidin and valinomycin (5 μg/ml) followed by a delay in the addition of Li (0, 5, 15, 20, 30, 60 and 120 min) reduced CFU‐GM; however, this reduction was less severe in the presence of gramicidin, indicating sodium transport pathways may be required for Li action. To further explore this hypothesis, studies were performed using the more specific sodium transport inhibitors, amiloride and phloretin. Both were effective in reducing the ability of Li to increase CFU‐GM. Studies incorporating the calcium ionophore, A23187, demonstrated that in the presence of Li, a further reduction in CFU‐GM was observed indicating that Li was unable to reverse this A23187 induced reduction in CFU‐GM. These data suggest that in the presence of active calcium transport, the ability of Li to increase CFU‐GM is restricted.
Binding of human thrombin to human factor VIII:RAg
Authors: Hau Lena , Firkin Barry G. , Howard Margaret A.
Published: March 2008

Summary. The binding capacity of radiolabelled human alpha‐thrombin (α‐thrombin), inactivated with phenyl‐methyl sulphonyl fluoride (PMSF), to factor VIII related antigen (VIII:RAg) multimers isolated by immune precipitation and resolved by sodium dodecyl sulphate (SDS) agarose gel electrophoresis has been examined. 125I‐PMSF α‐thrombin bound predominantly to four low molecular weight multimers (LMW) of VIII:RAg prepared from either normal or haemophilia A plasma. Two of these VIII:RAg multimers present in scrum had lost the capacity to bind thrombin. No bands were observed when either plasma or serum from three patients with severe von Willebrand's disease (vWd) was processed in an identical manner. This binding was not dependent on the presence of an intact catalytic site on the thrombin molecule. A similar pattern of binding to VIII:RAg multimers was observed with 125I‐labelled human anti‐FVIII antibodies as 125I‐α‐thrombin and appears to involve the same multimers.
Purine degradative enzymes and immunological phenotypes in chronic B‐lymphocytic leukaemia: indications that leukaemic immunocytoma is a separate entity
Authors: Ho Anthony D. , Dörken Bernd , Ma David D. F. , Pezzutto Antonio , Hunstein Werner , Hoffbrand A. Victor
Published: March 2008

Summary. Investigations of the purine degradative enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and ecto‐5′‐nucleotidase (5′NT) have been shown to be of value in defining subsets of lymphoid malignancies. We have studied the activities of these enzymes in the circulating malignant cells of 35 patients with chronic B lymphocytic leukaemia and have correlated the biochemical data with immunological phenotypes. Classification of the cases into those without evidence of secretory activity (‘true’ CLL, 14 patients) and those with cytoplasmic immunoglobulin (CIg) (‘immunocytoma’;2] patients) revealed that immunocytomas are phenotypically and biochemically associated with more mature features. Malignant cells without CIg were characterized by low activities of ADA, PNP and 5′NT. In malignant cells with evidence of secretory activity (immunocytoma), low activity of ADA was also observed, but the activities of PNP and 5′NT were relatively high and approached the range of normal B lymphocytes. The differences in PNP (P < 0.05) and in 5′NT (P< 0.01) between these two groups were significant. Phenotypically the cells without CIg were predominantly associated with IgM (+k light chains) as surface membrane immunoglobulin (Smlg) whereas expression of IgG was more often observed in the leukaemic cells with CIg. No correlation between enzyme patterns and the stage of the disease was apparent. Thus both biochemical and immunological criteria show that cases of CLL vary within a range of maturity and that those with CIg might be more mature in the B cell axis. The present study emphasizes the value of purine enzyme studies in defining subsets of B cell neoplasia.
A study of the haematological and haemorheological consequences of venesection
Authors: Challoner T. , Briggs C. , Rampling M. W. , Thomas D. J.
Published: March 2008

Summary A study has been made of a number of haematological and haemorheological factors following venesection therapy. Haematocrit was very effectively reduced by the venesection regimen and led to a fall in whole blood viscosity in spite of the microcytosis that was eventually induced in all the subjects. No clinically significant coagulation changes were observed. It is concluded that a simple venesection regimen without volume replacement is a safe procedure for improving blood rheology.
Monoclonal gammopathy with an insulin binding IgG(K) M‐component, associated with severe hypoglycaemia
Authors: Sluiter Wim J. , Marrink Jan , Houwen Berend
Published: March 2008

Summary A case is described in which a monoclonal gammopathy was associated with postprandial hypoglycaemia, resulting from insulin binding by the M‐component. The monoclonal immunoglobulin possessed a low affinity for insulin, but its binding capacity was very high. Cytotoxic chemotherapy and diazoxide treatment failed to prevent severe hypoglycaemic attacks.
Human urinary megakaryocyte colony‐stimulating factor in thrombopoietic disorders
Authors: Kawakita Makoto , Yamamoto Seiko , Asou Norio , Ishii Masao , Sakaguchi Mamoru , Takatsuki Kiyoshi
Published: March 2008

Summary The level of urinary Meg‐CSF activity in patients with various thrombopoietic disorders was studied. Five out of eight patients with idiopathic thrombocytopenic purpura had megakaryocytic hyperplasia in the marrow and increased Meg‐CSF activity in the urine. Urinary Meg‐CSF activity in patients with polycythaemia vera and essential thrombocythaemia was normal. There was a significant inverse correlation between urinary Meg‐CSF activity and peripheral blood platelet count but not bone marrow megakaryocyte mass. There was a significant increase of urinary Meg‐CSF activity during the period of thrombocytopenia after chemotherapy in patients with acute leukaemia who were in complete remission. The timing of maximal Meg‐CSF levels corresponded to the nadirs of platelet counts. These results support the concept that Meg‐CSF may play a significant role in the regulation of megakaryopoiesis and/or thrombopoiesis in vivo.
Immunologic and biochemical characterization of homozygous and heterozygous Glanzmann thrombasthenia in the Iraqi‐Jewish and Arab populations of Israel: comparison of techniques for carrier detection
Authors: Coller Barry S. , Seligsohn Uri , Zivelin Ariela , Zwang Ety , Lusky Ayala , Modan Michaela
Published: March 2008

Summary To define biochemically and immunologically the platelet defect in Iraqi‐Jews and Arabs with Glanzmann thrombasthenia in Israel, we tested the platelets of 32 thrombasthenics and 37 obligate carriers from 19 families with affected members. Thrombasthenic platelets were devoid of glycoprotein IIb (GPIIb) as judged by polyacrylamide gel electrophoresis and devoid of the GPIIb/IIIa complex as judged by radio‐electroimmunoassay. Binding of a murine monoclonal antibody directed at GPIIb and/or GPIIIa to intact thrombasthenic platelets averaged less than 2% of the control value. Evaluation of the number of molecules of antibody bound per platelet permitted discrimination between controls and obligate carriers with a high degree of accuracy (sensitivity = 91.9%, specificity = 92.3%). Obligate carriers could also be discriminated from controls by determining the ratio of GPIIb to GPIb by polyacrylamide gel electrophoresis and by quantifying the GPIIb/IIIa complex by radio‐electroimmunoassay. These studies indicate that the thrombasthenics in Israel have the severe form of the disease (type I) and that the platelets of heterozygotes have significantly reduced amounts of both total and surface‐exposed GPIIb and/or GPIIIa.
The Wiskott‐Aldrich syndrome: studies of platelets, basophils and polymorphonuclear leucocytes
Authors: Marone Gianni , Albini Fabrizio , Martino Lucio di , Quattrin Stefano , Poto Sergio , Condorelli Mario
Published: March 2008

Summary Platelets, basophils and neutrophils from a patient with the Wiskott Aldrich syndrome (WAS) were exposed to stimuli that activate specific membrane receptor or directly initiate biochemical events (e.g. the Ca2+ ionophore A23187 and ionomycin or arachidonic acid). Platelets from this patient did not aggregate in response to ADP, collagen, thrombin or adrenaline, which activate specific membrane receptors. Platelet aggregation, however, was normal in response to compound A23187, ionomycin or exogenous arachidonic acid. Histamine release from basophils of the WAS patient was normal in response to anti‐IgE, a formylated peptide (f‐met peptide), and to A23187. Similarly, the release of the lysosomal enzymes, β‐glucuronidase and lysozyme, from neutrophils of the WAS patient in response to serum treated zymosan (Zx), f‐met peptide, and A23187 was not significantly different from that of his parents and 13 normal donors. These results suggest that the primary defect in WAS is selectively present in platelets and is located in a biochemical step between receptor activation and Ca2+ influx and/or initiation of arachidonate metabolism.
In vivo differentiation of leukaemic blasts and effect of low dose Ara‐C in a marrow grafted patient with leukaemic relapse
Authors: Mittermüller J. , Kolb H. J. , Gerhartz H. H. , Wilmanns W.
Published: March 2008

Summary The possible induction of differentiation of leukaemic cells by low dose cytosine arabinoside (Ara‐C) was studied in an human chimaera with leukaemic relapse. A 35‐year‐old woman had been grafted with bone marrow from her HLA identical sister and developed a leukaemic relapse of the host type demonstrated by typing phosphoglucomutase 1 isoenzymes. Before treatment with low dose Ara‐C only leukaemic cells and a part of the mature granulocytes were of host type. Lymphocytes, platelets and erythrocytes were still of donor type. The leukaemic cells and the mature host type granulocytes disappeared and the patient went into complete remission after treatment with low dose Ara‐C. The results indicate that (a) the leukaemic blasts could differentiate in vivo into mature granulocytes, (b) leukaemic transformation was limited to granulopoiesis, and (c) low dose Ara‐C acted as a cytotoxic drug instead of inducing differentiation of host type granulopoiesis.
Haemoglobin gene frequencies in the Jamaican population: a study in 100,000 newborns
Authors: Serjeant G. R. , Serjeant B. E. , Forbes M. , Hayes R. J. , Higgs D. R. , Lehmann H.
Published: March 2008

Summary. The gene frequencies of abnormal haemoglobins have been determined in a group of 100 000 Jamaican newborns screened over a period of 8½ years. The population is predominantly of West African origin and the survey represents approximately one quarter of all island deliveries within the period of the study. The common β globin chain abnormalities βS and βc occurred with gene frequencies of 0.055 and 0.019 respectively; β thalassaemia was relatively rare. In contrast, α thalassaemia was quite common, occurring with a gene frequency of 0.183. In addition to these common abnormalities, the frequencies of 256 rare abnormal haemoglobins are described. This survey thus represents a complete and accurate documentation of the α and β globin variants that occur in the Jamaican population.
Interaction of five globin gene abnormalities in a Cambodian family
Authors: Simpkins H. , Hill A. V. S. , Derry S. , Clegg J. B. , Weatherall D. J.
Published: March 2008

Summary

Members of a Cambodian family with an undiagnosed hypochromic, microcytic anaemia were found by haemoglobin and DNA analysis to have five interacting globin gene abnormalities. One child has Hb E and typical Hb H disease, while his mother has the form of Hb H disease associated with Hb Constant Spring interacting with Hb E. Quantitation of Hbs E and A2 by globin chain separation and triton/urea gel electrophoresis support the concept that Hb H/Constant Spring disease is a more severe form of α thalassaemia than Hb H disease. This family illustrates how the remarkably high prevalence of globin gene abnormalities in Southeast Asians can give rise to a series of atypical thalassaemic phenotypes, and how they can be defined by direct globin gene analysis.
The influence of oxygen tension on the long‐term growth in vitro of haematopoietic progenitor cells from human cord blood
Authors: Smith Sydney , Broxmeyer Hal E.
Published: March 2008

Summary

Growth of low density human cord blood cells in long‐term suspension culture was evaluated at incubation conditions of 5% carbon dioxide in ∼ 20% (normal incubator) oxygen tension or in 5% (low) oxygen tension. During the first 5 weeks there was no difference in the numbers of morphologically recognizable cells grown at either oxygen tension, while growth was superior from weeks 5 to 8 at ∼ 20% oxygen tension. For the first 5 weeks, growth of granulocyte‐macrophage progenitors (CFU‐GM) was superior at 5% oxygen tension during the long‐term and semi‐solid culture phases and least impressive in the long‐term and semi‐solid cultures incubated at ∼ 20% oxygen. This trend was reversed after 5 weeks and after 8 weeks there were no detectable CFU‐GM in suspension cultures at 5% oxygen while steady state levels of CFU‐GM were maintained for greater than 12 weeks in suspension cultures at ∼20% oxygen. Semi‐solid cultures for erythroid (BFU‐E) and multipotential (CFU‐GEMM) progenitors were incubated at ∼ 20% oxygen only. During the first 4 weeks, the growth of BFU‐E and CFU‐GEMM was superior at 5% oxygen during long‐term culture. Numbers of these cells decreased by week 5 and at this time growth was better in the long‐term cultures grown at ∼ 20% oxygen. By week 6, no BFU‐E or CFU‐GEMM were detectable. Thus, growth of cord blood at low oxygen tension gives an initial enhancement in output of progenitor cells, but this appears to be at the expense of continued production.
A murine monoclonal IgM antibody specific for blood group P antigen (globoside)
Authors: Kr. vondem Borne A. E. G. , Bos M. J. E. , Joustra‐Maas N. , Tromp J. F. , Van Wijngaarden‐du Bois R. , Tetteroo P. A. T.
Published: March 2008

Summary

A murine monoclonal IgM erythrocyte antibody appeared to have anti‐P (anti‐globoside) specificity. The antibody was a relatively weak cold agglutinin, but a strong haemolysin and its reactivity with red cells was markedly enhanced by enzyme treatment. This antibody was used to study the cell and tissue distribution of globoside. Globoside was not only detectable on red cells and erythroblasts, but also on endothelial cells and on subsets of platelets, megakaryocytes and fibroblasts. It was not detectable on granulocytes, monocytes and most peripheral blood lymphocytes. Neither was it present on erythroblast precursors (CFU‐E, BFU‐E), pro‐erythroblasts or on the cells of the pro‐erythroblastic cell lines K562 and HEL. However, K562 cells expressed globoside when induced to mature into erythroblasts by sodium butyrate. Cells of patients with various leukaemias were also tested. A significant number of positively reacting cells was frequently (six out of 18) seen in cases with a CML blast crisis (CML‐BC) and rarely in AML (four out of 37 cases). In CML‐BC the P‐positive cells were probably erythroblasts and/or megakaryoblasts. Thus, globoside appeared to be an interesting marker in CML‐BC of the erythroblastic or mixed erythroblastic‐megakaryoblastic type.
Successful remission induction with deoxycoformycin in elderly patients with T‐helper prolymphocytic leukaemia
Authors: El’Agnaf M. R. , Ennis K. E. , Morris T. C. M. , Robertson J. H. , Markey Geraldine , Alexander H. D.
Published: March 2008

Summary

Two elderly patients with prolymphocytic leukaemia (PLL) of T helper phenotype were treated with the adenosine deaminase inhibitor— deoxycoformycin—and achieved remission. The first patient has remained in an unmaintained remission for over a year. The second patient, treated with a regime which produced less side effects, subsequently relapsed in skin and lymph nodes and died. In view of the rarity of this condition a multi‐centre assessment of the effectiveness of deoxycoformycin is indicated.

T‐lymphocyte colony formation in both cases was found to be reduced. Co‐culture of the patients’lymphocytes with nonadherent mononuclear cells from normal individuals also showed inhibition of T‐colony formation indicating that lack of nutrients or accessory cells was not responsible for low T‐colony forming capacity.
Lactoferrin binds to neutrophilic membrane DNA
Authors: Bennett Robert M. , Merritt Marcie M. , Gabor Gary
Published: March 2008

Lactoferrin (LF) binding to the surface of human neutrophils was shown to be dependent upon the presence of cell surface DNA by (i) an abrogation of LF binding after treatment of whole cells with DNAse; (ii) an abrogation of LF binding to a purified cell membrane suspension after DNAse digestion, (iii) a restoration of LF binding, after initial treatment of cells with DNAse, by the addition of exogenous DNA. Using a biotinylated LF probe, no other binding molecules were found after SDS PAGE of neutrophil cell membrane proteins. Further evidence of a DNA‐LF interaction was obtained by the co‐isolation of LF with DNA by both gel chromatography and affinity chromatography using Heparin Sepharose CL 6B. The interaction of LF with neutrophils was a saturable phenomenon with a Kd of 6.2 x 10‐6 M and a maximum binding of 9.2 x 106 molecules per cell. These results suggest that cell membrane DNA may have a novel role as a receptor for LF, and indicates the need for further experiments to determine whether the functional effects of LF are modified by the DNAse treatment of LF responsive cells.
Competitive survival/proliferation of normal and Ph1‐positive haemopoietic cells
Authors: Frassoni Francesco , Repetto Mario , Podestà Marina , Piaggio Giovanna , Raffo Maria Rita , Sessarego Mario , Bacigalupo Andrea , Marmont Alberto M.
Published: March 2008

Summary

Bone marrow (BM) cells from 10 patients with Ph1‐positive chronic granulocytic leukaemia (CGL) were placed in long‐term cultures in the presence of fetal calf serum (FCS) and horse serum (HoS), or in the presence of human AB serum. The long‐term cultures were started with three different cell combinations: (1) CGL BM cells (four cases), (2) CGL BM cells + normal BM cells (1:1 ratio) from an HLA identical sex‐matched sibling (five cases), (3) CGL BM cells + normal BM cells (1:1 ratio) from an HLA identical sex mismatched sibling (five cases). Cytogenetic studies were performed at weeks 0, 3, 4 and 5 of culture. The results of this study can be summarized as follows: (a) Ph1‐positive cells could be detected at any time of culture in all three of the described cell combinations; (b) a population of Ph1‐negative cells of patient origin could be detected after 3–5 weeks of culture; (c) there was a trend for a better survival of Ph1‐negative cells in cultures supplemented with FCS + HoS and, conversely, of Ph1‐positive cells in cultures containing human serum. These results warrant further studies on the possibility of manipulating survival and proliferation of normal and leukaemic cells by varying the culture conditions.
An association between chronic neutrophilic leukaemia and multiple myeloma with a study of cobalamin‐binding proteins
Authors: Lewis Michael J. , Oelbaum Maurice H. , Coleman Morton , Allen Steven
Published: March 2008

Summary

Two patients are described who had evidence of both multiple myeloma and chronic neutrophilic leukaemia at or near the time of presentation. Descriptions of five similar patients were found in the literature supporting an association between the two disorders. This association is further evidence of a link between myeloproliferative and lymphoproliferative disorders. Cobalamin‐binding studies of the plasma and neutrophils from one of these patients showed a gross elevation of plasma unsaturated TC I and abnormal neutrophils which contained TC I but not TC III.
Ph1 negative haematological chimaerism after marrow transplantation in Ph1 positive chronic granulocytic leukaemia
Authors: Vincent P. C. , Young G. A. R. , Singh S. , Atkinson K. , Biggs J. C.
Published: March 2008

Summary

A 17‐year‐old girl with Philadelphia chromosome (Ph1) positive chronic granulocytic leukaemia (CGL) who has undergone two bone marrow transplants from her HLA identical brother is described. Following the second transplant, cytogenetic analysis of her bone marrow cells showed haematological chimaerism with equal numbers of normal male cells and Ph1 negative female cells indicating the probable eradication of the Ph1 positive clones with the retention of normal host stem cells.
Purification of haemopoietic progenitor cells from patients with chronic granulocytic leukaemia using Percoll density gradients and elutriation
Authors: Martin Hans , Hibbin Jill A. , Dowding Charles , Matutes Estela , Tindle Robert , Goldman John M.
Published: March 2008

Summary

Purification of haemopoietic progenitor cells from chronic granulocytic leukaemia buffy coat preparations requires a multistep approach using complementary cell separation techniques. In this study Percoll density gradient centrifugation and centrifugal elutriation were used to isolate large numbers of immature progenitor cells. Percoll density gradients were valuable as a first separation step: CFU‐GM and CFU‐GEMM could be enriched 75‐fold in a light density fraction of d< 1mD056 g/ml and the technique could be adapted to cope with more than 1010 buffy coat leucocytes. Progenitors cells were concentrated 3‐fold by elutriation used as single method to separate buffy coat cells or when used to purify further light density Percoll fractions. When Percoll gradients and elutriation were used sequentially, undifferentiated mononuclear cells were enriched to more than 90% purity arid between 5% and 40% of these cells formed CFU‐GM or BFU‐E colonies consisting of more than 40 cells. The enriched fractions were further characterized with monoclonal antibodies. The density and elutriation profiles of these colony forming cells resembled corresponding profiles of cells that reacted with the monoclonal antibody BI‐3C5, which recognizes an antigen on primitive haemopoietic progenitor cells. Physical separation methods are a valuable first stage in the attempt to procure relatively pure myeloid progenitor cell populations, whose characteristics can then be further studied at a cellular or molecular level.