bluuebunny/crossref_metadata_2025 huggingface dataset:
Autologous bone marrow transplantation (ABMT) for acute leukaemia in complete remission: a pilot study of 33 cases
Authors: Cahn J. Y. , Herve P. , Flesch M. , Plouvier E. , Noik A. , Racadot E. , Montcuquet P. , Behar C. , Plgnon B. , Boilletot. A. , Lutz P. , Henon P. , Rozenbaum A. , Peters A. , L econte Des Floris R.
Published: March 2008

Summary

Thirty‐three leukaemic patients in CR were treated by high‐dose therapy followed by ABMT: 18 of them had acute non‐lymphoblastic leukaemia (ANLL) in first remission (CR1) with a mean age of 23.7 years (3–44). All but one of them were conditioned with a polychemotherapy regimen including 6‐thioguanine, Ara‐C, CCNU, and cyclophosphamide. The marrow cells were purged by chemical means in 16 cases.
Molecular pathology of haemoglobin H disease in Sardinians
Authors: Paglietti E. , Galanello R. , Moi P. , Pirastu M. , Cao A.
Published: March 2008

Summary

We investigated the molecular basis for haemoglobin H disease in 50 Sardinian patients by restriction endonuclease analysis. We found that the majority (78% of the cases) are due to gene deletion (–/ ‐ α). Among those with a combination of deletion and nondeletion defects (–/ααth), the most prevalent nondeletion lesion (70% of the nondeletion defects) was the initiation codon mutation of the α2 gene (αNcoα), previously discovered in this population. Of the remaining patients with the (–/ααth) genotype, two showed the IVS‐1 splice junction lesion and one a mutation in the α1 gene, removing the Nco I site within the 5’part of the α1 gene, which may arise from a process of gene conversion from the initiation codon mutant of the α2 gene. A single patient had the homozygous state for the initiation codon mutant of the α2 gene. Study of genotype‐phenotype correlations indicates that the (αNcoα) haplotype is associated with a more severe defect in the α‐globin chain output than that resulting from the (‐α) haplotype. We may conclude that restriction endonuclease analysis is a powerful method for the definition of the molecular heterogeneity of haemoglobin H disease.
Acute myeloid leukaemia in childhood: clinical features and prognosis
Authors: Chessells Judith M. , Ocallaghan Una , Hardisty Roger M.
Published: March 2008

Summary

Clinical and laboratory features at presentation were correlated with morphological (FAB) subclass of AML in a group of 112 children diagnosed between 1972 and 1982. Patients with a monocytic component of AML (M4, M5) had higher initial leucocyte counts, a higher incidence of extramedullary infiltration and of CNS involvement. In M4 AML CNS relapse occurred in patients with a high initial leucocyte count whereas in M5 AML CNS involvement tended to occur at presentation in children with low initial counts. Two‐thirds of patients treated achieved remission and most failures were due to inadequate chemotherapy, although haemorrhage, leucostasis or metabolic complications caused early death in patients with M4 and M5 AML. With a minimum follow up of 3 years only 12% of patients are alive; these figures have not improved in consecutive series despite increasing intensity of induction and more recent availability of bone marrow transplantation. No features predictive of long‐term survival were identified, but patients with myeloid differentiation (M1, M2, M3) did better than those with a monocytic or erythroid component (M4, M5, M6).
Red cell metabolism in hereditary pyrimidine 5′‐nucleotidase deficiency: effect of magnesium
Authors: Lachant Neil A. , Tanaka Kouichi R.
Published: March 2008

Summary

Since pyrimidine nucleotides avidly bind magnesium, we tested the hypothesis that the haemolytic anaemia in hereditary pyrimidine 5′‐nucleoti∼ dase (P5N) deficiency is due to a state of functional magnesium depletion in the red cell (RBC). In haemolysates from normal subjects, cytidine triphos‐phate (CTP) inhibited the activity of pyruvate kinase in a competitive manner for magnesium. The CTP Ki was 0.4 mmol/1. CTP inhibited the activity of hexokinase in a competitive manner for ATP (Mg‐ATP2‐) with a Ki of 4 mmol/1. The inhibitory effect of CTP on both enzymes was overcome by increasing the magnesium content of the test system. Since CTP appeared to inhibit enzymes which required magnesium as a cofactor or Mg‐ATP2‐ as a substrate, we tested the effect of exogenous magnesium on the metabolism of P5N deficient RBC. The autohaemolysis test, the incubated Heinz body assay and the rate of glucose oxidation by the pentose phosphate shunt were abnormal in the intact RBC from a patient with hereditary P5N deficiency. The addition of MgCl2 (6–10 mmol/1) did not improve these abnormal in vitro measures of metabolism in the P5N deficient RBC. This lack of effect of exogenous magnesium may be due to the slow uptake of magnesium by the human RBC.
Reduced risk of recurrent leukaemia in bone marrow transplant recipients after cytomegalovirus infection
Authors: Löunnqvist B. , Ringdegn O. , Ljungman P. , Wahren B. , Gahrton G.
Published: March 2008

Summary

The first 72 consecutive bone‐marrow transplant recipients with haematological malignancies (29 with acute nonlymphoblastic leukaemia, 31 with acute lymphoblastic leukaemia, nine with CML and three with myelofibro‐sis, IgA myeloma and T‐cell lymphoma, respectively) were investigated for the frequency of relapses 1 year or later after bone‐marrow transplantation. Seven relapses occurred from 30 to 850 d after transplantation (median 180 d). All relapses occurred in patients with acute leukaemia ≤ 18 years of age with a high risk for relapse, i.e. transplanted in second or later remission or with more than 10% blasts in the marrow before transplantation. Among all patients the probability of relapse was increased in patients without cytomegalovirus (CMV) infection (P= 0.001) and in patients without chronic GVHD (P= 0.049). Among leukaemic patients ≤ 18 years of age with a high risk of relapse all relapses occurred in patients (n= 11) without CMV infection, whereas no relapses were seen in patients (n= 13) with CMV infection (P= 0.006). Known risk factors for leukaemic relapse were comparable in both groups.
Autoimmune thrombocytopenia and neutropenia after remission induction therapy for acute leukaemia
Authors: Chapman J. F. , Murphy M. F. , Minchinton R. M. , Metcalfe P. , Lister T. A. , Waters A. H.
Published: March 2008

Summary

Patients with acute leukaemia who are exposed to intensive chemotherapy, with or without radiotherapy, may remain thrombocytopenic or neutropenic in remission. The incidence of these prolonged cytopenias was examined retrospectively in 46 patients in remission and prospectively in 14 patients. The patients were tested for the presence of autoantibodies to platelets and neutrophils using a fluorescent antihuman globulin technique with paraformaldehyde‐fixed cells. In the retrospective study nine patients (20%) had neutrophil autoantibodies and seven (15%) had platelet autoantibodies; only one of the former had neutropenia and one of the latter thrombocytopenia. In the prospective study three (21%) had neutrophil autoantibodies and seven (50%) had platelet autoantibodies. One of the patients with platelet autoantibodies had transient thrombocytopenia, and shortened platelet survival was demonstrated. None of the patients with neutrophil autoantibodies had neutropenia.

The rare occurrence of cytopenias in association with these autoantibodies was possibly due to bone marrow compensation for antibody‐mediated cell destruction, although other possible mechanisms are discussed.
Von Willebrand factor contaminating porcine factor VIII concentrate (Hyate: C) causes platelet aggregation
Authors: Altieri Dario C. , Capitanio Anna M. , Mannucci Pier Mannuccio
Published: March 2008

Summary

Severe thrombocytopenia has been reported to occur in some patients treated with a polyelectrolyte‐fractionated porcine factor VIII concentrate (Hyate:C) (Kernoff et al, 1984; Gatti & Mannucci, 1984). We report here that Hyate: C induces aggregation of platelet‐rich plasma in vitro accompanied by ATP release and thromboxane B2 formation, and is inhibited by EDTA and prostaglandin E1. Hyate:C induced platelet aggregation is affected by monoclonal antibodies anti‐glycoprotein Ib or anti‐glycoprotein IIb/IIIa and suppressed by polyclonal anti‐porcine von Willebrand factor antiserum. We conclude that the aggregating ability of Hyate :C is due to porcine von Willebrand factor present in the concentrate. The occasional thrombocytopenia described after Hyate: C infusions probably results from the interaction between platelets and the porcine von Willebrand factor contaminating the concentrate.
Canine platelet alloimmunization: the role of donor selection
Authors: Slichter Sherrill J. , O'Donnell Margaret R. , Weiden Paul L. , Storb Rainer , Schroeder Maria‐Louise
Published: March 2008

Summary

Five different platelet transfusion programmes were evaluated in a canine model to determine the most effective method of providing long‐term platelet support. When a weekly transfusion from a single unrelated donor was used, alloimmune platelet refractoriness developed in 95% of recipients after an average of 3.1 ± 0.7 transfusions, and donor platelets circulated for a total of 8.2 ± 2.3 d. When multiple unrelated donors were used, the percentage of refractory recipients was similar (60% versus 77%) whether platelets came from six single donors given sequentially or from a pool of the same six donors given repeatedly. There was, however, a significant difference in the number of transfusions given prior to immunization (14 ± 5 transfusions lasting 32 ± 12 d for sequential single donors as compared to 5.5 ± 1.0 transfusions lasting 13 ± 2 d for the pooled donors).
A stimulator of murine haemopoietic stem cell proliferation produced by human fetal liver cells
Authors: Cork Michael J. , Riches Andrew C. , Wright Eric G.
Published: March 2008

Summary

Normal murine bone marrow was incubated with medium conditioned by liver cells from human fetuses of 11–17 weeks gestational age. This treatment increased the proportion of murine haemopoietic stem cells (CFU‐S) which were synthesizing DNA from less than 10% to more than 30%. The human fetal liver cell supernatants were fractionated using Amicon filters to obtain nominal molecular weight ranges of 10–30 000, 30–50 000 and 50–100 000 daltons. The stimulator was present only in the 30–50 000 dalton fraction. When human fetal liver cells were separated by adherence to plastic, medium conditioned by the adherent, but not the non‐adherent, cells produced the stimulator. A population of non‐cycling GM‐CFC was not switched into cycle during incubation with this CFU‐S proliferation stimulator. Human fetal livers of all gestational ages tested contain a specific stimulator of CFU‐S proliferation. This appears to have properties similar to that demonstrated in murine fetal liver and regenerating murine bone marrow. Human and murine haemopoietic stem cell proliferation may therefore be regulated by similar mechanisms.
Prediction and prevention of relapse of acute lymphoblastic leukaemia after bone marrow transplantation
Authors: Barrett A. J. , Joshi R. , Kendra J. R. , Philips R. H. , Ashford R. , Shaw P. J. , Hugh‐Jones K. , Hobbs J. R.
Published: March 2008

Summary. Results of bone marrow transplantation (BMT) in 63 adults and children with ALL transplanted in the 5‐year period 1979–83 were analysed. Twenty‐one patients (33%) relapsed, 25% of the group died in relapse and 19% died from complications of BMT. The actuarial disease‐free survival at 6 years was 38%. Relapse after BMT could be predicted by standard prognostic diagnostic features such as age, sex, cell type and presenting blast cell count. Patients transplanted in first remission selected for their poor prognosis had a lower relapse risk than a similar group of poor prognosis patients transplanted in second or subsequent remission (P<0.05). Relapse following second and subsequent remission BMT was predicted by a score based on standard prognostic features or by the pace of the disease: patients with an interval of less than 2 years between diagnosis and first relapse having a 15% actuarial disease‐free survival, compared with 81% for patients with an interval greater than 2 years (P< 0.001). These results emphasize that ALL is a heterogeneous disease and establishes the importance of determining relapse risk when selecting BMT and other treatment schedules for ALL patients.
Healthy HTLV‐I carriers in Japan: the haematological and immunological characteristics
Authors: Yasuda Kaori , Sei Yoshitatsu , Yokoyama Mitchel M. , Tanaka Ken , Har Akinori
Published: March 2008

Summary. The haematological and immunological characteristics of 34 healthy anti‐HTLV‐I antibody‐positive individuals (HTLV‐I carriers) in southwestern Japan were examined. No significant difference was noted between carriers and the controls in counts of RBC, WBC and the absolute number of lymphocytes. The serum IgG in the carriers was higher than that of the controls. The percentages of OKT4, OKT8, OKIal and Bl‐positive cells were found to be normal in the peripheral blood of the carriers, whereas the percentages of OKT11 and anti‐Tac‐positive cells were significantly higher in the carriers than in the controls. A correlation was observed between the percentages of anti‐Tac‐positive cells and the titres of anti‐HTLV‐I antibody in the carriers. After a 72 h incubation of peripheral blood lymphocytes with medium alone, the percentage of anti‐Tac‐positive cells tended to decrease in the controls, but to increase in carriers, with the appearance of large blastoid cells resembling blastic transformed lymphocytes cultured with mitogen. Tac and la antigens were markedly expressed on these large blastoid cells.
Eosinophil production in human bone‐marrow cultures induced by 80–85 kDa serum component(s) of patients with eosinophilia
Authors: Kern P. , Horstmann R. D. , Dietrich M.
Published: March 2008

Eosinophil production from normal human bonemarrow cells was induced by plasma or serum of patients with eosinophilia. After 10–12 d of liquid suspension culture, 14–42% of bone‐marrow cells were identified as maturing eosinophil granulocytes. Upon molecular sieve chromatography, the activity eluted in a single peak corresponding to a substance of 80–85 kDa molecular weight. The activity did not stimulate colony formation in semi‐solid agar media, but was detected in liquid suspension cultures only. Furthermore, addition of exogenous colony‐stimulating factor (CSF) had a differential effect depending on the culture system used. In semi‐solid media, predominance of eosinophil colonies was only observed when CSF was added at a low concentration. In liquid suspension cultures, however, 30–40% of cultured cells were eosinophils independent of the CSF concentration used. These findings extend recent studies in an animal model and support the hypothesis that a differentiation factor is involved in the regulation of eosinophil production in man.
In vitro and in vivo effects of treatment by platelet‐activating factor on N‐formyl‐met‐leu‐phe‐mediated responses of polymorphonuclear leucocytes
Authors: Ingraham Leah M. , Lafuze Joan E. , Boxer Laurence A. , Baehner Robert L.
Published: March 2008

Two chemoattractants, the peptide N‐formyl‐met‐leu‐phe (FMLP), and the ether phospholipid, platelet activating factor (PAF), each stimulate a variety of in vitro responses in polymorphonuclear leucocytes (PMN). Because often more than one inflammatory mediator is active during inflammation, we determined the effect on PMN of sequential stimulation with these two agents. Before FMLP stimulation, human PMN were exposed to PAF, at concentrations which gave little or no response when administered alone. PAF enhanced FMLP‐elicited superoxide release in a dose‐dependent fashion. Likewise, release of granular lysozyme from the cells was increased in PAF treated cells. Similar treatment with other phospholipids, including the lyso derivation of PAF, failed to produce these effects. Incubation with nordihydro‐guaiaretic acid, an inhibitor of arachidonic acid metabolism, had little effect on the enhancement of lysozyme release by PAF. To determine if enhancing effects by PAF might occur also in vivo, we studied rabbits receiving PAF and/or FMLP intravenously. When rabbits received 0·01 μg PAF (a dose which does not elicit the sustained neutropenia observed with higher doses of PAF) followed by 0·05 μg FMLP the absolute granulocyte count (AGC) dropped at 1 min (46 ± 11% of original value), and continued to fall (24 ± 12% at 10 min). Controls, treated with the suspending fluid for PAF, and then 0·05 μg FMLP, had a similar 1 min AGC value, but at 10 min AGC returned to 65±6·1% (P<0·001 for comparison of 10 min values). Thus PAF pretreatment enhanced FMLP‐elicited granulocytopenia in vivo. Study of in vitro human PMN aggregation revealed that, at certain relative concentrations of PAF and FMLP. aggregation was enhanced. These studies show that both in vitro and in vivo responses of FMLP‐stimulated PMN may be exaggerated by pre‐exposure to PAF.
In vivo evidence of intravascular binding sites for coagulation factor IX
Authors: Stern David M. , Knitter Glen , Kisiel Walt , Nawroth Peter P.
Published: March 2008

Previous studies have demonstrated that factors IX/IXa bind to specific sites on the surfaces of cultured and native endothelium in vitro and that these sites should be occupied with factor IX in homeostasis. Since factor IX of different species binds to endothelium in a similar manner, we examined if infusion of heterologous factor IX into an animal should result in displacement of host factor IX antigen from its vessel wall site. Experiments were carried out in baboons with a large excess of bovine factor IX employing species‐specific radioimmunoassays. The results indicate that infusion of bovine factor IX or active site‐blocked factor IXa, but not prothrombin, resulted in a dose‐dependent rise in the plasma level of baboon factor IX antigen. This suggested that the infused factor IX was displacing the host clotting factor from some reservoir easily accessible to the intravascular space. Consistent with this hypothesis, infusion of 125I‐factor IX demonstrated accumulation in multiple organs. Radioiodinated factor IX comigrating with the initial tracer on SDS‐PAGE could be eluted from the luminal surface of pulmonary artery and aortic segments. 125I‐factor IX was not significantly associated with cellular elements of the blood. These results suggest that there is a pool of non‐circulating factor IX which is accessible to the intravascular space, widely distributed and involves endothelium.
Dysmegakaryocytopoiesis in acute leukaemias: its predominance in myelomonocytic (M4) leukaemia and implication for poor response to chemotherapy
Authors: Jinnai Itsuro , Tomonaga Masao , Kuriyama Kazutaka , Matsuo Tatsuki , Nonaka Hiroaki , Amenomori Tatsuhiko , Yoshida Yoshiharu , Kusano Miyuki , Tagawa Masuko , Ichimaru Michito
Published: March 2008

Megakaryocytopoiesis was morphologically investigated in 129 adults with de novo acute leukaemia. Three types were identified: type I (84 cases), no detectable megakaryocytes; type II (32 cases), quantitatively preserved megakaryocytes with normal morphology; type III (13 cases): quantitatively preserved megakaryocytes but with distinct dysplastic changes such as micromegakaryocytes and megakaryocytes with multiple small separated nuclei. Type III was found in M1 (one out of 21 cases), M2 (one out of 20 cases), M4 (eight out of 24 cases), M6 (two out of four cases) and hypoplastic leukaemia (one out of 13 cases). M3 cases were all classified into type I. Most of acute lymphoid leukaemia cases (21 cases) belonged to type II. Among AML cases, the complete remission (CR) rate by intensive chemotherapy with daunorubicin and cytosine arabinoside was significantly lower in type III (11%) than in types I (87%) and II (71%). Among M4 cases, CR rates in type III (14%) was also significantly lower than those in type I (75%) and II (100%). Thus, the present study indicates the importance of recognizing dysmegakaryocytopoiesis in AML for clariflcation of the heterogeneous biology or pathophysiology of acute leukaemias and formulation of an appropriate therapeutic strategy.
Cytogenetics of bone marrow fibroblastic cells in idiopathic chronic myelofibrosis
Authors: Greenberg Bernard R. , Woo Linda , Veomett Iris C. , Payne Claire M. , Ahmann Frederick R.
Published: March 2008

To determine the nature of the fibroblastic proliferation in myelofibrosis, we studied the cytogenetic composition of a pure population of in‐vitro bone marrow fibroblastic cells (FC) from a patient with idiopathic chronic myelofibrosis who had trisomy 8 in her unstimulated peripheral blood cells. This cytogenetic abnormality was absent in all the FC studied supporting the premise that in the myeloproliferative disorders the myelofibrosis represents a secondary or non‐neoplastic reaction to the clonal proliferation of haematopoietic stem cells.
Production of factor VIII deficient plasma by immunodepletion using three monoclonal antibodies
Authors: Takase Toshio , Rotblat Frances , Goodall Alison H. , Kernoff Peter B. A. , Middleton Sarah , Chand Sadhna , Denson K. W. , Austen Denis E. G. , Tuddenham Edward G. D.
Published: March 2008

Factor VIII deficient plasma was made from pooled, HIV antibody and hepatitis B antigen screened, normal human plasma by cryoprecipitation and immunodepletion, using three different monoclonal antibodies bound to Sepharose columns, in series. These monoclonal antibodies are specific respectively for von Willebrand factor, factor VIII heavy chain and factor VIII light chain.

The immunodepleted plasma contained less than 0·002 u/ml factor VIII coagulation activity (VIII:C) less than 0·0001 u/ml von Willebrand factor antigen and 1–2 g/l fibrinogen, while the levels of other clotting factors were unchanged. This immunodepleted plasma was compared with commercial factor VIII deficient plasma obtained from a severe haemophilia A patient as substrate in the one‐stage factor VIII assay. Plasmas obtained from 20 normal subjects and 28 patients with von Willebrand's disease or haemophilia A were assayed for VIII:C using the two substrates. The results were very highly correlated (r = 0·96). The columns have high capacity and can be regenerated at least 10 times. Large‐scale production of a substrate for factor VIII assays free of virus contamination is now feasible.
An abnormal antithrombin III (AT III) with low heparin affinity: AT III Clichy
Authors: Aiach M. , Franclois D. , Priollet P. , Capron L. , Roncato M. , Alhenc‐Gelas M. , Fiessinger J.‐N.
Published: March 2008

We have identified an inherited qualitative deficiency of antithrombin III (AT III) in a family with apparently no increased incidence of venous thrombosis. Plasma antithrombin and anti‐Xa activities were normal, but the interaction with heparin, heparan sulphate and low molecular weight heparin was uniformly decreased. An immunoblotting technique performed in plasma showed normal complex formation with thrombin. By using heparin‐Sepharose affinity chromatography and crossed immunoelectrophoresis, the variant could be separated: at least two fractions of low affinity AT III were obtained. A minor one had no antiprotease activity; the other one was further purified to homogeneity and found to have normal specific activity in absence of heparin and a 50% decreased activity in presence of heparin. We propose to call this new variant AT III Clichy.
Antithrombin III Glasgow: a variant with increased heparin affinity and reduced ability to inactivate thrombin, associated with familial thrombosis
Authors: Lane D. A. , Lowe G. D. O. , Flynn A. , Thompson E. , Ireland H. , Erdjument H.
Published: March 2008

A functional antithrombin III (AT III) deficiency has been identified in two generations of a family with a high incidence of thrombosis. The deficiency presented as 50% reduction in heparin cofactor activity compared to its antigen concentration. No abnormality was detected by crossed immunoelectrophoresis in the presence or absence of heparin. Plasma from the propositus was precipitated with dextran sulphate, applied to heparin‐Sepharose and the AT III stepwise eluted with NaCl. The AT III had a reduced ability to inactivate thrombin, when this was monitored by substrate hydrolysis or by SDS polyacrylamide gel electrophoresis. Its mobility was normal by the latter technique using 10–20% gradient gels under reducing and non‐reducing conditions. AT III from the patient was reapplied to heparin‐Sepharose and eluted with a NaCl gradient. An active pool eluted in the same NaCl concentration range used to purify normal AT III, while predominantly inactive AT III eluted at higher NaCl concentrations. It is concluded that this variant, designated AT III Glasgow, has increased affinity for heparin but reduced ability to inactivate thrombin.
Immunoreactive prolyl hydroxylase in patients with primary and secondary myelofibrosis
Authors: Wang Jen C. , Winston Ching WonC , Kao Whei‐Yang
Published: March 2008

Summary. Prolyl hyroxylase (PH) is an important enzyme in collagen synthesis. It is required for the hydroxylation of prolyl residues in peptide chains in collagen synthesis. Serum PH activity was measured in patients with primary myelofibrosis (agnogenic myeloid metaplasia and myelofibrosis with prior history of polycythaemia Vera), in patients with secondary myelofibrosis (in association with carcinoma metastasis), in patients with other myeloproliferative disorders and in controls (anaemia patients and normal volunteers). Both primary and secondary myelofibrosis had significantly elevated PH values. while other myeloproliferative disorders did not differ from normal controls. Increased PH levels in patients with primary and secondary myelofibrosis may signify increased collagen synthesis and may serve as an indicator for the development of fibrosis in the course of myeloproliferative disease.
Androgen therapy in haemodialysis patients.
Authors: Solomon L. R. , Hendler E. D.
Published: March 2008

Summary. Activities of the red cell enzymes hexokinase, glucose 6‐phosphate dehydrogenase, 6‐phosphogluconate dehydrogenase, lactic dehydrogenase and aspartate amino‐transferase were measured in 17 chronic haemodialysis patients receiving androgen therapy, 15 untreated chronic haemodialysis patients and 15 normal subjects. Compared to normal subjects, untreated haemodialysis patients had similar reticulocyte counts but significantly increased levels of all five enzymes studied. This finding suggests the presence of a younger red cell population in the peripheral blood and is consistent with the shortened red cell survival known to occur in this clinical setting. Red cell enzyme activities in untreated haemodialysis patients were significantly corre‐lated with one another and with the serum phosphate level. Moreover, in this population, red cell DPG content was directly related to hexokinase and glucose 6‐phosphate dehydrogenase activities while haemoglobin‐oxygen affinity (P50) was inversely related to all five enzyme activities. In contrast, in androgen‐treated haemodialysis patients, despite higher reticulocyte counts, red cell enzyme activities were the same or lower than those in the untreated haemodialysis group and only slightly higher than those in normal subjects, suggesting an overall older red cell population. Moreover, relationships of red cell enzymes to one another, to serum phosphate‐ levels and to both red cell DPG content and haemoglobin‐oxygen affinity were significantly different in androgen‐treated subjects than in the untreated haemodialysis group. These changes are consistent with a direct effect of androgens on red cell metabolism and an improved red cell survival during androgen therapy.
Development of the phagocytic and cidal capacity during maturation of myeloid cells: studies on cells from patients with chronic myelogenous leukaemia
Authors: Segel Erik K. , Ellegaard Jørgen , Borregaard Niels
Published: March 2008

Summary. Myeloid cells from peripheral blood of patients with chronic myelogenous leukaemia were isolated and fractionated by density gradient centrifugation using Lymphoprep gradient followed by discontinuous Percoll gradients. Six fractions were obtained, each enriched in one of the morphologically identifiable types of myeloid cells from myeloblasts to polymorphonuclear neutrophils. Each of these cell types were functionally and biochemically characterized.

The development of the capacities for phagocytosis and killing of yeast cells and the ability to generate a respiratory burst of phagocytosis correlated closely with the content of cytochrome b and vitamin B12‐binding protein, a marker of specific granules. These results support the notion that the specific granules provide the developing neutrophil with components which are essential for its microbicidal activity.
Absence of functional activity of tissue plasminogen activator in patients with severe forms of von Willebrand's disease
Authors: Jeanneau Ch. , Bachouchi N. Ounhoughene , Gorin I. , Meimon G. , Sultan Y.
Published: March 2008

Summary. Some patients with von Willebrand's disease do not respond to stimuli such as venous occlusion and infusion of a vasopressin analogue DDAVP. In these patients, fibrinolytic activity is not enhanced and von Willebrand's factor is not released into the blood. Skin biopsies and cryostat sections were used to study the fibrinolytic activity of skin vessels and localization of tissue plasminogen activator (t‐PA) in three patients with severe form of von Willebrand's disease. On fibrin films, no fibrinolysis developed around the skin vessels of the patients; however, using specific polyclonal and monoclonal antibodies to t‐PA, and peroxidase coupled specific IgG, presence of t‐PA antigen was demonstrated in endothelial cells (EC) of all of them. In plasma no t‐PA activity was detected either before or after venous occlusion although t‐PA inhibitor activity was in a normal range. Small amounts of t‐PA antigen was measured in blood by ELISA. From these results, it is concluded that in patients with severe forms of von Willebrand's disease, t‐PA present in EC is not functional and can not transform plasminogen into plasmin.
Factor XIII: a marker of mono‐ and megakaryocytopoiesis
Authors: ÁDány Róza , Kiss Attila , Muszbek László
Published: March 2008

Previous studies have shown that human platelets, megakaryocytes and peripheral blood monocytes contain the subunit α of plasma factor XIII (FXIII) which plays a key role in fibrin stabilization. To study the expression of FXIII subunit α during differentiation of the mono‐and megakaryocytic cell lines, bone marrow smears were examined by immunomorphological methods. In addition to megakaryocytes, FXIII was detected in a large number of cells by a highly sensitive immunoperoxidase staining. Characterization of these cells was carried out by double immunofluorescent labelling in which the detection of FXIII subunit α was combined by the labelling of either Leu M3 monocyte/macrophage surface antigen or platelet factor 4 (PF4) a marker for megakaryocytic cells. On the course of differentiation from early precursors to mature megakaryocytes all phenotypic form of the megakaryocytic cell line expressed FXIII subunit α though there were considerable changes in its subcellular distribution pattern. Leu M3 positive cells of monocytopoiesis, i.e. monocytes and promonocytes and in all probability monoblasts as well, were also labelled for FXIII. On this basis FXIII subunit α could be considered as a common marker of megakaryo‐ and monocytopoiesis and its immunomorphological detection might provide a useful diagostic tool for identifying normal and perhaps also malignant differentiation forms of these cell lines.
Splenic platelet‐sequestration following routine blood transfusion is reduced by filtered/washed blood products
Authors: Bareford D. , Chandler S. T. , Hawker R. J. , Jackson N. , Smith M. , Boughton B. J.
Published: March 2008

The mean fall in the platelet count following 23 routine transfusions of 3–5 units packed cells for anaemia was 32.5%. This was significantly reduced to 12.5% in 15 similar transfusions through a 40 μm microaggregate filter (P<0.01) and to 4.6% following five transfusions through a polyester fibre filter (P<0.005). In 10 transfusions with frozen or polyester fibre filtered, washed red cells, the decrease in platelet count was 4.2% (P<0.001). A study of 111In‐oxine labelled autologous platelets in nine patients indicated that the fall in platelet count was due to increased splenic sequestration. Since thrombocytopenia following routine transfusion is reduced by procedures which filter the packed cells, the decrease in platelets is probably caused by their adherence to infused microaggregate debris causing their premature removal from the circulation. Patients with preexisting thrombocytopenia who receive red cell transfusions for anaemia, should therefore receive blood products depleted of microaggregate debris in order to avoid exacerbation of thrombocytopenia and haemorrhagic complications.
A new unstable haemoglobin variant: Hb Shanghai [β131 (H9)Gl→Pro] found in China
Authors: Zeng Y. T. , Ren Z. R. , Chen M. J. , Zhao J. Q. , Qiu X. K. , Huang S. Z.
Published: March 2008

A 34–year‐old woman patient was found to have a chronic hereditary haemolytic anaemia. No abnormal haemoglobin band was detected by conventional electrophoresis, but a slow β chain could be separated on urea‐carboxymethyl cellulose chromatography. Investigations of the patient's haemoglobin revealed an unstable component. Analyses of chemical structure, including isolation and TPCK trypsin digestion of the abnormal globin chain, HPL chromatography, amino acid composition as well as sequence determination of the abnormal peptide, indicated that a glutamine was replaced by a proline at position β 131 (H9). Biosynthesis studies demonstrated a normal rate of synthesis but relatively fast degradation of the mutant β chain. The new variant is named as Hb Shanghai according to the place where it was discovered.
The purification of human basophils: their immunophenotype and cytochemistry
Authors: Bodger Michael P. , Newton Lesley A.
Published: March 2008

An IgM class monoclonal antibody Bsp‐1 that selectively reacts with human basophils was used to label basophils in normal and leukaemic haemopoietic samples. The immunophenotype of Bsp‐1+ basophils was determined using a panel of 21 IgG subclass monoclonal antibodies in two‐colour immunofluorescence assays. Basophils expressed the leucocyte common antigen, HLA‐ABC antigens and antigens defined by CD1 1 and CD1 3 monoclonal antibodies. Other myeloid cell (granulocyte‐monocyte) associated antigens, lymphoid cell surface determinants and HLA‐DR antigens were not detected. Basophil preparations of 95–98% purity were obtained from the peripheral blood of patients with CML and umbilical cords using fluorescence activated cell sorting techniques. Purified basophils exhibited metachromatic staining with toluidine blue, alcian blue and astra blue. PAS staining was observed in 7% of cord Bsp‐1+ cells and of 55% CML Bsp‐1+ cells. Between 5% and 10% of basophils were chloroacetate esterase‐positive which suggests that some Bsp‐1+ cells are immature basophils.
Autoantibodies against platelet GPIIb/IIIa in chronic ITP react with different epitopes*
Authors: Tsubakio T. , Tani P. , Woods V. L. , McMillan R.
Published: March 2008

Some patients with chronic immune thrombocytopenic purpura have autoantibodies to the platelet glycoprotein IIb/IIIa (GPIIb/IIIa) complex. To determine whether these autoantibodies are directed towards the same or different epitopes, we evaluated the ability of four murine monoclonal anti‐GPIIb/IIIa antibodies specific for different epitopes to block autoantibody binding. We noted a variation in blocking patterns among autoantibodies from patients with chronic ITP. In addition, we were able to map the relative epitope locations of both the autoantibodies and the monoclonal antibodies. These data show that the anti‐GPIIb/IIIa monoclonal autoantibodies in chronic ITP are directed towards different epitopes.
Establishment of three permanent human leukaemia cell lines producing immunoreactive calcitonin
Authors: Koeppler H. , Pflueger K. H. , Knapp W. , Havemann K.
Published: March 2008

Summary. Three permanent human leukaemia cell lines, designated EW2, LG3 and MS6, were established from bone marrow aspirates of a patient with acute myelomonocytic leukaemia (EW2), acute monocytic leukaemia (LG3) and acute myeloblasts leukaemia (MS 6). In vitro all lines exhibited a myelomonocytoid marker profile in terms of classical staining reactions and cell‐surface antigen structure. In supernatants and extracts of EW2 and MS 6 cells immunoreactive human calcitonin was found in raised levels. No significant levels of immunoreactive human calcitonin were found in supernatants of LG3 cells, while extracts of LG3 cells and the patients sera at diagnosis contained high levels, suggesting the in vitro selection of a non‐secreting clone of leukaemic cells. Gel‐chromatography showed several peaks of ihCT with higher molecular weight than normal human calcitonin, suggesting the production of precursor molecules. In addition to ihCT raised levels of immunoreactive calcitonin gene related peptide (ICGRP) was found in supernatants of EW2. These data support the concept of ectopic immunoreactive human calcitonin production by leukaemic cells.
Serum erythropoietin in rheumatoid arthritis and other inflammatory arthritides: relationship to anaemia and the effect of anti‐inflammatory treatment
Authors: Birgegård Gunnar , Hallgren Roger , Caro Jaime
Published: March 2008

Summary. Serum erythropoietin (s‐Epo) was measured with a sensitive radioimmunoassay method in 58 patients with classical rheumatoid arthritis (n= 41) or seronegative spondyloarthropathies (n= 17). Epo was significantly (P<0‐001) increased and on an average two times higher than in a healthy population. A correlation was found between Hb and s‐Epo (r= ‐0·46, P<0·005), indicating that these patients respond to anaemia with an increase in s‐Epo. In order to investigate if inflammation has a direct influence on s‐Epo levels a short period of corticosteroid treatment was given to rapidly decrease inflammatory activity. No increase in s‐Epo was seen after 1 week. Furthermore, there was a correlation between s‐Epo and ESR in all patients (r=0·59, P<0·01). These results indicate that s‐Epo is directed by the Hb level, which in turn is influenced by the inflammatory activity: a higher inflammatory activity gives a lower Hb and an increase in s‐Epo. In comparison to previously published figures for the relation between Hb and s‐Epo these patients seem to have an ordinary Epo response. We conclude that the anaemia of patients with chronic inflammatory joint disease is not caused by a diminished Epo production.
Autoimmune haemolytic anaemia in childhood associated with non‐complement binding IgM autoantibodies
Authors: Salama A. , Mueller‐Eckhardt C.
Published: March 2008

Red cell bound IgM autoantibodies free of associated IgG autoantibodies and unable to activate complement have not yet been considered as the cause of autoimmune haemolytic anaemia (AIHA). We report on warm type AIHA resulting from the action of IgM autoantibodies on circulating RBC. Twelve children (eight of whom were infants) with relatively severe haemolytic anaemia were studied. Whereas the clinical findings and courses of all children appeared to be compatible with warm type AIHA, the serological findings during the haemolytic phase were atypical in that the direct antiglobulin test (DAT) was negative in 11, and positive in one case due to C3d only. The use of radiolabelled antihuman globulin antibodies showed, however, that the RBC of all the patients were sensitized with warm IgM antibodies. Elevated values of IgA and/or IgG immunoglobulins on the patients’ RBC were found to be present in only two cases. Complement activation by IgM autoantibodies could not be detected in all other cases neitherin vivonorin vitro.Thus, we conclude that non‐complement binding IgM autoantibodies were responsible for the AIHA in the majority of these children possibly representing an as yet unrecognized variant of warm type AIHA.
Differential pattern of oncogene and beta‐actin expression in leukaemic cells from AML patients
Authors: Blomberg J. , Andersson M. , Fäldt R.
Published: March 2008

The presence of mRNA hybridizing to five oncogene (fos, mos, myc, sis, abl) and a beta‐actin probe(s) was studied with a semiquantitative dot‐blot procedure in eight AML patients. Around 107 leucocytes, corresponding to 1–10 ml blood, sufficed for the analysis. Each patient, regardless of the FAB group, exhibited a distinct pattern of oncogene or beta‐actin expression. Especially strong signals were obtained with the beta‐actin probe in two patients and with the fos probe in three patients. The findings underscore the heterogeneity of AML, either present from the first step towards malignancy or arising during the progression of the disease. The pattern of oncogene expression in leukaemic cells studied in routine blood samples may become an additional means for classification and follow‐up of AML patients.
The effect of anticoagulant on platelet associated IgG
Authors: Lucas G. F. , Holburn A. M.
Published: March 2008

Blood anticoagulated with solid K2EDTA had platelet associated IgG (PAIgG) levels five‐fold greater than citrated blood when the platelets were harvested by differential centrifugation but identical PAIgG levels when the platelets were harvested by Percoll density gradient. Furthermore, blood anticoagulated with increasing amounts of solid K2EDTA demonstrated a proportionate increase in both PAIgG and haemolysis whereas the same blood sample anticoagulated with aqueous K2EDTA exhibited neither of these effects. Coulter Counter analysis of platelet poor plasma (PPP) prepared from blood anticoagulated with solid K2EDTA revealed particles with a mean volume greater than normal RBC; such particles were not found in the PPP prepared from citrated blood. While some of these particles could be sedimented through plasma, those remaining were sufficiently buoyant to resist sedimentation unless the plasma was diluted with buffer. Studies with FITC conjugated antisera revealed that the particles could be classified into two types. Type I particles reacted strongly with a monoclonal anti‐RBC antibody, weakly with anti‐IgG antibody and had the appearance of stroma. Type II particles lacked any apparent biological structure, reacted strongly with anti‐IgG antibody but not with monoclonal anti‐RBC antibody. Thus, the buoyant IgG‐rich particles found in blood anticoagulated with solid K2EDTA appear to contribute to the high PAIgG levels associated with this anticoagulant. Consequently, solid K2EDTA should not be used to anticoagulate blood destined for PAIgG measurement.
The effect of total body irradiation and bone marrow transplantation during childhood and adolescence on growth and endocrine function
Authors: Leiper A. D. , Stanhope R. , Lau T. , Grant D. B. , Blacklock H. , Chessells J. M. , Plowman P. N.
Published: March 2008

Summary. Seventeen children (11 M, 6 F) with acute leukaemia and myeloproliferative disorders were investigated for growth and endocrine dysfunction. All had undergone bone marrow transplantation prepared with cyclophos‐phamide and single fraction total body irradiation (900–1000 cGy) between 1.5 and 3.8 (mean 2.2) years previously. The majority of children exhibited growth failure, which was of multiple aetiology. Ten patients, of whom eight had had previous prophylactic cranial irradiation, had evidence of growth hormone deficiency based on the reduced growth hormone response to insulin induced hypoglycaemia. Three patients had evidence of hypothalamic damage as shown by their growth hormone response to 200μg GHRH (1–29) NH2 intravenously. Gonadal failure was common, assessed clinically, and biochemically by basal gonadotrophin and sex steroid concentrations. All four girls of adolescent age (10.6–14.1 years) had ovarian failure requiring sex steroid replacement. Of the eight boys of adolescent age (12.3–18.3 years), two had testicular failure requiring sex steroid supplements. Both of these had had previous testicular irradiation. Five others had compensated gonadal failure, and one had normal Leydig cell function.

Abnormalities of the TSH response to TRH occurred in 10 patients but only three had overt hypothyroidism. Unlike growth hormone deficiency, gonadal and thyroid dysfunction showed no correlation with previous cranial radiotherapy.
In vitro multimerization of human von Willebrand factor from its subunits
Authors: Rand Jacob H. , Chu Sadie V. , Potter Barry J.
Published: March 2008

Summary. von Willebrand factor (vWF) is a multimeric glycoprotein composed of subunits having an estimated molecular mass of 220–240 kDa which are linked by disulphide bonds. Intact, unreduced vWF, has ristocetin cofactor activity, which is lacking in the reduced subunits. To determine whether reduced vWF subunits are capable of being re‐multimerized vWF was purified, reduced and then subsequently dialysed in the absence of reducing agents. (1) Purified vWF, (2) reduced vWF, and (3) reduced and dialysed vWF were compared for size composition and for ristocetin co‐factor activities. SDS‐PAGE showed that the purified vWF failed to enter the gels, while the reduced vWF migrated to a position consistent with a mass of approximately 240 kDa. Reduced and subsequently dialysed vWF subunits multimerized into larger forms, most of which did not enter the gels. No residual 240 kDa material was detectable following remultimerization. vWF multimer analysis confirmed the loss of monomer in the reduced and dialysed material and the appearance of new multimers with a size range smaller than the native material. Reduced vWF showed no detectable ristocetin co‐factor activity, while the re‐multimerized material regained some activity. Thus, we deduce that vWF can spontaneously re‐multimerize from its reduced subunits and regain a small but measurable quantity of ristocetin co‐factor activity.
Functional analysis of a clonal expansion of Leu 11 positive NK active lymphoid cells
Authors: Hart D. N. J. , Leahy M. F. , McKenzie J. L. , Furley A. J. , Carter J. , Beard M. E. J.
Published: March 2008

A female patient with an unusual lymphoproliferative disease associated with marked neutropenia has been observed for 36 months. The expanded cell population consists of large lymphocytes, many of which contain large azurophilic granules with acid phosphatase activity. These cells were T3, T8, Tl1 and Leu 11 positive but lacked the Ml, T10, IL‐2 receptor and HLA.DR antigens. The majority of these cells (60‐70%) were also Leu 7 (HNK‐1) positive. Strong natural killer (NK) activity was found in both the Leu 7 positive and negative cell populations. This cytotoxic activity was inhibited by monoclonal antibodies known to inhibit NK activity but was unaffected by antibodies which block T cell and T/NK cell cytotoxicity. Further functional analysis indicated that these cells suppressed normal T cell responses to mitogens, MLC responses and PWM induced B cell immunoglobulin synthesis. No effect on bone marrow progenitor cell growth was demonstrated. Antibody dependent cellular cytotoxic (ADCC) activity was barely detectable despite the presence of the Leu 11 antigen. Southern blot DNA analysis demonstrated clonal rearrangement of the T cell receptor β gene thereby confirming that this variant of Ty lymphoproliferative disease was a neoplastic condition.