bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of human neutrophil glycoproteins expressing the CD15 differentiation antigen (3‐fucosyl‐N‐acetyllactosamine)
Authors: Albrechtsen Merete , Kerr Michael A.
Published: March 2008

Summary. The expression of the CD15 antigen. 3‐fucosyl N‐acetyllactosamine, on neutrophil glycoproteins has been studied by SDS gel electrophoresis and immunoblotting. The antigen is expressed on several glycoproteins, both intracellularly and on the cell surface. Each subcellular compartment appears to contain a specific antigen. A soluble, granule glycoprotein (Mr 80–90K) probably accounts for most of the intracellular staining detected immunohistochemically. Membrane glycoproteins of Mr. 85–90K and 25K are associated with granule membranes, the latter being an integral membrane protein. The CD15 antigen is expressed on several cell surface glycoproteins with Mr in the range of 165K and 105K. These antigens are also contained in an intracellular pool which is brought to the surface on activation of the cells with chemotactic peptides.

The 165K and 105K antigens show identical electrophoretic mobility to two of the major glycoproteins detectable by PAS or protein staining of gels of detergent extracts of cell membranes. These glycoproteins include the complement receptor, CR3. The beta chain of CR3 (105K) and to a lesser extent the alpha chain (165K) express CD15; however, most of the CD15 antigen is associated with other glycoproteins of these molecular masses.
Effects of verapamil on the cellular accumulation of daunorubicin in blast cells and on the chemosensitivity of leukaemic blast progenitors in acute myelogenous leukaemia
Authors: Maruyama Yasuo , Murohashi Ikuo , Nara Nobuo , Aoki Nobuo
Published: March 2008

Summary. Verapamil, a calcium channel blocker, was studied for its effects on the cellular daunorubicin (DNR) accumulation in blast cells and on the sensitivity of the blast progenitors to DNR in 30 acute myelogenous leukaemia (AML) patients. Using flow cytometry, verapamil was shown to increase the accumulation of DNR in blast cells. The effect was more prominent in the patients who showed poorer response to chemotherapy including DNR. The per cent increases of DNR content by verapamil were 6·4 ± 6·3% and 19·5 ± 23·1% in the 16 responders and the 14 nonresponders, respectively (P < 0·05). The data suggest the presence of enhanced efflux of DNR in nonresponders. Marked variation in the effects of verapamil among nonresponders suggests the heterogeneity of the mechanisms of drug resistance involved. Verapamil also enhanced the sensitivity of blast progenitors to DNR. The degree of increase of cellular DNR accumulation by verapamil correlated with the degree of increase in chemosensitivity of blast progenitors (nonresponders, P < 0·005; responders, P < 0·05). We conclude that enhanced efflux of DNR in blast progenitors may be related to remission induction failure in at least some of resistant AML patients.
Factor VII and extrinsic pathway inhibitor in acute coronary disease
Authors: Sandset Per Morten , Sirnes Per Anton , Abildgaard Ulrich
Published: March 2008

Summary. This report describes studies on the activation of coagulation factor VII (FVII) and the inhibition of the extrinsic coagulation pathway in acute ischaemic heart disease. FVII and the inhibitor of the tissue thromboplastin‐FVII complex, called extrinsic pathway inhibitor (EPI), were determined in plasma from 68 patients and compared to findings in 3 7 normal individuals. The mean FVII amidolytic activity, the mean FVII clotting activity, as well as the FVII clotting/FVII amidolytic ratio were not significantly different in the patient groups as compared to the controls. The fraction of FVII clotting activity that is sensitive to phospholipase C, ‘the FVII‐phospholipid complex’, was 8% in controls, 19% (P <0·05) in patients with acute myocardial infarction, 1 5% (n.s.) in angina pectoris and 13% (n.s.) in heart failure/arrhythmia patients. The ‘FVII‐phospholipid complex’ was highly significantly correlated to triglycerides in plasma in patients with acute myocardial infarction (r= 0·88, P < 0·001) and angina pectoris (r=0·89, P < 0·001). The mean EPI levels were significantly increased in patients with acute myocardial infarction (132%), angina pectoris (134%), and heart failure (150%) as compared to the control population (110%). The FVII clotting/EPI ratio was significantly decreased both in patients with acute myocardial infarction and heart failure, whereas the FVII amidolytic/EPI ratio was significantly decreased only in the heart failure group. Apparently, in patients with acute ischaemic heart disease, a moderate increase in the procoagulant activity is accompanied by a marked increase in the anticoagulant activity of the extrinsic coagulation pathway, suggesting a balanced activation system.
Correlation of membrane anisotropy with function in subpopulations of human blood platelets*
Authors: Raha S. , Opper C. , Wesemann W.
Published: March 2008

Summary. Human blood platelets were fractionated on a discontinuous Percoll gradient into high density (HD), intermediate (ID), and low density (LD) platelets. The subpopulations were characterized with regard to [14C]serotonin uptake and release, cAMP content, aggregation, and membrane anisotropy, which is inversely related to membrane fluidity. Membrane anisotropy, which was high in LD platelets, was found to decrease with increasing density (LD> ID > HD). LD platelets showed significant lower cAMP levels and [14C]serotonin uptake than the total platelet population (TPP) and ID and HD platelets. Upon ADP and serotonin stimulation the cAMP content was reduced in all platelet populations with the exception of HD platelets in which cAMP was unchanged. Upon thrombin stimulation the cAMP content was reduced only in TPP and LD platelet population and it was increased in HD platelet population. Thrombin activation changed the anisotropy only in LD platelets. Thrombin at a concentration of 0·001 U/ml reduced whereas 0·01 and 0·05 U thrombin/ml increased the membrane anisotropy significantly. As compared with TPP and the other subpopulations, LD platelets were most sensitive upon ADP and thrombin stimulated [14C]serotonin release as well as upon ADP, serotonin and thrombin induced aggregation. The findings suggest that the differing functional abilities of the platelet subpopulations are correlated to the various membrane anisotropies observed in these fractions.
Immunoblot analysis of platelet glyeoprotein IIb in patients with Glanzmann thrombasthenia in Israel
Authors: Seligsohn Uri , Coller Barry S. , Zivelin Ariela , Plow Edward F. , Ginsberg Mark H.
Published: March 2008

Summary. Recent studies have indicated that severe (‘type I’) Glanzmann thrombasthenia is a heterogeneous hereditary disorder caused by quantitative and/or qualitative abnormalities of platelet membrane glycoproteins (GP) IIb and IIIa. Immunoblot analysis of sodium dodecyl sulphate (SDS)‐solubilized platelets was carried out on controls and 18 patients (12 Iraqi‐Jews, two Iranian Jews and four Arabs) employing three antibodies (one monoclonal and two poly‐clonal) directed at different sites on GPIIb. Nonreduced control platelet samples contained a major Mr∼ 140k immunoreactive protein that was split into an Mr∼ 120k (α) and an Mr∼25k (β) band after reduction with mercaptoethanol. The nonreduced samples from all 18 patients tested had trace amounts of Mr∼ 140k band corresponding to normal GPIIb; the intensity of this band was estimated to be < 1% of the normal amount. Unlike the control samples, however, this Mr∼ 140k band did not change electrophoretic mobility following reduction. Since GPIIb originates from a single chain precursor molecule of Mr∼ 140k that comprises both the α and β chains, and which does not change mobility with reduction, our data suggest that the platelets of these patients contain small amounts of this precursor.
Protein S in the first year of life
Authors: Sthoeger Dalia , Nardi Michael , Karpatkin Margaret
Published: March 2008

Summary. Total protein S, free protein S and C4b binding protein were measured in healthy term infants in order to establish the normal levels of these proteins during the first year of life. Total protein S rose from 36·5% of the adult mean level on day 1 of life to 90% between 6 and 12 months of age. Free protein S was 54·2% of the adult mean on day 1 of life, all values were in the adult range by 2 months and the mean value was no different from the adult at 4 months. This relatively high level of free and presumably active protein S reflects low levels of C4bBP at birth (28·8% of the adult mean) and a slow postnatal rise.
Should anti‐Rh immunoglobulin be given to D variant women?
Authors: Lubenko Anatole , Contreras Marcela , Habash Janet
Published: March 2008

Summary. D variant women occasionally form anti‐D during or following pregnancy with a D‐positive fetus. It is not known whether Rh immunization could be suppressed by using anti‐Rh immunoglobulin (Rh Ig), or whether the injected antibodies would be absorbed by the woman's D variant cells. In order to predict the likely outcome, three anti‐Rh Ig preparations were absorbed independently with five examples of D variant red cells: R1VIr (n = 3), R1Br(n= 2), and with cells of common Rh‐positive and Rh‐negative phenotypes: R0, r′r, r″r or rr cells (n= 1 each). The titres of the Ig preparations against all these cells were compared after three, six, nine and 12 absorptions.

The titres of the unabsorbed anti‐Rh Igs against R0 cells ranged from 215 to 218. Anti‐D activities were hardly affected by absorption with r′r, r″r or rr cells, but were completely exhausted after three to six absorptions with R0 cells. Most of the D variant cells were as ineffective as rr cells in reducing anti‐D activity against R0 cells, but one R1VIr variant produced a fall in titre against R0 cells, equivalent to four or five doubling dilutions after three to six absorptions, which could not be reduced further by subsequent absorptions. Therefore, a substantial proportion of the anti‐D in the immunoglobulin preparation does not bind to D variant red cells.

The component of the anti‐D that does not bind to D variant cells would be expected to be present in the circulation of D variant individuals injected with anti‐Rh immunoglobulin, and should be effective in suppressing Rh immunization. An increased dose of the anti‐Rh Ig might have to be administered in order to allow for the fraction that can bind to D variant red cells and which would not be available for immunosuppression.
Fetal haemoglobin and pregnancy in homozygous sickle cell disease
Authors: Dunn David T. , Poddar Damayanti , Serjeant Beryl E. , Serjeant Graham R.
Published: March 2008

Summary. The pregnancy related changes in fetal haemoglobin (HbF) have been observed in 152 pregnancies in 125 women with homozygous sickle cell (SS) disease and related to steady state levels in the same individual. Statistically significant increases in the first and second trimesters, were followed by significant falls below steady state levels in the third trimester and postpartum period. Although these corresponded to a mean increase of 0·7% and a mean decrease of 1·6%, much greater changes occurred in some individuals. Mean levels had not returned to steady state values 1 year after delivery. The hormonal changes in pregnancy appear to have profound effects on HbF level in SS disease, the mechanisms of which require further study.
Hereditary stomatocytosis: consistent association with an integral membrane protein deficiency
Authors: Eber S. W. , Lande W. M. , Iarocci T. A. , Mentzer W. C. , Höhn P. , Wiley J. S. , Schröter W.
Published: March 2008

Summary. We studied the RBC membrane proteins of four patients, including a mother and daughter, with hereditary stomatocytosis. One‐ and two‐dimensional gel electrophoresis revealed that a 28 kDa integral protein, present in normal RBC membranes, was absent in all four patients. This abnormality, reported once previously (Lande et al. 1982), appears to be a characteristic feature of hereditary stomatocytosis, and may be related to the underlying permeability defect in this disorder.
A non‐human primate model for the study of oral iron chelators
Authors: Wolfe Lawrence C. , Nicolosi Robert J. , Renaud Melanie M. , Finger Janice , Hegsted Mark , Peter Heinrich , Nathan David G.
Published: March 2008

Summary. Recent studies have demonstrated that parenteral deferroxamine can prolong life in patients with iron overload. We have developed a non‐human primate model of iron overload and have accurately determined negative iron balance in parenteral and oral studies of deferroxamine and a new chelator, desferrithiocin. Cebus monkeys were loaded with iron dextran (10 mg/kg twice weekly) until their serum contained a transferrin saturation >75%, and (in two animals) liver biopsies showed iron loading. When complete iron balance studies were performed at this time, basal iron balance was −53 ± 11 μg (N=4), providing a low back ground for provocative studies. Iron balance was determined for intramuscular (N= 2) and oral (N=3) deferroxamine, as well as intramuscular (N= 1) and oral (N= 4) desferrithiocin. The pattern of iron excretion after parenteral deferroxamine strongly resembled that of the iron‐loaded, transfused human. Desferrithiocin was found to have significant activity as an oral chelator. This Cebus monkey model accurately determines negative iron balance and readily permits precise comparison of iron chelators given parenterally or orally. This model may offer an important step between rodent and human trials of promising new iron chelators.
Secretion of coagulant factor VIII activity and antigen by in vitro cultivated rat liver sinusoidal endothelial cells
Authors: Hellman Lars , Smedsröd Bård , Sandberg Helena , Pettersson Ulf
Published: March 2008

Different types of liver cells and a few extrahepatic cell types were analysed for the presence and production of factor VIII activity (VIII: C). Only freshly prepared suspensions of rat liver sinusoidal cells and pure monolayer cultures of rat liver endothelial cells (LEC) were found to contain and secrete detectable amounts of the coagulation factor. Secretion of VIII: C by cultured LEC was inhibited by cycloheximide and by monensin. Constant levels of VIII: C were produced for at least 48 h suggesting continuous synthesis rather than a burst release of stored material. VIII: C, as measured spectro‐photometrically by conversion of X to Xa, was inhibited by anti‐human VIII: C antiserum. Indirect immunocytochemistry using this antiserum gave positive staining only with LEC. Immunoprecipitation of metabolically labelled proteins in conditioned rat LEC medium with the anti human VIII: C antiserum revealed the presence of proteins of similar sizes to those reported for human VIII:C. These results indicate that rat LEC are an important site for production and secretion of procoagulant factor VIII and are not only a site for storage and release of the factor. The established conditions for synthesis of VIII:C in in vitro cultivated rat LEC should provide the means to study the regulation of VIII:C synthesis.
The effect of near‐normoglycaemic control on plasma levels of coagulation factor VII and the anticoagulant proteins C and S in insulin‐dependent diabetic patients
Authors: Schernthaner Guntram , Vukovich Thomas , Knöbl Paul , Hay Ulrike , Müller Mathias M.
Published: March 2008

The present prospective follow‐up study was made to study the effect of glycaemic regulation on levels of factor VII, protein C and protein S in 15 insulin‐dependent diabetic patients without manifestations of vascular disease. Patients were tested before and after 8 weeks of ‘metabolic’ intervention, whereby a near‐normoglycaemic state was achieved. At baseline, values of cross‐linked fibrin degradation products (XL‐FDP) and levels of ‘total’ protein S were significantly increased and protein C values were decreased in the diabetic patients when compared to control subjects, whereas levels of factor VII and ‘free’ protein S were near normal. After ‘metabolic’ intervention a decrease of all haemostatic parameters were recorded, however XL‐FDP levels did not decline to control levels and the imbalance of factor VII and protein C persisted. When patients with newly diagnosed diabetes (n= 8) were compared to those with long‐term disease (n= 7) higher levels of factor VII, protein C and protein S were recorded in the latter group before and after metabolic intervention; at baseline the differences reached statistical significance for factor VII and protein S, and remained significant for factor VII after metabolic intervention. Before and after intervention XL‐FDP levels were higher in patients with newly diagnosed disease than in patients with long‐term diabetes. The correlation analysis revealed positive correlations of factor VII, protein C and protein S to cholesterol and triglycerides, of protein S to all glycaemic control parameters, negative correlations of protein C to glucose, and of XL‐FDP to factor VII, protein C and protein S. The results indicate an imbalance of haemostasis towards thrombophilicity in insulin‐dependent diabetic patients, not completely correctable by glycaemic control.
Platelet membrane abnormalities in myeloproliferative disorders: decrease in glycoproteins Ib and IIb/IIIa complex is associated with deficient receptor function
Authors: Mazzucato Mario , Marco Luigi De , Angelis Vincenzo De , Roia Dina De , Bizzaro Nicola , Casonato Alessandra
Published: March 2008

The number and functional activity of membrane glycoproteins (GP) Ib and IIb/IIIa were investigated in platelets from 11 patients with myeloproliferative disorders (MPD). Three patients had essential thrombocythaemia, two had chronic myeloid leukaemia and six had polycythaemia vera. The numbers of GPIb and GPIIb/IIIa molecules were detected on the platelet surface using different 125I‐labelled monoclonal antibodies. The functional properties of GPIb and GPIIb/IIIa were evaluated using purified 125I‐labelled asialo von Willebrand factor (vWF) and purified 125I‐labelled fibrinogen, respectively, in a binding assay, Binding of the anti‐GPIIb/IIIa antibody was decreased by 40% in almost all patients studied and, when measured, it was accompanied by decreased fibrinogen binding to activated platelets. Binding of anti‐GPIb antibodies to platelets was also slightly decreased or virtually the same in eight out of 11 patients. The decrease correlated with decreased binding of asialo vWF. The increased plasma glycocalicin levels, measured in four patients, depended on the high platelet count. Scatchard analysis revealed normal receptor binding affinity for all ligands tested in all but one patient. In this report we demonstrate that abnormalities in the concentrations of GPIIb/IIIa membrane proteins are commonly present in patients with MPD, while a decrease in GPIb concentration is also seen, although in fewer patients. These abnormalities are accompanied by a concurrent decrease in the respective receptor functions. These findings may explain part of the haemorrhagic tendency often encountered in MPD.
Factors influencing the transfusion response to HLA‐selected apheresis donor platelets in patients refractory to random platelet concentrates
Authors: McFarland Janice G. , Anderson Alfred J. , Slighter Sherrill J.
Published: March 2008

Immune and nonimmune causes of platelet refractoriness were evaluated in a group of patients receiving HLA‐selected single‐donor platelet transfusions. During a 1‐year observation period, 1 h and 24 h platelet recoveries were determined after 522 single‐donor platelet transfusions given to 43 patients persistently refractory to pooled random‐donor platelet transfusions. 72% of patients tested ultimately developed lymphocytotoxic antibodies suggesting they were alloimmunized. When significant lymphocytotoxic antibodies were demonstrable in these patients, HLA well‐matched platelet transfusions consistently produced good transfusion responses. In contrast, patients without lymphocytotoxic antibodies had clinical factors that adversely affected transfusion outcome (P < 0.0001). Fever and splenomegaly markedly reduced 1 h post‐transfusion platelet recoveries, while sepsis compromised the 24 h platelet recovery. Overall, the presence of any clinical factor was most likely to reduce 1 h platelet recovery, while donor‐recipient HLA incompatibilities correlated best with poor 24 h posttransfusion platelet recovery.

A platelet crossmatch test predicted the transfusion response when non‐immune clinical factors were absent.
Antibody‐dependent red cell removal during P. falciparum malaria: the clearance of red cells sensitized with an IgG anti‐D
Authors: Lee S. H. , Looareesuwan S. , Wattanagoon Y. , Ho M. , Wuthiekanun V. , Vilaiwanna N. , Weatherall D. J. , White N. J.
Published: March 2008

The clearance of autologous red cells sensitized with an IgG anti‐D has been studied in patients during and after an attack of P. falciparum malaria, and in 11 uninfected control subjects. In most patients with P. falciparum malaria there was evidence of increased clearance of sensitized red cells, compared to controls.

Clearance half‐times of IgG sensitized red cells were significantly decreased (P < 0.01) in 16 patients with acute falciparum malaria (median 21 min, range 11–53 min) compared to 11 control subjects (median 37 min, range 20–60 min). This difference was independent of the degree of IgG sensitization of red cells. In patients with acute falciparum malaria, clearance half‐times were positively correlated with venous haematocrit (P < 0.05). In 11 patients studied between 1 and 9 weeks after the attack, clearance half‐times (median 17 min, range 11–56 min) were also significantly decreased compared to controls (P < 0.01). In the majority of acute and convalescent patients, there was further evidence for early destruction of an additional substantial proportion of sensitized red cells.

Our data indicate that in most patients with uncomplicated P. falciparum malaria, IgG sensitized red cells are rapidly removed from the peripheral circulation by the spleen, and that rapid clearance persists during recovery.
Expression of the multidrug transporter, P‐glycoprotein, in chronic myelogenous leukaemia cells in blast crisis
Authors: Kuwazuru Yasuo , Yoshimura Akihiko , Hanada Shuichi , Ichikawa Misako , Saito Takeshi , Uozumi Kimiharu , Utsunomiya Atae , Arima Terukatsu , Akiyama Shin‐Ichi
Published: March 2008

The overexpression of a cell‐surface glycoprotein termed P‐glycoprotein (P‐gp) is frequently associated with multidrug resistance (MDR) in cell lines in vitro. To evaluate the implications of P‐gp expression in clinical drug‐resistance, we examined the expression of P‐gp in fresh leukaemia cells from chronic myelogenous leukaemia (CML) patients in blast crisis. By using immunoblotting with a monoclonal antibody against P‐gp, C219, we showed that leukaemia cells from three CML patients in blast crisis were P‐gp negative at the stage when these patients were in complete remission, and that the cells showed high levels of P‐gp expression at times when the same patients had relapsed and had not responded to chemotherapy. Six out of 11 patients (nine in the refractory state) were P‐gp positive and they rarely responded to chemotherapy. These data suggest that the expression of P‐gp is closely associated with drug‐resistance in CML.
Effect of recombinant human granulocyte‐macrophage colony stimulating factor on progenitor cells in patients with advanced malignancies
Authors: Villeval Jean‐Luc , Dührsen Ulrich , Morstyn George , Metcalf Donald
Published: March 2008

Haemopoietic progenitor cell levels were determined in the blood and marrow of 37 patients with advanced malignancies undergoing a phase I/II clinical trial of 0.3–30 μg/kg/d recombinant human granulocyte‐macrophage colony stimulating factor (rGM‐CSF). After injection of rGM‐CSF, the absolute number of circulating progenitor cells fell initially but after 4 d of infusion a dose‐dependent increase was observed in progenitor cells of all lineages with a slight bias favouring granulocyte‐macrophage progenitors. A mean 8.4‐fold increase in GM‐CFC and a 3.3‐fold increase in BFU‐E were observed at a dose level of 20 μg/kg/d of rGM‐CSF. Patients with malignant lymphoma showed a greater response than other patients at the same dose level and the CFU‐E rise correlated with the haematocrit. This study suggests that GM‐CSF may be of value in elevating circulating progenitor cells for subsequent autografting.
Interstitial pneumonitis after total body irradiation: effect of partial lung shielding
Authors: Weshler Z. , Breuer R. , Or R. , Naparstek E. , Pfeffer M. R. , Lowental E. , Slavin† S.
Published: March 2008

The potential beneficial effects of partial lung shielding were investigated in 44 patients conditioned by whole body radiotherapy as part of the conditioning for bone marrow transplantation (BMT). The first 23 patients were irradiated with fractionated total body irradiation (TBI) to a total dose of 12 Gy. In the second series of 21 patients a 50% transmission lung shield was utilized following 6 Gy TBI. This treatment was combined in the second group with four fractions of 1.5 Gy total lymphoid irradiation (TLI), given in 2 d. The rate of idiopathic interstitial pneumonitis (IP) was 26% in the first group, compared to none in the second.
Molecular heterogeneity of β‐thalassaemia in the Japanese: identification of two novel mutations
Authors: Fucharoen Supan , Katsube Takanori , Fucharoen Goonnapa , Sawada Hiroyoshi , Oishi Hiroyuki , Katsuno Makoto , Nishimura Junji , Motomura Seiji , Miura Yasusada , Fukumaki Yasuyuki
Published: March 2008

Five unrelated Japanese β‐thalassaemia genes, from one homozygote and four heterozygotes, have been systematically characterized using DNA polymorphism analysis, polymerase chain reaction, dot‐blot hybridization and direct sequencing of amplified genomic DNA. Four different molecular defects were observed on three different β‐globin gene frameworks. One of these, the A→G mutation in the TATA box, a previously described mutation, was detected by dot‐blot hybridization in one homozygote and one heterozygote with the β‐globin gene of framework 2. The second mutation is a C→T substitution at position 654 of IVS‐2, the mutation commonly found in Chinese, which was associated with the framework 1 gene. Another two mutations, both associated with framework 3 genes, are novel ones; an amber mutation in codon 90 (GAG to TAG) and a frameshift (+G) insertion in codon 54, both of which cause a β0‐thalassaemia phenotype by premature termination of the β‐globin chain synthesis.
Reactivation of HbF synthesis in normal adult erythroid bursts by IL‐3
Authors: Gabbianelli M. , Pelosi E. , Labbaye C. , Valtieri M. , Testa U. , Peschle C.
Published: March 2008

Reactivation of HbF synthesis has been reported in normal adult erythroblast colonies (‘bursts’) generated by erythroid progenitors (BFU‐E) after seeding peripheral blood mononuclear cells (PBMC) in fetal calf serum‐supplemented (FCS+) semisolid cultures stimulated by erythropoietin (Ep). Reactivation is almost totally suppressed when: (i) PBMC are grown in optimized FCS‐ culture or (ii) PBMC are first stringently depleted of monocytes and then plated in FCS+ medium (i.e. BFU‐E growth in FCS+Mo‐ culture). In either case, addition of biosynthetic granulocyte‐macrophage colony stimulating factor (GM‐CSF) induces a dose‐related increase of relative HbF synthesis up to the level in FCS+ culture.

We report that, in FCS‐ culture of partially purified adult blood BFU‐E, treatment with biosynthetic interleukin 3 (IL‐3) causes a dose‐related rise of relative HbF production in the bursts. A similar phenomenon is observed in FCS+ culture of highly purified BFU‐E. The rise of HbF synthesis is seemingly mediated, at least in part, by a direct effect of IL‐3 at BFU‐E level.

It is tentatively concluded that reactivation of HbF in vitro. as well as in a variety of in vivo conditions (i.e. stress erythropoiesis, marrow regeneration), may be at least in part mediated by IL‐3 and GM‐CSF.
The effects of interleukin 3 (IL‐3) on cells responsive to macrophage colony‐stimulating factor (CSF‐1) in liquid murine bone marrow culture
Authors: Breen Frances N. , Hume David A. , Weidemann Maurice J.
Published: March 2008

We have examined variation between inbred mouse strains in the proliferative response of bone marrow cells in liquid culture to macrophage colony‐stimulating factor (CSF‐1) and interleukin 3 (IL‐3). In all mouse strains, thymidine incorporation was stimulated by CSF‐1 and, after an initial lag period, it reached a peak on day 5. In contrast, two mouse strains, A/J and Balb c, had much lower proliferative responses to IL‐3 than did the other strains. In A/J there was no increase in thymidine incorporation above the initial baseline, although the fall in incorporation seen in the absence of any added growth factor was prevented. IL‐3 also presented the loss of CSF‐1 responsiveness observed when A/J bone marrow cells were incubated in medium alone. The lag phase in the response to CSF‐1 was progressively abolished following IL‐3 pre‐treatment. Thus, the data with A/J mice separate two distinct activities of IL‐3 and show that proliferation is not required for the synergistic effect exerted by IL‐3 on CSF‐1‐stimulated macrophage generation from bone marrow.

In strains in which IL‐3 alone was able to stimulate proliferation, its action was not additive with that of CSF‐1, and addition of both factors together did not overcome the lag phase. This suggests that the two factors act on the same cell population, and that the known synergistic effect of IL‐3 on macrophage colony formation in soft agar does not result from an increase in the initial rate of proliferation. The possibility that the combination of factors might alter the duration of the growth response in vivo is discussed.
CD30‐positive large cell lymphomas (‘Ki‐1 lymphoma’) are associated with a chromosomal translocation involving 5q35
Authors: Bastard C. , Rimokh Ruth , Dastugue Nicole , Huret J‐L. , Kristoffersson U. , Magaud J‐P. , Nezelof C. , Tilly H. , Vannier J‐P. , Hemet J. , Warnke R.
Published: March 2008

A chromosomal translocation involving a breakpoint on the long arm of chromosome 5 at position q35 has been reported previously in 17 cases of neoplasia. In 14 of these cases the translocation involves exchange of material between chromosome 2 p23 and chromosome 5. Most cases had been diagnosed histologically as malignant histiocytosis but it was suggested recently, following the study of three cases in one of the author's laboratories, that such tumours are in reality lymphoid tumours. In the present paper we report on 12 further neoplasms with a translocation involving the 5q35 breakpoint and show that all were large cell lymphomas expressing the CD30 (Ki‐1) antigen, often classifiable histologically as ‘Ki‐1 lymphoma’. In five cases there was evidence, based on antigen expression and/or genotypic studies, that the neoplasm was of T lymphoid derivation. These findings provide further evidence that translocations involving 5q35 are associated not with histiocytic malignancy, but with large cell lymphoid neoplasms, including typical cases of ‘Ki‐1 lymphoma’or anaplastic large cell lymphoma’. Since cell lines have been established from five of these cases it may be possible in the future to clone the breakpoint on chromosome 5 and to investigate whether there is a gene in its vicinity with oncongenic potential.
Initial treatment in multiple myeloma: no advantage of multidrug chemotherapy over melphalan‐prednisone
Authors: Hjorth Martin , Hellquist Louise , Holmberg Erik , Magnusson Bengt , Rödjer Stig , Westin Jan ,   for the Myeloma Group of Western Sweden
Published: March 2008

From October 1983 until December 1986, 164 patients with multiple myeloma stage II–III were included in a prospective randomized multi‐centre study comparing melphalan‐prednisone (MP) with multidrug chemotherapy (MDC). The patients comprised 77% of all newly diagnosed myeloma stage II–III cases reported from 18 hospitals covering the entire Health Care Region of Western Sweden (1.5 million inhabitants). Patients randomized to MP (29 stage II and 55 stage III patients) were given oral melphalan and prednisone every 6 weeks. For patients randomized to MDC, stage II patients (n= 25) were given VMCP every 4 weeks and stage III patients (n= 53) VBAP and VMCP alternately every 4 weeks.

For stage II patients, the response rate for MP compared to VMCP was 69 versus 56% and the median survival 46 versus 33 months. For stage III the response rate for MP compared to VBAP/VMCP was 58 versus 57% and the median survival 26 versus 24 months. No statistically significant differences were seen. The time to response and remission duration were also similar in both treatment arms.

The dose intensity index (cumulative given/planned dose of myelosuppressive drugs) was 0.8 in 89% of the MP patients and 81% of the MDC patients. Patients with index values <0.8 had the same response rate as patients with index 0.8.

This study has not demonstrated any advantage of MDC over traditional MP in multiple mayeloma stage II‐III.
Alloimmunization to the PIA1 platelet antigen: results of a prospective study
Authors: Blanchette Victor S. , Chen Lily , De Friedberg Zofia Salomon , Hogan Victoria A. , Trudel Elise , Décary Francine
Published: March 2008

The natural history of alloimmunization to the PlA1 platelet antigen is uncertain. We followed 50 PlA1‐negative pregnant women during pregnancy and for 6 months post‐partum in order to determine this natural history. The cohort of PlA1‐negative women was obtained by PlA1 typing 5000 women. Three PlA1‐negative women formed anti‐PlA1 antibodies during this prospective study, two in pregnancy and one in the immediate post‐partum period. All three PlA1 antibody producers were HLA‐DR3 positive, a histocompatibility phenotype that is strongly associated with alloimmunization to the PlA1 antigen. One of the three infants delivered to these mothers was thrombocytopenic (platelet count 9 x 109/1). The remaining two infants had normal platelet counts at birth (160 and 174 x 109/1). The HLA‐A1, ‐B8, ‐DR3 and ‐DRw52 phenotype frequencies in the group of PlA1‐negative women who did not form PlA1 antibodies (n= 47) was similar to that found in their husbands, and that expected in a normal Caucasian population. From our data we estimate that alloimmunization to the PlA1 antigen occurs in approximately one out of every 1000 pregnancies in a Caucasian population. It is important to recognize that not all pregnancies in which a mother has formed PlA1 alloantibodies will result in the delivery of a thrombocytopenic infant. These findings are relevant to programs designed to either prevent alloimmunization to the PlA1 antigen (through passive administration of anti‐PlA1 immunoglobulin to at‐risk PlA1‐negative mothers), or to identify women at risk of delivery of thrombocytopenic infants (by antenatal screening to detect women alloimmunized to the PlA1 antigen).
Infant acute lymphoblastic leukaemia with t(11;19)
Authors: Gibbons B. , Katz F. E. , Ganly P. , Chessells J. M.
Published: March 2008

Seven cases of infant acute lymphoblastic leukaemia with t(11:19) (q23;p13) are described. They are characterized by a high white cell count, organomegaly, early central nervous system (CNS) disease, and a poor prognosis. Blasts are usually of an immature early B‐cell lineage although monocytoid features are present in some cases. The characteristics of infant acute leukaemia with t(11;19) are very similar to those found with t(4;11), and the presence of t(11;19) may indicate the same poor prognosis.
Monoclonal antibody ratios in malignant myeloid diseases: diagnostic and prognostic use in myelodysplastic syndromes
Authors: Kristensen Josrgen Schosler , Hokland Peter
Published: March 2008

Based on a 6 1/2 year study of bone marrows (BM) from 74 consecutive adult patients with newly diagnosed primary myelodysplastic syndromes (MDS), we have evaluated the role of immunophenotyping (IP) for subclassification purposes with a selected panel of monoclonal antibodies (Mab). Compared to normal BM IP revealed increased numbers of Mab‐defined immature myeloid cells (defined by Mab MY7 [CD13] and MY9 [CD33] (P<0.05). Furthermore, decreased numbers of mature myeloid cells (defined by Mab NAT‐9 II:3F‐6F [NAT‐9])(P<0.001) were demonstrated in all French‐American‐British (FAB) subtypes except refractory anaemia with sideroblasts (RA‐S).

Since expression of CD13 and CD33 antigens on immature myeloid cells is variable, IP based on a single Mab was often found to be non‐informative. However, the construction of myeloid antibody ratios (MAR), designed to give higher figures with increasing numbers of immaturfe myeloid cells and/or decreasing numbers of mature myeloid cells in the BM, disclosed increasing mean ratio values with progressing FAB subtypes. More significant however, was that different prognostic subgroups based on the MAR could be identified independently of the FAB classification. We conclude that the use of IP—especially when MAR is included—is useful in prognostic scoring systems in MDS.
Molecular characterization of β‐thalassaemia in 174 Greek patients with thalassaemia major
Authors: Kattamis C. , Hu H. , Cheng G. , Reese A. L. , Gonzalez‐Redondo J. M. , Kutlar A. , Kutlar F. , Huisman T. H. J.
Published: March 2008

The mutations producing β‐thalassaemia in 174 Greek patients with thalassaemia major were investigated by dot‐blot hybridization of oligonucleotide probes to genomic DNA amplified by the polymerase chain reaction procedure, by direct sequencing of amplified DNA, and by gene mapping. β‐thalassaemia in Greeks was found to be very heterogeneous at the molecular level as 17 different mutations were observed; 86.6% of the β‐thalassaemic genes, however, could be identified with five probes: IVS‐I‐110 (G→A) (42.5%), codon 39 (C→T) (17%), IVS‐I‐1 (G→A) (13.2%), IVS‐I‐6 (T°C) (7.2%) and IVS‐II‐745 (C→G) (6→9%). Several mutations which had not previously been reported in the Greek population and which occurred at an incidence of 2% or lower were observed in this study. The information obtained will facilitate the prenatal diagnosis of β‐thalassaemia in Greece.
Non‐invasive quantitation of liver iron‐overload by magnetic resonance imaging
Authors: Kaltwasser J. P. , Gottschalk R. , Schalk K. P. , Hartl W.
Published: March 2008

A standard magnetic resonance imaging (MRI) system allowing spin echo times of 10 ms was used to quantitate liver iron concentration in nine healthy normal subjects and 13 patients with various grades of iron overload. Body iron status was estimated by measuring the serum ferritin concentration. In 11 subjects (two normal healthy controls, eight patients with HLA‐related hereditary haemochromatosis and one patient with thalassaemia major) non‐haem hepatic iron concentration was determined chemically in biopsy specimens (dry weight), in parallel to serum ferritin and MRI‐T2 relaxation times. A moderate correlation (r=0.79) was obtained for the correlation of the T2‐relaxation rate (1/T2) and serum ferritin of the 22 subjects investigated. A much closer correlation (r= 0.98) was observed for the 1/T2 liver iron relationship in the 10 subjects analysed by liver biopsy. It is concluded from these preliminary observations, that MR‐imaging may provide a useful non‐invasive tool for the quantitative determination of liver iron in iron overload‐syndromes.
Aggressive natural killer cell leukaemia/lymphoma: report of four cases and review of the literature POSSIBLE EXISTENCE OF A NEW CLINICAL ENTITY ORIGINATING FROM THE THIRD LINEAGE OF LYMPHOID CELLS
Authors: Imamura Nobutaka , Kusunoki Yoichiro , Kawa‐Ha Keisei , Yumura Keiko , Hara Junichi , Oda Kenji , Abe Kazuhiro , Dohy Hiroo , Inada Tominari , Kajihara Hiroki , Kuramoto Atsushi
Published: March 2008

The morphologic, immunologic, genotypic and functional properties of peripheral blood and bone marrow cells or cultured cells from four patients with a clinically aggressive non‐T, non‐B natural killer cell leukaemia/lymphoma (ANKL/L) are described. The leukaemic cells possessed medium to large granules in the cytoplasm, antigens against CD38, CD2, OKIa1 and NKH‐1 (CD56) monoclonal antibodies on their cell‐surface, and also showed natural killer (NK) activity. In addition, these ANKL/L belonged to neither T‐ nor B‐cell lineage, proved by studying clonal gene rearrangement for the Tβ, Tγ and Tδ receptors, and immunoglobulin.

After comparing them with the seven cases of ANKL/L reported in other institutions, with regard to immunophenotype, genotype and function, we conclude that ANKL/L originating from a third lineage of lymphoid cells is a distinct clinical entity.
Enhanced growth of clonogenic cells from acute myeloblastic leukaemia by erythropoietin
Authors: Motoji Toshiko , Hoshino Shigeru , Ueda Masatsugu , Takanashi Minoko , Masuda Michihiko , Nakayama Keiko , Oshimi Kazuo , Mizoguchi Hideaki
Published: March 2008

The effect of erythropoietin (Epo) on colony formation by blast progenitors in acute myeloblastic leukaemia other than erythroleukaemia was studied using a blast colony assay. Epo alone did not induce colony formation, but when it was used together with phytohaemagglutinin‐stimulated leucocyte‐conditioned medium (PHA‐LCM) the number of leukaemic colonies significantly increased in nine out of 12 cases studied. Preincubation of Epo with anti‐Epo antibody completely abolished this enhancement, indicating that the increase in colony numbers was caused by Epo itself. Cell surface phenotype analysis of colonies produced by Epo plus PHA‐LCM showed no increase in percentages of erythroid and megakaryocyte lineages. The addition of Epo also increased the self‐renewal capacity of leukaemic blast cells. Fresh leukaemic cells did not express Epo receptors, but they were induced after incubation with PHA‐LCM. The present study thus showed that the proliferative response to Epo is not restricted only to the erythroid lineage, but also extends to AML blast cells other than those in erythroleukaemia in the presence of colony stimulating factors.
Demonstration of platelet antigens that bind platelet‐associated autoantibodies in chronic ITP by direct immunoprecipitation procedure
Authors: Tomiyama Y. , Take H. , Honda S. , Furubayashi T. , Mizutani H. , Tsubakio T. , Kurata Y. , Yonezawa T. , Tarui S.
Published: March 2008

Platelet antigens that bind platelet‐associated autoantibodies in chronic idiopathic thrombocytopenic purpura (ITP) were demonstrated using a direct immunoprecipitation procedure. ITP platelets, with bound autoantibodies, were radiolabelled and solubilized, and then platelet antigen‐antibody complexes adsorbed to protein A‐bearing Staphylococcus aureus were analysed by 7.5% sodium dodecyl sulphate, polyacrylamide gel electrophoresis (SDS‐PAGE). Direct immunoprecipitation demonstrated the presence of platelet‐associated autoantibodies against glycoprotein (GP) IIb/IIIa in four of six ITP patients with an intensive band corresponding to platelet‐associated IgG. These results were confirmed by indirect immunoprecipitation using ether eluates from two ITP patients. In addition, only direct immunoprecipitation demonstrated the presence of autoantibodies against an unidentified protein having a molecular mass of 56 kDa in three of the six patients. These three ITP patients having autoantibodies against GP IIb/IIIa and against the 56 kDa protein were studied after splenectomy. Two patients, showing disappearance of autoantibodies against these antigens, attained a complete remission, and one patient, with autoantibodies against the 56 kDa protein despite splenectomy, attained only partial remission. These data suggest that autoantibodies against GP IIb/IIIa and against the 56 kDa protein may play a role in platelet destruction in some ITP patients.
Activation and inhibition of fibrinolysis in septic patients in an internal intensive care unit
Authors: Voss R. , Matthias F. R. , Borkowski G. , Reitz D.
Published: March 2008

Disseminated thrombotic processes in the microcirculation are considered to be an important cause of multiple organ failure in septic patients. Fibrinolysis is one endogenous mechanism protecting the circulation from overwhelming thrombosis. Therefore, we looked for alterations of fibrinolytic parameters (tissue plasminogen activator (t‐PA), tissue plasminogen activator inhibitor (PAI), D‐dimer, euglobulin‐clot‐lysis‐time (ECLT), plasminogen, α2‐antiplasmin) and of some coagulation parameters (prothrombin time, fibrinogen, platelets, antithrombin III, protein C, factor XII) in clearly defined septic patients and for the relations of these values to the severity of the disease (APACHE II‐score).

An increase in D‐dimer and t‐PA‐antigen was registered in all patients, while factor XII and plasminogen were decreased, indicating an activated fibrinolysis. In contrast the systemic fibrinolytic capacity of the blood was strongly inhibited: t‐PA‐activity was not detectable, PAI‐function was elevated, the ECLT was prolonged and α2‐antiplasmin was normal. Coagulation was moderately activated: the platelets, antithrombin III and protein C were decreased, the prothrombin time was prolonged and fibrinogen was normal. The changes in t‐PA‐antigen, PAI‐function, factor XII, prothrombin time and antithrombin III were significantly related to the APACHE II‐score of the patients.

We conclude that the activation of coagulation is accompanied by an activation of fibrinolysis in the microcirculation, but that systemically the increased inhibitors of fibrinolysis (PAI, α2‐antiplasmin) induce a decrease of the fibrinolytic capacity of the blood. The severity of the disease determines the extent of the alterations.
Protein C and protein S levels in two patients with acquired purpura fulminans
Authors: Madden Renee M. , Gill Joan Cox , Marlar Richard A.
Published: March 2008

Purpura fulminans (PF) is a cutaneous manifestation of a dramatic and deadly syndrome of systemic disseminated intravascular coagulation (DIC). It is characterized by microvascular thrombosis in the dermis followed by perivascular haemorrhage. Since two other related syndromes involve the protein C (PC) system, we undertook a serial study to investigate the levels of PC and protein S (PS) in two patients with acquired PF. Laboratory findings were consistent with DIC, and both patients were treated with blood replacement and heparin therapy. The levels of PC activity were very low during the initial 24–36 h after onset and gradually increased until returning to normal levels. The total and ‘free’PS were also abnormal during the initial onset of PF. The total and free PS increased to normal after 4–6 d. Although the pathogenesis is not fully understood, the infection and sepsis appears to consume PC and PS selectively during the PF and DIC phase. Acquired PF appears to selectively involve the PC system in a similar fashion to two other syndromes of PF‐like lesions.