bluuebunny/crossref_metadata_2025 huggingface dataset:
Monoclonal antibodies to crosslinked fibrin degradation products (XL‐FDP) I. CHARACTERIZATION AND PRELIMINARY EVALUATION IN PLASMA
Authors: Gaffney P. J. , Creighton L. J. , Perry M. J. , Callus M. , Thorpe R. , Spitz M.
Published: March 2008

Summary Monoclonal antibodies (mabs) were raised against X‐oligomers, the earliest soluble fragments released from crosslinked fibrin (XL‐FN), by the action of plasmin. Two of the mabs (NIBn 52 and NIBn 123) were monospecific for X‐oligomers in that they showed no binding to fibrinogen, the plasmic fragments of fibrinogen (D and E) and non‐cross‐linked fibrin (X, Y, D and E), or the terminal digestion product of XL‐FN, fragment DD‐E. One other mab (NIBn 178) was panspecific for X‐oligomers in that it exhibited a weak affinity for fibrinogen. The mabs were used to develop a two‐site immunoradiometric assay (IRMA) and an enzyme‐linked immunospecific assay (ELISA) which permitted the specific measurement of X‐oligomers directly in plasma, rather than in serum. This immunoassay is a true assay of fibrinolysis as distinct from fibrinogenolysis and may be a potential aid in the diagnosis and evaluation of thrombosis. In preliminary studies, the assay detected low levels of X‐oligomers in normal plasma and elevated levels in patients with disseminated intravascular coagulation.
Monoclonal antibodies to crosslinked fibrin degradation products (XL‐FDP) II. EVALUATION IN A VARIETY OF CLINICAL CONDITIONS
Authors: Gaffney P. J. , Creighton L. J. , Callus M. , Thorpe R.
Published: March 2008

Summary Plasmas from patients with a wide variety of thrombotic and presumed prethrombotic conditions were examined for high molecular weight crosslinked fibrin degradation products (known as X‐oligomers) using a two‐site enzyme‐linked immunospecific assay (ELISA). This assay employed a catcher‐tag principle using two monoclonal antibodies (mabs) directed towards different epitopes on the complex X‐oligomer fraction. In general, thrombotic events (pulmonary embolism, PE, myocardial infarction, MI, peripheral vascular disease, PVD, and disseminated intravascular coagulation, DIC) were accompanied by elevated levels of X‐oligomers in the plasma. During pregnancy the value of Xoligomer assays was demonstrated to be a clear‐cut marker for pre‐eclampsia. Patients following a variety of forms of surgery present with heterogeneous plasma levels of X‐oligomers and this may merely reflect the formation and lysis of the fibrin formed during and after surgery. The possible value of this ELISA procedure in monitoring thrombolytic therapy is discussed with a critical analysis of the data presented herein. While the assay of X‐oligomer was demonstrated to be a valuable marker of fibrinolysis in plasma, more extensive data are required in order to assess whether such an assay is of diagnostic value in thrombosis‐related conditions.
Immunostimulatory effects of different antilymphocyte globulin preparations: a possible clue to their clinical effect
Authors: Kawano Yoshifumi , Nissen Catherine , Gratwohl Alois , Speck Bruno
Published: March 2008

Summary Antilymphocyte globulin (ALG) and antithymocyte globulin (ATG) have an established role in the treatment of severe aplastic anaemia. The response rate ranges from 40% to 80%. Its mode of action is believed to be complement dependent lysis of immunocompetent cells which inhibit haemopoietic maturation. This might not be the sole mechanism. We have tested four different preparations of ALG/ATG for their mitogenic effect on normal peripheral blood cells and on enriched T‐cells in vitro by 3H‐thymidine incorporation. We found marked differences between the four preparations. One was strongly mitogenic and able to induce profound release of haemopoietic growth factors. This mitogenic effect could be detected in the serum of patients during ALG treatment. Clinical response rates of this preparation are about 80%. Three other preparations were of lower or no stimulatory effect. Clinical response rates with these preparations vary between 40% and 60%. From our results, we postulate that the beneficial effect of ALG could be partially due to its ability to stimulate release of haemopoietic growth factors. The mitogenicity of different ALG/ATG preparations should be tested as an in vitro parameter of clinical efficacy.
Heterogeneity of CLL: high CD23 antigen and αIFN receptor expression are features of favourable disease and of cell activation
Authors: Dadmarz R. , Cawley J. C.
Published: March 2008

Summary The relationship between α‐interferon receptor (αIFNR) numbers, B‐cell‐antigen expression and clinical stage was determined in 35 cases of typical chronic lymphocytic leukaemia (CLL). αIFNR numbers were shown to be low in patients with advanced disease and high in those with a more favourable prognosis. The B‐cell activation antigen CD 23 was similarly related to stage, being high in more favourable disease. Also, αIFNR expression was directly related to CD 23 positivity, but αIFN binding was not inhibited by CD 23 monoclonal antibody. There was no correlation between CD 19, 20, 22 and 24 antigen expression and either αIFNR numbers or clinical stage.

Since CD23 antigen expression is a feature of B‐cell activation, we suggest that high CD23 and αIFNR positivity are manifestations of an activated cell phenotype and that cell activation in CLL is a feature of favourable disease.
Transient platelet and HLA antibody formation in multitransfused patients with malignancy
Authors: McGrath K. , Wolf M. , Bishop J. , Veale M. , Ayberk H. , Szer J. , Cooper I. , Whiteside M.
Published: March 2008

Summary Fifty‐nine patients receiving platelet transfusions for bone marrow failure secondary to malignancy were screened at regular intervals for the presence of antibodies to human leucocyte (HLA) and platelet specific antigens. HLA antibodies occurred in 19 patients, 10 of whom also developed platelet specific antibodies. The HLA antibodies disappeared in 10 of 15 patients followed for periods of 2–14 months. In two patients this occurred whilst still receiving platelet transfusions. Antibody reappeared in only two of six patients subsequently transfused. Antibodies to platelet specific antigens were detected in 28 patients. They were transient, often appeared in association with infection, and in 50% of cases tested demonstrated autoantibody activity. There was no association with antibiotic drug therapy, or PFA/EDTA‐dependent cryptantigens. Platelet recovery at 1 h or 20 h post transfusion was not significantly reduced in the presence of platelet specific antibodies. These findings have important implications for the selection of platelet donors for alloimmunized recipients.
Effects of recombinant human H‐subunit and L‐subunit ferritins on in vitro growth of human granulocyte—monocyte progenitors
Authors: Dezza Laura , Cazzola Mario , Bergamaschi Gaetano , Stella Carmelo Carlo , Pedrazzoli Paolo , Recalde Helios R.
Published: March 2008

Summary We have studied the influence of purified recombinant human H‐subunit (rHF, acidic) and L‐subunit (rLF, basic) ferritins on in vitro colony formation by normal human granulocyte‐macrophage progenitor cells (CFU‐GM). Whereas rLF had no significant effect, rHF produced significant decrease in colony formation: mean inhibition of CFU‐GM was 38%±13% at 10‐8m and 22%±13% at 10‐9m. The inhibitory activity of rHF was lost at 10‐10m, and was inactivated with a monoclonal antibody recognizing the H subunit, but not with a monoclonal antibody recognizing the L subunit. Although H‐type isoferritins were found in normal and leukaemic cells, their concentration in peripheral blood plasma and bone marrow plasma from normal subjects and patients with different haematological disorders including acute leukaemia were 10‐11m or lower, i. e. levels showing no activity in vitro. We conclude that: (i) acidic H‐subunitrich isoferritins have inhibitory effects on in vitro growth of granulocyte‐macrophage progenitors; (ii) levels of these isoferritins in peripheral blood and bone marrow plasma are 2–3 orders of magnitude lower than the effective concentrations in vitro, indicating that these molecules do not behave as circulating regulatory or suppressive factors in vivo.
Killing of K562 cells with conjugates between human transferrin and a ribosome‐inactivating protein (SO‐6)
Authors: Bergamaschi Gaetano , Cazzola Mario , Dezza Laura , Savino Eufemia , Consonni Luigi , Lappi Douglas
Published: March 2008

Summary Cellular iron uptake is mediated by binding of transferrin with specific surface receptors and internalization of the Fe‐transferrin‐receptor complex. This has been examined as a possible pathway for carrying into leukaemic cells a ribosome‐inactivating protein (RIP), SO‐6, derived from Saponaria officinalis. Purified human differic transferrin was conjugated with SO‐6 and a pool of proteins was obtained, with variable numbers of SO‐6 molecules linked to a single transferrin molecule. Human erythroleukaemic K562 cells were grown in the presence of human transferrin, SO‐6 and human transferrin conjugated with SO‐6. The conjugate was found to be internalized via binding with transferrin receptor. Whereas the presence of unconjugated human transferrin and SO‐6 in the medium did not significantly influence K562 cell growth, the conjugated proteins displayed an inhibitory activity on cell proliferation. This was maximal after 72 h at a transferrin concentration of 10‐9m, with about 50% of cells being killed. Bovine transferrin, present in fetal calf serum, did not appear to compete with human diferric transferrin in binding to K562 cells in suspension culture. In a clonogenic assay, colony formation by leukaemic cells was not influenced by free SO‐6 or transferrin, whereas the conjugated proteins were markedly inhibitory (about 100% at 10‐9m). Our findings indicate that SO‐6 can be efficiently carried into mammalian cells via the transferrin‐transferrin receptor cycle and exert its ribosome inactivating activity. This is in keeping with the existence of an alternative pathway of transferrin endocytosis in addition to the classic acidic endosome pathway. From a practical viewpoint, conjugates between transferrin and SO‐6 can be useful tools for studying the expression of transferrin receptors, and deserve also to be investigated for a possible use in cancer therapy.
Myelodysplastic relapse of de novo acute myeloid leukaemia with trilineage myelodysplasia: a previously unrecognized correlation
Authors: Brito‐Babapulle F. , Catovsky D. , Galton D. A. G.
Published: March 2008

Summary We describe the occurrence of an unusual mode of relapse in six of 24 patients who presented with de novo acute myeloid leukaemia (AML) associated with trilineage myelodysplasia (TMDS). After the induction of complete remission (CR) by intensive chemotherapy in five patients and following bone marrow transplantation (BMT) in one, the myelodysplastic state, but not overt AML, recurred. Relapse of myelodysplasia occurred at a median of 147 weeks (50–520) from presentation and in two instances was followed a year later by AML. In five cases, myelodysplastic relapse was treated with low‐dose cytosine arabinoside given alone or with other chemotherapeutic agents. Three patients remain in CR after 1,2 and 5 years. The reappearance of myelodysplastic features in these six patients was strongly correlated with the presence of TMDS at presentation of the AML. It was not observed once in the 136 AML patients, treated similarly, who did not have associated TMDS at presentation (P<0.001). Thus, relapse with myelodysplasia is not an effect of chemotherapy as has been previously postulated.
TdT positive B‐cell acute lymphoblastic leukaemia (B‐ALL) without Burkitt characteristics
Authors: Michiels J. J. , Adriaansen H. J. , Hagemeijer A. , Hooijkaa H. , Van Dongen J. J. M. , Abels J.
Published: March 2008

Summary The leukaemic cells in a 23‐year‐old man were small to medium‐sized lymphoblasts with no cytoplasmic vacuoles and negative with PAS as well with peroxidase and acid phosphatase staining. Cytogenetic analysis showed ‐6,+12,‐22,+mar(6p::22q), resulting in a trisomy 12 and monosomy of the long arm of chromosome 6. Immunological marker analysis revealed that the majority of the blasts was positive for terminal deoxynucleotidyl transferase (TdT) as well as surface membrane immunoglobulin (SmIg, ü, λ), although B‐ALL are supposed to be negative for TdT. The blasts were also positive for HLA‐DR, CD9 (BA‐2), CD10 (VIL‐A1) and CD24 (BA‐1), but negative for the B‐cell markers CD20 (B1) and Y29/55. Double immunofluorescence staining confirmed that almost all TdT+ cells were also positive for Smμ, Smλ, HLA‐DR and CD10. We thus made a diagnosis of TdT+ B‐ALL without Burkitt characteristics. Since we could not detect SmIg+/TdT+ cells in bone marrow samples from adult healthy volunteers and from 10 children with ALL in complete remission, we conclude that TdT+ B‐ALL cells may not have a normal counterpart in bone marrow or represent a malignant counterpart of a very rare cell in an intermediate differentiation stage between the pre‐B‐cell and the early B lymphocyte.
Prospective study of hepatitis after factor VIII concentrate exposed to hot vapour
Authors: Mannucci P. M. , Zanetti A. R. , Colombo M.
Published: March 2008

Summary A factor VIII concentrate prepared from large plasma pools and then exposed to hot vapour to inactivate blood‐borne viruses was evaluated in 28 factor‐VIII deficient patients (14 vaccinated against the hepatitis B virus, HBV) who had not been treated with any blood product and hence were highly susceptible to the development of post‐transfusion hepatitis. Tests for aminotransferases and HBV markers were made every 2 weeks in the first 4 months and at 5, 6 and 12 months, Twenty‐four patients were considered not to have developed hepatitis, either because they had no elevations of aminotransferases or did not become seropositive for HBV markers. The four remaining unvaccinated patients (three treated with the same batch and the fourth with a different one) developed HBV infection 8–24 weeks after the first concentrate infusion. Hence, this method of viral inactivation did not afford complete protection from hepatitis B.
Increased levels of platelet associated IgG in patients with thrombocytopenia are not confined to any particular size class of platelets
Authors: Holme S. , Heaton A. , Kunchuba A. , Hartman P.
Published: March 2008

Summary Patients with immune thrombocytopenia have an increased percentage of microthrombocytes/platelet fragments and megathrombocytes. It has been suggested that increased levels of platelet associated IgG (PA‐IgG) found in these patients might be related to the presence of this abnormal platelet size distribution. In this study we used flow cytometry to investigate the distribution of PA‐IgG within a population of platelets and, in particular, we examined the relationship between platelet size and PA‐IgG determined simultaneously on individual platelets. Platelet samples from 10 normals and 31 thrombocytopenic patients were studied. PA‐IgG was estimated using immunofluorescent FITC anti‐IgG antibody. Binding of FITC anti‐IgG to the platelets was quantitated in the flow cytometer as relative mean fluorescence (RMF) which was calibrated against values (in fg/plt of FITC anti‐IgG) obtained by spectrofluorometry after solubili‐zation of the platelets. A high correlation (r0.89) was found between flow cytometric RMF value and spectrofluorometric FITC anti‐IgG values. The flow cytometric studies showed that platelet samples with abnormally elevated levels of FITC anti‐IgG (>1.7 fg/plt) not only have a higher percentage of platelets with elevated FITC anti‐IgG, but that these platelets also have increased levels of FITC anti‐IgG as compared to platelets from normal samples. Platelet size was measured by the amount of forward light scatter in the flow cytometer. A low but significant correlation (r=0.33±0.12) was found between size (FALS) and fluorescent signals in samples with elevated FITC anti‐IgG. The contribution of 10% of the smallest platelets by FALS and 10% of the largest platelets by FALS to the total levels of flow cytometer platelet fluorescence in these samples was only 4.4% and 19.4% respectively which was not higher than obtained with samples with normal levels of FITC anti‐IgG. In conclusion. this study showed that increased levels of PA‐IgG found among thrombocytopenic patients were not confined to any particular size class of platelets.
Plasma concentrations of C4b‐binding protein and vitamin K‐dependent protein S in term and preterm infants: low levels of protein S‐C4b‐binding protein complexes
Authors: Malm Johan , Bennhagen Rolf , Holmberg Lars , Dahlbäck Björn
Published: March 2008

Summary We have determined the plasma concentrations of protein S and C4BP in 25 term and 26 preterm infants by radioimmunoassay. Both the total concentration and the concentration of free protein S were quantified. The concentration of C4BP was very low in preterm infants (mean 6% of the adult level). In term infants, the level had increased to a mean of 18%. Total protein S was decreased both in preterm and term infants, 4.0 mg/l and 6.8 mg/l respectively, as compared to the mean adult concentration, 20.6 mg/l. In preterm infant plasma, free protein S was the predominant (85%) form, probably due to the very low C4BP level. In plasma from term infants, free protein S represented 68% of the total protein S, the corresponding value in adult controls being 37%. The plasma concentration of free protein S in preterm and term infants was 3.3 mg/l and 4.6 mg/l, respectively (mean adult value 7.6 mg/l). These results demonstrate that, while the total protein S concentration in preterm and term infants was very low in comparison to the adult level, the difference in the concentration of the anticoagulant, active, free form of protein S between infants and adults was less pronounced.
IgG subclass identification of inhibitors to factor IX in haemophilia B patients
Authors: Örstavik Karen Helene , Miller Connie H.
Published: March 2008

Summary Inhibitors to factor IX in haemophilia B are IgG antibodies. It is of interest to know the subclass type of these antibodies since the biological activity of the four IgG subclasses are different. Subclass identification has been difficult due to the lack of specific antibodies to IgG subclasses. Monoclonal hybridoma antibodies to these subclasses are now available. Here we present the results on the subclass identification of inhibitors from six patients with severe haemophilia B (titres 0.8–800 u/ml). Plasma from one patient was obtained before inhibitor could be detected by coagulation assays. He had circulating complexes between factor IX and inhibitor. The technique used was crossed immunoelectrophoresis combined with an enzyme‐linked immunoassay. This technique allows the subclass determination of both low‐titre and high‐titre inhibitors. All inhibitors gave a very strong reaction with antibody to IgG4, and four inhibitors also reacted with antibody to IgG1. Three of these inhibitors gave a faint reaction with antibody to IgG2 also, whereas no inhibitor contained any detectable IgG3. The finding of IgG4 as the main subclass is important since IgG4 antibodies do not bind complement. The presence of more than one subclass in the inhibitors demonstrates the heterogeneous nature of the inhibitors. This was demonstrated also at a stage of inhibitor development when the inhibitor could not be detected by coagulation assay.
Four base‐pair DNA deletion in human Aγ globin‐gene promoter associated with low Aγ expression in adults
Authors: Oilman John G. , Johnson Mary E. , Mishima Noboru
Published: March 2008

Summary Fetal haemoglobin (α2γ2) is predominant in red cells of the fetus and newborn baby, and is largely replaced after birth by adult haemoglobin (α2β2). The two types of γ chains (Aγ and Gγ) are generally less than 1% of total β‐like chain in adults, and the Gγ:Aγ ratio is typically 40:60. Higher Gγ values (> 50% of γ chain) are frequently associated with a T for C nucleotide substitution 158 base pairs 5’of the Gγ Cap site (‐158). The first exception to this rule was a β ‐thalassaemia in a Black family that was associated with about 60%Gγ in heterozygotes. A DNA fragment containing the Gγ and Aγ genes of the high Gγ haplotype of this case has now been cloned. DNA sequencing from ‐ 383 to the Cap site showed no differences from normal for the Gγ gene, except for C at ‐ 158. For the Aγ gene, however, a deletion of four base pairs (AGCA) at ‐ 222 to ‐ 225 was found. It is hypothesized that this deletion causes reduced Aγ globin gene expression in adults, which suggests that promoter elements important for the regulation of fetal haemoglobin expression in adults extend upstream at least to ‐225.
Carbimazole‐induced immune haemolytic anaemia: role of drug‐red blood cell complexes for immunization
Authors: Salama A. , Northoff H. , Burkhardt H. , Mueller‐Eckhardt C.
Published: March 2008

Summary We report on a patient who developed acute intravascular immune haemolysis while receiving carbimazole. Serological studies revealed a strongly (3+) and a discretely positive (1+) direct antiglobulin test due to C3d and IgG respectively, and a very weak IgG autoantibody in the eluates. Serum from the patient contained specific carbimazole‐dependent red blood cell (RBC) antibodies which reacted with all normal human RBC in the presence of free carbimazole as well as with RBC coated with the drug either in vitro or in vivo, although carbimazole itself is not detectable in plasma after oral administration. The results provide direct evidence for the sequence of the drug‐RBC‐antibody interaction and show that the RBC (and not plasma proteins) function as ‘carrier‐like’macromolecules in the immune response.
Carrier mediated iron transport through erythroid cell membrane
Authors: Egyed A.
Published: March 2008

Summary The transport of iron through erythroid cell membrane was studied in a model system, measuring ferrous iron uptake by reticulocytes. It was found that these cells were able to take up ferrous iron and to incorporate it into haem at a rate similar to that observed when diferric transferrin was the iron donor. No comparable iron uptake could be measured when the metal was provided as Fe3+ citrate or when reticulocytes were replaced by mature erythrocytes.

The involvement of end geneous transferrin in the Fe2+ uptake by reticulocytes could be excluded, since proteolytic treatment of the cells had no significant effect on the process.

Fe2+ uptake by reticulocytes followed saturation kinetics, characteristic to carrier mediated transport processes. Kinetic analysis of the data revealed the following apparent transport parameters: Km= 8.8±3.8 μm; Vmax= 1.1±0.2 ng/108 reticulocytes/ min.

These results indicate that a high affinity, carrier mediated iron transport system is present in the reticulocyte membrane, ensuring the efficient translocation of the metal through the membrane barrier between the site of its release from transferrin and the site of its utilization.
Elevated red cell adenosine deaminase activity: a marker of disordered erythropoiesis in Diamond‐Blackfan anaemia and other haematologic diseases
Authors: Glader Bertil E. , Backer Karen
Published: March 2008

Red‐cell adenosine deaminase (ADA) activity in children with Diamond‐Blackfan anaemia is significantly increased (1·91±0·90 U/g Hb) compared to that seen in transient erythroblastopenia of childhood (0·80±0·16 U/g Hb) or normal individuals (0·61±0·13 U/g). These data thus further support that measurement of this purine metabolic enzyme is useful in diagnosing the cause of pure RBC aplasia in children. Of interest, however, elevated RBC‐ADA activity also is seen in some children with acute leukaemia and other haematologic disorders. In children with acute lymphoblastic leukaemia (ALL), the increase in RBC‐ADA activity is proportional to the degree of anaemia. However, the elevated RBC‐ADA activity in this leukaemic population is not related to the fetal haemoglobin concentration. These data suggest increased RBC‐ADA activity may be a non‐specific manifestation of abnormal erythroid stem cell function, an alteration distinct from that seen with reactivation of fetal erythropoiesis. However, since almost all patients with Diamond‐Blackfan anaemia manifest elevated RBC‐ADA activity, this chemical alteration yet may reflect the specific erythroid differentiation lesion in this disorder.
Electron‐microscopic characterization of mixed granulated (hybridoid) leucocytes of chronic myeloid leukaemia
Authors: Schmidt U. , Mlynek M.‐L. , Leder L.‐D.
Published: March 2008

In chronic granulocytic leukaemia, hybridoid leucocytes can regularly be found. Light microscopically they contain a mixture of eosinophilic, basophilic and naphthol AS‐D chloroacetate esterase‐positive granules. The present study was done to clarify the ultrastructural composition of these cells. It could be clearly shown that in some leukaemic granulocytes primary and secondary eosinophilic as well as basophilic granules occur side by side. There were also basophils with additional tissue mast cell granules. Since normal mast cell granules as well as granules of normal eosinophilic promyelocytes are naphthol AS‐D chloroacetate esterase‐positive, it would appear possible that mastocytoid as well as primary eosinophilic granules within the leukaemic basophils are responsible for the atypical, granular naphthol AS‐D chloroacetate esterase‐positivity of these cells. The existence in chronic myeloid leukaemia both of mixed basophilic and eosinophilic granulated leucocytes and of mixed basophilic and mastocytoid granulated leucocytes may suggest a common myeloid precursor of eosinophils, basophils and tissue mast cells. In addition, the hybridoid granulocytes may be considered an expression of a neoplasiarelated lineage infidelity.
Allogeneic bone marrow transplantation versus intensification chemotherapy for acute myelogenous leukaemia in first remission: a prospective controlled trial
Authors: Conde E. , Iriondo A. , Rayon C. , Richard C. , Fanjul E. , Garijo J. , Hermosa V. , Coma A. , Bello C. , Carrera D. , Baro J. , Zubizarreta A.
Published: March 2008

In 1982 we began a prospective controlled trial to assess the effectiveness of allogeneic bone marrow transplantation and intensive post‐remission chemotherapy for patients with acute myelogenous leukaemia in first complete remission. Fourteen patients, 3–45 years of age, who had an HLA‐identical sibling donor, received bone marrow transplantation. Twenty‐five patients who either lacked an HLA‐identical sibling or were over 45 years of age received intensive consolidation chemotherapy including high‐dose cytosine arabinoside with or without adriamycin.

The actuarial rate of continued complete remission (CCR) at 3 years was significantly higher in the transplantation group than in the chemotherapy group: 70% (95% confidence interval 35–91%) compared with 10% (95% confidence interval 2–30%); P = 0·01. However, the actuarial rate of CCR was not significantly different between the transplantation group and patients under 45 years in the chemotherapy group: 70% (95% confidence interval 35‐91%) compared with 17% (95% confidence interval 4–45%), 0·1>P>0·05. The actuarial probability of leukaemia relapse was significantly lower in the transplantation group than in the chemotherapy group: 10% (95% confidence interval 4–21%) compared with 88% (95% confidence interval 70–96%), 0·005 >P>0·001. There was no significant difference between both groups if we compare only the patients who died of non‐leukaemic causes: 22%, (95% confidence interval 9–42%) versus 25% (95% confidence interval 7–59%), P = NS.

In summary, this study shows that allogeneic bone marrow transplantation is a better anti‐leukaemic treatment than is intensive consolidation chemotherapy in patients with AML in first complete remission.
Abnormal membrane phospholipid organization in Plasmodium falciparum‐infected human erythrocytes
Authors: Joshi P. , Gupta C. M.
Published: March 2008

The membrane phospholipid organization in Plasmodium falciparum‐infected human erythrocytes was analysed by employing phospholipase A2 and Merocyanine 540 as external membrane probes. Both bee venom and pancreatic phospholipases A2 failed to hydrolyse phosphatidylserine in uninfected human red cells isolated from in vitro P. falciparum cultures. However, these enzymes under identical conditions readily degraded this aminophospholipid in P. falciparium‐infected erythrocytes. Phosphatidylethanolamine hydrolysis also increased in parasitized cells. The degree to which these aminophospholipids were cleaved by the enzymes in intact infected cells depended on the developmental stage of the intracellular parasite, and was maximum at the schizont stage. This was consistent with the finding that the ‘fluid‐sensing’ fluorescent dye, Merocyanine 540, readily labelled both the schizont and trophozoite‐infected cells but not the fresh, uninfected or ring‐infected erythrocytes. These results demonstrate that P. falciparum produces stage‐dependent changes in the membrane phospholipid organization of its host erythrocyte.
Effects of platelet derived growth factor, epidermal growth factor and transforming growth factor‐β on the growth of human marrow fibroblasts
Authors: Kimura Akiro , Katoh Osamu , Kuramoto Atsushi
Published: March 2008

Summary. The effects of three growth factors contained in platelets on human bone marrow fibroblasts as well as fibroblast colony‐forming cells were investigated to clarify the pathogenesis of marrow fibrosis frequently associated with myeloproliferative disorders. Two growth factors, platelet derived growth factor (PDGF) and epidermal growth factor (EGF), cooperatively stimulated the growth of passaged fibroblasts as well as fibroblast colony‐forming cells. Transforming growth factor‐β augmented the growth of fibroblasts at low concentrations in the presence of EGF and/or PDGF, but inhibited that of colony‐forming cells in the presence of PDGF or EGF. These results suggest that the growth of fibroblasts causing marrow fibrosis might be finely regulated, depending on the concentrations of growth factors released or leaked from platelets or megakaryocytes into marrow environment.
Hypereosinophilic syndrome human eosinophil degranulation induced by soluble and particulate stimuli
Authors: Henderson William R. , Harley John B. , Fauci Anthony S. , Chi Emil Y.
Published: March 2008

Summary. Eosinophil degranulation induced by the calcium ionophore A23187 and opsonized zymosan particles was examined ultrastructurally in peripheral blood cells obtained from patients with the hypereosinophilic syndrome. Unstimulated hypereosinophilic syndrome eosinophils contained altered, vacuolated cytoplasmic granules and large cytoplasmic crystalloid structures not seen in normal cells. By morphometric analysis of transmission electron micrographs, the hypereosinophilic syndrome eosinophils contained a larger percentage of smaller sized cytoplasmic granules than normal cells. The hypereosinophilic syndrome eosinophils, however, were capable of undergoing noncytotoxic degranulation after stimulation with either A23187 or opsonized zymosan. Hypereosinophilic syndrome eosinophil degranulation was characterized by fusion of the perigranular membranes of adjacent cytoplasmic granules and vesiculation of the fused granules. Granule contents were released intracellularly into vacuoles after ionophore stimulation and into phagosomes containing the ingested zymosan particles. Noncytotoxic extracellular release of eosinophil peroxidase (EPO) was also observed after cell stimulation by either A23187 or zymosan. The capacity of hypereosinophilic syndrome eosinophils to degranulate after appropriate stimulation with release of toxic granule constituents such as EPO and other basic proteins may be important in the tissue injury observed in this syndrome.
Immunochemical specificity of autoanti‐Gerbich from two patients with autoimmune haemolytic anaemia and concomitant alteration in the red cell membrane sialoglycoprotein β
Authors: Reid Marion E. , Vengelen‐Tyler Virginia , Shulman Ira , Reynolds Marilyn V.
Published: March 2008

Summary. Blood samples from two patients with autoimmune haemolytic anaemia, whose autoantibodies failed to agglutinate certain examples of red cells that lack Gerbich blood group antigens, were studied using immunochemical analyses. One of these autoantibodies differed from all other anti‐Ge in that it showed a unique β sialoglycoprotein (SGP) specificity. It reacted with normal β but not with the abnormal β‐related SGPs associated with Gerbich‐negative red cells of the Gerbich and Yus types. Red cells from this patient had an alteration of β SGP, while the α, γ and δ SGPs appeared to be normal. The autoantibody from the other patient did not show this unique characteristic. Its immunochemical specificity was similar to alloanti‐Ge3 in that it reacted with both β and γ SGPs from normal red cell membranes and with the abnormal β‐related SGPs found in red cell membranes from individuals with Gerbich‐negative red cells of the Yus type. Red cells from this patient could not be analysed because she had recently received a massive transfusion of red cells.
Comparative histology of malignant lymphomas in lymph node and bone marrow
Authors: Bartl R. , Hansmann M.‐L. , Frisch B. , Burkhardt R.
Published: March 2008

Summary. Lymph node and bone marrow biopsies of 226 patients with malignant lymphomas were available for evaluation and comparison. 120 of the 226 lymphoma patients had classifiable infiltration in both lymph node and bone marrow and these were used for comparison. Congruence in the histologic subtypes in lymph node and bone marrow was found in 91 patients (76%). The majority of the 29 cases with divergent histologies showed ML of low grade malignancy in the bone marrow even with long time intervals between the two biopsies. The most notable in this group were seven cases who had ML centroblastic/centrocytic in the lymph node biopsy and ML immunocytic in the bone marrow, although none had a serum M‐component. 65 of the 106 patients excluded from comparison had involvement in only one organ, the others had unclassifiable or equivocal histologies in either lymph node or bone marrow or in both. The findings are discussed with respect to the histologic behaviour of the lymphomas. The results show that comparison between lymph node and bone marrow findings in patients with ML is feasible and clinically relevant.
A comparison of the effects of two triphasic oral contraceptives on haemostasis
Authors: Cohen H. , Mackie I. J. , Walshe K. , Gillmer M. D. G. , Machin S. J.
Published: March 2008

Summary. The effects of two cyclically administered, triphasic, combined low dosage oestrogen and progestogen oral contraceptives on haemostasis have been compared in a longitudinal study, over 6 months, in 26 healthy females aged 16–30 years. Subjects received either Logynon (ethinyl oestradiol and Levonorgestrol, n= 14) or SHD 415G (Schering U.K., n= 12), which contains a similar dosage of ethinyl oestradiol, but in combination with a new progestogen, gestodene. Both groups showed increases in biological activities of procoagulant factors fibrinogen, X and XII and decreased activity of the naturally occurring coagulation inhibitor antithrombin III (AT‐III). The majority of these changes were statistically significant (P<0.05 to < 0.001), apparent after one cycle and maintained over the six cycle period. FVII activity increased in both groups, achieving statistical significance (P < 0.01) by cycle 6 in the SHD 415G group but not in the females receiving Logynon. Protein C activity remained unchanged in both groups. Between‐group comparisons showed no differences in the procoagulant factor changes, but protein C was lower (P < 0.05) in the SHD 415G group after three cycles of therapy. These findings indicate that both triphasic oral contraceptives Logynon and SHD 415G induce increases in procoagulant factor activities which are not balanced by increased biological levels of the two most important physiological coagulation inhibitors AT‐III and protein C. These prothrombotic changes are not modified by the new progestogen, gestodene.
Acetaldehyde‐modified serum proteins inhibit mitogen‐stimulated human lymphocyte transformation
Authors: Wickramasinghe S. N. , Barden G.
Published: March 2008

Summary. Sera obtained from healthy volunteers immediately before and 8 h after the rapid consumption of 1.2 g ethanol/kg body weight were dialysed against RPMI 1640 and added ab initio to microcultures of normal human lymphocytes containing 1–3 μg phytohaemagglutinin (PHA)/ml or 2–8 μg pokeweed mitogen (PWM)/ml. When compared with the pre‐alcohol sera, the post‐alcohol sera inhibited lymphocyte transformation after 48 h incubation with either mitogen. In other experiments, acetaldehyde‐albumin complexes were generated by reacting solutions of human serum albumin with 45–720 μm acetaldehyde, and the same quantity of either unmodified albumin or acetaldehyde‐modified albumin was included in freshly‐prepared lymphocyte microcultures containing 3 μg PHA/ml or 8 μg PWM/ml. When compared with unmodified albumin, acetaldehyde‐modified albumin inhibited lymphocyte transformation after 48 h of culture with either mitogen. The inhibition of lymphocyte transformation caused by post‐alcohol sera and acetaldehyde‐modified albumin was partially corrected after treatment of the proteins with 1.55 mm sodium borohydride at a pH of 9.5. The data indicate that post‐alcohol sera contain a non‐dialysable activity which inhibits mitogen‐stimulated lymphocyte transformation in vitro and that at least part of this activity may reside in acetaldehyde‐modified serum albumin.
Circulating erythropoietin in patients with myelodysplastic syndromes
Authors: Jacobs A. , Janowska‐Wieczorek A. , Caro J. , Bowen D. T. , Lewis T.
Published: March 2008

Summary. Serum concentration of erythropoietin (Epo) has been measured by radioimmunoassay in 46 patients with myelodysplastic syndromes. There is an overall inverse relationship between the level of Epo and the degree of anaemia but a wide range of Epo response between patients with similar haemoglobin concentrations. No differences were found between the different FAB groups, but the highest Epo levels were found in those patients with erythroid hypoplasia in the bone marrow. It is suggested that the intensity of erythroid activity in the marrow, as well as the degree of anaemia, may be a factor determining serum Epo concentration.
Cytogenetics and their prognostic value in de novo acute myeloid leukaemia: a report on 283 cases
Authors: Fenaux P. , Preudhomme C. , Laï J. L. , Morel P. , Beuscart R. , Bauters F.
Published: March 2008

Summary. Cytogenetic analysis was successfully performed at diagnosis in 283 patients with de novo acute myeloid leukaemia (AML), including eight children aged 6–15 and 275 adults. Mean age was 50.3 (range 6–86) and the M/F ratio was 1.23. 214 patients were treated by intensive chemotherapy and 75.2% achieved complete remission. Patients with inv(16) and t(8;21) had very high CR rates whereas those with complex cytogenetic abnormalities (with or without involvement of chromosomes 5 and/or 7) had a poor response to therapy. Other karyotypic abnormalities and normal karyotypes were associated with intermediate CR rates. In a multivariate analysis, cytogenetics were the best prognostic factor of the achievement of CR, followed by age and platelet count. Median actuarial disease‐free survival (DFS) of the patients who achieved CR was 19 months. No significant differences in actuarial DFS were found according to karyotype. However, no relapses were seen in patients with inv(16) and t(9;11) or its variants. Conversely, the median DFS was only 12.5 months in patients with t(15;17). Median actuarial DFS was 24 months in patients with t(8;21), but a high incidence of early relapses were seen, and the actuarial DFS was only 54% at 12 months. Patients with trisomy 8 also had a median actuarial DFS of 24 months. Our findings do not support the previously reported ‘favourable’ prognosis of t(15;17) translocations. They suggest that, although it is characterized by a high CR rate, t(8;21) might be associated with a high incidence of early relapses and that, finally, inv(16) might be the only ‘favourable’ cytogenetic rearrangement in AML. Furthermore, the prognosis associated with t(9;11) or its variants (at least in adults), and trisomy 8 might be less severe than suggested in other studies.
IgG and IgM antibodies to platelet membrane glycoprotein antigens in acute childhood idiopathic thrombocytopenic purpura
Authors: Winiarski Jacek
Published: March 2008

Summary. In contrast to the situation in chronic idiopathic thrombocytopenic purpura (ITP) it has not been known whether acute childhood ITP has an autoimmune aetiology and to what extent autoantibodies directed to platelet autoantigens appear during the transient course of this disease. In the present study of 21 children with acute ITP an immunoblot technique was applied to detect serum antibodies to electrophoretically (SDS‐PAGE) separated normal donor platelet membrane proteins. Platelet antigen binding antibodies detected by alkaline phosphatase conjugated protein A or anti‐IgM antibody were observed in 13 out of 21 (62%) patients while controls were negative. In nine children antibodies were directed to antigens localized on the three major membrane glycoproteins, GPIb (four), GPIIb (one) and GPIIIa (five). Antibodies to a 250 kDa antigen were noted in one case and to smaller 25–52 kDa proteins in 12 patients. Four patients had IgG as well as IgM platelet antibodies while in nine only IgM was found. The reactions were eliminated after absorptions of sera with fresh platelets. In all of three tested patients the glycoprotein antigen specific antibodies could be detected in acid eluates prepared from the absorbing platelets. The presence of antibodies, predominantly IgM, to platelet surface antigens in a majority of the children with acute ITP, may suggest an autoimmune process, albeit transitory, similar to that in chronic ITP.
Comparison of the in vitro characteristics of von Willebrand factor in British and commercial factor VIII concentrates
Authors: Lawrie A. S. , Harrison P. , Armstrong A. L. , Wilbourn B. R. , Dalton R. G. , Savidge G. F.
Published: March 2008

Summary. Qualitative/quantitative analysis of von Willebrand factor antigen (vWf:Ag) in either heat or solvent/detergent treated factor VIII concentrates, used for haemophilia replacement therapy, was undertaken to assess their suitability for the treatment of vWD. For the first time immunoaffinity purified vWf:Ag (Monoclate by‐product) was also evaluated by in vitro assessment.

Potencies of vWf: Ag varied considerably but were consistently higher (28.9–420.5 iu/ml) than factor VIII:C (one‐stage) activity (8.13–42.44 iu/ml). The functional activity of vWf was assessed by either Ristocetin Cofactor (vWf:RCo) or collagen binding methods (vWf:CBA) with typical vWf:RCo/vWf:Ag ratios ranging from 0.08 to 0.94. Multimeric analysis confirmed that in vitro biological activity was dependent on the presence of the high molecular weight forms of vWf: Ag. A significant correlation (r=0.95) between vWf:RCo activity and collagen binding was observed in all of the concentrates with the exception of the immunopurified product.

The data suggest that either NHS 8Y (mean vWfRCo/vWf:Ag=0.94), Haemate P (mean vWf:RCo/vWf:Ag=0.69) and high purity Octapharma V.I (vWf:RCo/vWf:Ag=0.82) which contain medium/high MW vWf:Ag multimers are likely to be most cost‐effective in the treatment of symptomatic severe vWD patients than other currently available concentrates.
An intrinsic progenitor defect in Diamond‐Blackfan anaemia
Authors: Tsai Philip H. , Arkin Steven , Lipton Jeffrey M.
Published: March 2008

Summary. To determine whether the erythropoietin (epo) insensitivity of erythroid progenitor differentiation in congenital pure red cell aplasia or Diamond‐Blackfan anaemia is intrinsic to the progenitor itself or is due to defective accessory cell function or active suppression, progenitors from normals and two patients (one steroid resistant and one spontaneously remitting), separated from all known accessory cells using sequential negative selection techniques (adherence, E‐rosetting, and direct and indirect immune‐panning), were studied. Initially, we evaluated three patients with DBA using unfractionated bone marrow mononuclear cells. Progenitors from two steroid non‐responsive patients showed insensitivity to crude epo (c‐epo) while one steroid responsive patient demonstrated normal in vitro sensitivity to c‐epo. When recombinant epo (r‐epo) was used in place of c‐epo, the two steroid non‐responders continued to demonstrate in vitro progenitor epo insensitivity. However, sensitivity of progenitors from the steroid responder, which was normal in the presence of c‐epo, became abnormal when recombinant epo (r‐epo) was substituted. Thus, using unfractionated bone marrow, the abnormal response to epo of progenitors from some patients with DBA appears to be obscured by stimulating factors termed erythroid burst‐promoting activity (BPA) which are present in c‐epo. Using fractionated highly enriched progenitors, from normals and a steroid responsive patient a final 3–10‐fold enrichment of progenitors was achieved, but no such enrichment was seen when marrow from a steroid resistant patient was cultured. The epo sensitivities of normal and of patient erythroid progenitors were similar. However, at sub‐optimal epo concentrations in both patients CFU‐E responsiveness to crude BPA was abnormal compared to the three controls. We conclude from these studies that in DBA: (a) the failure of erythropoiesis is due to an intrinsic progenitor defect; (b) this defect involves progenitor insensitivity to factors in addition to erythropoietin; and (c) there exists a spectrum of disease reflected in the degree of the in vitro abnormality observed.
The use of DNA probes to monitor minimal residual disease in childhood acute lymphoblastic leukaemia
Authors: Katz Fay , Ball Lynne , Gibbons Barbara , Chessells Judith
Published: March 2008

DNA probes to both the joining region (JH) of the immunoglobulin heavy chain gene (IgH) and to the β chain of the T‐cell antigen receptor complex (TCR) have been used as tumour‐specific markers to monitor the rearrangements of the IgH chain gene and the TCR β gene in the blast cells of children presenting with acute lymphoblastic leukaemia (ALL) of B or T cell origin. Blast cells from 68 children with early B cell ALL and eight with T‐ALL were examined at presentation, at day 28 after commencement of therapy and at varying times thereafter. An additional 43 patients (42 with B cell ALL, one with T‐ALL) were studied both at presentation, at completion of their 2‐year treatment course and 3 months later. Twelve patients, drawn from both these groups, were studied at relapse as were a further eight patients in whom an extramedullary relapse had occurred. Persistence of clonally‐derived cells as a predictor of early relapse was seen in the day 28 bone marrows of 11/76 newly‐diagnosed children (nine early B and two T‐ALL) followed by rapid, overt relapse in four of the early B ALL cases. No minimal residual disease (MRD) was detected in bone marrows from any of the 43 patients completing their 2‐year treatment course, but six of these subsequently relapsed at varying time periods thereafter. Identical patterns of rearrangement at both presentation and relapse were seen in most cases. Oligoclonality, or multiple IgH chain gene rearrangements was seen in the blast cells of 15% of patients with early B cell ALL. No correlation between oligoclonality, high white count, unfavourable phenotype, or abnormal karyotype could, however, be ascertained.