bluuebunny/crossref_metadata_2025 huggingface dataset:
Alloantibodies to Factor IX in Haemophilia B Characterized by Crossed Immunoelectrophoresis and Enzyme‐conjugated Antisera to Human Immunoglobulins
Authors: ØRstavik K. H.
Published: July 2010

Summary. Incubation of factor IX with non‐precipitating alloantibodies to factor IX gives rise to soluble complexes between factor IX and the alloantibodies. These complexes appear as a factor IX molecule with a reduced mobility in crossed immunoelectrophoresis against a rabbit antiserum to factor IX. This factor IX immunoprecipitate was used for the study of alloantibodies to factor IX from five patients with severe haemophilia B (antibody titres 0·1–800 units/ml plasma). Incorporation of antiserum to κ light chains or λ light chains of human immunoglobulin G in an intermediate gel in crossed immunoelectrophoresis gave a reduction of the factor IX precipitin arc, indicating the presence of immunoglobulin G alloantibodies containing both κ and λ light chains in complex with factor IX. Incubation of the factor IX immunoprecipitate with peroxidase‐conjugated antisera to the same immunoglobulins, and staining for peroxidase activity, confirmed the presence of immunoglobulin G containing both types of light chains in the factor immunoprecipitate. It is concluded that all five alloantibodies were polyclonal immunoglobulin G antibodies. The technique had the advantage that the light chain types of low‐titre antibodies could be determined, and it may be suitable for further characterization of alloantibodies to factor IX if antibodies to immunoglobulin subclasses are available.
A chronic myeloproliferative disorder associated with monosomy 7 in the bone marrow cells; normal karyotype in acute transformation
Authors: Linch D. C. , Walker H. , Roberts P. , McKinnon J. , Goldstone A. H. , Huehns E. R.
Published: July 2010

Summary.

A chronic myeloproliferative disorder associated with monosomy 7 of the bone marrow cells is described in a male infant. After nearly 4 years, during which time no drug therapy was given, the disease transformed to a ‘blast crisis’ and at this stage the karyotypes of the marrow and unstimulated blood cells were shown to be normal. It is proposed that the development of the myeloproliferative disorder associated with monosomy 7 is a multistage process and the acquisition of the chromosomal abnormality is a secondary event.
The Evans syndrome: characterization of the responsible autoantibodies
Authors: Pegels J. G. , Helmerhorst F. M. , VanLeeuwen E. F. , van de Plas‐van Dalen C. , Engelfriet C. P. , Kr. vondemBorne A. E. G.
Published: July 2010

Summary.

The immunofluorescence test on paraformaldehyde‐fixed cells was used for the detection of antibodies bound to the platelets and granulocytes and present in the sera of 24 patients with Evans syndrome and a further 29 patients with both idiopathic thrombocytopenia (ITP) and idiopathic neutropenia (INP), but without autoimmune haemolytic anaemia (AIHA). The direct immunofluorescence test on platelets and/or on granulocytes was positive in all patients with a cytopenia, but the sera of only 17 patients with the Evans syndrome and 15 of the other patients contained platelet‐ or granulocyte‐specific autoantibodies.

From absorption and elution experiments, it appeared that the autoantibodies were directed against antigens specific for the various peripheral blood cells, i.e. erythrocytes, platelets and granulocytes and that they were not cross‐reacting.
Studies on the haemopoietic toxicity of nitrous oxide in man
Authors: Skacel P. O. , Hewlett A. M. , Lewis J. D. , Lumb M. , Nunn J. F. , Chanarin I.
Published: July 2010

Summary. Nitrous oxide inactivates vitamin Bi2 and in man can produce a megaloblastic anaemia. Haematological and biochemical changes were studied in nine surgical patients ventilated with 70% N20 for up to 24 h and in three control patients. There was a rise in the numbers of hypersegmented neutrophils in peripheral blood following N2O. Serial bone marrow aspirates showed gross megaloblastic change after 24 h of N20 which had reverted to normoblastic but dyserythropoietic haemopoiesis by 1 week. Giant forms of early myeloid precursors were also seen after 24 h ventilation with N20 but by 1 week abnormalities were evident in more mature cells, metamyelocytes and segmented neutrophils. Megalo‐blastosis was associated with abnormal dU suppression which showed a correction pattern similar to that seen in vitamin Bi2 deficiency. Administration of N20 was also associated with a progressive rise in serum folate and fall in serum methionine levels. No similar patterns were seen in the three control patients.
Quantitative cytochemistry of the toxic granulation blood neutrophil
Authors: Schofield K. P. , Stone P. C. W. , Beddall A. C. , Stuart J.
Published: July 2010

Summary. A quantitative cytochemical study has been made, using scanning‐integrating microdensitometry, of 1000 toxic granulation blood neutrophils from 20 infected patients, in comparison with 1250 normal blood neutrophils. Myeloid precursor cells in 10 normal marrows were also studied. Normal bone marrow granulocyte maturation was associated with a progressive decrease in azurophilic granule enzymes (myeloperoxidase, β‐glucuronidase, acid phosphatase, chloroacetate esterase), and also Alcian blue staining for acid mucosubstance, but an increase in the specific granule marker lactoferrin. Toxic granulation blood neutrophils showed minor changes in the enzyme content of their azurophilic and specific granules, consistent with cell immaturity, and an increase in acid mucosubstance in azurophilic granules. Abnormal maturation of azurophilic granules, with persistence of acid mucosubstance, is the likely explanation for the intense Romanowsky dye staining of the toxic granulation neutrophil.
Immunoelectrophoretic evidence of a thrombin‐induced abnormality in a new variant of hereditary dysfunctional antithrombin III (AT III ‘Vicenza’)
Authors: Barbui T. , Finazzi G. , Rodeghiero F. , Dini E.
Published: July 2010

Summary. An Italian family with thrombosis and hereditary dysfunctional AT III, i.e. reduced biological activity and normal levels of the immunoreactive protein is presented. The abnormal molecule, called AT III ‘Vicenza’, was characterized by two‐dimensional crossed immunoelectrophoresis either in the absence or presence of heparin. In the presence of heparin, a normal pattern was found in plasma, but abnormality was evident in serum. A similar abnormality was found in plasma artificially clotted with thrombin. The role of thrombin in inducing the immunoelectrophoretic abnormality of AT III ‘Vicenza’ is discussed.
Testosterone, erythropoietin and anaemia in patients with disseminated bronchial cancer
Authors: Schreuder W. O. , Ting W. C. , Smith S. , Jacobs A.
Published: July 2010

Summary. In 21 newly diagnosed male patients with disseminated bronchial cancer, anaemia was associated with a reduced serum concentration of testosterone. Erythropoietin levels were increased in anaemic patients.

The results suggest a normal erythropoietin response to anaemia in this condition. The reduced testosterone concentration is associated with normal levels of luteinizing hormone and its significance in the pathogenesis of the anaemia is uncertain.
A pathway of coagulation on bovine capillary endothelial cells
Authors: Nawroth Peter P. , Stern David M. , Kisiel Walter , Dietrich Manfred
Published: July 2010

Summary. In this report cultured bovine capillary endothelial cells are demonstrated to specifically bind factors IX and X and also their activated forms. Bound factor IXa and cell‐associated factor VIII can activate factor X. The product of this reaction, factor Xa, can then interact with a factor V‐like molecule expressed by capillary endothelial cells promoting thrombin formation. The thrombin formed can cleave fibrinogen leading to release of fibrinopeptide A and clot formation. Endotoxin‐treatment of capillary endothelial cells leads to induction of tissue factor activity which, in the presence of factor VIIa, promotes activation of factors IX and X. The amount of factor Xaformed endotoxin‐treated endothelial cells incubated with factors VIIa, IX, VIII and X, is 8 times greater than cells incubated with factors VIIa and X alone. This indicates that on the perturbed endothelial cell surface factors VIII and IX do play an important role in factor X activation by the tissue factor pathway. The perturbed capillary endothelial cell can thus provide a model of the thrombotic state promoting initiation and propagation of a procoagulant pathway leading to thrombin formation. This pathway of coagulation is endothelial cell‐dependent, since it requires expression of tissue factor and factor V by capillary endothelial cells, as well as interaction of coagulation factors with the surface of capillary endothelial cells.
Platelet‐derived growth factor(s) mitogenic activity in patients with myeloproliferative disease
Authors: Bernabei P. A. , Arcangeli A. , Casini M. , Grossi A. , Padovani R. , Ferrini P. Rossi
Published: July 2010

Summary. Platelet‐derived growth factor has been invoked in the pathogenesis of medullary fibrosis during myeloproliferative disorders. In this study we compared the mitogenic activity of heat‐stable platelet‐derived growth factor(s) from 13 patients suffering from myeloproliferative disorders with that of a normal group. The test was carried out on G0 growth arrested Balb/c 3T3 fibroblasts incubated with various concentrations of platelet extracts, determining the entrance into the S phase by means of [14C]thymidine uptake. The incorporation curves of [14C]thymidine by the fibroblast culture, under the effect of pathological extracts, were consistently lower than the control curve, indicating a lower level of PDGF(s) in platelets from patients. The greatest depression of this activity was found to be associated with highest degree of medullary fibrosis (agnogenic myeloid metaplasia patient group), in agreement with the hypothesis that fibroblast activation within bone marrow during myeloproliferative disorders might be correlated with a PDGF(s) release in the bone marrow environment.
Hereditary monocyte esterase deficiency
Authors: Markey Geraldine M. , Alexander H. D. , Mcconnell Robyn , Kyle Anne , Morris T. C. M. , Robertson J. H.
Published: July 2010

Summary. An inherited cytochemical staining abnormality of monocytes is described, affecting members of three generations of one family. a‐Naphthyl acetate and a‐naphthyl butyrate esterase staining reactions have been consistently negative in 9 5% of the monocytes of the propositus and her son and in 60‐70% of the monocytes in two of four grandchildren.

Observation of persistently esterase‐negative monocytes in a patient with non‐Hodgkin's lymphoma led to us to investigate her family and we here report the findings.
Whole blood viscosity in polycythaemia: the effect of iron deficiency at a range of haemoglobin and packed cell volumes
Authors: Van de Pette J. E. W. , Guthrie D. L. , Pearson T. C
Published: July 2010

Summary. The effect of iron deficient red cells changes (MCH range 20‐0‐32‐6 pg) on whole blood viscosity (WBV) at high and low shear rates (94‐5 and 0‐51 s_1), has been assessed at a wide range of standardized PCV (0‐30‐0‐65) and standardized Hb values (10‐0‐22‐0 g/dl). Particular attention was given to the precision of PCV measurement. At all PCV values there was no correlation between MCH and WBV, whereas at all Hb values there was a significant negative correlation between MCH and WBV.

If the treatment of polycythaemia is by venesection, iron deficient red cell changes occur frequently. These changes are unimportant if control of the WBV is the objective of treatment, as long as the PCV is accurately measured. Alternatively, in the secondary polycythaemias, where oxygen saturation is reduced, the induction of iron deficient red cell changes could be regarded as disadvantageous since at a standard PCV of 0‐50 the Hb and hence oxygen carrying capacity is reduced by approximately 11% between MCH values of 30 and 20 pg.
Frequent lymphocytes infection by hepatitis B virus in haemophiliacs
Authors: Lauré F. , Chatenoud L. , Pasquinelli C. , Gazengel C. , Beaurain G. , Torchet M.‐F. , Zagury D. , Bach J. F. , Bréchot C.
Published: July 2010

Summary. We have tested the different mononuclear blood cell populations of seven patients with severe haemophilia and one patient with F VII deficiency for the presence of HBV DNA. These subjects were all polytransfused with non‐heated coagulation factors; three were HBsAg positive, five HBsAg negative but anti‐HBc and anti‐HBs positive; HBV DNA sequences were detected in six subjects including three without detectable serum HBsAg. Furthermore the viral DNA sequences were identified in the T lymphocyte subpopulations (OKT4+ and/or OKT8+ cells). This observation suggests that HBV infection of lymphocytes might be related to the immunological disorders observed in these patients.
Human serum ferritin G‐peptide is recognized by anti‐L ferritin subunit antibodies and concanavalin‐A
Authors: Santambrogio Paolo , Cozzi Anna , Levi Sonia , Arosio Paolo
Published: July 2010

Summary. Ferritin was purified from serum of patients with idiopatic haemochromatosis. Analysis on SDS electrophoresis showed that it is composed of two major bands of 19 000 and 23000 Mr. The smaller peptide has an electrophoretic mobility and immunochemical reactivity similar to that of tissue L subunit. The larger, previously named G subunit, is recognized by concanavalin‐A and by anti ferritin L‐subunit, but not by anti‐H, monoclonal antibodies. All of the antibodies show higher affinity for the L than for the G subunit. Therefore, the G chain appears immunochemically similar, but not identical, to ferritin L chain, and is responsible for serum ferritin binding concanavalin‐A.
Spectrum and frequency of autoimmune derangements in lymphoproliferative disorders: analysis of 637 cases and comparison with myeloproliferative diseases*
Authors: Dührsen Ulrich , Augener Wiprecht , Zwingers Thomas , Brittinger Günter
Published: July 2010

The records of 637 patients with lymphoproliferative disorders and 346 patients with myeloproliferative disorders were retrospectively analysed for the presence of additional autoimmune derangements. The frequency of autoimmune perturbations in lymphoproliferative diseases (51 cases; 8.0%) was significantly higher than in myeloproliferative diseases (six cases; 1–7%: P<0.0001). Rheumatic disorders, autoallergic haematological manifestations and other organ‐specific autoimmune derangements were responsible for about one third each of the observed disturbances.

Autoimmune diseases which preceded the onset of malignancy, occurred in lymphoproliferative and myeloproliferative disorders with a comparable frequency without significant differences between individual subgroups of lymphoproliferative diseases. In contrast, autoimmune complications developing in the course of the neoplastic disease were significantly more frequent in lymphoproliferative (4.9%) than in myeloproliferative disorders (0.3%; P<0.0005). Here marked differences were observed between individual lymphoma entities, the rate of concomitant autoimmune derangements ranging from zero to over 15%. With the exception of centroblastic‐centrocytic lymphoma which in no case was associated with secondary autoimmune complications, the proportion of patients with autoimmune perturbations increased with improving prognosis of the lymphoproliferative diseases. Possible pathogenetic mechanisms involved in the manifestation of autoimmune complications in malignant lymphomas are discussed.
Does histologic grading of inflammation in bone marrow predict the response of aplastic anaemia patients to antithymocyte globulin therapy?
Authors: Sale George E. , Rajantie Jukka , Doney Kristine , Appelbaum Frederick R. , Store Rainer , Thomas E. Donnall
Published: July 2010

We tested the hypothesis of teVelde and Haak that the degree of bone marrow inflammation in aplastic anaemia might correlate with an immunological process responsive to immunosuppressive therapy. 120 patients with aplastic anaemia but no suitable marrow donor were treated with horse antithymocyte globulin (ATG) and 53 who had matched sibling donors with bone marrow transplants. Pretreatment bone marrow histology in methacrylate and paraffin specimens was graded by degree of inflammatory infiltrate in a four‐tiered system. High grade (II‐III) was compared to low (0–1) as a correlate of response to ATG. Complete and partial response to ATG was seen in 50% of patients with high grade marrow and 31% of patients with low grade marrow (P= 0.099). Only one of four patients with grade III inflammation responded significantly to ATG treatment. Median survival following ATG therapy was similar in both groups as well. There was a significantly lower median age in the patients with low grade (24.5 years) versus high grade (37.5 years) inflammation (P= 0.016). Grade also had no prognostic value in the marrow transplant group.
Type IIB von Willebrand's disease with probable autosomal recessive inheritance and presenting as thrombocytopenia in infancy
Authors: Donnear Mikael , Holmberg Lars , Nilsson Inga Marie
Published: July 2010

von Willebrand's disease (vWD) is a congenital bleeding disorder that exists in two main forms. In the classic form, type I, the concentration of the von Willebrand factor (vWF) in plasma is decreased. In type II vWD, the vWF is structurally altered. Type II can be further divided into at least six subtypes (A, B, C, D, E and F). In type IIB the vWF, in contrast to other variants of vWD, shows an increased affinity for platelets. IIB vWD is generally believed to be inherited in an autosomal dominant manner. We describe two families with three affected children in whom an autosomal recessive inheritance is more likely. Thrombocytopenia, constant or variable, was present from early infancy in all three cases. Type IIB vWD should thus be included in the differential diagnosis of congenital thrombocytopenia.
DNA haplotype heterogeneity of β‐thalassaemia in Greece: feasibility of prenatal diagnosis
Authors: Athanassiadou A. , Zarkadis I. , Papahadjopoulou A. , Maniatis G. M.
Published: July 2010

We have carried out DNA haplotype analysis of 69 β‐thalassaemia patients in Greece and 42 of the parents using seven standard polymorphic sites. Our data show a high degree of heterogeneity of the chromosomal background in which β‐thalassaemia occurs in Greece, suggesting a high degree of heterogeneity in the β‐thalassaemia mutations involved. Haplotype I is found here to represent 45% of total β‐thalassaemia mutations detected, a proportion well below the 67% reported in earlier studies with Greek‐American patients. Nine different haplotypes are detected and the ones carrying β+ mutations are the majority, including those which are linked to β+ mutations associated with a thalassaemia intermedia phenotype, and which constitute 11% of all haplotypes. One of these haplotypes (—‐ + + +) has never before been reported to occur in non‐Africans, whether in βthal or βA chromosomes, and it is found here to be of African origin rather than the product of recombination. In 21 families haplotype analysis showed that prenatal diagnosis for a second child was feasible in 81% of the cases. Use of the AvaII‐Ψβ polymorphic site as well as the seven standard ones brought this proportion up to 90%.
The development of non‐responsiveness to immunotherapy with monoclonal anti‐idiotypic antibodies in a patient with B‐CLL
Authors: Allebes Wil A. , Preijers Frank W. M. B. , Haanen Clemens , Capel Peter J. A.
Published: July 2010

A patient having a B cell chronic lymphocytic leukaemia was treated with a monoclonal anti‐idiotypic antibody (MoAb anti‐id). Up to 24.5 g of MoAb anti‐id has been administered to the patient over a period of 1 year without serious side effects. Despite a substantial amount of serum idiotype (id = 100 μg/ml) and a low expression of id on the tumour cells (±6000 molecules per cell) clearance of serum id and a marked tumour reduction was obtained. Therapy resistance developed and coincided with a decreased clearance rate of circulating id–anti‐id immune complexes and an increased modulation of cellular id expression, in vivo. This suggests that a decreased clearance rate of anti‐id coated tumour cells provided more time for id modulation in vivo. resulting in therapy resistance. Therefore, the overall capacity of the natural effector system may have an important influence on the ultimate therapeutic effect of immunotherapy with MoAb anti‐id. Although the partial remission obtained was not long‐lasting, this study shows that MoAb anti‐id therapy can be effective even when id expression on the tumour cells is low and a substantial amount of serum id is present.
Relationship between ex vivo anti‐proteinase (factor Xa and thrombin) assays and in vivo anticoagulant effect of very low molecular weight heparin, CY222
Authors: Tew C. J. , Lane D. A. , Thompson E. , Ireland H. , Curtis J. R.
Published: July 2010

There is uncertainty as to which activities of unfractionated heparin (UFH) and low MW heparin are responsible for their anticoagulant and antithrombotic properties. We have sought to answer this question by examining plasma samples taken during a recently conducted dose‐finding study of the low MW heparin, CY222, in haemodialysis for chronic renal failure. In this study, in vivo anticoagulant effect was assessed by measurement of plasma FPA levels. UFH was administered as a dose of 5000 iu bolus + 1500 iu/h maintenance infusion, while the effects of three doses of CY222 were studied (10000, 15000 and 20000 Institute Choay anti‐factor Xa u bolus, all with 1500 Institute Choay anti‐factor Xa u/h maintenance infusion). Anti‐factor Xa levels were determined by chromogenic substrate assay. Anti‐thrombin levels were determined by chromogenic substrate assay and by quantitation of catalysed thrombin–inhibitor complexes (using autoradiography). Analysis of the results indicate that plasma fibrinopeptide A (FPA) levels correlate with anti‐factor Xa (r= 0.45) and anti‐thrombin (substrate) (r= ‐0.63) levels of UFH, but only with the anti‐factor Xa levels (r= ‐0.41) of CY222. These results suggest that the anti‐factor Xa assay is currently the most suitable assay for monitoring low MW heparins such as CY222 in humans.
A detailed morphological evaluation of the evolution of the haemostatic plug in normal, factor VII and factor VIII deficient dogs
Authors: Velden Peter Vander , Giles Alan R.
Published: July 2010

A standardized injury of the nail cuticle of normal, factor VII and factor VIII deficient dogs was used to study the evolution of the morphological changes occurring within the forming and formed haemostatic plug at the site of vascular injury. The morphological changes occurring were documented by light and transmission electron microscopy (TEM). Randomized measurements were made of the distances between adjacent platelets as a function of platelet interdigitation or compaction and the degree of dilatation of the open canalicular system (OCS) was used as an indicator of the degree of platelet activation. Fibrin deposition was noted both in terms of its location and the point in time at which it first appeared. TEM demonstrated major differences between the factor VIII deficient and the normal and factor VII deficient groups. In the normal animals the intermembrane distance showed noticeable changes with the platelets becoming tightly interdigitated at the time bleeding stopped. During the same period the OCS became dilated. These changes, which were not seen in the factor VIII deficient animals, continued until many platelets lost their intracellular content and became balloon cells or ghosts and fibrinous transformation became prominent. Although those events did occur in the factor VII deficient state, each was delayed and resulted in significant differences between the factor VII and normal animals suggesting that the extrinsic pathway may play an important role in initiating the changes noted. The results suggested that the generation of thrombin and/or factor Xa is essential to promote the initial stabilization of the platelet plug as well as initiating its subsequent consolidation by fibrinous transformation.
Factor × Roma: a congenital factor × variant defective at different degrees in the intrinsic and the extrinsic activation
Authors: Stefano V. De , Leone G. , Ferrelli R. , Hassan H. J. , Macioce G. , Bizzi B.
Published: July 2010

A factor × molecular variant was identified in a 13‐year‐old girl affected by a bleeding tendency. Factor × antigen levels and activation by Russel's viper venom (tested both by clotting and amidolytic assays) were normal. Factor × crossed immunoelectrophoresis was found to be identical to that of the control plasma. Factor × functional activity was low (3% of the normal) if tested by PTT‐derived assays, whereas it was found at intermediate levels (about 30–50% of the normal) if measured by prothrombin time‐derived assays. The defect in the extrinsic activation was more clearly disclosed using as activating agent thromboplastin from ox brain. The factor × of the patient was completely adsorbed by aluminium hydroxide. The parents of the propositus (first degree cousins) showed factor × functional levels compatible with a condition of heterozygosity for the abnormality. This factor × molecular variant appears different from the other ones so far described and was named ‘Factor × Roma’.
The spectrum of β‐thalassaemia mutations in Sicily
Authors: Marzo Rosalba Di , Dowling Carol E. , Wong Corinne , Maggio Aurelio , Kazazian Haig H.
Published: July 2010

To characterize β‐thalassaemia genes among the Sicilian population we have previously determined the DNA haplotypes in the β‐globin gene cluster of 99 β‐thal chromosomes. We found seven haplotypes, although 95 of 99 β‐thal chromosomes contained framework 1 and framework 3 β genes. We have now determined the mutation in all 99 of these β‐thal genes by the use of oligonucleotide hybridization, PCR‐amplification and direct genomic sequencing, and direct restriction analysis. Our results indicate that (1) the β0‐39 mutation is most frequent (35%): (2) β0‐39, IVS‐1 nt 110 and IVS‐1 nt 6 mutations account for 90% of β‐thal genes: (3) the IVS‐1 nt 6 mutation is more frequent in thalassaemia intermedia (77%) than in Cooley's disease (34%): (4) the association between haplotypes and specific mutations is imperfect, but mutation spread has occurred within haplotypes containing the same β‐gene framework: (5) the β0‐39 and the IVS‐1 nt 6 mutations, with a mutation spread to two major haplotypes, may be older than the IVS‐1 nt 110 mutation: (6) these data make possible first‐trimester prenatal diagnosis in many families (85%) in Sicily using only three pairs of oligonucleotides. In addition, a new mutation, a frameshift at codon 76 due to loss of a C residue, was found in a single β‐thal chromosome.
Increased proteolytic activity of erythrocyte membrane in spur cell anaemia
Authors: Olivieri O. , Guarini P. , Negri M. , Bassi A. , Corrocher R. , Biffanti S. , De Matteis M. C. , Vettore L.
Published: July 2010

In a case of ‘spur cell anaemia’(SCA) a reduced esterified/free cholesterol ratio was found in plasma, in LDL and HDL fractions, and an increased cholesterol/phospholipid (C/PL) molar ratio in erythrocyte membrane. Cation transport was normal with the exception of Li‐Na counter‐transport which was decreased. An increased intrinsic membrane proteolytic activity (IMPA) was demonstrated by the generalized reduction or, sometimes, disappearance of protein bands on SDS‐PAGE in patient ghosts when the proteolysis was allowed. This characteristic was found to be transferable to normal cells by incubation in SCA‐plasma; moreover membrane C/PL molar ratio was augmented after incubation. Normal plasma was not able to normalize IMPA of SCA cells ‘in vitro’, even if it induced a remarkable decrease of membrane C/PL molar ratio. Nevertheless IMPA normalization did occur ‘in vivo’, when the SCA cells were exposed to therapeutic ‘plasma exchange’(3.3 litre/week). The results suggest the following conclusions: (a) in our SCA patient there is an increased IMPA; (b) this feature, as well as membrane lipid alteration, is transferable to normal erythrocytes; (c) this case seems to demonstrate, for the first time in our knowledge, a modulating effect of plasma on IMPA in erythrocytes.
Granulocyte macrophage colony stimulating factor induced changes in cellular adhesion molecule expression and adhesion to endothelium: in‐vitro and in‐vivo studies in man
Authors: Devereux S. , Bull H. A. , Ccmpos‐Costa D. , Saib R. , Linch D. C.
Published: July 2010

Summary The administration of recombinant human granulocyte‐macrophage colony‐stimulating factor (rhGM‐CSF) causes a transient leucopenia. Radionuclide labelling studies showed this to be due to margination of neutrophils and monocytes predominantly in the pulmonary vasculature. No evidence of complement activation was found. A rapid in‐vivo rise in neutrophil cellular adhesion molecule (CAM) expression was observed paralleling the development of the neutropenia. Neutrophils exposed to rhGM‐CSF in‐vitro showed similar rapid increases in CAM expression. The adherence of chromium‐labelled neutrophils to endothelial cell cultures was modestly but highly significantly increased by rhGM‐CSF, an effect that was reduced by the binding of a monoclonal antibody to the beta chain of neutrophil CAM. The margination of phagocytic cells induced by rhGM‐CSF administration is therefore likely to be due at least in part to increased expression of adhesion promoting glycoproteins. The demargination, however, occurred at a time when neutrophil CAM expression was still high, suggesting that dissociation of the neutrophil‐endothelial cell interaction depends on factors other than downregulation of CAM expression. In‐vivo modulation of phagocyte CAMS and adhesive properties by GM‐CSF may be of importance in the normal inflammatory response.
Decreased interleukin 1 production in aplastic anaemia
Authors: Nakao Shinji , Matsushima Kouji , Young Neal
Published: July 2010

Summary. Interleukin 1 (IL‐1) is an important regulator of immune system function. IL 1 also affects baematopoiesis in vitro: it causes release of colony stimulating factors from fibroblasts and endothelial cells and can directly act on primitive haeniatopoietic stem cells. We investigated IL 1 production in vitro by stimulated peripheral blood mononuclear cells of patients with aplastic anaemia (N = 17), patients with other haematologic diseases (N = 27), and normal individuals (N = 22) using a bioassay for IIL 1 activity. Ten aplastic patient s showed markedly decreased IL 1 production. IL 1 production by fibronectin‐affinity purified monocytes was decreased in six of seven of these patients: in three other cases, in which IL 1 mononuclear cell production was undetectable, sufficient monocytes could not be isolated. IL lα and IL 1β precursor molecules were also absent or much decreased when mononuclear cell lysates from these patients were analysed by immunoblot using specific polyclonal sera. Aplastic patients with low IL 1 production were distinguished by the severity of their disease and the degree of neutropenia. Patients with rnyelodysplasia with comparable degrees of pancytopenia had normal IL 1 production. This is the first example of deficient haematopoietic growth factor production in a bone marrow failure syndrome. Decreased IL I production may contribute to the pathogenesis of some cases of aplastic anaemia and to susceptibility to infection.
Effect of warfarin on plasma concentrations of vitamin K dependent coagulation factors in patients with stable control and monitored compliance
Authors: Kumar S. , Haigh J. R. M. , Tate G. , Boothby M. , Joanes D. N. , Davies J. A. , Roberts B. E. , Feely M. P.
Published: July 2010

There is a discrepancy in the results of reported studies of levels of vitamin K dependent coagulation factors in patients on warfarin therapy. This may have arisen partly because of the problem of assuring compliance with therapy in outpatients. The plasma concentrations of the vitamin K dependent clotting factors II, VII, IX and X were studied in 23 outpatients whose adherence to prescribed warfarin therapy was determined using a pharmacological indicator of complicance. In these patients, who were shown to have consistently good compliance and stable anticoagulant control over a period of 3–6 months, the activities in plasma of the four coagulation factors were not equally suppressed. Factor IX levels were significantly greater than those of factor VII (P<0.0001) which in turn were significantly greater than the levels of factor II (P<0.0001) or factor X (P<0.0001). There was no significant difference between the levels of factors II and X which were depressed to a similar extent. The proportion of variability of the International Normalized Ratio (INR) explained by linear regression was 51–77% and a model was derived to predict the INR from the mean of the levels of the four clotting factors. The concentrations of the coagulation factors II, VII, IX and X are likely to be highly dependent on the degree of compliance with warfarin therapy which should be taken into account when investigating the behaviour of these factors.
Japanese B cell chronic lymphocytic leukaemia: a cytogenetic and molecular biological study
Authors: Asou Hiroya , Takechi Miho , Tanaka Kimio , Tashiro Satoshi , Dohy Hiroo , Ohno Ryuzo , Kamada Nanao
Published: July 2010

Summary. Clinical, cytogenetic, and molecular genetic studies were performed to clarify the pathophysiology of Japanese B cell chronic lymphocytic leukaemia (B‐CLL), since the incidence of B‐CLL in Japan is significantly lower than in western countries. The clinical and laboratory features of 55 Japanese patients with B‐CLL in this study did not differ from those of Americans or Europeans with B‐CLL. In the chromosome analyses, suitable metaphases with good band quality were obtained from 48 patients (87.2%), of whom 22 patients (45.8%) showed clonal chromosome aberrations and 14 (29.2%) had non‐clonal aberrations. Trisomy 12 and abnormalities of 14q and 13q were found in four (18.2%), two (9.1%) and six patients (27.2%). respectively. There were no particular chromosome abnormalities or specific breakpoints in Japanese B‐CLL. However, complex karyotype was found in higher incidence than in western countries. In the Southern blot analyses, rearranged band patterns were observed in the major breakpoint region (mbr) of the bcl‐2 gene in one case, in the 5′‐breakpoint region (5′‐bcl‐2) in two, and bcl‐3 in one. Of the two patients with 5′‐bcl‐2 rearrangements, one had a normal karyotype and the other had t(2:18)(p12:q21). The incidence of rearrangements of the bcl‐1, bcl‐2 and bcl‐3 genes in Japanese B‐CLL was similar to that in western countries. These findings suggest that the biological characteristics of B‐CLL in Japan are almost the same as those in western countries, although the incidence of B‐CLL in Japan is quite different: this may be related to racial differences, which seem to be an important factor in the development of B‐CLL.
Multiple myeloma: ‘early’ plasma cell phenotype identifies patients with aggressive biological and clinical characteristics
Authors: Omedè Paola , Boccadoro Mario , Fusaro Antonella , Gallone Gabriele , Pileri Alessandro
Published: July 2010

Summary. The immunological phenotype of bone marrow myeloma cells and peripheral blood lymphocytes was evaluated in 38 untreated myeloma patients. A striking increase of monotypic cells expressing the same light chain as the M component was observed in bone marrow from 18/38 (47%) patients. A two‐colour analysis clarified that the majority of myeloma cells co‐expressed plasma cell and B lymphocyte markers (cyIg, CD38, CD56 and sIg), and were regarded as early‐plasma cells (early‐PC). In the remaining patients, myeloma cells expressed plasma cell markers only (late‐PC). Phenotype corresponded to a distinct morphological pattern: early‐PC showed a lympho‐plasmocytoid feature with significantly lower diameters than late‐PC (12.1 v 14.8 μm, P<0.007). Moreover, the plasma cell labelling index was significantly increased in early‐PC patients (1.2 v 0.5%, P<0.04). In peripheral blood from patients with early‐PC, monotypic cells co‐expressing sIg and CD38, CD56, but not CD19, were also detected. These data suggest a recirculation of early‐PC. Myeloma cells maintained their phenotypic pattern during the course of the disease. This observation suggests that the degree of maturation is an intrinsic characteristic of the myeloma cell population in individual patients. The evaluation of prognostic factors, such as β2‐microglobulin, C‐reactive protein and neopterin, showed a statistically significant increase in the early‐PC patients, suggesting a poor outcome. In conclusion, myeloma cell phenotype allows identification of a myeloma variant with aggressive biological and clinical characteristics.
Aprotinin reduces cardiopulmonary bypass‐induced blood loss and inhibits fibrinolysis without influencing platelets
Authors: Orchard M. A. , Goodchild C. S. , Prentice C. R. M. , Davies J. A. , Benoit S. E. , Creighton‐Kemsford L. J. , Gaffney P. J. , Michelson A. D.
Published: July 2010

Summary. Cardiopulmonary bypass (CPB) induces a bleeding defect which leads to enhanced blood loss. A double‐blind study was carried out comparing aprotinin with placebo in patients undergoing re‐operation for heart valve replacement. The results confirm that aprotinin is effective at reducing such loss.

In the placebo treated group, significant increases were observed, during CPB, in the plasma concentrations of fibrinolytic activity, tissue plasminogen activator antigen. D‐dimer, and β‐thromboglobulin. Platelet counts fell within 5–10 min of the patients going onto CPB, but this could be accounted for by the dilutional effect of the extracorporeal circuit. Inhibition of responsiveness of platelets, as judged by aggregometry. was significant only at the end of bypass when collagen was the agonist and after protamine reversal when ristocetin was the agonist. CPB did not enhance the release, into the circulation, of glycocalicin (a proteolytic fragment of glycoprotein Ib).

In the aprotinin‐treated group, the formation of fibrin degradation products as measured by D‐dimer was inhibited. However, aprotinin did not influence the change in platelet count, suppress β‐thromboglobulin release from platelets, prevent the inhibition of platelet function or influence the concentration of plasma glycocalicin during the study period.

These observations confirm that CPB leads to a fibrinolytic state and less responsive platelets. This study also indicates that aprotinin‐induced reduction in blood loss is associated with inhibition of plasmin‐mediated fibrin digestion and that the mechanism by which aprotinin reduces blood loss is not via protection of platelets during CPB.
Adhesion of flowing leucocytes to immobilized platelets
Authors: Lalor Patricia , Nash Gerard B.
Published: July 2010

Adhesion of neutrophils, lymphocytes and promyelocytic HL60 cells was compared in a flow‐based model in which a monolayer of activated platelets formed the adhesive substrate. Each type of leucocyte formed P‐selectin‐mediated rolling attachments on the platelet surface under physiologically relevant flow conditions. Lymphocytes adhered less, and HL60 in similar numbers, compared to neutrophils, whereas the lymphocytes and HL60 cells rolled much more rapidly. Sulphated, sialylated saccharide(s) were implicated as ligand(s) for P‐selectin for all leucocytes, but L‐selectin (borne by neutrophils and lymphocytes, but not HL60 cells) appears to be a major presenter of ligand for neutrophils alone. T cells enriched from peripheral blood lymphocytes adhered in greater numbers than B cells. Differentiation of HL60 cells to neutrophil‐like cells (induced by DMSO) caused cell volume to decrease and surface expression of integrin adhesion molecules to increase, but only a small percentage of cells were converted to an L‐selectin‐bearing phenotype. Differentiated cells showed evidence of stabilization of adhesion with increasing stress and a marked reduction in rolling velocity. These studies indicate that cell differentiation may be accompanied by alteration of adhesive behaviour, resulting from changes in physical characteristics as well as surface properties. Moreover, results suggest that P‐selectin could promote lymphocyte attachment to endothelium in acute inflammatory conditions and possibly mediate lymphocyte‐platelet interaction during thrombosis.
Interpretation of measured red cell mass and plasma volume in adults: Expert Panel on Radionuclides of the International Council for Standardization in Haematology
Authors: Pearson T. C. , Guthrie D. L. , Simpson J. , Chinn S. , Barosi G. , Ferrant A. , Lewis S. M. , Najean Y.
Published: July 2010

Four European centres provided height and weight data on 202 males and 204 females undergoing red cell mass (RCM) and plasma volume (PV) measurements. For these populations, the RCM and PV predictions by the various published methods were compared. It was shown clearly that predictions based solely on body weight were inappropriate, particularly because approximately half of the male and female populations could be regarded as overweight or obese. Although there was reasonable agreement in the prediction values given by the formulae based on both height and weight, it was not possible to establish which formulae could be recommended. For that reason, the published literature containing normal RCM and PV measurements were re‐examined. RCM data for 283 males and 171 females and PV data for 100 males and 67 females were included. Measurements were standardized for variables such as trapped plasma in the PCV, exclusion of buffy coat in the PCV and calculation of PV at zero time. As a result of this analysis, prediction formulae based on surface area for RCM and PV with 98/99% reference ranges have been established.
t(8;21) myelodysplasia, an early presentation of M2 AML
Authors: Taj A. S. , Ross F. M. , Vickers M. , Choudhury D. N. , Harvey J. F. , Barber J. C. K. , Barton C. , Smith A. G.
Published: July 2010

The reciprocal translocation of genetic material between chromosomes 8 and 21, t(8;21), is usually restricted to cases of acute myeloid leukaemia (AML). Cases of AML with t(8;21) exhibit characteristic dysplastic features in myeloid and erythroid lineages with reduction in megakaryoctes. We report details of three patients presenting with myelodysplastic features; two had a typical t(8;21), and the thir had a variant t(8;21) translocation. We discuss the signifcance of t(8;21) in the aetiology of myelodysplastic syndrome (MDS) and implications for the management of such patients.
Long‐term disease‐free survival after autologous bone marrow transplantation in a primary plasma cell leukaemia: detection of minimal residual disease in the transplant marrow by third‐complementarity‐determining region‐specific probes
Authors: Yeh Kun‐Huei , Lin Ming‐Tseh , Tang Jih‐Luh , Yang2 Chih‐Hsin , Tsay Woei , Chen Yao‐Chang
Published: July 2010

Primary plasma cell leukaemia (PPCL) is a rare form of plasma cell neoplasm. Treatments of PPCL have been most disappointing. A patient with PPCL received high‐dose melphalan plus total body irradiation and autologous bone marrow transplantation (ABMT). By using third‐complementarity‐determining region (CDRIII)‐specific probes, minimal residual disease (MRD) was detected in remission marrow, collected 1 month before ABMT. MRD was no longer detected by CDRIII‐specific probes 6, 19 and 26 months after transplantation. The patient remained in complete remission up to 59 months after ABMT.
Osteoporosis in severe congenital neutropenia treated with granulocyte colony‐stimulating factor
Authors: Bishop N. J. , Williams D. M. , Compston J. C. , Stirling D. M. , Prentice A.
Published: July 2010

Recombinant human granulocyte colony‐stimulating factor (G‐CSF) has substantially improved life expectancy for children with severe congenital neutropenia (SCN). Severe osteoporosis, reported in this population, may relate to the disease process, or be a therapeutic side‐effect. This report details bone loss, quantitated absorptiometrically and histomorphometrically, in a child with SCN and vertebral collapse, and the positive response to anabolic steroid and bisphosphonate therapy.
Hydration of sickle erythrocytes using a herbal extract (Pfaffia paniculata) in vitro
Authors: Ballas Samir K.
Published: July 2010

Pfaffia paniculata (PP) is a perennial wild plant that grows in South America. Its root powder has been used by South American Indians for a variety of ailments and has been reported to have a salutary effect on sickle cell disease in Brazil. Its mechanism of action, however, is unknown. In this report, we present experimental data showing that PP improves the deformability of sickle cells, increases their Na+ content and their mean corpuscular volume (MCV). These findings indicate that PP functions as a sodium ionophore on sickle cells and improves their hydration status and rheological properties.
Allogeneic bone marrow transplantation from unrelated donors using in vivo anti‐T‐cell globulin
Authors: Finke Jürgen , Bertz Hartmut , Schmoor Claudia , Veelken Hendrik , Behringer Dirk , Wäsch Ralph , Kunzmann Regina , Heidecker Leonora , Lang Helmut , Meyer‐König Ursula , Mertelsmann Roland
Published: July 2010

Despite improvements in HLA typing, graft‐versus‐host disease (GVHD) continues to impair the results after volunteer unrelated donor bone marrow transplantation (VUD‐BMT) in adult patients compared with matched sibling BMT. Here, the outcome after VUD‐BMT using a specific regimen with high‐dose anti‐T‐lymphocyte globulin (ATG) was analysed. Fifty‐five adult patients, median age 34 years (range 17–55 years), with acute or chronic leukaemia or myelodysplastic syndrome (MDS) were transplanted in first complete remission (CR1)/first chronic phase (CP1) (early disease) (n = 21) or in advanced (CR2/CP2, no remission) disease (n = 34) from an unrelated marrow donor. GVHD prophylaxis consisted of ATG‐S (Fresenius) 60–90 mg/kg b.w. prior to transplantation, in addition to cyclosporin A and short‐course methotrexate. Graft failure did not occur and white blood cell count (WBC) > 1·0 × 109/l was reached at median day +16. The cumulative incidence of acute (a)GVHD grade II–IV was 15% [95% CI (8%, 28%)] and of chronic GVHD was 51% [95% CI (38%, 68%)]. The cumulative incidence of relapse within 1 year was 0% [95% CI (0%, 19%)] and 21% [95% CI (11%, 40%)] for patients with early and advanced disease respectively. With a median follow‐up of 28 months (range 16–45 months), 2‐year disease‐free and overall survival for patients transplanted in CR1/CP1 was 81% and 81% [95% CI (64%, 98%)], respectively, and for patients with advanced disease was 33% [95% CI (17%, 50%)] and 40% [95% CI (23%, 57%)] respectively. Complete and persistent donor chimaerism was seen in 77·5% of 40 patients evaluated. All 14 chronic myeloid leukaemia (CML)‐CP1 patients became bcr–abl negative within 250 d. High‐dose ATG pretransplant results in a low incidence of severe aGVHD without compromising donor chimaerism or elimination of minimal residual disease. Our results are similar to data obtained after matched sibling donor transplantation.