bluuebunny/crossref_metadata_2025 huggingface dataset:
Disulfide bonds are a requirement for Kell and Cartwright (Yta) blood group antigen integrity
Authors: Branch Donald R. , Muensch Helmut A. , Hian Anita L. Sy Siok , Petz Lawrence D.
Published: July 2010

Summary. We have investigated the effect of dithiothreitol (DTT) upon the Kell blood group system and other red cell antigens. All Kell blood group antigens studied (K, k, Kpa, Kpb, Jsa, Jsb and Ku) as well as the Cartwright (Yta) antigen were completely denatured after treatment with DTT. The Gerbich antigen was substantially weakened but not completely denatured. The Jsa and Jsb antigens appear to have an exquisite sensitivity to treatment with DTT and can be completely denatured using very low concentrations (≤2 mm) whereas other Kell system antigens require much higher concentrations of DTT for their denaturation (100–200 mm). Of 38 other blood group antigens investigated, only the Yta antigen was completely denatured using 200 mm DTT. Furthermore, the Yta antigen was denatured within the same concentration range as Kell and one can speculate that this indicates some biochemical relationship between these two blood group systems. From our results, we conclude that: (1) at least two distinct disulfide (S–S) bonds are required for maintenance of the Kell blood group antigen system; (2) Jsa and Jsb antigens are distinctly different from other Kell system antigens based upon sensitivity to treatment with DTT; these antigens may be located on a different antigenic domain; and (3) the Yta antigen requires at least one disulfide bond for its maintenance of antigen integrity. Although the Gerbich antigen was not completely denatured, results indicate that disulfide bonds may also be important structural determinants for these antigens.
Factors associated with graft rejection after HLA‐identical marrow transplantation for aplastic anaemia
Authors: Storb Rainer , Prentice Ross L. , Thomas E. Donnall , Appelbaum Fred R. , Deeg H. Joachim , Doney Kristine , Fefer Alexander , Goodell Brian W. , Mickelson Eric , Stewart Patricia , Sullivan Keith M. , Witherspoon Robert P.
Published: July 2010

Summary. One hundred and seventy‐five consecutive patients with severe aplastic anaemia were given high‐dose cyclophosphamide followed by marrow grafts from healthy, HLA‐identical family members, and 168 lived long enough to show engraftment. In 38 patients the graft was rejected and 29 of these died. This analysis, using a binary logistic regression model, was aimed at identifying factors that predicted marrow graft rejection. Five factors correlated with graft rejection: (1) previous blood transfusion; (2) a positive relative response in mixed leucocyte culture indicating sensitization of patient against donor; (3) a low number of marrow cells used for transplantation; (4) marrow grafts from male donors; and (5) lack of infusion of viable donor buffy coat cells in addition to the marrow for transfused patients. The findings confirm the importance of transplanting early before transfusion and indicate that the greatest possible amount of donor marrow (supplemented by stem cells/lymphoid cells derived from the peripheral blood) should be obtained.
Thymosin fraction 5 does not accelerate reconstitution of immunologic reactivity after human marrow grafting
Authors: Witherspoon R. P. , Hersman J. , Storb R. , Ochs H. , Goldstein A. L. , McClure J. , Noel D. , Weiden P. L. , Thomas E. D.
Published: July 2010

Summary. More than 1 year is required for immunologic function to recover following human marrow grafting. In an attempt to shorten the time required for immunologic reconstitution, 14 patients were treated with thymosin fraction 5 after transplantation. Two died before administration of thymosin could be completed. In the remaining 12 patients, immunologic studies were compared to those of patients who were transplanted but did not receive thymosin. While five patients had transient elevation of in vitro lymphocyte blastogenesis during thymosin treatment, results of other immunologic studies from patients treated with thymosin were similar to those from patients not treated. The subsequent development of graft‐versus‐host disease, major or minor infection, and leukaemic relapse was not different between the groups. Six patients are alive and five are well without problems; one has chronic graft‐versus‐host disease. We conclude that thymosin fraction 5 administered as described was not toxic. Although modifying some immunological parameters, thymosin did not appear to alter the incidence of graft‐versus‐host disease, infection or leukaemic relapse or to accelerate immunologic reconstitution.
Spontaneous regression of cytogenetic and haematologic anomalies in Ph ‐positive chronic myelogenous leukaemia
Authors: Smadja N. , Krulik M. , Audebert A. A. , de Gramont A. , Debray J.
Published: July 2010

Summary. We report a case of a 27‐year‐old man with Ph1‐positive chronic myelogenous leukaemia (CML). At the time of diagnosis 100% Phpositive cells were found with a trisomy 8 in 50% of them. In absence of therapy, his haematological status remained stable for 3 years. Subsequently a progressive regression of haematologic and cytogenetic data was observed. Eight years after diagnosis the karyotype showed only 37% Ph1‐positive cells and the trisomy 8 had disappeared.
Acquired folate deficiency in phytohaemagglutinin‐stimulated human lymphocytes
Authors: Matthews J. H. , Wickramasinghe S. N.
Published: July 2010

Summary. 3H‐thymidine (3H‐TdR) incorporation, deoxyuridine (dU)‐sup‐pressed values, modal cell volume and the relative DNA content of individual cells have been determined in phytohaemagglutinin (PHA)‐stimulated normal human lymphocytes cultured for varying periods in folate‐free medium and media supplemented with various concentrations of folic acid. The results indicate that when the intracellular folate content falls below 400‐600 pg/106cells, lymphocytes exhibit increased 3H‐TdR incorporation into DNA, raised dU‐suppressed values, an increase in cell volume and abnormal DNA synthesis. This critical concentration of folate is just present in resting lymphocytes, but falls on stimulation by PHA, and the effects of folate deficiency become increasingly evident beyond a culture period of 24 h unless the folic acid concentration in the medium is at least 0‐32 /ig/ml. Thus human lymphocytes cultured in folate‐free medium provide a model for folate‐deficient DNA synthesis, but their usefulness for the performance of dU suppression tests in the diagnosis of clinical folate deficiency is likely to be limited.
The effect of an adrenaline infusion on the splenic blood flow and intrasplenic platelet kinetics
Authors: Wadenvik Hans , Kutti Jack
Published: July 2010

The effect of an adrenaline infusion on the venous platelet count, splenic blood flow and intrasplenic platelet kinetics was investigated in seven healthy male volunteers by using111In‐labelled platelets and dynamic gamma camera scintigraphy. The infusions were administered in two different doses, 0.2 μg/kg/min and 0.1 μg/kg/min, respectively. Regardless of the given dose, adrenaline was found to markedly decrease the splenic blood flow, decrease the exchangeable splenic platelet pool size and to prolong the intrasplenic platelet transit time. In response to the higher dose of adrenaline the splenic blood flow was 1.3 ± 0.5 (SD) % of total blood volume per min and the intrasplenic platelet transit time 17.3 ± 2.3 (SD) min. In contrast, after termination of infusion, the splenic blood flow was 7.0 ± 2.1 (SD)% of total blood volume per min and the intrasplenic platelet transit time was 11.5 ± 1.5 (SD) min. The present results firmly demonstrate that the splenic blood flow is the immediate variable governing the size of the exchangeable splenic platelet pool, and that the adrenaline‐induced depletion of the splenic platelet pool is a consequence of the splenic blood flow reduction. Finally, the splenic perfusion appears to be a major determinant of the intrasplenic platelet transit time.
Evolution of coagulation abnormalities following Russell's viper bite in Burma
Authors: Than Than , EiHan Khin , Hutton R. A. , Lwin Myint , Swe Tin Nu , Phillips R. E. , Warrell D. A.
Published: July 2010

Summary. The evolution of coagulation abnormalities was studied in Russell's viper bite victims who, on admission to hospital, showed no clinical signs of systemic envenoming. Based on the laboratory results and subsequent clinical course, three groups were distinguished. The first group, consisting of five cases, showed no activation of coagulation at any stage. The second group. consisting of six cases, developed mild to moderate abnormalities in some tests, particularly in the aPTT and factor V assay. which corrected to normal without treatment. The third group, consisting of nine patients, developed haemostatic abnormalities as early as 1–2 h after the bite, which progressed to severe deflbrination 4–8 h later at which time antivenom was given. Comparison of the haemostatic abnormalities in the three groups suggested that serial monitoring of the serum FDP concentration may be of value in predicting the likelihood of systemic envenoming and progression to complete defibrination.
Abnormal phosphoinositide metabolism and protein phosphorylation in platelets from a patient with the grey platelet syndrome
Authors: Rendu F. , Marche P. , Hovig T. , Maclouf J. , Lebret M. , Tenza D. , Levy‐Toledano S. , Caen J.‐P.
Published: July 2010

Washed platelets isolated from one patient suffering from the inherited grey platelet syndrome were studied during thrombin‐induced activation. The agonist‐induced changes in (i) morphology, (ii) typical functional cell responses, (iii) membrane phospholipid metabolism and protein phosphorylation were studied and compared with the changes obtained with normal platelets. The morphology of the platelets as visualized by electron microscopy confirmed the almost total absence of intracellular alpha‐granules and marked vacuolization. During thrombin stimulation the morphological changes were clearly delayed as compared to normal platelets, the granule centralization and aggregation occurring only 15s after thrombin addition instead of 5 s in normal platelets. After 15 s, however, even though no alpha‐granules were observed, a ring‐like structure occurred centrally, indicating that they are not a prerequisite for this reaction. The whole release reaction, i.e. liberation of [14C]ser‐otonin from dense granules and β‐n‐acetylglucosaminidase activity from lysosomes, and the thromboxane synthesis were delayed and remained lower than in normal platelets. No thrombin‐induced phosphatidyl 4,5‐bisphosphate breakdown was measurable on 32P‐prelabelled platelets although [32P]phosphatidate formation occurred normally. Phosphorylation time courses of myosin light chain (P20) and of protein P43 (mol wt 43 000) markedly differed from those of controls, being less than half of the normal during the first 15 s and remaining subnormal even after complete aggregation. These results suggest that in platelets devoid of alpha‐granules a deficient transmembrane signalling system is likely responsible for the impaired physiological responses.
Rapid reappearance of large granular lymphocytes (LGL) with concomitant reconstitution of natural killer (NK) activity after human bone marrow transplantation (BMT)
Authors: Niederwieser D. , Gastl G. , Rumpold H. , Marth Ch. , Kraft D. , Huber Ch.
Published: July 2010

The frequency of large granular lymphocytes and their relationship to functional NK‐activity as assessed by the capacity to lyse the K562 tumour target was analysed in five allogenic and two autologous human bone marrow transplant recipients. Date revealed: (i) almost identical disappearence and reconstitution of both parameters further indicating that LGL represent effector cells of spontaneous lysis of K 562 targets; (ii) a long‐lasting suppression of the absolute numbers per ml of blood of both LGL and functional NK activity which we believe was not the consequence of reconstitution with immature effector cells but rather reflected immunosuppressive therapy; (iii) LGL exhibits the fastest reappearance rate subsequent to total body irradiation of all populations of circulating leucocytes.
Establishment and characterization of a permanent human IgA2/kappa myeloma cell line
Authors: Lohmeyer J. , Hadam M. , Santoso S. , Förster W. , Schulz A. , Pralle H.
Published: July 2010

A human myeloma cell line designated LOPRA‐1 has been established from ascites fluid containing malignant plasma cells of a patient with IgA2/kappa multiple myeloma. The cultured cells which are Epstein‐Barr virus (EBV) negative have retained the morphological, cytochemical, ultrastructural and immunophenotypical features of well‐differentiated plasma cells. They express the plasma cell antigen PCA‐1, the antigens CD28 (Kolt‐2) and CD38 (OKT10), the transferrin‐receptor (OKT9), and some epitopes of the CD24 antigen (HB8, VIB E3), but are negative for surface immunoglobulins, HLA class II antigens (HLA‐DP, ‐DQ, ‐DR) and other B‐cell markers such as CD10 (CALLA), CD19 (B4), CD20 (B1), CD21 (B2), CD22 (HD39), CD23 (MHM6), CD37 (BL14) and CD39 (G28‐8) as analysed by both flow cytometry and immunocytochemistry (PAP/APAAP). With respect to immunoglobulin synthesis, two stable clones were selected by single cell cloning: clone LOPRA‐1/5 synthesizes large amounts of alpha2 heavy and kappa light chains, but secretes only small amounts of these molecules, whereas clone LOPRA‐1/4 is clearly devoid of intracellular immunoglobulin heavy and light chains and thus appears to be a chain loss variant. Cytogenetic analysis revealed a pseudotriploid phenotype with several structurally abnormal marker chromosomes: 3n+ ‐, 70, XX, ‐X, ‐1, ‐4, ‐6, ‐8, ‐8, ‐13, ‐16, +7, +18, +21, +i(1q), +i(1q), +6q‐, +3mar.
Congenital macrothrombocytopenia, leucocyte inclusions, deafness and proteinuria: functional and electron microscopic observations on platelets and megakaryocytes
Authors: Heynen M. J. , Blockmans D. , Verwilghen R. L. , Vermylen J.
Published: July 2010

We report a patient with a variant of Alport's syndrome: macrothrombocytopenia, leucocyte inclusions, deafness and proteinuria. Ultrastructural studies revealed giant spheroid platelets with a high density of organelles and a disorganized microtubular system. In addition, Fechtner inclusions were observed in neutrophils of the patient and her mother. In platelet rich plasma platelets aggregated normally for the low platelet number, although no shape change was visible. Platelet studies in whole blood using impedance aggregometry gave supernormal aggregation curves; this is not in agreement with the abnormally long bleeding time, showing the limited usefulness of this technique in patients with such large platelets. The megakaryocytes (MK) showed two different distribution patterns of the demarcation membrane system (DMS), which may explain the production of few large platelets. The formation of platelets occurred by fragmentation of the granular zone of the MKs, which seemed to be followed by expulsion of platelets through openings of the peripheral zone. The involvement of cytoskeletal structures in the organization of the DMS and the expulsion of platelets is discussed.
Duration of first remission as an indicator of long‐term survival in chronic myelogenous leukaemia
Authors: Prischl Friedrich C. , Haas Oskar A. , Lion Thomas , Eyb Richard , Schwarzmeier Josef D.
Published: July 2010

Summary Approximately 31 patients with chronic myelogenous leukaemia (CML) are documented in the literature who survived more than 10 years after diagnosis. We present a CML‐patient whose survival of 27 years is probably the longest reported so far. The analysis of the course of disease in these patients revealed that the duration of unmaintained first remission after chemotherapy is of high prognostic significance. In 17 of 24 evaluable patients the remission lasted more than 1 year and in another five at least 6 months (mean 73.8 months, range 0‐240 months). In most patients busulfan was used as initial therapy. There was no correlation between the amount of drug given and the duration of remission or survival. Other parameters such as sex, age, initial leucocyte counts, differential count, haemoglobin, platelet count or spleen size seemed to have no prognostic relevance. While approximately 25% of CML patients with typical duration of survival exhibit a ‐Phl chromosome mosaicism only, this finding was present in nearly half of the long‐term survivers.
Burkitt cell acute leukaemia (L3 ALL) in adults: a report of 18 cases
Authors: Fenaux P. , Lai J. L. , Miaux O. , Zandecki M. , Jouet J. P. , Bauters F.
Published: July 2010

Summary Between 1981 and 1987, L3 ALL was diagnosed in 18 adult patients, with a median age of 26 (range 16‐66) and M/F ratio of 3.5. At diagnosis, 11 patients had splenomegaly, 11 had enlarged lymph nodes, and 15 patients had central nervous system (CNS) disease, of whom 10 had mental neuropathy. Anaemia was found in 13 patients, thrombocytopenia in 17 and the median white cell count was 25.5 × 109/1(range 8.6‐89). Surface immunoglobulins were found on the blasts of every patient. Seventeen patients had a t(8;14) (q24;q32) translocation. One had an apparently normal karyotype, but only six mitoses could be examined. During the period of the study different treatment protocols, which comprised increasingly intensive systemic and CNS chemotherapy, were used. Six patients died less than 3 weeks after admission, two of them of acute tumour lysis syndrome and two of CNS haemorrhage. In two other patients, rapid progression of CNS leukaemia was seen in spite of the treatment. Ten patients (56%) achieved complete remission (CR). Two were allografted and two were autografted early in CR. Four patients relapsed, three of the four relapses involving the CNS. A median actuarial disease‐free survival was not attained, and a plateau was achieved at 57% after 7 months, with no later relapse. Median actuarial survival of the 18 patients was only 6 months, but a plateau was obtained at 31% after 11 months. Prognosis seemed related to the intensity of chemotherapy: recent patients, treated more aggressively, achieved CR more often than earlier patients, treated with less intensive protocols, although the number of patients was too small to draw any firm conclusion. The initial white cell count was also a prognostic factor, as none of the patients with more than 30 × 109/1 leucocytes achieved CR. Our results suggest that the outcome of adult L3 ALL can be improved, as in children, by increased intensity of treatment, particularly with regard to CNS leukaemia therapy. Early deaths are still frequent, however, but their incidence can probably be reduced by better prevention and early management of the acute tumour lysis syndrome.
Lupus anticoagulant induces a selective defect in thrombin‐mediated endothelial prostacyclin release and platelet aggregation
Authors: Schorer Anna E. , Wickham Nicholas W. R. , Watson Kathleen V.
Published: July 2010

Summary A patient with microvascular thrombosis and thrombocytopenia was found to have a high‐titre lupus anticoagulant. The biological effects of the patient's lupus anticoagulant were studied using whole patient serum and plasma, Staph Protein A eluate, and affinity‐purified lupus anticoagulant. The latter was isolated by immunoadsorption of serum onto cardiolipin/phosphatidylserine/cholesterol liposomes. Each source of lupus anticoagulant demonstrated‘anticoagulant’activity, defined as prolongation of a modified kaolin clotting time, and contained antibody which bound to endothelial monolayers. Each interfered with thrombin‐mediated prostacyclin release from endothelial cells, but had no effect on arachidonate‐induced prostacyclin release. In addition, the lupus anticoagulant selectively blocked platelet aggregation in response to thrombin, but not in response to arachidonate, ADP or epinephrine. Lupus anticoagulant also reduced thrombin‐stimulated shifts in cytosolic calcium. Thrombin‐mediated membrane inositol metabolism and total thrombin binding to endothelium were unaffected by lupus anticoagulant, and another endothelial anticoagulant function related thrombin binding, Protein C activation by thrombomodulin, was not altered. We conclude that the binding of lupus anticoagulant to endothelial cells and platelets does not prevent all thrombin signalling events, but does interrupt prostacyclin production.
Acquired factor VII deficiency associated with aplastic anaemia: correction with bone marrow transplantation
Authors: Weisdorf Daniel , Hasegawa Duane , Fair Daryl S.
Published: July 2010

Summary. We report a patient with severe aiplastic anaemia found to have a prolonged prothrombin time due to acquired factor VII deficiency. No evidence for a factor VII inhibitor or inactivator was demonstrable. Laboratory studies identified deficiency both of factor VII activity and factor VII antigen. The factor VII deficiency persisted from clinical presentation until approximately 50 d after allogeneic marrow transplantation when restoration of factor VII activity;and antigen was noted. The patient's serum could be depleted of factor VII activity by in vitro incubation with Protein A bound to Sepharose, suggesting the presence of an IgCl or IgG containing complex able to bind factor VII, but not neutralize its procoagulant activity. A dual specificity solid phase immunoassay identified a factor VII binding immunoglobulin which was detectable throughout the course of factor VII deficiency. The concordant appearance of this factor VII reactive immunoglobulin and the factor VII deficiency suggested the pathologic role of this immunoglobulin in the aetiology of the factor VII deficiency. This factor VII binding immunoglobulin may have induced rapid plasma clearance of the factor VII molecule or, alternatively, may have modified factor VII synthesis. The immunosuppressive therapy and subsequent lymphohaematopoietic engraftment following allogeneic marrow transplant was accompanied by complete resolution of the factor VII deficiency.
Direct activation of platelets by heat is the possible trigger of the coagulopathy of heat stroke
Authors: Gader A. M. A. , Al‐Mashhadani S. A. , Al‐Harthy S. S.
Published: July 2010

The trigger of the coagulopathy that complicates heat stroke is obscure, but direct platelet activation by heat is a possibility we set out to study. Platelet rich plasma (PRP), prepared from blood donors, was incubated at increasing temperatures (38–45°C) and then platelet aggregation was undertaken in response to decreasing low doses of ADP (<2.0 μmol/l). Hyperaggregability was manifested when the incubation temperature reached 43°C and was maximum at 44°C before complete inhibition of responses at 45°C.

The platelet hyperactivity induced by heating at 44°C persisted after reincubating PRP samples at 37°C. These platelet responses could not be triggered in PRP samples prepared from subjects after the overnight ingestion of aspirin or after the addition of aspirin to PRP before starting the heating procedure. However, aspirin was less effective when added to PRP after the appearance of the heat‐induced hyperaggregability.

In conclusion, these results indicate that platelets can be activated directly by heat. This mechanism which may be operational in heat stroke, is unaffected by cooling (body cooling being basic in the management of heat stroke) but can be prevented by the early administration of aspirin.
Spontaneous platelet aggregation during pregnancy in a patient with von Willebrand disease type IIB can be blocked by monoclonal antibodies to both platelet glycoproteins Ib and IIb/IIIa
Authors: Pareti Francesco Innocente , Federici Augusto Bramante , And Marco Cattaneo , Mannucci Pier Mannuccio
Published: July 2010

Spontaneous platelet aggregation appeared in a patient with von Willebrand disease type IIB during the 37th week of pregnancy. This phenomenon was not associated with symptoms of thrombosis and the patient delivered by caesarian section with no complications. Her platelet‐poor plasma (PPP) aggregated normal platelet‐rich plasma (PRP) and washed platelets. Aggregation was inhibited by monoclonal antibodies with known specificity for the platelet receptors of von Willebrand factor (vWF), i.e. the glycoprotein Ib (GPIb) and the GPIIb/IIIa complex. A monoclonal antibody, which selectively inhibits the binding of vWF to the GPIIb/IIIa complex, did not block aggregation, suggesting that spontaneous aggregation is not dependent on the binding to GPIIb/IIIa of vWF from patient plasma. Aggregation induced by patient plasma could also be blocked either by two monoclonal antibodies raised against vWF or by a fragment derived from trypsin digestion of normal vWF which blocks the ristocetin‐induced binding of normal vWF to platelets. These findings indicate that the spontaneous platelet aggregation in this patient results from the binding of her vWF to GPIb but is independent from the binding of her vWF to GPIIb/IIIa.
Immortalization and characterization of human cell lines with mast cell and monocytic properties
Authors: Townsend Megan , Macpherson Janet , Krilis Steven , Reddell Roger , Symonds1 Geoff
Published: July 2010

Summary. We have previously derived a cell strain which had both mast cell and monocytic properties from the bone marrow of a child with diffuse cutaneous mastocytosis. This cell strain, termed HBM‐M, consisted of two cell populations both of which possessed certain ultrastructural, cytochemi‐cal and surface phenotypic features of degranulated mast cells. The cells also displayed cytochemical and surface phenotypic features of monocytes. These cells may represent a common bone marrow derived mast cell/monocyte precursor. Studies of human mast cells have been hindered by the fact that it is difficult to establish such cells in long‐term culture. Thus, we sought to immortalize HBM‐M cells by introducing Simian virus 40 large T‐antigen. Following transfection by the strontium phosphate technique, transformed cells were selected, expanded and passaged until the cells entered a non‐proliferative phase termed crisis. Certain clones passed through crisis 3 months later and by this means two immortal cell lines, HBM‐MI‐1 and HBM‐MI‐2, were obtained. The criterion for immortality was growth for greater than 100 population doublings. The immortal cell lines retained some, but not all. of the mast cell and monocytic properties of the original HBM‐M cell strain. The immortalization of the cell strain HBM‐M provides an opportunity to investigate the mast cell and monocytic properties of these cells, and the apparent relationship between mast cells and monocytes.
Interleukin 4 prevents the induction of tissue factor mRNA in human monocytes in response to LPS or PMA stimulation
Authors: Ramani Mohamed , Ollivier Veronique , Ternisien Catherine , Vu Thi , Elbim Carole , Hakim Jacques , Prost Dominique de
Published: July 2010

Summary.Increased expression of tissue factor (TF) procoagulant activity by blood monocytes and tissue macrophages is implicated in a number of thrombotic disorders, as well as in fibrin deposition associated with inflammatory lesions and immunological diseases. We found that interleukin 4 (IL‐4), a T lymphocyte‐derived cytokine known to regulate a number of monocyte functions, inhibited the production of TF by monocytes in response to endotoxin and phorbol myristate acetate (PMA)in vitro. IL‐4 had a concentration‐dependent inhibitory effect on functional TF procoagulant activity (PCA) and reduced the binding of an anti‐TF antibody, as assessed by flow cytometry analysis. Moreover, IL‐4 reduced LPS‐ and PMA‐induced TF mRNA levels. TF mRNA stability was not modified by IL‐4 after the arrest of transcription by actinomycin D. We thus conclude that mRNA suppression is mediated by an effect occurring at the transcriptional level. Our results also show that the suppressive effect of IL‐4 is independent of an increase in the intracellular concentration of cyclic AMP, another established inhibitor of TF production. Locally produced IL‐4 might thus contribute to limiting the consequences of monocyte activation.
Decreased contact factor mediated fibrinolysis in cirrhosis
Authors: Cohen H. , Hunt J. B. , Dixit M. , Kanwar S. , Thomas H. C.
Published: July 2010

Summary. We studied extrinsic and intrinsic fibrinolysis in 20 patients with cirrhosis (nine mild/moderate, group 1:11 severe, group 2) and 19 normal controls to define the role of intrinsic (contact factor mediated) fibrinolysis in cirrhosis.

Global plasma fibrinolytic activity (fibrin plate lysis) was similar in all groups. Dextran sulphate activated contact factor mediated fibrinolysis was decreased in group 2 (median 95.2%) compared with group 1 (121.0%) and controls (131.7%). Tissue plasminogen activator antigen (t‐PA Ag) levels were increased in group 2 (28.2 ng/ml) compared both with group 1(8.5 ng/ml) and controls (5.9 ng/ml). Plasma t‐PA activity was raised in group 2 (5.50 IU/ml) and group 1 (5.25 IU/ml) versus controls (0.82 IU/ml). Plasminogen activator inhibitor‐1 (PAI‐1 Ag) levels were raised in group 2 (28.0 IU/ml) versus controls (8.5 IU/ml) but PA1 activity was similar in all groups. Factor XII activity was decreased in group 2 (48.76 u/dl), but not group 1. versus controls (89.1 u/dl). Prekallikrein activity was decreased both in group 2 (27.27 u/dl) and group 1 (33.01 u/dl) versus controls (108.59 u/dl) and was lower in group 2 than group 1. C1‐esterase inhibitor chromogenic activity was decreased in group 1 (102.30 u/dl) and group 2 (58.76 u/dl) versus controls (116.24 u/dl).

The normal global fibrinolytic activity despite increased t‐PA activity may be due to a concomitant increase in PAI. The decreased intrinsic fibrinolysis in severe cirrhosis, unaccompanied by a rise in C1‐esterase inhibitor, may be explained by the decreased factor XII and prekallikrein activity. These changes are probably due to reduced liver cell mass.
Potentiation of lysis of leukaemia cells by a bispecific antibody to CD33 and CD16 (FcγRIII) expressed by human natural killer (NK) cells
Authors: Silla Lucia M. R. , Chen Jian , Zhong Rui‐kun , Whiteside Theresa L. , Ball Edward D.
Published: July 2010

Bispecific antibodies recognizing tumour‐associated antigens and trigger molecules expressed on immune effector cells have been shown to redirect cytotoxicity of several types of peripheral blood cells against relevant tumour targets. Among various effector cells, natural killer (NK) cells appear to play a role in defence against leukaemia. Here we report the successful chemical conjugation of monoclonal antibodies to CD33 and CD16 to create a bispecific antibody (BsAb 251x3G8). This bispecific antibody is capable of augmenting the killing of otherwise resistant leukaemia cells by peripheral blood lymphocytes (PBL), purified resting NK (R‐NK) cells, and activated NK (A‐NK) cells. BsAb 251x3G8 may play a role in the therapy of acute myeloid leukaemia (AML) through redirecting the cytotoxic activity of endogenous or adoptively transferred NK cells.
Absence of G‐CSF receptors and absent response to G‐CSF in childhood Burkitt's lymphoma and B‐ALL cells
Authors: Tomeczkowski Jörg , Yakisan Elif , Wieland Britta , Reiter Alfred , Welte Karl , Sykora Karl‐Walter
Published: July 2010

The expression of the granulocyte colony‐stimulating factor (G‐CSF) receptor in childhood Burkitt's lymphoma (BL) cells, and the mitogenic effect of G‐CSF on these cells, was studied in a panel of 13 Epstein‐Barr virus (EBV) positive and negative BL cell lines derived from nine children.

G‐CSF receptor mRNA expression was investigated by Northern blot analysis and reverse transcriptase polymerase chain reaction (RT‐PCR). Binding of G‐CSF to BL cell lines was measured by chemical crosslinking of 125I‐G‐CSF, and proliferation by thymidine incorporation. Inducibility of the G‐CSF receptor was studied by stimulation with interleukin‐1β, tumour necrosis factor‐α, Staphylococcus aureus Cowan A, anti‐human IgM, phorbol myristate acetate, calcium ionophore A23187, and by infection in vitro by immortalizing and non‐immortalizing strains of EBV.

BL cell lines, unstimulated or stimulated by biological reagents or EBV infection, did not bind radioionated G‐CSF in crosslinking experiments. No stimulation by recombinant human G‐CSF was observed in 3H‐thymidine incorporation assays. No G‐CSF receptor mRNA was detected by Northern blot analysis or RT‐PCR in BL cell lines.

It is concluded that G‐CSF plays no direct stimulatory role in the growth of these malignant B‐cells, making a deleterious influence of G‐CSF in the clinical treatment situation unlikely.
Factor V inhibitor associated with Sjögren's syndrome
Authors: Koyama Takatoshi , Saito Takako , Kusano Takayoshi , Hirosawa SHINSAKU
Published: July 2010

We report an unusual case of a 74‐year‐old male who developed a serum autoantibody reactive with human coagulation factor V (FV) in an activated form, as demonstrated by coagulation studies and immunoblotting analysis. Despite marked prolongation of a prothrombin time and an activated partial thromboplastin time in this patient, the inhibitor was not associated with clinical bleeding but with multiple cerebral infarctions. The patient had suffered from Sjögren's syndrome with polyclonal hypergamma‐globulinaemia. The patient's purified IgG, an immediately acting inhibitor to FV, reacted with a light chain of thrombin‐activated FV (FVa) and inhibited the procoagulant activity of FVa without affecting the cleavage of FVa by activated protein C. The FV inhibitor may arise from activation of FV with consequent exposure of neoantigen during the activation of coagulation cascade in the patient with an autoimmune disorder for the background.
AcSDKP serum concentrations vary during chemotherapy in patients with acute myeloid leukaemia
Authors: Liozon Eric , Volkov Leonid , Comte Lydie , Trimoreau Franck , Pradelles Philippe , Bordessoule Dominique , Frindel Emilia , Praloran Vincent
Published: July 2010

AcSDKP is a physiological negative regulator of cell proliferation in mammals. In Ara‐C‐treated mice its plasmatic concentrations decrease while the CFU‐S start cycling. Infusion of AcSDKP protects these animals from death by blocking the proliferation of primitive haemopoietic cells. We measured AcSDKP serum concentrations in 20 AML patients during the course of high‐dose cytoreductive treatment. We observed an early and sharp increase of AcSDKP during the induction treatment in 12 patients, reaching a peak during the initial 3 d of treatment in nine of them. These results are contrary to those observed in mice treated with high doses of Ara‐C. They encourage further clinical investigation, and suggest that treatments with synthetic AcSDKP (Seraspenide) will perhaps have to be adjusted to the type of disease and the schedule of chemotherapy in order to optimize its myeloprotective effect.
Reversal of aberrant splicing of β‐thalassaemia allele (IVS‐2‐654 C→T) by antisense RNA expression vector in cultured human erythroid cells
Authors: Gong Lan , Gu Xiao‐Feng , Chen Yun‐Di , Ren Zhao‐Rui , Huang Shu‐Zhen , Zeng Yi‐Tao
Published: July 2010

The antisense fragment targeting the aberrant splice sites of the β‐thalassaemia allele, IVS‐2‐654 C→T (β654), pretranscript was cloned into the mammalian expression vector, pcDNA3. The recombinant construct, pCMVA, was then used to repair the defective splicing of the β654 mutant pretranscript in cultured β654 erythroid cells by the lipofectin‐mediated DNA transfection method. The total RNA was extracted at given time points after transfection and the effect of antisense RNA was studied by reverse transcription polymerase chain reaction (RT‐PCR)‐mediated mRNA quantitative assay, as well as globin chain microbiosynthesis. The antisense fragment transcribed from pCMVA effectively improved the β654 splicing pattern in cultured erythroid cells. The level of correctly spliced transcript increased from 0·19 (day 0 after transfection) to 0·58 (day 8) in β654/β654 homozygous erythroid cells, and from 0·45 (day 0) to 0·83 (day 8) in β654/βA heterozygous erythroid cells, as determined by the ratio of normally spliced β‐globin transcript over total β‐globin transcript. Correspondingly, the ratios of globin chain biosynthesis (β/α) increased from 0·16 (day 0) to 0·52 (day 8) in β654/β654 erythroid cells, and from 0·39 (day 0) to 0·84 (day 8) in β654/βA erythroid cells. Antisense RNA had no significant effect on the splicing pattern in βA/βA erythroid cells. The splicing pattern in transfected cells with pCMVA showed significant changes compared with that in untransfected cells and that in transfected cells with the control antisense fragment (human SRY gene sequence). In addition, we did not observe side‐effects on cytological features after the introduction of pCMVA. All these results indicated that the antisense RNA transcribed from the mammalian expression vector pCMVA could efficiently and specifically suppress the aberrant splicing pattern of β654 mutant pretranscript and restore the correct splicing pathway in vivo, leading to the improvement of globin chain biosynthesis in thalassaemic cells.
The epidemiology of fusariosis in patients with haematological diseases
Authors: Girmenia Corrado , Pagano Livio , Corvatta Laura , Mele Luca , Del Favero Albano , Martino for the Gimema Infection Programme Pietro
Published: July 2010

Fusariosis has been considered an emerging infection in patients with haematological malignancies. In a multicentre retrospective study on filamentous fungi infections in patients with haematological diseases over a period of 10 years in Italy, fusariosis was documented in six patients from two of the 14 centres with a 0·06% incidence in acute leukaemia. A literature search yielded 177 cases of Fusarium infections in haematological diseases and acute leukaemia accounted for 71% of the underlying conditions. An increase from 0·5 to 3·8 cases per year was observed at the M.D. Anderson Cancer Center of Houston in the periods 1975–85 and 1986–95 respectively. Conversely, only 5·1 and 6·3 cases per year have been reported in the periods 1981–90 and 1991–96, respectively, from the other centres in the world. Half of the cases have been observed in the USA. In Europe, most of the cases have been observed in France and Italy. Invasive fusariosis is a rare complication in haematological diseases, as its overall frequency does not seem to be significantly increased within the last 20 years. However, its epidemiological distribution is not homogeneous and the possibility of local clusters of infections by this deadly pathogen should be carefully considered.
FLT3 internal tandem duplication mutations in adult acute myeloid leukaemia define a high‐risk group
Authors: Abu‐Duhier F. M. , Goodeve A. C. , Wilson G. A. , Gari M. A. , Peake I. R. , Rees D. C. , Vandenberghe E. A. , Winship P. R. , Reilly J. T.
Published: July 2010

Genomic DNA from 106 cases of adult de novo acute myeloid leukaemia (AML) was screened by polymerase chain reaction (PCR) and gel electrophoresis for FLT3 internal tandem duplication (ITD) mutations within the juxtamembrane (JM) domain. FLT3 mutations were detected in 14 cases (13·2%) and occurred in FAB types M1 (4 out of 14 cases), M3 (1 out of 10 cases), M4 (5 out of 37 cases) and M5 (4 out of 11 cases). Sequence analysis of four cases with abnormal PCR electrophoretic patterns revealed in frame duplications in the region of exon 11 of between 27 and 111 base pairs. Three are predicted to result in the tandem duplication of adjacent amino acid residues and one to result in a tandem duplication plus insertion of a novel amino acid motif. Statistical analysis showed the FLT3 mutations to be a strong prognostic factor, with patients lacking the mutation surviving significantly longer from diagnosis (mean 29·1 months) than those with an ITD (mean 12·8 months; P = 0·0002). Thirteen of the 14 patients with FLT3 mutations died within 18 months of diagnosis. FLT3 mutations were of prognostic significance in good risk disease (P = 0·04), as well as in patients with standard risk disease (P = 0·0096). This study demonstrates that the FLT3 ITD mutation occurs in a significant percentage of adult AML cases and is an important adverse prognostic factor that appears independent of conventional karyotypic findings.
The effects of the 32‐bp CCR‐5 deletion on HIV transmission and HIV disease progression in individuals with haemophilia
Authors: Pasi K. J. , Sabin C. A. , Jenkins P. V. , Devereux H. L. , Ononye C. , Lee C. A.
Published: July 2010

The chemokine receptor CCR5 is an important co‐receptor for cell fusion. A 32‐bp deletion of the CCR5 gene, leading to complete absence of functional CCR5 expression, has been associated with resistance to human immunodeficiency virus (HIV) infection in homozygotes and slower HIV disease progression in heterozygotes. The objectives of this study were to assess the effects of this 32‐bp deletion on transmission of HIV infection and on HIV disease progression in haemophilic individuals. Six HIV‐negative patients from our centre, known to have been exposed to infectious factor VIII concentrates, have been analysed. Three of these patients possess the CCR5 32‐bp deletion, two patients being homozygous. The presence of the CCR5 32‐bp gene deletion has also been analysed in 71 HIV‐positive patients. In this group of patients, there was a lower than expected incidence of the 32‐bp deletion. Those who possess the 32‐bp deletion progress to AIDS more slowly than those who do not (P = 0·05, log‐rank test). Rates of CD4 loss were slower in those heterozygous for the gene deletion. We confirm that heterozygosity for the 32‐bp gene deletion in CCR5 is partially protective against initial infection with HIV. In those heterozygous patients who became infected with HIV, disease progression was slower.
p53 intronic point mutation, aberrant splicing and telomeric associations in a case of B‐chronic lymphocytic leukaemia
Authors: Bromidge Teresa , Lowe Christopher , Prentice Archie , Johnson Stephen
Published: July 2010

We report a case of chronic lymphocytic leukaemia (CLL) with telomeric associations and a p53 intronic point mutation. Karyotypic analysis revealed clonal and non‐clonal telomeric associations, accompanied by clonal cytogenetic abnormalities and also in isolation. The p53 mutation, which occurred at the invariant base pair −2 of the splice acceptor site in intron 7 resulted in the abolition of correct splicing of exon 7 to exon 8. Multiple aberrant splice products were characterized, all of which differed from wildtype in the DNA binding domain. Fluorescence in situ hybridization demonstrated that the clone retained two copies of the p53 gene and wild‐type p53 transcript was detected on cloning of reverse transcriptase polymerase chain reaction (RT‐PCR) product, indicating that one wild‐type allele remained. However, a plasmid clone with correct splicing at the exon 7/8 boundary, but with a 21 bp deletion in exon 8, was also found at low frequency. This finding indicates clonal evolution, resulting in complete loss of wild‐type p53. The intronic point mutation was not present in DNA extracted from cervical tissue indicating that it was a leukaemic phenomenon. This is the first case of an intronic point mutation to be reported in CLL. This mutation led to chaotic p53 expression and, interestingly, occurred in a case showing telomeric associations, a rare phenomenon in B‐CLL.
Intracellular function in rehydrated lyophilized platelets
Authors: Fischer Thomas H. , Merricks Elizabeth P. , Russell Karen E. , Raymer Robin A. , White Gilbert C. , Bode Arthur P. , Nichols Timothy C. , Read Marjorie S.
Published: July 2010

This study aimed to evaluate the effect of cross‐linking and lyophilization on intracellular signalling processes in rehydrated, lyophilized (RL) platelets, which are under development as a platelet substitute for transfusion. Exposure of RL platelets to thrombin resulted in enhanced phosphorylation of several proteins, including 18 kDa and 42 kDa kinase substrates that were shown to be the substrates of myosin light chain and protein kinase C respectively. Cross‐linking and lyophilization depleted the platelets of free cytoplasmic ADP and ATP, but had less effect on protein‐bound nucleotides. The surface membrane of RL platelets was found to be permeable to poly dT probes less than approximately 3 kDa in size; larger nucleotide probes and proteins did not penetrate the surface membrane. Taken together, our results indicate that RL platelets retain some of the haemostatic stimulus‐response functions of fresh platelets and are capable of feedback amplification in coagulation.
Spontaneous regression of congenital leukaemia with an 8;16 translocation
Authors: Weintraub Michael , Kaplinsky Chaim , Amariglio Ninette , Rosner Esther , Brok‐Simoni Frieda , Rechavi Gideon
Published: July 2010

Congenital leukaemia (CL) is a rare disorder that presents with extramedullary infiltrates and a myeloid phenotype. CL can progress rapidly without adequate treatment, but, paradoxically, may also remit spontaneously. Because of the significant toxicity involved in delivering chemotherapy to newborns, it is important to identify those newborns who may not require treatment. We describe an infant who presented at 1 week of age with congenital myeloid leukaemia. Cytogenetic analysis revealed a t(8;16)(q11;p13) translocation. The infant's leukaemia underwent a spontaneous regression. This case further confirms the possibility of spontaneous remission in congenital leukaemia. Moreover, it suggests that the presence of a clonal cytogenetic aberration does not preclude the possibility of a spontaneous regression in CL.
Autologous T lymphocytes may specifically recognize leukaemic B cells in patients with chronic lymphocytic leukaemia
Authors: Rezvany Mohammad Reza , Jeddi‐Tehrani Mahmood , Rabbani Hodjattallah , Lewin Nongnit , Avila‐Cariño Javier , Österborg Anders , Wigzell Hans , Mellstedt Håkan
Published: July 2010

This study analysed a naturally occurring specific cellular immunity against tumour cells in chronic lymphocytic leukaemia (CLL) patients. Five out of eight patients had blood T lymphocytes able to recognize spontaneously and specifically the autologous tumour B cells (proliferation assay). In these five patients, detection of cytokines by real‐time reverse transcription polymerase chain reaction (RT‐PCR) revealed that granulocyte–macrophage colony‐stimulating factor (GM‐CSF) was the most abundant cytokine gene expressed by the T cells that recognized the autologous tumour B cells. Other activated cytokine genes were γ‐interferon (IFN), interleukin (IL)‐2 and tumour necrosis factor (TNF)‐α, but not IL‐4. This profile suggests a type 1 anti‐B‐CLL T‐cell response. CD80 and CD54 were relatively downregulated on the native tumour B cells compared with control normal B cells. Upregulation of CD80 on the leukaemic cells was mandatory for the induction of such a specific T‐cell response. CD80 and CD54 monoclonal antibodies inhibited the specific T‐cell DNA synthesis proliferation. The proliferative T‐cell response was either MHC class I or class II restricted (inhibition by monoclonal antibodies). The specific cytokine gene expression could be found in isolated CD4, as well as CD8, T‐cell subsets. This study demonstrated the presence of a potential natural specific CD4, as well as a CD8 type 1 T‐cell immunity against the leukaemic CLL tumour B cells in CLL. A further detailed analysis of the spontaneous anti‐CLL T‐cell immunity is warranted that may facilitate the development of effective anti‐tumour vaccines in CLL.