bluuebunny/crossref_metadata_2025 huggingface dataset:
The storage pool deficiency in platelets from humans with the Chédiak‐Higashi syndrome: study of six patients
Authors: Apitz‐Castro Rafael , Cruz Maria Rosa , Ledezma Eliades , Merino Fernando , Ramirez‐Duque Pedro , Dangelmeier Carol , Holmsen Holm
Published: March 2008

Summary Functional and biochemical studies of platelets from human Chédiak‐Higashi syndrome (CHS) are scarce and/or incomplete. In the present report, the aggregation response to a variety of inducers of platelet aggregation, the content of the dense granule constituents ATP, ADP, serotonin and calcium, the secretion of ATP, ADP, and calcium induced by thrombin, the total content of magnesium, the incorporation of 14C‐adenine in the cytoplasmic pool of adenine nucleotides, as well as the content of intracellular cyclic‐AMP, have been quantitated in six patients with CHS. Furthermore, data is presented on the kinetics of uptake of radiolabelled serotonin and its storage in human CHS platelets. An abnormal aggregation behaviour was found in all patients. However, the response of CHS platelets to the different inducers studied did not show a uniform pattern. The total content and the maximal amounts of the dense granule constituents secretable by thrombin were greatly decreased in all six patients. Total magnesium content was similar to that of normal platelets. The ATP/ADP ratio was higher than in controls. Uptake of radiolabelled serotonin by CHS platelets closely followed the uptake by normal platelets; during the first 2–3 min, however, incorporation of the amine by CHS platelets came rapidly to a plateau which contrasts with the steady, linear increase in uptake found in controls. CHS platelets loaded with radiolabelled serotonin and gel‐filtered, showed a spontaneous release of radioactivity not observed in normal platelets under the same conditions. The cyclic‐AMP content of CHS platelets was similar to that of normals. In contrast to platelets from patients with storage pool disease, the secretable calcium from CHS platelets represents a 67% of total platelet
Immuno‐enzymatic detection of fibronectin in normal and pathological haematopoietic tissue
Authors: Reilly J. T. , Nash J. R. G. , Mackie M. J. , McVerry B. A.
Published: March 2008

Summary Cryostat sections of bone marrow biopsies were stained immuno‐enzymatically for fibronectin using the alkaline phosphatase: anti‐alkaline phosphatase (APAAP) technique. In normal marrow fibronectin was present only in megakaryocytes and the walls of blood vessels. In contrast, myelofibrotic bone marrow possessed an extensive distribution of fibronectin which did not correspond to the distibution of reticulin. Infiltrated and hypercellular marrows showed an increase in fibronectin which appeared to be related to increased marrow vascularity. Again no correlation with the reticulin pattern was evident. In contrast to platelets, other circulating blood cells had no demonstrable fibronectin. This study suggests that fibronectin does not act as a mediator of haemopoiesis in vivo as has been suggested by the results of experiments in non‐human models.
Megakaryocytic cells in mixed haemopoietic colonies (CFU‐GEMM) from the peripheral blood of normal individuals
Authors: Ganser Arnold , Elstner Elena , Hoelzer Dieter
Published: March 2008

Summary The cellular composition of mixed haemopoietic colonies CFU‐GEMM from the peripheral blood of normal individuals was investigated using both an improved in vitro culture assay and a monoclonal antibody reactive with the earliest megakaryocytic precursor cells. The frequency of CFU‐GEMM was found to be 22± 7/ml blood (mean± SE; n = 8) and 4.9± 1.8/106 mononuclear cells. Double‐labelling of single colonies with monoclonal antibodies against megakaryocytic as well as granulocytic cells revealed that only 18% of mixed colonies from the peripheral blood contained megakaryocytic cells as compared to 87% of mixed colonies from the bone marrow, while no difference existed with regard to granulocytic cells. It is concluded that circulating multilineage progenitor cells either require different culture conditions to express their megakaryocytic component or, alternatively, that the circulating pluripotent stem cell pool mainly consists of bipotent rather than multipotent progenitor cells.
Staging systems for multiple myeloma: a comparison
Authors: Gassmann W. , Pralle H. , Haferlach T. , Pandurevic S. , Graubner M. , Schmitz N. , Löffler H.
Published: March 2008

Summary In 152 patients with multiple myeloma who had been treated with cytostatic agents the prognostic value of seven staging systems was evaluated: (1) Carbone et al (1967); (2) Acute Leukemia Group B (ALGB) and Eastern Cooperative Oncology Group (ECOG) (Costa et al, 1973); (3) Southeastern Cancer Study Group (SECSG) (1975); (4) Durie & Salmon (1975); (5) Alexanian et al (1975); (6) Merlini et al (1980); (7) British Medical Research Council (1980).

The staging systems of the ALGB (Costa et al, 1973) and SECSG (1975), both dividing patients into ‘good risk’ and ‘poor risk’ groups, showed significantly different survival curves. Nevertheless, despite statistical significance the observed differences were rather small. In the systems of Carbone et al (1967), Merlini et al (1980), Alexanian et al (1975) and Durie & Salmon (1975) some of the differences in the survival curves were statistically significant while others were not. Our data best fitted into the British Medical Research Council (1980) staging system, the survival curves of all three stages showing significant differences, with median survival time dropping from 83 months in stage A to 52 months in stage B and 26 months in stage C. Nevertheless, none of those systems was clearly superior to single risk factors, especially creatinine and haemoglobin.
Non‐benign sickle cell anaemia in western Saudi Arabia
Authors: Acquaye Joseph K. , Omer Awad , Ganeshaguru Kanagasabai , Sejeny Saud A. , Hoffbrand A. Victor
Published: March 2008

Summary

Seventy‐one Saudi and Yemeni Arabs with sickle cell anaemia from western Saudi Arabia aged between 11/2 and 42 years were studied. The mean steady state haemoglobin concentration of 8.1 g/dl was lower than that of 10.7 g/dl reported previously for sickle cell anaemia in eastern Saudi Arabia. The patients were divided into an SSLF group with fetal haemoglobin (HbF) of 10.0% or below (44 patients) and an SSHF group having HbF above 10.0% (27 patients). No significant differences were found in the haemoglobin concentrations, haematological indices and incidences of bone changes of the two groups. SSLF patients were significantly more prone to infections (P<0.01), however. Also, there was an overall high incidence of hepatomegaly (69.0%) and splenomegaly (54.9%) and hepatomegaly was significantly more common in the SSLF group (P<0.02).

Many of the patients, even with HbF levels over 10.0%, did not follow a benign course and suffered from severe anaemia, infections of the respiratory and urinary tracts, bone pains and infarcts, or bossing of the skull. Rarer complications included hepatic crisis, chest syndrome, retinal haemorrhage, epistaxis and hemiplegia. It is therefore apparent that Saudi Arabian sickle cell anaemia, even in patients with raised haemoglobin F levels, may be as clinically severe as in African patients.
Colony formation by primitive haemopoietic progenitors in cocultures of bone marrow cells and stromal cells
Authors: Gordon M. Y. , Hibbin J. A. , Kearney L. U. , Gordon‐Smith E. C. , Goldman J. M.
Published: March 2008

Summary

Human bone marrow contains a class of human haemopoietic progenitor cells that adhere to cultured marrow stromal cells and form colonies of blast cells. These progenitor cells are found in the non‐adherent mononuclear fraction of normal human bone marrow. They are not in active cell cycle and do not express Ia‐like (HLA‐DR) antigens but appear to be capable of self‐renewal in vitro. These properties indicate that they should be classified as members of the primitive haemopoietic progenitor cell compartment.
ADCC lysis of human erythrocytes sensitized with rhesus alloantibodies IV. CHARACTERIZATION OF ANTI‐D SERA WHICH ARE INACTIVE IN ADCC
Authors: Barclay G. R. , Forouhi P. , McCann Moira C. , Greiss M. Ayoub , Urbaniak S. J.
Published: March 2008

Summary

Certain anti‐D sera, selected on the basis of their agglutination characteristics in vitro, fail to induce lysis of Rh(D) positive red cells by lymphocyte mediated antibody dependent cell mediated cytotoxicity (ADCC). Further investigation revealed that the non‐lytic anti‐D blocked in an antigen specific manner the effect of other anti‐D sera which were normally lytic in ADCC. Absorption selection studies and fractionation of a non‐lytic anti‐D serum showed that the blocking effect was associated with IgG anti‐D. Antigen binding and lymphocyte Fc‐receptor binding studies indicated that the non‐lytic anti‐D was bound to Rh(D) positive red cells and enabled them to be bound by lymphocytes, but failed to mediate ADCC.
Platelet membrane glycoprotein abnormalities in patients with myeloproliferative disorders and secondary thrombocytosis
Authors: Clezardin P. , McGregor J. L. , Dechavanne M. , Clemetson K. J.
Published: March 2008

Summary

Carbohydrate‐specific surface labelling and 125I‐labelled lectin binding techniques, in combination with one‐ or two‐dimensional (non‐reduced/reduced) SDS‐polyacrylamide gel electrophoresis have been used with platelets from patients with myeloproliferation disorders and secondary thrombocytosis and from healthy donors. In essential thrombocythaemia platelet membrane glycoproteins were significantly less sialylated than in normals (particularly GP Ib and IIIa). Increased binding of125I‐labelled Lens culinaris lectin to thrombospondin and GP IIIa indicated a defect in the glucose/mannose glycosylation of the platelet glycoproteins in essential thrombocythaemia. In polycythaemia vera and in chronic myeloid leukaemia the terminal sialic acid of glycoprotein IIIb was labelled slightly more than normal. In chronic myeloid leukaemia there was increased labelling of the penultimate galactose/N‐acetylgalactosamine residues of GP Ib, IIb, IIIa and IIIb. In comparison to myeloproliferative disorders, platelets from patients with secondary thrombocytosis showed no significant changes, except for platelets from two patients with idiopathic thrombocytopenic purpura which showed an increased sialylation of all surface glycoproteins.
Book Reviews
Authors: Shinton N. K. , Lewis S. M. , Brozovic Milica , Marcus Robert
Published: March 2008

Book Reviews in This Article:

Practical Haematology, 6th edition. By J. V. Dacie and S. M. Lewis.

Blood Cells in Nuclear Medicine, Part II. Migratory Blood Cells. Edited by G. F. Fueger.

L'Héparine—Fabrication, Structure, Propriétés, Analyses. By J. P. DuClos.

The Leukaemias—Epidemiological Aspects. By Martha S. Linet.
B‐cell activation in human plasma cell dyscrasias
Authors: Levinson A. I. , Dziarski A. , Blankenhorn E. , Schreiber A. D. , Negendank W. G.
Published: March 2008

Summary

We studied the abnormal in vitro polyclonal B‐cell activity observed in patients with multiple myeloma and Waldenström's macroglobulinaemia. Numbers of cells spontaneously secreting immunoglobulin (Ig) in freshly isolated suspensions of peripheral blood mononuclear cells and pokeweed mitogen (PWM) stimulated cultures of blood mononuclear cells were determined with a protein. A reverse haemolytic plaque assay. These data were correlated with both the clinical and laboratory parameters of disease. Furthermore, Ig secreted into supernatants of PWM‐stimulated cultures was examined by a light chain radioimmunoassay for evidence of in vitro activation of malignant B‐cells. The mean level of circulating immunoglobulin secreting cells (IgSC) in patients was elevated when compared to that of normal subjects. The highest values were observed in those patients with the highest levels of serum paraprotein. However, experiments with cycloheximide suggested that such increased circulating IgSC values were often caused by the detection of Ig passively adsorbed to blood mononuclear cells. The studies with PWM stimulation of blood mononuclear cells were particularly revealing. Cultures of patient blood mononuclear cells with PWM showed depressed IgSC responses as a group compared to cultures of normal blood mononuclear cells: nevertheless, approximately half the patients demonstrated a sizeable response to PWM. No evidence for preferential activation of Ig secretion by the malignant B‐cell clone was observed. Impaired PWM induced responses were associated with advanced or progressive clinical disease.
Low‐dose cytosine arabinoside regimen induced a complete remission with normal karyotypes in a case with hypoplastic acute myeloid leukaemia with No. 8‐trisomy: in vitro and in vivo evidence for normal haematopoietic recovery
Authors: Tagawa M. , Shibata J. , Tomonaga M. , Amenomori T. , Yoshida Y. , Kuriyama K. , Matsuo T. , Sadamori N. , Ichimaru M.
Published: March 2008

Summary

Complete remission was achieved in a case of hypoplastic acute myeloid leukaemia with chromosomal abberation of No. 8‐trisomy by giving very low dose of cytosine arabinoside (ara‐C), 10 mg (7.5 mg/m2)/d, by 24 h continuous intravenous infusion for 20 d. We observed a definite cytoreduction phase in the bone marrow (BM) before normal haematopoiesis resumed. The remission BM showed only normal karyotypes in all metaphases examined. Granulocyte/macrophage colonies and erythroid bursts recovered sufficiently in numbers and cytogenetic study on single colonies and bursts revealed only normal karyotypes. These observations provide evidence that a low dose ara‐C regimen can induce remission by cytoreduction which diverts the growth advantage from the leukaemic clone to the normal clones.
Cyclical neutropenia and T8 lymphocyte mediated stimulation of granulopoiesis
Authors: Smith J. G. , Seenan A. K. , Smith M. A. , Galloway E. , Lesko M. J. , Lucie N. P. , Robertson M. R. I. , Rowan R. M.
Published: March 2008

Summary

A 33‐year‐old female with cyclical neutropenia and a reciprocally cycling T8 (suppressor/cytotoxic) lymphocytosis was investigated. T8 lymphocytes ranged between 1.4 and 5.6 × 109/1 and a significant proportion (50–75%) were preactivated (1a+). Fcy receptors were detected in only a minority (7–10%). Functional studies on the lymphocytes indicated that despite their phenotype, little natural killer and reduced suppressor activities were present. Anti‐granulocyte antibodies were not detectable in the serum.

Production of colony stimulating activity (CSA) was assessed in the patient and control subjects’lymphocytes. Using a methylcellulose marrow culture system, the CSA production by the patient's lymphocytes was markedly increased compared with the control. Monoclonal antibody cytotoxic experiments confirmed that the T8 lymphocytes were responsible.

As peaks of circulating T8 lymphocytes were synchronous with granulopoietic activity in the marrow, the above findings may represent a homeostatic mechanism which is attempting to compensate for an underlying stem cell defect.
The iron‐chelating agent picolinic acid enhances transferrin receptors expression in human erythroleukaemic cell lines
Authors: Testa U. , Louache F. , Titeux M. , Thomopoulos P. , Rochant H.
Published: March 2008

Summary

Picolinic acid, a metal chelating molecule, was administered to human erythroleukaemic cell lines (K 562 and HEL) that were grown in serum‐containing media. Picolinic acid inhibited both iron uptake and cell growth. Furthermore, picolinic acid was shown to markedly decrease the level of ferritin in the cells. In spite of the inhibition of cell growth, picolinic acid induced a marked increase in the transferrin‐binding capacity of the cells. This phenomenon was due to a two‐fivefold enhancement of the rate of transferrin receptor biosynthesis. Other iron‐chelating compounds, capable of reducing the level of intracellular iron, also elicited a marked enhancement of the transferrin‐binding capacity of the cells. However, the addition of iron, as ferric ammonium citrate, in the culture medium elicited a marked increase in the level of ferritin and a strong decrease in the transferrin‐binding capacity of the cells. On the basis of these data we propose that a feed‐back mechanism is involved in the regulation of transferrin receptors: when the cells accumulate iron they decrease the number of transferrin receptors in order to prevent further accumulation of iron; when no or low iron is available to the cells, the number of transferrin receptors markedly increases as a compensatory mechanism.
Serum immunoreactive erythropoietin and erythropoiesis in protein‐energy malnutrition
Authors: Wickramasinghe S. N. , Cotes P. Mary , Gill D. S. , Tam R. C. , Grange Adenike , Akinyanju O. O.
Published: March 2008

Summary

Immunoreactive erythropoietin was estimated in the sera of 23 Nigerian children with protein‐energy malnutrition (PEM) and 14 healthy Nigerian children of similar age attending a well baby clinic. The geometric mean estimate for this parameter was 262 mlU/ml (observed range 39–1340 mIU/ml; 95% confidence range 25–1738 mIU/ml) in the children with PEM and 80 mIU/ml (observed range 43–257 mIU/ml; 95% confidence range 27–241 mIU/ml) in the healthy children. Erythropoietin levels were above the 95% confidence range for the healthy children in 14 of the cases of PEM. There was a statistically significant inverse correlation between the haemoglobin levels of the children with PEM and the logarithm of immunoreactive serum erythropoietin estimates (r=–0.73; P<0.001). By contrast, statistically significant correlations were not found between the logarithm of erythropoietin estimates and either the percentage of erythroblasts in the marrow, the M/E ratio or the logarithm of the absolute blood reticulocyte count. These data suggest that there is no abnormality of erythropoietin production in PEM and that the anaemia seen in this condition results from an impairment of erythropoiesis. A stepwise multiple regression analysis revealed a positive correlation between the logarithm of the erythropoietin level and the logarithm of the concentration of circulating neutrophil metamyelocytes plus myelocytes and we speculate on the aetiology of this finding.
Absence of ABH antigens on neutrophils
Authors: Dunstan R. A. , Simpson M. B. , Borowitz M.
Published: March 2008

Summary

Many investigators have concluded that polymorphonuclear leucocytes (PMN) express ABH antigens in parallel to red cells (RBC). We have examined human PMN for ABH antigens using human isoantibodies and mouse monoclonal antibodies with three highly sensitive and specific two‐stage assay systems: (1) fluorescence flow cytometry, (2) immunofluorescence microscopy, and (3) avidinbiotin immunoperoxidase microscopy. In all three assays the ABH antigens could not be detected on the surface of PMN. Previous reports alleging that ABH antigens occur on PMN probably represent false positive reactions due to inherent technical problems.
Deficient natural killer (NK) cells in paroxysmal nocturnal haemoglobinuria (PNH): studies of lymphoid cells fractionated by discontinuous density gradient centrifugation
Authors: Y oda Yasuhiro , Abe Tsukasa
Published: March 2008

Summary

Non‐adherent, non‐B lymphoid cells from six patients with PNH and six healthy subjects were fractionated by Percoll discontinuous density gradient centrifugation (DDGC). The cell distribution pattern, NK cell activity (NKA), large granular lymphocytes (LGL) count and surface marker phenotypes were studied. The distribution patterns of patients’cells did not significantly differ from the controls. The peak of the NKA was found in low density fractions where the maximum counts of LGL, Leu‐7+ 2‐ cells and Leu‐11+ cells were present. The NKA and the proportion of Leu‐7+2‐ cells and Leu‐11+ cells were significantly lower in patients with PNH (P<0.001 for NKA and surface phenotypes; P<0.02 for LGL counts). NKA in the Percoll fractions was correlated with the counts of LGL (r= 0.69, P<0.001), Leu‐7+2‐ cells (r= 0.75, P<0.001) and Leu‐11+ cells (r= 0.89, P<0.001). Therefore, we concluded that NKA is deficient in PNH because of decreased NK cell counts.
DNA and RNA determination in 111 cases of childhood acute lymphoblastic leukaemia (ALL) by flow cytometry: correlation of FAB classification with DNA stemline and proliferation
Authors: Suarez Carlos , Miller Denis R. , Steinherz Peter G. , M elamed Myron M. , Andreeff Michael
Published: March 2008

Summary

Flow cytometry with acridine orange was used in 111 cases of childhood acute lymphoblastic leukaemia (ALL) to determine simultaneously DNA index, cell cycle distribution and cellular RNA content in bone marrow samples. The overall incidence of DNA aneuploidy in the population was 40%. Significantly higher incidence of DNA aneuploidy was present in the L2 (70.6%) as compared to the L1 (34.1%) group (P= 0.007), using the FAB classification. No difference in the frequency of normal and abnormal DNA stemlines was found between newly diagnosed and relapsed patients. The L2 group had significantly higher proliferation (S phase = 12.4%) than L1 (S phase = 5.8%) (P=0.01), but RNA content was not significantly different. The aneuploid group had significantly more (P=0.01) cases with L2 morphology (28.6%) compared to the diploid group (8%) and proliferation was higher in DNA aneuploid (S‐phase = 9.5%) compared to DNA diploid (S‐phase = 4.7%) leukaemias. Likewise, RNA content was significantly higher in aneuploid than in diploid ALL (P= 0.006). These correlations between morphology, cell kinetics and DNA index elucidate biological properties of leukaemic cells with potentially prognostic value.
Relationship between erythropoiesis and the enhanced intestinal uptake of ferric iron in hypoxia in the mouse
Authors: Raja K. B. , Pippard M. J. , Simpson R. J. , Peters T. J.
Published: March 2008

Summary. An in vitro technique was used to determine the unidirectional Fe3+ uptake rates in mouse duodenal fragments. In animals in which erythropoiesis had been stimulated by hypoxia, Fe3+ uptake by the duodenal fragments was enhanced due to an increase in Vappmax However, when erythropoiesis was increased by injections of erythropoietin, intestinal Fe3+ uptake rates were unaffected. Mice subjected to sub‐total nephrectomy showed an increased Vappmax for Fe3+ after exposure to hypoxia, despite the absence of an erythropoietic response. When the bone marrow was obliterated by treatment with 89Sr, a small increase in Vappmax for Fe3+ was found: permeability studies and morpho‐metric analyses demonstrated no apparent irradiation damage to the duodenal mucosa of these animals. Exposure of 89Sr‐treated mice to hypoxia caused a further significant increase in Vappmax. These results indicate that the increase in intestinal mucosal iron uptake which follows hypoxia is not mediated by erythropoietin or other factors associated with increased erythropoiesis.
Nicotine alters fibronectin and factor VIII/vWF in human vascular endothelial cells
Authors: Blaes N. , Piovella F. , Samaden A. , Boutherin‐Falson O. , Ricetti M.
Published: March 2008

Summary. Primary cultured human endothelial cells derived from umbilical cord vein were exposed during the growth of the culture to medium containing nicotine at various concentrations (0.5‐200 μg/ml). Patterns of cellular fibronectin and factor VIII/vWF were compared to control by immunofluores‐cence technique. The levels of glycoproteins released in the culture medium were quantified by ELISA method. Treated cells showed an important decrease in fibronectin content with fragmentation of the fribronectin pericellular filaments, whereas the levels of secreted fibronectin were reduced in a dose‐dependent manner. This reduction of fibronectin availability was correlated with an elongation of cell shape as revealed with phase contrast microscopy. By immunofluorescence, factor VIII/vWF cytoplasmic granules appeared drastically reduced whereas the secretion of the protein was significantly increased. As shown by electron microscopy, there was a concomitant reduction in the number and size of Weibel‐Palade bodies. These studies indicate that nicotine modifies fibronectin and factor VIII/vWF distributions but in different ways.
Respective role of antithrombin III and heparin cofactor II in the in vitro anticoagulant effect of heparin and of various sulphated polysaccharides
Authors: Sie P. , Ofosu F. , Fernandez F. , Buchanan M. R. , Petitou M. , Boneu B.
Published: March 2008

Summary. The in vitro anticoagulant effects of standard heparin (SH) and of seven other sulphated polysaccharides (SPS) were investigated by measuring activated partial thromboplastin time (APTT) prolongation of normal plasma and of plasmas selectively depleted of antithrombin III (AT III), of heparin cofactor II (HCII) and of both heparin cofactors. This allowed the determination of the relative contribution of each of the two heparin cofactors to the SPS anticoagulant effect. The SPS varied in their relative activities as catalysts of thrombin inhibition by purified AT III or HC II. The anticoagulant activities of heparin and dermatan sulphate were primarily attributable to their ability to enhance thrombin inhibition by AT III and HC II respectively. Heparin had an additional minor anticoagulant activity which was independent of both AT III and HC II. Pentosan polysulphate, high molecular weight dextran sulphate, heparin with low affinity for AT III and a sulphated heparin derivative had weaker anticoagulant activities in normal plasma than standard heparin. The anticoagulant activities of these last four SPS in plasma depleted of both AT III and HC II were similar to their respective activities in normal plasma. This suggests that these SPS act by directly preventing thrombin generation rather than by enhancing thrombin inhibition.
Interferon responses of peripheral blood mononuclear cells from normal and leukaemic children
Authors: Mohammed G. S. , Taylor C. E. , Dickinson A. M. , Craft A. W. , Kernahan J. , Reid M. M. , Proctor S. J. , Toms G. L.
Published: March 2008

Summary. Human peripheral blood mononuclear cell cultures (PBMC) stimulated with Sendai virus or K562 cells produce a mixture of interferons. Temperature and pH stability characteristics and reactions with monospecific antibodies indicate that PBMC cultures from adults produce interferons alpha and gamma in approximately equal proportions. PBMC cultures from children produce lower levels of interferons with a higher proportion of type alpha.

The ability of PBMC cultures from children with acute lymphoblastic leukaemia (ALL) to produce interferon was determined. Little or no gamma interferon was induced by either Sendai virus or K562 cells. Cultures from some children with ALL produced alpha interferon but mean levels were significantly lower than from normal children. A group of older children with ALL who had completed their course of therapy and were off treatment produced levels of interferon indistinguishable from those of normal children. This in vitro deficit, possibly induced by chemotherapy, may reflect an in vivo deficit and may contribute to the impaired handling of viruses seen in children being treated for ALL.
The bone marrow histological pattern has independent prognostic value in early stage chronic lymphocytic leukaemia
Authors: Geisler C. , Ralfkiær E. , Hansen M. Mørk , Hou‐Jensen K. , Larsen S. Olesen
Published: March 2008

Summary.The initial diagnostic bone marrow specimens from 90 consecutive, untreated patients with chronic lymphocytic leukaemia were examined for the pattern of lymphocytic infiltration in relation to clinical stage (International Workshop System) and survival. Three non‐diffuse (interstitial, nodular, mixed nodular–interstitial) and one diffuse pattern were recognized. Generally, the bone marrow patterns correlated well with clinical stage: a non‐diffuse pattern prevailed in early, and a diffuse pattern in later stages. However, with a Cox analysis of the covariate effect of clinical stage and bone marrow pattern on survival, the bone marrow pattern was shown to have independent prognostic significance in the early stage A in which a diffuse pattern carried a fourfold increase in death rate as compared to a non‐diffuse pattern. These high risk patients could not be identified by the Rai substages of the International Workshop System.
Phorbol ester‐induced production of beta‐2‐microglobulin in B‐CLL cells: relation to IgM secretory response and disease activity
Authors: Tötterman Thomas H. , Nilsson Kenneth , Simonsson Bengt
Published: March 2008

Summary. Earlier studies have indicated that serum β2m levels correlate with the disease activity, estimated tumour cell mass and prognosis of CLL. We have therefore analysed the spontaneous and phorbol ester (TPA)‐induced capacity of CLL cells to produce β2m in vitro in relation to disease activity. Cell cultures (>90% B cells) from 15 patients with active disease contained significantly higher (P<0·001) levels of β2m (0·22·0±09 mg/l) than cultures from 17 patients with inactive CLL (0·08·0±06). When TPA‐induced CLL cell cultures were compared, this difference was even more striking (0·48·0±18 versus 0·13·0±09). The capacity of TPA‐treated B‐CLL cells to export β2m was positively correlated with their capacity to secrete monoclonal IgM (r=0·73;P<0·001). Cell depletion experiments showed that the β2m export by B‐CLL cells is enhanced by accessory T cells. Four individual CLL patients were followed for 14 months and repeatedly investigated for signs and symptoms of active disease, and the CLL cells were tested for in vitro production of β2m. Fluctuations in the clinical activity of the leukaemia were paralleled by similar switches in CLL cell production of β2m.
Reduced natural killer T‐cells in B‐cell chronic lymphocytic leukaemia identified by three monoclonal antibodies: Leu‐11, A10, AB8.28
Authors: Foa R. , Fierro M. T. , Lusso P. , Raspadori D. , Ferrando M. L. , Matera† L. , Malavasi F. , Lauria F.
Published: March 2008

Summary. The proportion of E‐rosette positive T‐lymphocytes capable of reacting with three monoclonal antibodies (MoAbs)—Leu‐11, A10, AB8.28—‐ which appear to recognize specifically natural killer (NK) cells, was assessed in a series of untreated patients with B‐cell chronic lymphocytic leukaemia (B‐CLL). Irrespective of the clinical stage of the disease, the capacity of B‐CLL T‐lymphocytes to react with all three MoAbs was significantly reduced compared with that of normal circulating T‐cells (Leu‐11: 2·5%±1·9SD; A10: 2·3% 1·3; AB8.28: 7%±6·6 v Leu‐11: 13·5%±4·5; A10: 8·5%±4·6: AB8.28: 12% 5·5) Furthermore, a marked difference was demonstrated between the reactivity with the Leu‐11, A10, AB8.28 MoAbs and the proportion of Leu‐7 positive T‐cells, which in B‐CLL is significantly higher than in normal blood (23%±12·1 v 11·9%±5·9).

These findings are in agreement with previous evidence of a discrepancy in B‐CLL between the phenotypic expression, assessed by Leu‐7 positivity, and the true functional activity of NK T‐cells, and suggest that the Leu‐11, A10 and AB8.28 MoAbs correlate well with the depressed NK function found in this disease.
Cellular levels of mRNA from c‐myc, c‐myb and c‐fes onc‐genes in normal myeloid anderythroid precursors of human bone marrow: an in situ hybridization study
Authors: Emilia Giovanni , Donelli Amedea , Ferrari Sergio , Torelli Umberto , Selleri Licia , Zucchini Patrizia , Moretti Luigi , Venturelli Donatella , Ceccherelli Giovanni , Torelli Giuseppe
Published: March 2008

Summary. The expression of three onc‐genes, c‐myc, c‐myb and c‐fes, has been evaluated at the cellular level in myeloid and erythroid precursors of normal human bone marrow, by ‘in situ’ hybridization with tritium‐labelled probes. A relatively large amount of m‐RNA from the three onc‐genes was detected in myeloblasts and promyelocytes, but whereas the expression of c‐myb and c‐myb decreased in more advanced stage of maturation of the myeloid lineage, c‐fes mRNA remained at a relatively high level until the granulocyte stage, c‐myc and c‐myb were expressed at a fairly high level in basophilic erythroblasts, which also showed low levels of c‐fes mRNA. No expression of these onc‐genes was detectable in more mature erythroblasts, Megakaryocytes showed high levels of m‐RNA from all three onc‐genes.

Our results suggest that c‐myc and c‐myb expression is related in some way to the cellular proliferation of myeloid and erythroid precursors, whereas c‐fes expression is more restricted to myeloid differentiation.
Enzyme linked immunosorbent assay (ELISA) for the measurement of factor VIII coagulant antigen (CAg) using haemophilic antibodies
Authors: Ingerslev Jørgen , Stenbjerg Stener
Published: March 2008

Summary.A method for the quantitation of factor VIII clotting antigen (VIII:CAg) has been developed based on a micro enzyme linked immunosorbent assay (ELISA) principle employing antibodies from two polytransfused haemophilia A patients. Solid polystyrene support bound IgG fraction of inhibitor plasma extracted VIII:CAg from normal plasma and samples. Bound VIII:CAg was detected by peroxidase labelled F(ab′)2fragment of the IgG used for solid phase. Two assays, each based on its particular inhibitor antibody, were set up. The F VIII clotting antigen in plasma of 30 healthy persons was found identical with the two VIII:CAg assays (r= 0.97) and closely correlating with clotting activity (VIII:C) (r= 0.84). Serum VIII:CAg was 67% (±14.5%) of the corresponding plasma value. In severe haemophilia A, 17 out of 19 had VIII:CAg values < 1 U/dl. Two patients with cross‐reactive material (CRM) were found. In some milder cases of haemophilia A, higher values of VIII:CAg than VIII:C was recorded. The sensitivity of the method was 0.08 U/dl. Inter assay coefficient of variation at the 100 U/dl level was 9.5% (CV%), at the 2 U/dl level 16.4% (CV%). Mainly due to the great stability of enzyme conjugated antibody compared to the natural decay of radioiodinated material and subsequent loss of detecting material, ELISA was found superior to immunoradiometric assay (IRMA).
Influence of graft‐versus‐host disease prophylaxis on early T‐lymphocyte regeneration following allogeneic bone marrow transplantation
Authors: Gratama J. W. , Würsch A. M. , Nissen C. , Gratwohl A. , D'Amaro J. , de Gast G. C. , van Rood J. J. , Speck B.
Published: March 2008

Summary. An early decrease in the ratio between T4 + and T8 + T lymphocytes has been shown to correlate with the development of grade II‐IV GVHD in allogeneic bone marrow transplant (BMT) recipients receiving methotrexate (MTX) as prophylaxis for acute graft‐versus‐host disease (GVHD). This study compares the onset of T‐cell regeneration in patients receiving cyclosporin A (CyA) with those receiving MTX. Firstly, lymphoid recovery occurred at a significantly faster rate in the patients on CyA. Secondly, in those patients, the repopulation of T4+ and T8+ T cells started simultaneously, whereas in patients on MTX the repopulation of the T8+ subset lagged about a week behind that of the T4+ subset. Thirdly, the decrease in the T4/T8 ratios as a function of the lymphocyte counts occurred at a significantly slower rate in the patients on CyA than in those on MTX. Thus, the differences in the onset of T‐cell regeneration in BMT recipients on CyA as compared with those on MTX abolished the correlation of the T4/T8 ratio changes with grades II‐IV GVHD as described in patients receiving MTX.
Use of leucocyte‐poor blood components and HLA‐matched‐platelet donors to prevent HLA alloimmunization
Authors: Murphy M. F. , Metcalfe P. , Thomas H. , Eve J. , Ord J. , Lister T. A. , Waters A. H.
Published: March 2008

Summary. Recent studies have shown that the incidence of alloimmunization due to repeated platelet transfusions from random donors may be reduced by the use of leucocyte‐poor blood components. These results were confirmed by this study, where 16% of patients with acute leukaemia undergoing initial chemotherapy and receiving leucocyte‐poor blood components developed lymphocytotoxic antibodies, compared with 48% of patients in a control group receiving standard (non‐leucocyte‐depleted) blood components. In a third group, who received leucocyte‐poor blood components and HLA‐matched platelets, none of the patients developed lymphocytotoxic antibodies. There was a low incidence of platelet‐specific antibodies (8%) but no difference between the three groups. Improved methods of removing leucocytes from blood components appear to offer the best approach for minimizing HLA alloimmunization, as the provision of HLA‐matched platelet donors for prophylactic platelet support of all patients is not feasible.
A relatively high frequency of severe (type III) von Willebrand's disease in Israel
Authors: Berliner Shlomo A. , Seligsohn Uri , Zivelin Ariela , Zwang Ety , Sofferman Gideon
Published: March 2008

Summary. Sixty patients with von Willebrand's disease belonging to 34 unrelated families were classified into the various types of the disease by analysis of the multimer patterns of von Willebrand factor. Type I disease was observed in 62% of the families, a finding similar to the recently published British and Swedish series of patients. Type III (severe) von Willebrand's disease was observed more frequently in the Israeli group (29%) than in the other two series. Of the 16 patients with type III disease eight were Arabs living in Israel, the West Bank and the Gaza Strip (total population about 1·5 × 106). Thus, it appears that the frequency of type III disease is very high among Arabs (5·3/106). Sibship analysis of all families affected by the type III mutant gene was compatible with an autosomal recessive mode of in heritance. An attempt was made to define carriers of type III disease and to distinguish them from type I patients and from healthy subjects. In 15 obligate carriers of type III disease mean levels of factor VIII clotting activity, of von Willebrand factor and of ristocetin cofactor were significantly higher than the corresponding mean values observed in 31 symptomatic and 12 asymptomatic type 1 patients, and in turn significantly lower than the corresponding mean values observed in 30 healthy subjects. Ristocetin cofactor was the best criterion for discrimination among carriers of type III disease, normal subjects and type I patients.
Blood cell phosphogluconolactonase: assay and properties
Authors: Beutler Ernest , Kuhl Wanda , Gelbart Terri
Published: March 2008

Summary. 6‐Phosphogluconolactonase (6‐PGL) catalyses the second reaction of the hexosemonophosphate pathway. Although the ‐Iactone of 6‐phosphogluconic acid is the natural substrate for this enzyme, the more stable γ‐lactone may also be used. We prepared the γ‐lactone of 6‐phosphogluconic acid from 6‐phosphogluconate. When stored in dimethylsulfoxide, this material was found to be stable in liquid nitrogen for several months. A method for measuring 6‐phosphogluconolactonase (6‐PGL) using the γ‐lactone as substrate has been developed, after defining conditions under which spontaneous hydrolysis of the lactone is relatively slow and the enzymatic velocity is relatively rapid.

The enzyme had no divalent cation requirement and was not significantly inhibited by a 50‐fold excess of gluconolactone. It was distinct, therefore, from gluconolactonase. At 25C normal human red cells were found to contain approximately 50 IU of 6‐phosphogluconolactonase/g Hb. The activity of the enzyme was independent of red cell age. Based on protein content, human lymphocytes, monocytes, granulocytes and platelets, contained approximately 10 times the activity of red blood cells. The activity of 6‐PGL was stable for at least 6 d in red cells stored at 22C and for at least 20 d in red cells stored at 4C.
The expression of ABH antigens during in vitro megakaryocyte maturation: origin of heterogeneity of antigen density
Authors: Dunstan Robert A.
Published: March 2008

Summary. An indirect immunofluorescence technique with single and double labelling has been used to examine cultured human megakaryocytes for ABH antigens. This technique demonstrated the presence of these antigens on megakaryocytes and a population of small mononuclear cells that probably represent the differentiated precursors of megakaryocytes. In contrast to the intense homogeneous labelling with human anti‐PlA1, the labelling with human anti‐A, mouse monoclonal anti‐A and anti‐type 2H is light and heterogeneous, with many cells staining weakly or not at all. This variability in blood group ABH surface antigen expression appears to occur at the level of cell proliferation within colonies. While cells within an individual colony are homogeneous in fluorescence intensity, there is considerable variation between colonies. The progeny of individual megakaryocytes also appear uniform in ABH antigen expression.
A monoclonal antibody reactive with a subset of human plasma cells
Authors: Gonchoroff Nick J. , Katzmann Jerry A. , Garton John P. , Ruiz‐Argüelles Guillermo J. , Eilers Carol R. , Greipp Philip R. , Kyle Robert A.
Published: March 2008

Summary A mouse monoclonal antibody (Rl‐3) raised against a human plasmacytoma xenograft and reactive with human plasma cells is described. The antibody reacts with a subset of plasma cells and lymphocytes, but is nonreactive with granulocytes, normoblasts, thymus, spleen or T lymphocytes. Flow cytometer double labelling experiments showed that approximately 50% of Rl‐3 positive bone marrow cells expressed surface immunoglobulin, but no Rl‐3 positive cells also expressed B1 or Leu 4. In a myeloma patient the R1‐3 reactive cells were found to have a higher rate of DNA proliferation and were aneuploid as determined by flow cytometry. The R1‐3 antigen is expressed on late B lymphocytes and early plasma cells.
Transient activation of the acetyltransferase necessary for paf‐acether biosynthesis in thrombin‐activated platelets
Authors: Coëffier E. , Ninio E. , Le Couedic J. P. , Chignard M.
Published: March 2008

Summary Thrombin‐activated platelets formed paf‐acether (1‐0‐alkyl‐2‐acetyl‐sn‐glyceryl‐3‐phosphorylcholine), a molecule susceptible to play a role in haemostasis and thrombosis, and its deacetylated analogue, lyso paf‐acether, a biologically inactive molecule. We presently show the presence in human and rabbit platelet lysates of an acetyltransferase which transfers the acetyl moiety of acetyl‐coenzyme A (acetyl‐CoA) onto synthetic lyso paf‐acether, yielding the fully active paf‐acether molecule. Under our optimal standard conditions, 0.36 ± 0.23 nmol paf‐acether/10 min/mg proteins was formed by the acetyltransferase from resting human platelets. Upon thrombin stimulation, the acetyltransferase activity doubled within 30 s, reaching a maximum at 2 min (1.17 ± 0.31 nmol paf‐acether/10 min/mg proteins) and decreased progressively. Similar results were obtained using rabbit platelets. In addition we demonstrated that the activation and deactivation of the acetyltransferase correlated with the kinetics of paf‐acether formation by thrombin‐activated rabbit platelets. It is hypothesized that this enzyme may represent one of the regulating mechanism of paf‐acether formation by platelets.
Platelet‐associated IgG in idiopathic glomerulonephritis and the nephritis of systemic lupus erythematosus
Authors: Bennett A. , Frampton G. , Cameron J. S.
Published: March 2008

Summary We studied platelet‐associated IgG (PAIgG) in patients with the nephritis of systemic lupus erythematosus (SLE) and with idiopathic glomerulonephritis (IGN), and found no increase in PAIgG in the patients with IGN, and an increase in only a minority of patients with SLE, all of whom had active disease. Patients with IGN and a nephrotic syndrome had both a low serum IgG and a low PAIgG. The increase in PAlgG in the patients with SLE correlated with the titres of antibody against dsDNA in the serum, but not with the platelet‐agglutinating immune complexes also present. Intraplatelet serotonin, however, was reduced in both groups, and this correlated with the amounts of platelet‐agglutinating complexes in the serum of the SLE patients. Immune complexes may associate with platelets in vivo to cause this release, but if so this must be a reversible phenomenon (‘hit and run’immune platelet injury); alternatively, the Fc binding to the platelet surface may be weak and insufficient to survive the ex vivo washing procedures.
Myelofibrosis in chronic granulocytic leukaemia: clinicopathologic correlations and prognostic significance
Authors: Lazzarino M. , Morra E. , Castello A. , Inverardi D. , Coci A. , Pagnucco G. , Magrini U. , Zei G. , Bernasconi C.
Published: March 2008

Summary. We performed 221 marrow trephine biopsies in 139 patients with Ph1‐positive (Ph1‐+) chronic granulocytic leukaemia (CGL) in order to assess the incidence, degree and prognostic significance of marrow fibrosis (MF) at various stages of the disease. We also attempted to elucidate the relationship between development of MF and the various clinical and haematological features of CGL. A significant correlation was found between the amount of fibrosis (graded from 0 to 3) and the stage of CGL, indicating that major fibrotic changes are associated with accelerated or blastic disease. Survival studies performed to assess the prognostic significance of the various degrees of MF, showed a progressively worse life‐expectancy from grade 0 to grade 3 fibrosis. Multivariate regression analysis indicated Hb level, age, number of marrow megakaryocytes (MKs), time from diagnosis as the features most significantly correlated with the degree of MF. This study demonstrates that the natural history of CGL involves a progressive increase in reticulin deposition towards severe MF, although the rate of this progression varies widely. Monitoring changes of fibrosis with sequential biopsies could give a measure of the rate of progression of the disease and help in prognostic assessment of CGL patients. Our findings also confirm that among marrow features the number of MKs is the cytological variable most significantly correlated with MF.
Coated pit formation: a membrane function involved in the regulation of cellular iron uptake
Authors: Egyed A. , Fodor I. , Lelkes G.
Published: March 2008

Summary. Diferric‐transferrin induces a marked increase in the number of coated pits on the reticulocyte membrane. This increase is followed by a decline, often to below the initial number. Since a good correlation was found between the rate of iron uptake and the number of coated pits, but not between the rate of transferrin recycling and the coated pit count, it is likely that coated pit formation is necessary for the removal of iron from transferrin.

The decline in the number of transferrin‐induced coated pits was observed only when haem synthesis was undisturbed, indicating that the accumulation of intracellular haem inhibits coated pit formation.

Based on these results we suggest that haem regulates the rate of iron uptake by inhibiting iron removal rather than receptor recycling.
Disappearance of factor VIII:C antibodies in patients with haemophilia A upon frequent administration of factor VIII in intermediate or low dose
Authors: van Leeuwen E. F. , Mauser‐Bunschoten E. P. , van Dijken P. J. , Kok A. J. , Sjamsoedin‐Visser E. J. M. , Sixma J. J.
Published: March 2008

Summary. In 18 haemophilia A patients with antibodies against factor VIII:C (F VIII:c) the effect of regular treatment with factor VIII (F VIII) in intermediate or low dose was studied. All patients with previous maximal F VIII:c antibody levels between 5 and 60 Bethesda Units per millilitre (BU/ml) showed a decrease of antibody level and normal F VIII recovery within 1‐2 months. From nine patients with previous maximal antibody levels above 60 BU/ml four showed a decrease of antibody level within 2‐26 months. In four young patients F VIII prophylactic therapy was started or continued as soon as there was evidence of F VIII:c antibody activity. In three of these patients F VIII recovery normalized within a few months.