bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin‐1 producing ability of leukaemia cells and its relationship to morphological diagnosis
Authors: Furukawa Yusuke , Ohta Masatsugu , Miura Yasusada , Saito Masaki
Published: March 2008

Interleukin‐1 (IL‐1) is a low molecular weight polypeptide produced by monocyte–macrophage lineage cells. IL‐1 production by primary‐cultured leukaemic cells of several FAB subtypes was estimated and compared with in vitro and in vivo lysozyme production.

The results indicate that IL‐1 production by monocytic leukaemia cells (M4 and M5) is significantly higher than that of myelocytic leukaemia cells (M1, M2 and M3). On the other hand, the serum lysozyme level was not correlated with the FAB subtypes and in vitro lysozyme production by monocytic leukaemia cells was higher than that of myelocytic leukaemia cells, but the M2 subtype was indistinguishable from monocytic leukaemia cells solely on the basis of lysozyme production.

We concluded that measurement of IL‐1 production by leukaemic cells, as a marker of monocytic leukaemias, was convenient and reliable, and might be useful for the diagnosis of morphologically or cytochemically atypical cases.
Mitozantrone‐induced toxicity and DNA strand breaks in leukaemic cells
Authors: Ho Anthony D. , Seither Eva , Ma David D. F. , Prentice H. Grant
Published: March 2008

By applying the fluorometric analysis of DNA unwinding (FADU) the in vitro effect of mitozantrone on DNA strand breaks was studied in seven different human leukaemic/lymphoma cell lines and in fresh leukaemic samples from seven patients with acute myeloid leukaemia refractory to conventional treatment. Pulse exposure to mitozantrone for 30 min invariably caused strand breaks. In the cell lines JM 1, KM3 and RPM I 8420 DNA strand breakage was progressive upon further incubation in drug free medium. These cell lines were killed by pulse exposure to mitozantrone. In the cell lines Molt 4, Daudi, Raji and HL‐60, the DNA strand breaks induced by mitozantone were only moderate and these cell lines were also resistent to killing by mitozantrone in vitro. The leukaemic cells of one of the seven patients behaved also like the cell lines that were sensitive and a complete remission was achieved in this patient using mitozantrone as single agent therapy. The other patients with a pattern similar to the resistent cell lines proved to be clinically refractory. Thus mitozantrone induces rapidly progressive DNA strand breaks as early as 30 min in leukaemic cells that are sensitive. The measurement of DNA strand breaks by the fluorometric analysis of DNA unwinding is a rapid method which might predict response to drugs whose major effect is on the induction of strand breaks.
Detection of cells of megakaryocyte lineage in haematological malignancies by immuno‐alkaline phosphatase labelling cell smears with a panel of monoclonal antibodies
Authors: Erber W. N. , Breton‐Gorius J. , Villeval J. L. , Oscier D. G. , Bai Y. , Mason D. Y.
Published: March 2008

Immuno‐alkaline phosphatase staining (by the APAAP technique) has been used to identify promegakaryo‐blasts in cell smears from 10 cases of leukaemia (three acute leukaemia, seven blast transformations). In all cases pro‐megakaryoblasts were labelled by at least two anti‐platelet glycoprotein (gp) antibodies, the highest percentages being obtained with anti‐gp IIIa (antibody C17). HLA‐DR was expressed by a variable percentage of neoplastic cells in all cases, the T11 (CD2) antigen (sheep red cell receptor) in four of seven cases tested and the p150,95 antigen in three of the six cases tested. In some cases of acute myeloid leukaemia APAAP staining of blood smears revealed circulating promegakaryoblasts and micromegakaryocytes (which superficially resemble small lymphoid cells). It is concluded that immuno‐alkaline phosphatase staining of cell smears offers a convenient means of diagnosing acute megakaryoblastic leukaemia in the routine laboratory.
Chronic and subacute myelomonocytic leukaemia in the adult: a report of 60 cases with special reference to prognostic factors
Authors: Fenaux Pierre , Jouet Jean Pierre , Zandecki Marc , Lai Jean Luc , Simon Marc , Pollet Jean Paul , Bauters Francis
Published: March 2008

We report 60 cases of chronic and subacute myelomonocytic leukaemias (CMML and SMML) in the adult, using the FAB group criteria. The M/F sex ratio was 3·3 and the mean age 67·5 years. Splenomegaly was found in 32% of cases, hyperleucocytosis in 52% of cases and mean blood monocytosis was 4·3 × 109/l. Marrow smears showed an excess of blasts in 57% of patients, a moderate increase in monocytes in most cases and frequent myelodysplastic features. An increase in serum lysozyme and polyclonal hypergammaglobulinaemia were usual and clonal cytogenetic anomalies found in about half of the patients tested. Treatment was usually palliative and the median survival was 28 months, a blastic transformation being responsible for a third of the deaths.

Prognostic factors at diagnosis were analysed retrospectively in the 46 patients who had sufficient follow up. Percentage of marrow blasts haemoglobin level and blood monocytosis at diagnosis, were subject to multivariate analysis, resulting in a discriminant ‘score’. This allowed assignment of each patient into one of two prognostic subgroups (10·9% probability of error): a poor prognosis one, with a life expectancy of less than 1 year and a high risk of acute transformation (subgroup termed SMML) and a better prognosis subgroup (termed CMML), with some CMML patients surviving over 5 years.
HLA class II antigen associated invariant chain gene expression in malignant lymphoma
Authors: Gondo Hisashi , Narni Franco , Lee Ming‐Sheng , Buck Mark B. , Cabanillas Fernando , Saunders Grady F.
Published: March 2008

Summary. Expression of the gene encoding HLA class II antigen‐associated invariant chain was studied in several types of fresh human malignant lymphoma by Northern blot analysis or slot‐blot analysis. The invariant chain mRNA levels decreased with the stage of B‐lymphocyte differentiation to plasma cells such as in immunoblastic lymphoma (IBL) or multiple myeloma (MM). The invariant chain gene (In‐gene) was expressed in diffuse large cell lymphoma (DLCL), while its expression was hardly detected in IBL. It is suggested that the difference in In‐gene expression between DLCL and IBL could be useful in distinguishing these two morphologically similar diseases.
Electroblotting and immunohistochemical staining for identification of platelet antibodies
Authors: Rock G. , Decary F. , Tittley P. , Fuller V.
Published: March 2008

Summary. The Western blot procedure has been adapted for use with a biotinylated antiglobulin reagent and a horseradish peroxidase complex (Vectastain ABC) to determine specific sites of antigen binding and permit discrimination between various types of platelet antibodies.

Monospecific anti‐PlA1 antisera or sera containing mixtures of multispecific HLA and unidentified platelet specific antibodies were tested with PlA1 phenotyped platelets. Using monospecific anti‐PlA1, one intense band with relative molecular weight (Mr) of 88000 and corresponding to glycoprotein IIIa was seen with the PlA1+ platelets. With mixtures of antibodies, reactions were seen with platelet specific antibodies without interference from the HLA antibodies; with one serum a band of Mr approximately 135000 was identified with Baka+, but not Baka‐ platelets, indicating the presence of an anti‐Baka in the serum. The location of the Baka antigen corresponded to the area for GP IIb. With another serum, a band of Mr 88000 was revealed with PlA1‐and some PlA1+ platelets suggesting the presence of an anti‐PlA2.

Two additional bands of Mr 160000 and 200000 were present on all preparations including autologous controls, probably due to the presence of non‐specific IgG. Thus, immunohistochemical staining is readily adaptable to the Western blot technique, and antibodies to platelet‐specific antigens can be easily differentiated from HLA antibodies.
Red‐cell GSH regeneration and glutathione reductase activity in G6PD variants in the Ferrara area
Authors: Anderson Barbara B. , Carandina Gianni , Lucci Marco , Perry Gillian M. , Vullo Calogero
Published: March 2008

Summary. Red‐cell studies were carried out on three groups of G6PD‐deficient subjects with different G6PD variants from the Ferrara area of Northern Italy. Red‐cell GSH and activities of G6PD, glutathione reductase (GR), glutathione peroxidase (GPx) and superoxide dismutase (SOD) were measured. A method was developed to measure red‐cell GSH regeneration after oxidation of endogenous GSH in whole blood by diamide and only this clearly distinguished the variants from each other and from normal. Regeneration by 1 h was lowest in the Mediterranean variant, 0–10.2% in contrast to 93–98% in normal. A predisposition to a haemolytic crisis after ingestion of fava beans was not clearcut, but subjects appeared to be at risk if GSH regeneration at 1 h was less than 30% of the endogenous level, and red‐cell FAD+ was very high indicated by high in vitro GR activity and inhibition by added FAD+.

It is suggested that the most informative tests in G6PD deficiency are measurements of GSH regeneration in intact red cells plus GR activity and/or red‐cell flavin compounds.
Effect of human recombinant erythropoietin on human marrow megakaryocyte colony formation in vitro
Authors: Dessypris Emmanuel N. , Gleaton John H. , Armstrong Otho L.
Published: March 2008

The effect of human recombinant erythropoietin (EP) on the growth of human marrow megakaryocyte colony forming units (CFU‐M) in vitro was investigated by the use of a plasma clot assay. EP as a single stimulating factor or as an additional factor to optimal concentration of leucocyte conditioned medium (PHA‐LCM) had no effect on the number of CFU‐M derived colonies. However, addition of EP (O·5‐1 U/ml) to cultures with suboptimal concentrations of PHA‐LCM increased megakaryocyte colony formation by 50‐90% but had no effect on the number of granulocytic‐monocytic colonies (CFU‐GM). Exposure of marrow cells to EP for 24‐48 h in liquid suspension cultures, followed by removal of the hormone and assaying the cells for CFU‐M in plasma clots, resulted in a 50‐100% increase of megakaryocyte colony formation in vitro. The augmenting effect of EP on CFU‐M growth in vitro was abolished when EP was added to the medium after the third day of culture. The presence of factors in human serum and in PHA‐LCM was an absolute requirement for the hormone to exert its potentiating effect on human CFU‐M growth in vitro. Recombinant EP potentiates the growth of human marrow CFU‐M and this effect seems to be exerted during the early stages of CFU‐M development in vitro.
An improved acidified glycerol lysis test (AGLT) used to detect spherocytes in pregnancy
Authors: Acharya J. , Ferguson I. L. C. , Cassidy A. G. , Grimes. A. J.
Published: March 2008

The acidified glycerol lysis test (Zanella et al, 1980) had proved unreliable in our hands. We now report the test to be stable and reliable when conducted at 37°C rather than at ambient laboratory temperature as used by earlier workers. With this modification, blood samples were tested from 167 pregnant subjects. Forty‐eight were less than 26 weeks pregnant and gave normal AGLT values (T50 > 1800 s) while of 119 more than 26 weeks pregnant 57 (48%) gave shortened T50 values indicative of the presence of spherocytes.
The role of intensive remission induction and consolidation therapy in patients with acute myeloid leukaemia
Authors: Tricot G. , Boogaerts M. A. , Vlietinck R. , Emonds M. P. , Verwilghen R. L.
Published: March 2008

Sixty‐one patients with AML, 59 adults and two children, were treated with intensive remission induction and consolidation therapy. The median age was 36 years. Forty‐four (72%) patients entered complete remission (CR): 11 patients received a bone marrow transplantation. The median survival of complete remitters was 26.5 months; the probability of remaining in CR at respectively 1 and 2 years was 75% and 62%. The only factor significantly correlated with the outcome of remission induction, survival and duration of CR was age. Patients < 30 years fared significantly better than those 30 years or older; no difference in outcome was observed between patients aged 30‐50 and those over 50 years. In patients < 30 years the CR rate was 95%; 75% of them were still alive at 2 years and only one (5%) has relapsed. In contrast, in patients 30 years or older the CR rate was 60% and the median survival only 11.5 months. 50% of the complete remitters in this age group have relapsed. Morbidity from intensive consolidation therapy was considerable; more than 50% of consolidation courses were complicated by high fever, needing urgent admission; only four (3%) courses had a fatal event. It is concluded that intensive consolidation therapy may be considered as a major advance in the treatment of younger patients with AML. while its role in older individuals remains questionable. A possible explanation for the completely different outcome in younger and older patients with AML is discussed.
A randomized trial of 13‐cis retinoic acid with or without cytosine arabinoside in patients with the myelodysplastic syndrome
Authors: Clark R. E. , Ismail S. A. D. , Jacobs A. , Payne H. , Smith S. A.
Published: March 2008

Ninety‐eight consecutive patients with myelodysplastic syndrome were randomized to a treated or a control group, both receiving conventional supportive therapy. The treated group were given 13‐cis‐retinoic acid 20 mg/d if marrow blasts were ≦ 5% or cytosine arabinoside 10 mg/d subcutaneously on 6 d /week if marrow blasts were 6‐30%, to which retinoic acid was added after 12 weeks. Serum levels of the drugs in the treated group were similar to those that would produce inhibition of CFU‐GM growth in vitro. In patients in the low blast group receiving retinoic acid. myeloid surface antigens reverted from an abnormal to a normal pattern.

Log rank analysis carried out after 25 months showed no significant difference in survival between the treated and control group, either in the total patient population or in the high and low blast groups considered separately. However, analysis of 39 non‐sideroblastic patients with ≦ 5% blasts showed an increase in survival in the treated group.
Aberrant pattern of red cell membrane and cytosolic proteins in a case of congenital dyserythropoietic anaemia
Authors: Pothier Brigitte , Morlea Laurette , Alloisio Nicole , Ducluzeau Marie‐Thearegse , Feao Claude , Blanchard Dominique , Cartron Jean‐Pierre , Garbarz Michel , Chaveroche Isabelle , Boivin Pierre , Favre‐Gilly Jean , Delaunay Jean
Published: March 2008

We report an unusual case of congenital dyserythropoietic anaemia (CDA). The propositus is a 25‐year‐old gipsy female presenting with a recessively inherited haemolytic anaemia. The diagnosis of CDA was based on erythrokinetic data and the morphological appearance of the erythroid precursors. The direct assay of HEMPAS antigen was negative. In peripheral blood there were 15% dacryocytes. The red cell membrane protein pattern was dramatically altered, with four major aberrant bands. Band a (mol wt 86 000) was at the lower edge of band 3, band b (mol wt 82 000) was below band 3, band c (68 000) and band d (67000) were below band 4.2. In addition, there was an array of aberrant minor bands below band d. Gel densitometric determinations and immunological characterization showed that these bands did not derive from any of the major components of the membrane. In fact, membrane proteins appeared normal in many respects, although periodic acid‐Schiff staining revealed an apparent decrease of sialoglycoproteins. The major aberrant bands a, b and c occur in very low amounts in controls. These bands, as well as band d, also exist in normal cytosol and were strongly increased in the propositus.
The lectin leucoagglutinin binds specifically to human granulocytes, monocytes and tissue mast cells: further evidence for a common origin of the three cell types
Authors: Schumacher U. , Horny H. P. , Welsch U.
Published: March 2008

The lectin leucoagglutinin (LA) binds to the granules of all forms of granulocytes as well as mast cells and monocytes, which is considered to be a further indication for a common origin of these cells.
Beneficial effect of intravenous gammaglobulin in a patient with complement‐mediated autoimmune thrombocytopenia due to IgM‐anti‐platelet antibodies
Authors: Brand A. , Witvliet M. , Claas F. H. J. , Eernisse J. G.
Published: March 2008

Intravenous gammaglobulin (IV‐IgG) was administered to a patient with chronic idiopathic thrombocytopenic purpura due to an unusual IgM platelet autoantibody causing in vitro complement‐dependent thrombocytoxicity and in vivo intravascular platelet destruction. After IV‐IgG infusion the peripheral platelet count temporarily increased to normal values, the mean platelet survival time increased and the platelet sequestration pattern changed from intravascular to predominantly hepatic destruction. In vivo and in vitro observations in this patient illustrate a transient beneficial effect of gammaglobulin infusion due to interference with the complement‐fixing autoantibodies against platelets.
A prospective study of the incidence of delayed haemolytic transfusion reactions following peri‐operative blood transfusion
Authors: Hewitt Patricia E. , Macintyre Elizabeth A. , Devenish A. , Bowcock Stella J. , Contreras Marcela
Published: March 2008

Delayed haemolytic transfusion reactions (DHTRs) are a recognized sequel of blood transfusion. The true incidence and importance of this complication have been difficult to estimate due to the lack of any prospective studies. We have carried out such a study by testing 530 patients who were transfused during cardiac surgery. 2% of the patients had new red cell alloantibodies detectable 1 week following transfusion. Despite this finding, and the fact that at the time the study was performed pre‐transfusion antibody screening of recipients was not routine practice, no DHTRs were diagnosed on clinical or laboratory criteria. These results indicate that the reported incidences, based on retrospective recognition of DHTRs, are not a serious underestimate of the frequency of the complication.
A specific radioimmunoassay for 5′‐deoxyadenosyl cobalamin in serum
Authors: Quadros Edward V. , Rothenberg Sheldon P. , Polu Sailaja
Published: March 2008

A specific radioimmunoassay for 5′‐deoxyadenosyl cobalamin (ado‐Cbl) has been developed using an antiserum raised to this cobalamin (Cbl). At a 1:100 or greater dilution the antiserum did not bind radiolabelled methyl‐hydroxo‐, sulphito‐ or cyano‐Cbl and these Cbl analogues did not compete in the radioimmunoassay even at 100‐fold higher concentration.

The serum concentration (range and mean ± SD) of ado‐Cbl in 30 normal subjects was 47–134 and 81 ± 16 pg/ml. The corresponding values for total Cbl in these sera were 189–610 and 355 ± 144 pg/ml, and the computed values for methyl‐Cbl were 142–476 and 274 ± 127 pg/ml. The coefficient of variation was substantially greater for methyl‐Cbl than for ado‐Cbl (46% v. 21%, respectively). The ado‐Cbl concentration was in the normal range (75–95 pg/ml) in five healthy subjects with a low normal concentration (189–217 pg/ml) of total Cbl. In two subjects with low total Cbl (118 and 170 pg/ml) and without any clinical evidence of Cbl deficiency, ado‐Cbl was normal (63 and 95 pg/ml. respectively). Thus, in this group, low methyl‐Cbl accounted for the lower total Cbl.

The concentration (mean ± SD) of serum ado‐Cbl and methyl‐Cbl in six patients with low total Cbl (57 ± 25 pg/ml) and clinical evidence for Cbl deficiency was 35 ± 12 pg/ml and 22 ± 22 pg/ml, respectively. This difference from the normal mean for each cofactor was highly significant (P<0.001). The decrease in the concentration of methyl‐Cbl in Cbl deficiency was relatively greater than the decrease in ado‐Cbl (92% v. 57%, respectively) which raised the relative concentration of ado‐Cbl in Cbl deficiency to 61% of the total Cbl. Although a low serum methyl‐Cbl is a sensitive indicator of Cbl deficiency, it may not be as specific as a decrease in serum ado‐Cbl. Factor(s) other than tissue stores of Cbl may lower serum methyl‐Cbl below the 95% confidence limit in the absence of clinical Cbl deficiency.
Interaction of acidic isoferritins with human promyelocytic HL60 cells
Authors: Covell Ann M. , Cook James D.
Published: March 2008

We have used undifferentiated human promyelocytic HL60 cells to study the binding of radioiodinated human ferritin in vitro. Specific binding of human heart ferritin could be demonstrated at 37°C, whereas no binding of liver ferritin could be found. The uptake of labelled heart ferritin was abolished by incubation at 4°C, by prior treatment of the HL60 cells with pronase and by the addition of human plasma to the medium. On the other hand, the addition of excess unlabelled human liver or rat liver ferritin had no effect on the uptake of labelled human heart ferritin. Dissociation studies showed that about 55% of the bound heart ferritin radioactivity could be released by incubation with medium alone and at least 90% with excess unlabelled heart ferritin. Over 70% of the dissociated ferritin could be precipitated with polyclonal anti‐ferritin serum or trichloroacetic acid. More than two‐thirds of the radioactivity which could not be released after washing in medium alone was recovered in the soluble intracellular fraction following cell lysis. Almost all of the soluble radioactivity could be precipitated with the polyclonal antiserum, indicating that very little lysosomal degradation of internalized heart ferritin had occurred. The present studies demonstrate a proteinmediated binding mechanism for acidic isoferritins on HL60 cells. These observations agree with published evidence that ferritin is often associated with cell membranes and are consistent with a possible role for the protein in the regulation of haematopoiesis or in iron transfer.
Effect of phytohaemagglutinin on the synthesis and secretion of ferritin in peripheral blood lymphocytes
Authors: Pattanapanyasat K. , Hoy T. G. , Jacobs A.
Published: March 2008

Stimulation of lymphocytes with phytohaemagglutinin (PHA) causes an increase in the rate of ferritin and total protein synthesis which is not enhanced by supplementing the medium with iron. There is an increase in intracellular ferritin content which is amplified by the presence of iron but this is also observed in the absence of PHA stimulation. The increased cellular ferritin is greater for heart type ferritin than for spleen type ferritin. Ferritin secreted into the medium from the cells shows a relative preponderance of spleen type ferritin compared to the intracellular protein. The data shows a dissociation between ferritin synthesis, cellular accumulation and secretion for which the mechanisms have still to be elucidated.
Upper and lower time limits in the decision to recommend marrow transplantation for patients with chronic myelogenous leukaemia
Authors: Simon William , Segel George B. , Lichtman Marshall A.
Published: March 2008

Long‐term survival of patients with chronic myelogenous leukaemia (CML) requires marrow transplantation from a histocompatible donor. The optimal timing of the transplant is difficult to determine because of the high peritransplant mortality of 20‐35% and the existence of a group of patients who can have the disease controlled by drug treatment for prolonged periods. We have developed a mathematical model implemented with a computer program which calculates lower and upper time limits for the timing of marrow transplantation. The lower time limit for transplantation is derived from the loss of life expectancy with delay, and the upper time limit is calculated by comparing the transplant survival probability with the probability of surviving an additional year without a transplant. Thus, an objective basis is provided for bracketing the most appropriate time for transplantation. This analysis suggests that the decision to transplant can be postponed in some patients for periods longer than may generally be recommended.
Cytokine‐induced enhancement of the susceptibility of hairy cell leukaemia lymphocytes to natural killer cell lysis
Authors: Cordingley F. T. , Hoffbrand A. V. , Brenner M. K.
Published: March 2008

In seven patients with HCL we have investigated how the cytokines TNF, α and γ IFN and IL2 modify the interactions between HCL lymphocytes and MHC unrestricted cytotoxic effector cells [natural killer (NK) and lymphokine activated killer (LAK), cells]. We find that IL2 and αIFN increase patient NK activity against K562, though killing remains subnormal. IL2 and αIFN also induce normal levels of LAK activity, measured against an NK resistant B lymphoblastoid cell line. In contrast, γIFN has no effect on patient effector function. However, promotion of cytotoxic effector activity is unlikely to produce therapeutic benefit since HCL cells themselves are entirely resistant to NK/LAK killing. Susceptibility of HCL cells can be induced by culturing the cells in the presence of both γIFN and TNF, but not by culture with either cytokine alone. This synergy is not mediated by a γIFN induced increase in TNF receptors on HCL lymphocytes, and therefore occurs at a post‐receptor level.
Comparison of the mouse spleen cell assay and a radioimmunoassay for the measurement of serum erythropoietin
Authors: Lappin Terence R. J. , Elder G. Elizabeth , Taylor Timothy , McMullin Mary Frances , Bridges John M.
Published: March 2008

The mouse spleen cell assay (MSCA) has been compared with a radioimmunoassay for the measurement of serum erythropoietin (Ep). In 20 normal subjects the serum values ranged from 15 to 73 mU/ml for the MSCA compared with 5‐30 mU/ml for the RIA. For normal sera there was no correlation between the results of the two assays. In 37 patients with anaemias of differing aetiologies and at various stages of treatment values ranged from 10 to 3645 mU/ml for the MSCA and 13‐10 000 mU/ml for the RIA. Although patient values from the two assays were highly correlated (r=0.98, P < 0.001), the MSCA results were generally lower. These discrepancies can be largely accounted for by two factors. Firstly the MSCA is sensitive to non‐specific matrix effects. Secondly, heat inactivation of serum, a prerequisite for the MSCA, but not for the RIA, destroys a variable and unpredictable proportion of the Ep in the test sera leading to an underestimation of Ep in the MSCA. We conclude that the RIA is more reliable than the MSCA which, in its present form, cannot be recommended for the accurate measurement of serum erythropoietin.
Characterization of two deletions that remove the entire human ζ‐α globin gene complex (—THAI and —FIL)
Authors: Fischel‐Ghodsian N. , Vickers M. A. , Seip M. , Winichagoon P. , Higgs D. R.
Published: March 2008

We have fully characterized two α thalassaemia mutants that occur in Southeast Asia, —THAI and —FIL. Each mutant is due to a deletion that removes the entire ζ‐α‐globin gene complex. Localization of the 5’breakpoints described here, allows the identification of unique fragments that are specific for each of the two mutations. This information can be used to assess the frequency of these mutants in Southeast Asia and will be of value in prenatal testing for α thalassaemia in this area.
Occurrence of hereditary spherocytosis and β thalassaemia in the same family: globin chain synthesis and visco diffractometric studies
Authors: Pautard B. , Feo C. , Dhermy D. , Wajcman H. , Baudin‐Chich V. , Delobel J.
Published: March 2008

Hereditary spherocytosis and β thalassaemia are rarely inherited together. We have studied a large family of Caucasian extraction in whom these two diseases segregate independently over four generations. The diagnosis rested on specialized laboratory findings and in a number of subjects on the measurement of α and β globin chain synthesis. In addition, a viscometric method (osmotic gradient ektacytometry) was used to evaluate the rheological function of the erythrocytes. In patients inheriting both diseases, the results indicate that the clinical and biological expression of hereditary spherocytosis is modulated by the degree of imbalanced globin chain synthesis. The opposite properties of spherocytes (decreased surface/volume ratio, increased haemoglobin concentration) and thalassaemic red cells (increased surface/volume ratio, decreased haemoglobin concentration) may explain the antagonistic influence of each genotype.
Serum immunoreactive erythropoietin in healthy normal children
Authors: Hellebostad M. , Hågå P. , Cotes P. Mary
Published: March 2008

To provide reference data for normal children, serum immunoreactive erythropoietin (siEp) was estimated by radioimmunoassay in samples from 130 healthy children, 57 girls and 73 boys, with ages between 1 month and 16 years. In 128 of the children the (geometric) mean siEp was 15.8 miu/ml (iu: international units) with 95% range (the range within which 95% of the observations are predicted to fall) 9.1–27.6 miu/ml. There was no relation between siEp and the variables haemoglobin (Hb), PCV, age and sex. There were two outliers, both girls, aged 9.5 and 9.8 years. in whom siEp was >256 miu/ml. In both, Hb and PCV were normal and we are unable to account for these atypical findings.

Estimates of siEp in the 128 children were not significantly different from those in 22 healthy adults investigated simultaneously (mean 16.2 miu/ml, with 95% range 11.2–23.3 miu/ml).
Haemopoiesis in long‐term cultures of liver, spleen and bone marrow of pre‐ and postnatal mice: CFU‐GM production
Authors: Van Den Heuvel Rosette L. , Schoeters Greet E. R. , Vanderborght Oscar L. J.
Published: March 2008

The CFU‐GM yield in confluent long‐term cultures (LTC) derived from liver, spleen and bone marrow cells at different gestational and postnatal ages has been studied after the stromal adherent layer reached confluency. The stromal cell compartment of fetal and neonatal haemopoietic organs is able to sustain haemopoiesis in vitro. Moreover, the granulocyte‐macrophage stem cell (CFU‐GM) yield of these LTC reflects the CFU‐GM content of the haemopoietic organs from which the cultures are originated. LTC from the liver produce high numbers (between 100 and 150 CFU‐GM per well) of CFU‐GM if the cultures are derived from fetal livers between 13 d of gestation and birth. Cultures from spleens just before and after birth, give maximal CFU‐GM numbers (between 50 and 100 CFU‐GM per well). The CFU‐GM yield in long‐term bone marrow cultures increases 10 times from 17‐day‐old fetus towards adult life (between 700 and 1000 CFU‐GM per well).
Relapse after allogeneic bone marrow transplantation for acute leukaemia: a survey by the E.B.M.T. of 117 cases
Authors: Frassoni F. , Barrett A. J. , Grañena A. , Ernst P. , Garthon G. , Kolb H. J. , Prentice H. G. , Vernant J. P. , Zwaan F. E. , Gratwohl A.
Published: March 2008

A multi‐centre retrospective analysis on 117 patients relapsing after bone marrow transplantation (BMT) for acute leukaemia was carried out by the Leukaemia Working Party of the European Group for Bone Marrow Transplantation (E.B.M.T.). Forty‐one patients had acute myeloid leukaemia (AML) and 76 had acute lymphoblastic leukaemia (ALL). Relapse ocurred between 3 and 30 months after BMT and where investigated the leukaemia was found to have relapsed in recipient cells. In 10 cases the relapse was associated with new cytogenetic abnormalities. 74 patients received further treatment for leukaemia. Of these 21 out of 50 with ALL and 11 out of 24 with AML achieved a complete remission and had a median survival of 12 months compared with a median survival of 4 months for untreated patients or patients not achieving complete remission (P < 0.001). Factors predictive for successful remission induction were a long interval between bone marrow transplant and relapse in ALL patients; and isolated extramedullary relapse. Presenting blast count, karyotype and remission status and number at the time of BMT were not predictive. Donor bone marrow was shown to be responsible for haemopoietic recovery occurring in the 21 out of 31 patients tested who achieved remission using donor karyotype or red blood cell antigens as markers. Nine patients received a second bone marrow transplant but only one became a long‐term survivor. The results show that chemotherapy can usually prolong survival in selected patients with acute leukaemia relapsing after BMT but further BMT has a poor outlook.
The adaptation of Plasmodium falciparum to oxidative stress in G6PD deficient human erythrocytes
Authors: Roth Eugene , Schulman Seymour
Published: March 2008

It has recently been found that growth of P. falciparum in human G6PD deficient red cells is impaired in vitro; however, the inhibition is overcome after two or three growth cycles. There is evidence to suggest that the parasite can produce its own G6PD enzyme which may compensate for the lack of host enzyme and could account for the resumption of normal growth in G6PD deficient host cells. It is unclear whether the parasite enzyme can enable the host cell to resist oxidative stress as normal cells do. To answer this question, P. falciparum was grown in vitro in: (a) normal red cells, (b) G6PD deficient red cells for one growth cycle only, (c) G6PD deficient cells for a minimum of five cycles. All groups were then challenged with acetylphenylhydrazine (APH) which served as an oxidative stress. Both G6PD (A ‐) and Mediterranean deficient types were studied. The results show a two‐fold increase in resistance to oxidative stress by parasites adapted to G6PD‐Mediterranean deficient host cells as compared to unadapted ones, but the parasite‐red cell system remains 4 times more sensitive to APH than normal infected cells. In parasitized G6PD (A ‐) red cells, evidence of adaptation could be seen in the growth curves, but no detectable increase in resistance to APH was found in adapted parasites. It is concluded that the role of the parasite G6PD is not likely to be mainly related to oxidative stress resistance and therefore other functions of this enzyme should be investigated.
Irreversible erythrocyte volume expansion induced by tellurite
Authors: Kurantsin‐Mills Joseph , Klug Robert K. , Lessin Lawrence S.
Published: March 2008

Tellurite (K2TeO3) has been suggested as a potential anti‐sickling compound because it causes a selective increase in the water content of RBC. To investigate the conditions underlying the increase in RBC volume due to tellurite, normal RBCs were incubated with the compound in a physiological medium and the cells washed with a 10‐fold volume of the medium. The washed cells were then incubated at 24°C for periods up to 4 h and the following parameters were determined: MCV, MCH, MCHC and supernatant haemoglobin concentration by standard methods, the density distribution profile by phthalate esters and cell morphology by scanning electron microscopy (SEM). The effect of hypertonic PBS on the tellurite‐treated cells was also tested. K2TeO3 induced concentration and time dependent increases in MCV and decreases in MCHC without any apparent change in MCH. The median density and the transitional 60% density range of the cell distribution profile respectively decreased and increased in proportion to [K2TeO3] and time. Hypertonic PBS did not inhibit or reverse the tellurite‐induced changes in MCV and MCHC, SEM and photovolumetric measurements demonstrated tellurite‐induced large vesicles ranging in size from 24 to 32 μ3. The proportion of these vesicles increased with time and K2TeO3 concentration. Since tellurite is an oxidant, these findings suggest that its influx into the red cell results in irreversible reactions that disrupt the ion and water regulatory properties of the membrane.
Maternal antibodies against fetal blood group antigens A or B: lytic activity of IgG subclasses in monocyte‐driven cytotoxicity and correlation with ABO haemolytic disease of the newborn
Authors: Brouwers H. A. A. , Overbeeke M. A. M. , Ouwehand W. H. , Keuning K. , Ertbruggen I. van , Van Leeuwen E. F. , Stoop J. W. , Engelfriet C. P.
Published: March 2008

IgG antibodies against blood group antigens A or B (anti‐A/B) are able to sensitize erythrocytes for destruction in an antibody‐dependent cell‐mediated (ADCC) assay with monocytes as effector cells. The activity of maternal IgG anti‐A/B in this test was compared with clinical signs of haemolytic disease of the newborn (HDN). When the ADCC was negative (less than 10% of the sensitized cells lysed), signs of increased red‐cell destruction in the children were never observed. In three cases with a strongly positive ADCC (>45% lysis), the children were severely affected and needed more than one exchange transfusion. In the cases with >10% but less than 45% lysis in the ADCC, there was no clear correlation between the result of the ADCC and the degree of lysis in the newborn infants. In these cases, the degree of lysis of the red cells of the infant was shown to be strongly influenced by the number of A/B antigens per red cell. There was a direct correlation between the degree of lysis in the ADCC and the titre of IgG3 anti‐A/B in the sera. There was comparable activity of maternal IgG anti‐A/B in the ADCC test in the 32nd week of pregnancy and at the moment of delivery.
Effect of iron on polymorphonuclear granulocyte phagocytic capacity: role of oxidation state and effect of ascorbic acid
Authors: Hoepelman Ilja M. , Jaarsma Emmy Y. , Verhoef Jan , Marx Joannes J. M.
Published: March 2008

It has been shown that iron (III) impairs the function of polymorphonuclear granulocytes (PMN). We have studied the effect of iron (II), on the membrane function of PMN, by assessing the uptake of radiolabelled Staphylococcus aureus by these cells. Iron (II), significantly impaired PMN phagocytic function. Addition of ascorbic acid reduced uptake further. Ferrous ascorbate, molar ratio 1:20, impaired phagocytic capacity of PMN significantly at iron concentrations as low as 1–10 μM. The toxic effect of iron (II) was not observed when desferrioxamine or transferrin was present in the incubation medium. The oxygen‐free radical scavengers thiourea, mannitol and catalase prevented toxicity mediated by ferrous ammoniumsulphate but not by ferrous ascorbate (molar ratio of 1:20). Although high concentrations of ascorbic acid inhibited the generation of .OH and also the formation of the DMPO‐.OH adduct by zymosan stimulated PMN, toxicity of iron increased. Iron (II) impaired the uptake of S. aureus by PMN of a patient with chronic granulomatous disease while iron (III) did not. Iron mediated impairment of PMN function is not only a result of the generation of toxic oxygen metabolites but also of direct interaction of iron (II) or an iron (II)–oxygen intermediate with molecules of the cell membrane.
Prognostic features of chronic myelomonocytic leukaemia: a modified Bournemouth score gives the best prediction of survival
Authors: Worsley A. , Oscier D. G. , Stevens J. , Darlow S. , Figes Ann , Mufti G. J. , Hamblin T. J.
Published: March 2008

Summary. We have studied the clinical and laboratory features of 53 cases of chronic myelomonocytic leukaemia (CMML) defined according to the FAB criteria. A granulocyte count of < 16 x 109/l or a monocyte count of < 2.6 x 109/l correlated with a poorer survival, but the best predictor of poor survival was the modified ‘Bournemouth’ score in which one point each was allocated for Hb < 10 g/dl, neutrophils < 2.5 x 109/l or > 16 x 109/l, platelets < 100 x 109/l and bone marrow blasts > 5%. A score of 2 or more was predictive of poor survival (Chi2= 10.25; P < 0.001). Those patients with low monocyte and neutrophil counts and a low modified Bournemouth score tended to have a clinical course similar to refractory anaemia (RA) with an indolent course not requiring treatment and long survival whereas those with high monocyte and neutrophil counts and a high modified ‘Bournemouth’ score have a clinical course resembling that of refractory anaemia with excess of blasts (RAEB).
Lack of evidence for the presence of neutrophil autoantibodies in the serum of patients with Felty's syndrome
Authors: Goldschmeding R. , Breedveld F. C. , Engelfriet C. P. , Von Dem Borne A. E. G. Kr.
Published: March 2008

Summary. Sera of 22 patients with Felty's syndrome and 14 patients with rheumatoid arthritis were tested in assays routinely used for the detection of neutrophil antibodies (i.e. immunofluorescence, agglutination and cytotoxicity tests) as well as in the antibody‐dependent lymphocyte‐mediated granulocytotoxicity test, One or more of the routinely used assays were positive in a high percentage of the sera (77% and 64%, respectively). The antibody‐dependent lymphocyte‐mediated granulocytotoxicity test was only positive with sera of three patients with Felty's syndrome. Positive results in the immunofluorescence and agglutination tests could be attributed to the presence of immune complexes in the sera, whereas positive antibody‐dependent lymphocyte‐mediated granulocytotoxicity tests were probably due to the presence of HLA alloantibodies. It is concluded that serological tests routinely used for the detection of neutrophil autoantibodies should be interpreted with caution in patients with Felty's syndrome. Our results also indicate that the neutropenia in Felty's syndrome is rarely, if ever, due to neutrophil‐specific autoantibodies.
Complement inhibitor S protein is associated with membranes of red blood cells from patients with paroxysmal nocturnal haemoglobinuria
Authors: Salama A. , Preissner K. T. , Göttsche B. , Müller‐Berghaus G. , Mueller‐Eckhardt C.
Published: March 2008

Summary. S protein is a plasma glycoprotein (Mr= 78 000) which binds to nascent C5b‐7 complexes upon complement activation in the fluid phase in whole serum. It thereby protects innocent bystander cells from complement mediated lysis. It is unknown whether S protein also functions as complement inhibitor on cell surfaces. We here report that S protein is recognized on red blood cells (RBC) from patients with paroxysmal nocturnal haemoglobinuria (PNH), but not on normal RBC. RBC from eight PNH patients showed 12–48% haemolysis subsequent to complement activation in the fluid phase, while normal RBC did not respond. Preincubation of the PNH cells with affinity‐purified antibodies against human S protein resulted in a three‐ to five‐fold increase of haemolysis, while preincubation of these cells with S protein decreased haemolysis by 40%. In contrast, haemolysis remained unaffected by other unrelated antibodies, i.e. IgG anti‐Rh(D) and anti‐A. If PNH RBC, normal RBC pretreated with 2‐amino‐ethylisouronium bromide (AET), or untreated normal RBC, respectively, were incubated with purified S protein in vitro, the uptake of antibodies against S protein was significantly enhanced with PNH and with AET‐treated, but not with untreated normal RBC. Additionally, while normal RBC did not respond to reactive lysis initiated by purified C5b‐6 and C7, PNH as well as AET‐RBC showed significant haemolysis that could be inhibited by S protein in a dose‐dependent fashion. These findings strengthen the assumption that the increased sensitivity of PNH cells towards reactive complement lysis is either due to the lack of an inhibitor of the terminal complement sequence and/or enhanced insertion of the membrane attack complex. These defects of PNH RBC may partly be overcome by the fluid phase complement inhibitor S protein which binds to PNH RBC and may thereby suppress homologous cytolysis.
Effect of commercial immunoglobulin G preparation on human monocyte Fc‐receptor dependent binding of antibody coated platelets
Authors: Saleh Mansoor , Court Wayne , Huster William , Shaw Denise , LoBuglio Albert
Published: March 2008

Summary. This study examined the in vitro effect of a commercial immunoglobulin preparation on human monocytes and the Fc‐receptor dependent binding of antibody coated platelets. Monocytes were exposed to Sandoglobulin in vitro and subsequently examined for membrane surface bound IgG. Dramatic increments of surface IgG were found which were maximal with 18 h exposure and somewhat higher at 4°C than 37°C. Ultracentrifugations of Sandoglobulin immediately prior to monocyte exposure reduced the monocyte membrane IgG by 75%. The 18 h exposure at 37°C produced dramatic impairment of monocyte Fc‐receptor binding of IgG coated platelets (P < 0.001) while exposure for 18 h at 4°C produced a modest impairment of Fc‐receptor function. These studies indicate that Sandoglobulin contains IgG aggregates which are able to firmly bind to the monocyte surface in a time and temperature dependent fashion. The dramatic impairment of Fc‐receptor function at 37°C and not at 4°C suggests that Fc‐receptor modulation, as well as competitive inhibition/steric hindrance, contribute to impairment of monocyte Fc‐receptor function.
Heterogeneous defects of platelet secretion and responses to weak agonists in patients with bleeding disorders
Authors: Lages Bruce , Weiss Harvey J.
Published: March 2008

Summary. Eleven patients with mild bleeding disorders had as a common abnormality, impaired platelet aggregation and secretion with low concentrations (0.5–1.0 μg/ml) of collagen and, in most cases, an absence of second phase aggregation with epinephrine. Platelet granule contents were normal, ruling out storage pool deficiency. To characterize further the platelet abnormalities, we measured aggregation, 14C‐5HT secretion, and TxB2 formation induced by a variety of platelet agonists. In eight of the 11 patients we observed decreased initial rates as well as extents of aggregation with one or more weak agonists (ADP, epinephrine, thromboxane A2 and the endoperoxide analogue U44069), i.e. agonists which induced secretion only as a result of aggregation, but normal responses to strong agonists such as arachidonate and high (10 μg/ml) concentrations of collagen, which can induce secretion in the presence or absence of aggregation. In all of these patients. TxB2 formation with arachidonate and all concentrations of collagen was normal. The platelet defects in these eight patients have been designated as weak agonist response defects (WARDs). In contrast, the initial aggregation responses to all weak agonists were normal in the three other patients, while secretion and TxB2 formation induced by strong agonists were impaired. Thus, in contrast to the eight patients above, the platelet defects in these three patients were characteristic of defects in the secretion response per se. The results obtained in the 11 patients studied indicate that these types of platelet disorders, previously referred to as primary secretion defects, include defects in the initial platelet responses which precede secretion (WARD) as well as defects in the secretory mechanism per se. Both groups of defects appear to be heterogeneous in nature.
High‐dose intravenous immunoglobulin and splenectomy for the treatment of HIV‐related immune thrombocytopenia in patients with severe haemophilia
Authors: Beard J. , Savidge G. F.
Published: March 2008

Summary Four patients with severe haemophilia A and one patient with severe Christmas disease developed severe immune thrombocytopenia (platelet count < 20 × 109/1). All five patients were HIV‐antibody positive and one was HIV‐antigen positive. Four patients were treated initially with prednisolone, but with only a transient platelet response in three and no response in the fourth. All patients were treated with high dose intravenous immunoglobulin (0.4 g/kg daily for 5 d) resulting in a rise in platelet count in all cases (range 138–300 x 109/1) and then proceeded to splenectomy. Three remain in complete remission after 6–14 months, and one showed a good response with platelet counts ranging from 103 to 187 x 109/1. The fifth patient achieved a normal platelet count for 3 months post‐splenectomy, but suffered a relapse with platelet counts ranging from 25 to 108 x 109/1 over the next 3 years. However, following a severe Varicella infection 10 months ago, during which he developed a marked transient thrombocytosis, he has also maintained a normal platelet count.