bluuebunny/crossref_metadata_2025_split huggingface dataset:
Decay‐accelerating factor (DAF) on the blood cell membranes in patients with paroxysmal nocturnal haemoglobinuria (PNH): measurement by enzyme‐linked immunosorbent assay (ELISA)
Authors: Ninomiya Haruhiko , Abe Tsukasa , Shichishima Tsutomu , Terasawa Takashi , Fujita Teizo 
Published: March 2008

Summary. We developed a quantitative enzyme‐linked immunosorbent assay (ELISA) for decay‐accelerating factor (DAF) on blood cell membranes using monoclonal anti‐DAF antibodies. DAF is an integral membrane protein of several blood cells. It regulates the C3 and C5 convertase of the classical and alternative pathways of complement activation. It is partially or completely deficient in the membranes of blood cells of patients with paroxysmal nocturnal haemoglobinuria (PNH).The ELISA we developed for DAF measurement indicated a reliable range of measurement from 2.25 to 11.25 ng of DAF. In particular, ELISA proved to be a technically simple method and its sensitivity was enhanced by using avidin‐biotin complex. In this study, DAF levels in extracts of erythrocytes from 30 healthy donors and in extracts of PMN and platelets from 15 healthy donors were measured by ELISA. The DAF content of blood cells from eight patients with PNH and erythrocytes from 13 patients with anaemia were also measured. The DAF levels of normal erythrocytes, PMN and platelets were 3110.960, 28000.13900 and 3100.1370 (mean±SD) molecules/cell, respectively. In general, the DAF content of PNH cells was below the normal range, although it was within the normal range in some cases of PNH. The DAF levels of PNH‐I and ‐II erythrocytes were estimated from the ratio of PNH‐I, ‐II and ‐III erythrocytes. And, in four cases of PNH, the DAF levels of PNH‐I erythrocytes separated by acidified serum lysis were measured by ELISA. In most cases of PNH, DAF was found to be partially deficient in PNH‐I and PNH‐II erythrocytes.
5‐Aminolaevulinic acid synthase activity in developing human erythroblasts
Authors: Fitzsimons E. J. , May A. , Elder G. H. , Jacobs A. 
Published: March 2008

Summary. 5‐Aminolaevulinic acid (ALA) synthase activity was measured in highly purified preparations of age‐matched human erythroblasts. Enzyme activity in immature normoblasts was four‐fold higher than that found in late orthochromatic normoblasts. ALA synthase activity in the immature erythroblasts in primary acquired sideroblastic anaemia (PASA) was reduced and remained unchanged during further erythroid differentiation. The pattern of erythroblast ALA synthase activity in two patients with congenital dyserythropoietic anaemia (CDA) and in one patient with β‐thalassaemia intermedia was similar to that found in PASA.This study has clearly demonstrated reduced erythroblast ALA synthase activity in PASA but has also found reduced enzyme activity in conditions in which ring sideroblasts are not prominent. This would suggest that haem synthesis is abnormal in PASA but that reduced erythroblast ALA synthase activity does not inevitably lead to ring sideroblast formation.
Abnormal megakaryopoiesis in patients with myelodysplastic syndromes: analysis of cellular and humoral defects
Authors: Geissler D. , Zwierzina H. , Pechlaner C. , Gaggl S. , Schmalzl F. , Konwalinka G. , Braunsteiner H. 
Published: March 2008

Summary. In 13 patients with myelodysplastic syndrome (MDS) mature and immature erythropoietic (CFU‐E, BFU‐E), granulopoietic (CFU‐GM) and megakaryopoietic (CFU‐Meg) colony formation from human bone marrow mononuclear cells was evaluated in a microagar culture system. All but three patients exhibited abnormal CFU‐Meg. The defect of CFU‐Meg paralleled the reduction of BFU‐E, whereas CFU‐GM number declined to a lesser extent. Not only the CFU‐Meg number, but also the number of megakaryocytes (Mk) per colony was reduced suggesting an additional functional CFU‐Meg defect.Megakaryocytic growth factor (Meg‐CSF) abnormalities in MDS patients were detected using normal nonadherent T‐lymphocyte depleted bone marrow cells as target cells for serum testing. Even for sera from patients with a reduction of platelets and bone marrow megakaryocytes Meg‐CSF levels were not increased.No cellular or humoral inhibition could be detected in an MDS patient with a 5q – karyotype, who had an isolated defect of the megakaryocytic cell lineage at presentation.Some patients revealed a spontaneous formation of mixed erythrocytic, granulocytic and megakaryocytic clusters in the presence of fetal calf serum or autologous patient serum, probably representing autonomous proliferation of the malignant clone. In conclusion, both cellular and humoral factors can cause abnormalities of the megakaryocytic cell lineage in MDS patients.
Plasma from patients with severe Lassa fever profoundly modulates f‐met‐leu‐phe induced superoxide generation in neutrophils
Authors: Roberts Pamela J. , Cummins David , Bainton Adrian L. , Walshe Karen J. , Fisher‐Hoch Susan P. , McCormick Joseph B. , Gribben John G. , Machin Samuel J. , Linch David C. 
Published: March 2008

SummaryA recurrent theme in studies of the pathology of fatal Lassa fever in man is the lack of histological lesions to explain disordered cell function and death. Recently, we demonstrated the existence of a factor in the plasma of patients with Lassa fever which markedly inhibits the aggregation responses of normal platelets in vitro. To assess whether this factor could mediate more global cellular dysfunction, we studied the effects of Lassa plasma on the respiratory burst of neutrophils.Thirteen of 15 samples from patients in the acute phase of Lassa fever profoundly inhibited the amount of superoxide generated by normal neutrophils in response to the chemotactic peptide, f‐met‐leu‐phe (FMLP) (mean superoxide generated = 54.7 ± 6.1% of control). In contrast, eight of nine samples from patients who had infections other than Lassa fever enhanced the neutrophil response to the peptide. All Lassa samples which inhibited the ADP‐induced aggregation responses of normal platelets inhibited the neutrophil response to FMLP. Unlike the effect on platelets, however, the inhibition of neutrophils was only apparent when the cells were stimulated within 5 min of exposure to the plasma. The inhibition of neutrophils is not due to either interference with FMLP‐neutrophil binding or an effect on the NADPH‐oxidase, suggesting a suppression of signal transduction. Our data suggest the inhibitory factor in Lassa plasma has global effects on cellular function, and may play a central role in the pathogenesis of this often fatal illness.
Immunochemical analysis of platelet autoantibodies in HIV‐related thrombocytopenic purpura: a study of 68 patients
Authors: Bettaieb Ali , Oksenhendler Eric , Fromont Patricia , Duedari Najib , Bierling Philippe 
Published: March 2008

SummarySerum antiplatelet IgG and platelet‐associated IgG (PAIgG) were studied in 68 AIDS‐free human immunodeficiency virus (HIV)‐infected patients with severe immunologic thrombocytopenic purpura (ITP), for the presence of platelet autoantibodies. Serum IgG with antiplatelet activity was found in 72% of the sera. However, the presence of autoantibodies against platelet surface glycoproteins was not found in these sera by means of Western blot and immunoprecipitation procedures. Nevertheless, an immunoblot immuno‐assay and an indirect immunofluorescence test against semi‐permeabilized platelets demonstrated the presence of antibodies in the patient sera, that reacted with intracyto‐plasmic platelet components, and which might participate in the elimination of platelet fragments. Direct immunofluorescence tests demonstrated an increased amount of PAIgG in 75% of the patients; the bound antibodies could be eluted with ether in 44% of the cases. These eluates were found to bind to normal platelets but not to Glanzmann type I platelets. Finally, immunoprecipitation procedures demonstrated the presence of platelet autoantibodies in six of the 35 eluates studied. These antibodies recognized GPIIb in two cases. GPIIIa in one case, and an unidentified platelet protein of 150 kDa in the three other cases. The discrepancy between sera and platelet eluates was interpreted as being due to the low titre of the antibodies and to their dilution in polyclonal hypergammaglobulinaemia.The present study provides direct evidence that isolated ITP in some HIV‐positive patients is due to the presence of platelet autoantibodies. These results, however, do not exclude either direct or indirect involvement of HIV in the platelet destruction.
Anticardiolipin antibodies and lupus anticoagulants comprise separate antibody subgroups with different phospholipid binding characteristics
Authors: McNeil H. Patrick , Chesterman Colin N. , Krilis Steven A. 
Published: March 2008

SummaryAutoantibodies to phospholipid antigens can be characterized using solid phase immunoassays to detect anticardiolipin antibodies (ACA) or in phospholipid‐dependent clotting tests where lupus anticoagulant (LA) activity can be demonstrated. It has not been established whether each activity is due to the same or separate antibody subgroups. Plasma from two patients with high levels of both activities were used for purification of ACA and LA using sequential ion‐exchange, gel‐filtration, and anti‐Ig affinity chromatography. Plasma could be separated into fractions containing each activity in the absence of the other. In these fractions, antibodies responsible for LA activity do not bind to isolated phospholipids in solid phase immunoassays, and conversely antibodies binding in these assays (ACA) do not possess LA activity, suggesting LA are directed against a more complex lipid epitope. In addition, in one patient ACA was of IgG isotype only, whilst LA was due to IgG and IgM isotypes. In this patient, the IgG‐ACA was heterogenous with three peaks clearly separated on ion‐exchange chromatography. Affinity purified antiphospholipid antibodies have been previously prepared from a number of patients using a phospha‐tidyl‐serine column and antibodies purified in this manner possess ACA but not LA activity. Taken together, these data indicate that tests for ACA and LA define separable subgroups of phospholipid binding antibodies, thus explaining the discordance often seen between the two activities.
The rapid and decremental change in haem oxygenase mRNA during erythroid differentiation of murine erythroleukaemia cells
Authors: Fujita Hiroyoshi , Sassa Shigeru 
Published: March 2008

SummaryChanges in mRNA for haem oxygenase (HO), the rate‐limiting enzyme for haem catabolism, were examined in murine Friend‐virus transformed erythroleukaemia (MEL) cells while they were induced to undergo erythroid cell differentiation by treatment with dimethyl sulfoxide (DMSO). When MEL cells were treated with 1.5% (v/v) DMSO, a rapid decrease in HO mRNA content was observed (< 12 h) which reached the lowest value at 18 h (18% of the untreated control). HO mRNA levels remained at substantially lower levels (< 50%) than those in untreated controls thereafter. A rapid decline in HO mRNA may be involved in the cellular events that determine the onset of erythroid differentiation.
The beneficial effects of αIFN in CGL are probably not mediated by NK cells
Authors: Galvani D. W. , Owens W. , Nethersell A. B. W. , Cawley J. C. 
Published: March 2008

Summary Natural killer (NK) cells in CGL were measured phenotypically (by Leu7 and CD16 Mab staining) and functionally (by a standard chromium‐release cytotoxicity assay) in eight patients before and during α‐IFN therapy.Before αIFN therapy, phenotypic NK cells were normal in relative and absolute numbers but were consistently defective functionally; this defect was partially corrected by in vitro exposure to αIFN.During αIFN therapy, there was no change in NK function in five patients and enhanced (two patients) or reduced (one patient) activity was observed in the other three. Cold‐target inhibition experiments showed no evidence of NK binding to normal or CGL myeloid progenitors.It is concluded that αIFN‐enhanced NK function, a known anti‐tumour mechanism in animal models, is probably not the basis of the responsiveness of CGL to αIFN therapy.
Erythropoiesis following bone marrow transplantation from donors heterozygous for β‐thalassaemia
Authors: Galanello Renzo , Barella Susanna , Maccioni Liliana , Paglietti Elisabetta , Melis Maria A. , Rosatelli Maria C. , Argiolu Francesca , Cao Antonio 
Published: March 2008

Summary This study shows a marked and protracted activation of HbF synthesis in homozygous β°‐thalassaemia patients transplanted from HLA identical siblings heterozygous for β‐thalassaemia, as compared to patients transplanted from normal donors. HbF synthesis in recipients was much higher in relation to the corresponding bone marrow donor values either normal or heterozygous for β thalassaemia.γ‐chain synthesis and Gγ/Aγ ratio were also studied in peripheral blood BFU‐E from recipients and their donors. BFU‐E from donors heterozygous for β‐thalassaemia showed higher γ chain synthesis as compared to normal donors. Peripheral blood BFU‐E γ/β+γ ratios and Gγ percentage were higher in recipients than in their corresponding donors both normal or heterozygotes.The marked and protracted reactivation of HbF synthesis in recipients of heterozygous β‐thalassaemia bone marrow most likely results from an increased erythropoietic stress on erythroid progenitors. In order to obtain adequate Hb levels heterozygous β‐thalassaemia bone marrow should produce more red blood cells to compensate for the low MCH. The magnitude of activation of HbF synthesis was very variable. This variability may result from inherited differences in the capacity of reactivation of HbF synthesis of red cell progenitors from heterozygous β‐thalassaemia under stressed erythropoiesis.
Circulating activated suppressor T lymphocytes in hepatitis‐associated aplastic anaemia
Authors: Kojima Seiji , Matsuyama Kohji , Kodera Yoshihisa , Okada Junichi 
Published: March 2008

SummaryLymphocyte subpopulations were measured in the blood of 21 children with severe aplastic anaemia (SAA). Five children with hepatitis‐associated AA appeared to constitute a unique group. Compared with children with idiopathic AA, four of five children had a striking increase in percentage of activated suppressor T lymphocytes, which were defined by binding of both anti‐Leu 2a and anti‐HLA DR monoclonal antibodies (children with hepatitis‐associated AA, 21·8±19·9% [mean ±SD], children with idiopathic AA, 2·9±1·2%). However, there was no difference in the mean absolute number of the activated suppressor T lymphocytes between the two groups, because the mean number of lymphocytes was markedly less in children with hepatitis‐associated AA (0·28±0·06 × 109/1) than in those with idiopathic AA (1·64±0·74 × 109/1). The mean helper: suppressor T lymphocyte ratio was significantly lower in children with hepatitis‐associated AA (0·58±0·74) than in those with idiopathic AA (1·22±0·44). On the other hand, we did not find such a remarkable alteration of lymphocyte subpopulations in children with non‐A non‐B hepatitis who did not develop AA. These findings suggested that an imbalance of lymphocyte subpopulations and T lymphocyte activation may have pathogenetic relevance in some of the children with hepatitis‐associated AA.
N‐RAS mutations in T‐cell acute lymphocytic leukaemia: analysis by direct sequencing detects a novel mutation
Authors: Bar‐eli M. , Ahuja H. , And A. Foti , Cline M. J. 
Published: March 2008

Summary. A novel mutation of the N‐RAS gene of T‐ALL blast cells was detected by a direct sequencing of in vitro amplified exon‐1 of the N‐RAS gene. Threonine (ACA) was substituted for alanine (GCA) at codon 11. This mutation would have been overlooked by conventional probe hybridization techniques.A search for other mutations in N‐RAS exon‐1 in T‐ALL revealed a codon 13 mutation substituting aspartic acid (GAT) for glycine (GGT) in one of 18 patients. No mutations at codon 12 were detected.
Morphologic and morphometric light and electron microscopic studies of the spleen in patients with hereditary spherocytosis and autoimmune haemolytic anaemia
Authors: Ferreira J. A. , Feliu E. , Rozman C. , Berga Ll. , Bombi J. A. , Marti M. , Vives‐Corrons J. Ll. , Climent C. , Ferrán M. J. 
Published: March 2008

Summary.With the aim of contributing to a better understanding of the haemolytic function of the spleen, a morphologic and morphometric study of this organ fixed by arterial perfusion was performed in nine patients with hereditary spherocytosis (HS), three with autoimmune haemolytic anaemia (AHA) and six with Hodgkin's disease without splenic involvement (controls).The spleen weight in HS and AHA (621 ± 429 g. mean ± SD) was significantly increased with respect to controls (168 ± 3 6 g) (P= 0·003). In HS the red cell retention in the cords of Billroth was significantly increased (203 ± 68 per 104μm2) with respect to the cases with AHA(93 ± 35 per 104μm2) and to the controls (57 ± 28 per 104μm2) (P= 0·004). In HS and AHA the number of macrophages per 104μm2of red pulp was significantly increased (5·41 ± 1·10 and 7·52 ± 2·91, respectively) with respect to the controls (3·25 ± 0·58) (P> 0·003). There was no statistically significant difference between the number of macrophages in HS and AHA.The transmission (TEM) and scanning electron microscopy (SEM) studies demonstrated predominantly red cell retention in the cords of HS spleens, red cell phagocytosis by cordal macrophages in AHA spleens and in a lesser intensity in HS spleens, and phagocytosis of haematic corpuscles by sinus endothelial cells (SEC) in the cases of HS. These quantitative studies allow a better understanding of splenic red cell destruction in haemolytic syndromes.
In situ hybridization detection of light chain mRNA in routine bone marrow trephines from patients with suspected myeloma
Authors: Akhtar N. , Rupral A. , Pringle J. H. , Lauder I. , Durrant S. T. S. 
Published: March 2008

SummaryA method for the detection of immunoglobulin light chain mRNA by in situ hybridization has been applied to routine bone marrow trephines, from patients suspected of having multiple myeloma. The principle aim was to define monoclonal populations even in cases lacking a paraprotein or an obvious atypical plasma cell proliferation.The expression of light chain mRNA in plasma cells in routine bone marrow trephines from 14 patients with suspected myeloma was studied. Plasma cells in the marrow ranged from 4% to 99%. Clonal populations based on light chain restriction were delineated in 12 patients, using biotinylated probes to kappa and lambda mRNA and visualized using an alkaline phosphatase/fast red naphthol capture method. Normal tonsil and bone marrow trephines were used as controls. In situ hybridization was found to be a specific, safe and rapid technique. It could be applied to both fixed and fresh tissue. It was found to lack the background staining of interstitial immunoglobulin, particularly in sclerotic cases, which is so often encountered with immunocytochemistry.Additionally, this technique will type plasma cell neoplasms (undetected by routine immunocytochemistry) in post‐transcriptional block or non‐secretory myeloma.
Ultrastructural and ultracytochemical differences between transient myeloproliferative disorder and megakaryoblastic leukaemia in Down's syndrome
Authors: Eguchi Mitsuoki , Sakakibara Hitoshi , Suda Junko , Ozawa Takebumi , Hayashi Yasuhide , Sato Takeyuki , Kojima Seiji , Furukawa Toshiharu 
Published: March 2008

SummaryUltrastructural and ultracytochemical studies were performed on blast cells from 12 Down's syndrome neonates with transient myeloproliferative disorder (TMD) and 13 Down's syndrome patients with megakaryoblastic leukaemia (MKL), in order to clarify the cytological characteristics of these cells. Average platelet peroxidase‐positivity in blast cells of TMD patients was similar to that found in cases of MKL. Blast cells from subjects with TMD contained a number of different granules, namely, alpha granules, those that were myeloperoxidase (MPO)‐positive, electron‐lucent or basophil‐like, and those containing membrane components or ferritin particles. On the other hand, granules found in the blast cells of MKL patients with Down's syndrome included the electron‐lucent variety, those with membrane components and a few that were basophil‐like, but not alpha and MPO‐positive granules nor those containing ferritin particles. A demarcation membrane system was observed in blasts from the TMD group, but not in the MKL group.These findings suggest that blast cells in TMD patients differentiate to megakaryocytes, neutrophils, basophils and erythroblasts, while those in cases of MKL show limited differentiation to immature megakaryocytes, erythroblasts and, sometime, basophils. Such results correspond well with those of culture studies, in which TMD blasts were found to be precursors of various types of blood cell.
Desferrioxamine ototoxicity: evaluation of risk factors in thalassaemic patients and guidelines for safe dosage
Authors: Porter John B. , Jaswon Mervyn S. , Huehns Ernst R. , East Charles A. , Hazell Jonathan W. P. 
Published: March 2008

SummaryForty‐seven patients with thalassaemia have been studied to define risk factors for development of sensorineural hearing loss, and to establish guidelines for safe chelation. Sensorineural hearing loss was only present in patients who had previously received desferrioxamine (DFO). The two most significant risk factors were the maximum dose of DFO previously received (P < 0.01), and a serum ferritin of < 2000 μg/1 at that time (P < 0.001). A therapeutic index obtained from the ratio of the mean daily dose of DFO mg/kg divided by the serum ferritin identifies patients with a ratio of > 0.025 as at risk of sensorineural hearing loss (P < 0.001) and can be used as a guideline for safe DFO dosage. Follow‐up audiometry of the affected patients over a 2‐year period indicated that adjustment of the dose to a therapeutic index of < 0.025 resulted in the stabilization of hearing loss in seven patients and improvement in two.
The position of the M‐BCR breakpoint does not predict the duration of chronic phase or survival in chronic myeloid leukaemia
Authors: Jaubert Jerome , Martiat Philippe , Dowding Charles , Ifrah Norbert , Goldman John M. 
Published: March 2008

SummaryIt has been reported that patients with chronic myeloid leukaemia (CML) with 5’breakpoints within the major breakpoint cluster region (M‐BCR) of the BCR gene have somewhat better prognoses than those with 3’breakpoints. We studied the position of the breakpoint in 67 patients with CML in chronic phase using conventional Southern blotting. Using restriction enzymes BglII, BamHI and HindIII and two genomic probes, a 0.6 kb (3’M‐BCR) probe hybridizing to a part of the intron between exons b3 and b4 and a 2.0 kb (5’M‐BCR) probe hybridizing to sequences including exon b1, we localized the breakpoint in M‐BCR as occurring 5’(n= 38) or 3’(n= 28) of the HindIII restriction site located just downstream of exon b3. We failed to localize the breakpoint in one patient. The median durations of chronic phase (37 versus 44 months respectively) and of survival (50 versus 51 months respectively) for patients with 5’and 3’breakpoints were not significantly different. When we analysed only patients whose DNA was collected within 4 weeks of diagnosis (5’breakpoints, n= 30; 3’breakpoints, n= 19), there was again no significant difference in duration of chronic phase or survival. The median survivals of patients divided into good, intermediate and poor prognosis categories in accordance with the prognostic index developed by Sokal and colleagues were 54, 50 and 26 months respectively. This study confirms the value of the Sokal prognostic index but provides no support for the notion that the precise genomic position of the breakpoint in M‐BCR correlates with prognosis.