bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood neutrophil increment after a single injection of rhG‐CSF or rhGM‐CSF correlates with marrow cellularity and may predict the grade of neutropenia after chemotherapy
Authors: Hansen Per Boye , Johnsen Hans E. , Ralfkiaer Elisabeth , Hansen Niels Ebbe
Published: March 2008

Summary. The grade of neutropenia after chemotherapy seems to be correlated to the bone marrow cellularity as judged by biopsies. Prolonged blood neutropenia after sequential chemotherapy reduces dose intensity and increases the risk of severe infections. A predictive non‐invasive test for marrow cellularity is needed in the attempt to predict chemotherapy‐induced blood neutropenia.

Thirty‐one patients with haematological disorders were studied with measurements of blood absolute neutrophil counts (ANC) 24 h after a single subcutaneous injection of recombinant human granulocyte colony stimulating factor (rhG‐CSF) or granulocyte‐macrophage CSF (rhGM‐CSF). Before cytokine administration all patients had bone marrow biopsies performed.

The median increase in blood ANC 24 h after cytokine administration was 15·9 × 109/l (range 3·7–34·2) in 18 patients with normo‐ or hypercellular marrows and only 0·4 × 109/l (range 0·0–11·2) in 13 patients with hypocellular marrows (P<0·00001). An increase in ANC or more than 5 × 109/l was predictive for normo‐ or hypercellular bone marrows with a sensitivity and specificity of 94% and 84%, respectively.

A subsequent pilot study in selected patients with prolonged neutropenia was performed. The ANC increment in 12 cases before chemotherapy correlated to the grade of neutropenia and may predict the risk of febrile neutropenia.

It is suggested that blood responsiveness to myeloid growth factors correlates with marrow cellularity and may identify outpatients with risk for severe neutropenia after cyclic chemotherapy.
The effects of interleukin‐8 on neutrophil fMetLeuPhe receptors, CD11b expression and metabolic activity, in comparison and combination with other cytokines
Authors: Roberts Pamela J. , Pizzey Arnold R. , Khwaja Asim , Carver Julia E. , Mire‐Sluis Anthony R. , Linch David C.
Published: March 2008

Summary. The effect of the chemotactic cytokine, IL‐8, on neutrophil function was compared with that of other cytokines, GM‐CSF, G‐CSF TNFα and IFN‐γ. IL‐8 rapidly stimulated a three‐fold enhancement of the fMLP‐stimulated respiratory burst, but this priming effect was transient compared with the slower and sustained effects of GM‐CSF and IFNγ. Apart from G‐CSF, IL‐8 was the weakest priming agent and was weaker than GM‐CSF in priming arachidonic acid metabolism stimulated by calcium ionophore. When incubated in combination, IL‐8 and TNFα were highly synergistic in their effects on respiratory burst priming, whereas IL‐8 and GM‐CSF showed little synergy. In contrast, IL‐8 was as potent as GM‐CSF at increasing the expression of neutrophil chemotactic peptide receptors and the β2 integrin, CD11b. The latter was maximally upregulated within 5 min of stimulation with IL‐8, whereas the effect of GM‐CSF was much slower. The kinetics of neutrophil respiratory burst priming by IL‐8 were the same when measured in whole blood samples and in purified cell suspensions, and IL‐8 dose‐response curves were similar, showing that the low affinity IL‐8 receptors on erythrocytes do not rapidly sequester circulating IL‐8. The data suggest that IL‐8 plays a minor role in priming neutrophil function and that a more major activity is the regulation of neutrophil adhesion and migration.
Accumulation of 5‐methyltetrahydrofolic acid and folylpolyglutamate synthetase expression by mitogen stimulated human lymphocytes
Authors: Fort Daniel W. , Lark Richard H. , Smith Angela K. , Marling‐Cason Margaret , Weitman Steven D. , Shane Barry , Kamen Barton A.
Published: March 2008

Summary. The accumulation of 5‐methyl[3H]tetrahydrofolic acid (5CH3[3H]FH4) by phytohaemagglutinin stimulated lymphocytes (PHA‐L) cultured in folate free media was investigated to determine the mechanism of uptake of 5CH3FH4 and the requirement of the cells for this vitamin as assessed by monitoring de novo thymidine synthesis. When grown in 20 nm 5CH3[3H]FH4 PHA‐L accumulate radiolabel at a rate of 0·04 pmol/h/106 cells. This doubles the endogenous folate pool of unstimulated cells (0·6±0·16 pmol/106 cells) in about 15 h. Uptake proceeded via a saturable process, independent of a high affinity folate receptor as assessed by ligand binding and by Northern and Western blot analysis. However, transport was blocked by probenecid, which is consistent with an anion carrier mechanism. Unstimulated cells lacked folylpolyglutamate synthetase (FPGS) activity and did not express significant amounts of FPGS mRNA. After 48 h of mitogen stimulation there was a 4–10‐fold increase in FPGS mRNA and folylpolyglutamate formation (Glu≥5) was essentially simultaneous with 5CH3[3H]FH4 transport. Increasing extracellular folate to 2μm only increased intracellular folate 8‐fold, but the length of the folylpolyglutamates decreased. The increased folate did not increase de novo thymidine synthesis compared to cells grown in physiological folate. We conclude that mitogen stimulation activates the process(es) for folate accumulation, especially FPGS, and that physiological uptake (0·04 pmol/h/106 cells) is adequate for meeting the cells' need for the vitamin.
Detection of a molecular defect in 40 of 44 patients with haemophilia B by PCR and denaturing gradient gel electrophoresis
Authors: Tartary Michel , Vidaud Dominique , Piao Yingchao , Costa Jean‐Marc , Bahnak Bruce R. , Fressinaud Edith , Congard Brigitte , Laurian Yves , Meyer Dominique , Lavergne Jean‐Maurice , Vidaud Michel
Published: March 2008

Summary. Oligonucleotides were computer designed to amplify by the polymerase chain reaction (PCR) the coding region, splice junctions, 112 bp of the 5’flanking region and 279 bp surrounding the polyadenylation site of the factor IX gene for analysis by denaturing gradient gel electrophoresis (DGGE). Forty‐four unselected haemophilia B patients were studied of whom 24 had severe haemophilia and 20 had a mild to moderate form of the disease. Potential mutations were identified in 40 (91%) of the 44 cases. A defect could not be detected in three severe and one mild haemophiliac by DGGE analysis and direct sequencing of all the PCR fragments from these patients revealed no nucleotide alteration supporting the DGGE results. A total of 37 point mutations, two complete gene deletions and a duplication of 26 bp were found. The 37 point mutations included 35 single nucleotide substitutions, a deletion and an insertion of one nucleotide. The 35 single nucleotide substitutions included 26 missense mutations, seven nonsense mutations, a G (‐6) to A transition in the promoter region and a G (30154) to A transition within the donor splice site of the last intron. Fifteen of these nucleotide substitutions involved CpG di‐nucleotides. Fifteen point mutations were found at codons where nucleotide substitutions had not been detected before. An insertion of a single nucleotide T at position 6370 and deletion of a G at nucleotide 30845 resulted in frameshift mutations creating stop codons at amino acid positions −2 and 250, respectively. A duplication of 26 bp (17747–17772) in exon V was found in a severe haemophilia patient resulting in a termination codon in exon VI. The detection of the mutation by the combined use of PCR, DGGE and direct sequencing was important for carrier diagnosis of 20 families with no prior history of haemophilia B.
Concurrent label method with 111In and 51Cr allows accurate evaluation of platelet viability of stored platelet concentrates
Authors: Holme S. , Heaton A. , Roodt J.
Published: March 2008

Summary. The precision and reproducibility of 111In and 51Cr platelet radiolabel agents for in vivo kinetic studies of stored platelet concentrates (PC) were investigated. The objective was to develop a precise method with concurrent labelling of two platelet populations using different isotopes, which would allow identification of small differences in in vivo platelet quality. Identical labelling procedures were used to investigate the effects of PC storage age, different methods of red cell (RBC) and white cell (WBC) contamination correction, and label elution correction on the results of 111In and 51Cr kinetic studies.

111In and 51Cr platelet survival curves from the same PC, even when uncorrected for elution and RBC contamination, exhibited excellent correlation, irrespective of the age of the concentrate and its viability. However, slightly higher, but statistically significant, post‐infusion per cent recoveries with 51Cr labelled platelets were found. Two factors were identified as the cause for this difference. There was a higher affinity of contaminating RBC/WBC in PC for 51Cr than for 111In. With determination of RBC/WBC activity by centrifugation/density separation, RBC/WBC fractions from the injectate were found to contain 12.6 ± 3·8%v 7·1 ± 3·6% of total 51Cr and 111In activity, respectively, in 20 studies. In addition, there was a significantly higher 111In activity in plasma immediately post‐infusion than with 51Cr, 5·2 ± 1·3%v 2·8 ± 1·6%, respectively, suggesting more label elution or carryover. After correction for the activity of RBC/WBC and for elution or carryover, essentially identical 51Cr/111In platelet survival curves were found. In 31 stored PC studies, the absolute average difference between 51Cr and 111In per cent recoveries was only 4 ± 3% in a group of donors whose platelet recoveries ranged from 10% to 80%. Similarly, the average difference between 51Cr and 111In survival was only 8±4 h within a range of survivals from 40 to 220 h.

In conclusion, after correction for elution and contaminating RBC/WBC binding, these studies show that 51Cr and 111In may be used interchangeably for labelling of stored PC, and that small differences between test and control platelets could be reliably detected using concurrent labelling with simultaneous infusion.
Degradation of human erythrocyte surface components by human neutrophil elastase and cathepsin G: preferential digestion of glycophorins
Authors: Bykowska K. , Duk M. , Kuσnierz‐Alejska G. , Kopeć M. , Lisowska E.
Published: March 2008

Summary. Human erythrocytes treated with purified human neutrophil elastase (HNE) or cathepsin G (CathG) were analysed by serological methods and by SDS‐polyacrylamide gel electrophoresis followed by staining or immunoblotting with monoclonal antibodies. Both enzymes digested exhaustively glycophorins A, B and C, and HNE caused a partial digestion of band 3 protein. The degradation of other membrane proteins was not detectable by the methods used. Immunoblotting with the use of monoclonal antibodies against the defined epitopes of glycophorin A showed that HNE and CathG hydrolysed distinct peptide bonds in this antigen. The antibody PEP80, specific for the epitope in the cytoplasmic fragment of glycophorin A, gave patterns of bands which were characteristic for each of the two proteases. These bands could be distinctly identified in erythrocyte membrane samples containing only few percents of digested glycophorins. Therefore, the immunoblotting with this antibody may be useful as a sensitive assay for detecting the action of neutrophil proteases on red blood cells.
Improved cytogenetic analysis of bone marrow plasma cells after cytokine stimulation in multiple myeloma: a report on 46 patients
Authors: Facon T. , Lai J. L. , Nataf E. , Preudhomme C. , Zandecki M. , Hammad M. , Wattel E. , Jouet J. P. , Bauters F.
Published: March 2008

Summary. Cytogenetic analysis was successfully performed in 46 consecutive myeloma patients (40 newly diagnosed and six relapsed patients). Karyotype was performed on bone marrow cells after long‐term cultures (6 d) stimulated by GM‐CSF, GM‐CSF + IL6 or GM‐CSF + IL6 + IL3. Nineteen patients (41%) had cytogenetic abnormalities including 17/40 patients at diagnosis (42·5%) and 2/6 patients at relapse. Hyperdiploidy was found in 12 patients and hypodiploidy in four patients. Of the 17 newly diagnosed patients with cytogenetic abnormalities, five died from myeloma after 1–14 months and three other patients had primary drug resistance. Our results suggest that cytogenetic analysis after stimulation of cultures by cytokines detects clonal abnormalities in 40–50% of newly diagnosed myeloma patients and that these patients often have a short survival and/or primary drug resistance.
Haemoglobin Manukau β67[E11] Val→Gly: transfusion‐dependent haemolytic anaemia ameliorated by coexisting alpha thalassaemia
Authors: Fay Keith C. , Brennan Stephen O. , Costello Jack M. , Potter Howard C. , Williamson David A. , Trent Ronald J. , Ockelford Paul A. , Boswell D. Ross
Published: March 2008

Summary Haemoglobin Manukau (β67 Val→Gly) is a novel haemoglobin variant presenting in two brothers as non‐spherocytic haemolytic anaemia which became transfusion dependent by 6 months of age. The severity of clinical expression seems to be modulated by coexisting alpha thalassaemia: the severely affected children have a normal complement of alpha globin genes with an unusual genotype (‐α3,7/ααα3,7), while their father, who carries the abnormal gene with minimal symptoms, has homozygous α+ thalassaemia (‐α3,7/ ‐α3,7) Another unusual feature of this case is the association of the β67 Val→Gly mutation with modification of β141 Leu to a residue (believed to be hydroxyleucine) that is not detected by standard amino acid analysis. This finding offers an explanation for the previous report of an association of another mutation at this site (Hb Sydney β67 Val→Ala) with Hb Coventry (deletion of β141 Leu).
Image analysis studies of the degree of irreversible deformation of sickle cells in relation to cell density and Hb F level
Authors: Horiuchi Kazumi , Stephens Monica J. , Adachi Kazuhiko , Asakura Toshio , Schwartz Elias , Ohene‐Frempong Kwaku
Published: March 2008

Summary We analysed, quantitatively by image analysis, the degree of irreversible deformation of red cells (SS cells) from patients with homozygous sickle cell disease, and studied the relationships among the degree of irreversible cell deformation, cell density, and Hb F level. SS cells from 2 5 patients (aged 1–36 years) whose Hb F levels ranged from 2.5%) to 40.0%. were fully oxygenated and then were separated into four fractions by density centrifugation. Every fraction was studied for morphology and Hb F content. We found that the irreversible deformation of SS cells from the circulation occurred mainly by elongation and that the degree of elongation was extremely variable. We also found that in the cells of patients with Hb F levels < 20% the degree of irreversible elongation increases as cell density increases, suggesting that dehydration occurs concomitantly with irreversible elongation. Statistical analysis (Student t test) indicated that there were significant differences (P= 0.008 or < 0. 001) in the degree of elongation among density‐fractionated SS cells from patients with Hb F < 20%. although there was no significant difference (P>0.1) among those from patients with Hb F ≥20%. We also found that cell density increased as Hb F level of the density fraction decreased in all patients with Hb F < 20% but not always in those with Hb F ≥20%. This suggests that cells with lower Hb F levels are selectively susceptible to dehydration. Furthermore, we found that the mean degree of irreversible elongation decreases linearly with increasing levels of Hb F and reaches the normal range at 21–24%. Since the degree of irreversible deformation of SS cells quantified by image analysis is directly related to cell density, and inversely to Hb F levels, mechanical stress or membrane damage caused by Hb S polymerization may be an important factor in the formation of dense cells in viva
Subcellular distribution of desferrioxamine and hydroxypyridin‐4‐one chelators in K562 cells affects chelation of intracellular iron pools
Authors: Hoyes Katherine P. , Porter John B.
Published: March 2008

Summary The interactions of iron chelators with intracellular iron pools have been examined by measuring the subcellular distribution of radiolabelled desferrioxamine (DFO) and the orally active hydroxypyridinone (HPO) chelator 1, 2‐diethyl‐3‐hydroxypyridin‐4‐one (CP94), as well as the ability of these chelators to modify the subcellular distribution of 59Fe delivered by the receptor mediated endocytosis of transferrin. K562 cells were pulsed with 59Fe transferrin and challenged with DFO or CP94 (100 μm IBE) for 20 or 240 min and then subjected to subcellular fractionation. At 20 min there was a significant decrease (P <0.45) in both lysosomal/particulate 59Fe (75% of control) and cytosolic 59Fe ferritin (50% of control) in cells incubated with CP94, unlike cells treated with DFO where no decrease was observed. By 240 min, in addition to the above, 59Fe accumulation was significantly decreased in the nuclear, mitochondrial, and low molecular weight cytosolic fractions with CP94 (P < 0.05). With DFO a significant decrease in 59Fe in only the lysosomal/particulate and cytosolic ferritin compartments was observed at 240 min (P <045). At this time, however, there was a significant accumulation of both cytosolic low molecular weight 59Fe and cytosolic DFO. The relatively rapid decrease of 59Fe within intracellular compartments seen with CP94 compared to DFO was paralleled by a significantly higher accumulation of CP94 than DFO in nuclear, lysosomal/particulate and low molecular weight cytosolic compartments at 20 min (P <0 05). These results suggest that transferrin derived endosomal iron may be chelated by HPOs, unlike DFO, due to their faster uptake into these organelles. The more rapid access of HPOs than DFO to certain intracellular iron pools may explain the greater possibility of HPOs to inhibit proliferation of cells in vivo.
Reversing of chlorambucil resistance by ethacrynic acid in a B‐CLL patient
Authors: Petrini M. , Conte A. , Caracciolo F. , Sabbatini A. , Grassi B. , Ronca G.
Published: March 2008

Summary We evaluated the reversing activity of ethacrynic acid in a B‐CLL patient resistant to chlorambucil. The glutathione S‐transferase (GST) activity, measured in peripheral blood lymphocytes, resulted extremely elevated. Etha crynic acid, at pharmacological concentrations, partially reversed chlorambucil resistance and this result appeared related to the increased GST levels.
Contamination of peripheral blood by monoclonal B cells following treatment of multiple myeloma by high‐dose chemotherapy
Authors: Dreyfus François , Melle Josiane , Quarre Marie‐Catherine , Pillier Catherine
Published: March 2008

Summary Peripheral blood samples obtained from five patients with multiple myeloma, after high‐dose chemotherapy, were studied for monoclonal B plasma cell contamination. We used the technique of immunoglobulin heavy chain gene ‘fingerprinting’ at the time of diagnosis and during apheresis. The level of sensitivity of this technique is between 041% and 0.001% in two patients in whom a monoclonal population was detected in peripheral blood.
Fechtner syndrome: report of a third family and literature review
Authors: Rocca Bianca , Laghi Ferdinando , Zini Gina , Maggiano Nicola , Landolfi Raffaele
Published: March 2008

Summary We observed macrothrombocytopenia with leucocyte inclusions in 10 out of 14 members of a four‐generation family. Morphological features of leucocyte inclusions and the presence of Alport‐related symptoms supported the diagnosis of Fechtner syndrome. Compared to the two previously reported Fechtner families, our kindred showed reduced expression of Alport manifestations. These, in members aged less than 50, were represented by clinically silent ocular abnormalities. Due to the frequent non‐recognition of macrothrombocytopenia, Fechtner variants with low penetrance might be difficult to diagnose. In addition, Sebastian syndrome, recently distinguished from Fechtner disease in lacking Alport manifestations, might be one of these variants.
Two chronic myelogenous leultaemia cell lines which represent different stages of erythroid differentiation
Authors: Endo Kazuyasu , Harigae Hideo , Nagai Tadashi , Fujie Hiromi , Meguro Kuniaki , Watanabe Norimichi , Furuyama Kazumichi , Kameoka Junichi , Okuda Mitsutaka , Hayashi Norio , Yamamoto Masayuki , Abe Keishi
Published: March 2008

Summary We established two cell lines, YN‐1 and Y‐1K. from the peripheral blood of two chronic myelogenous leukaemia patients in blastic crisis. Characterization of the YN‐1 and Y‐1K cells revealed that these cells expressed erythroid lineage markers. However, there was a marked difference in the level of γ‐globin mRNA and haenioglobin in YN‐1 and Y‐1K cells. YN‐1 contained approximately 1–5% benzidine‐positive staining cells, whereas no benzidine‐positive cells were observed in Y‐1K cells. Haemoglobin production in YN‐1 cells was markedly increased with various chemical inducers of erythroid differentiation, but was not in Y‐1K cells. In contrast, Y‐1K cells expressed CD34 stem cell antigen and CD41 megakaryocyte‐specific antigen. These observations suggested that, although both cell lines were committed to the erythroid lineage. each cell line represented a distinct differentiation stage in the erythroid differentiation programme. Y‐1K seemed to correspond to an early stage of cells in erythroid lineage, whereas YN‐1 represented a more advanced stage in human erythropoiesis.
Haem is necessary for a continued increase in ferrochelatase mRNA in murine erythroleukaemia cells during erythroid differentiation
Authors: Fukuda Yoshiaki , Fujita Hiroyoshi , Taketani Shigeru , Sassa Shigeru
Published: March 2008

Summary The level of mRNA encoding ferrochelatase (FeC) was examined in two murine erythroleukaemia (MEL) clones, DS and DR, a DMSO‐sensitive, and a DMSO‐resistant clone, respectively. DS cells undergo erythroid differentiation by DMSO treatment with a marked increase in haem synthesis, while DR cells fail to do so due to the lack of the erythroid‐specific δ‐aminolaevulinate synthase (ALAS‐E). Both DS and DR cells showed an increase in the level of FeC mRNA within 18 h of DMSO treatment. The level of FeC mRNA in DR cells was then decreased, while that in DS cells continued to increase for 72 h. Treatment with haemin significantly increased FeC mRNA in DR cells. When cells were treated with both DMSO and haemin, the level of FeC mRNA in DR cells increased to a level comparable to that in DS cells. These findings suggest that the failure to maintain increased FeC mRNA DR cells after DMSO treatment may be due to a deficiency of haem in these cells.
Clonal remissions in acute myeloid leukaemia are commonly associated with features of trilineage myelodysplasia during remission
Authors: Jowitt S. N. , Yin J. A. Liu , Saunders M. J. , Lucas G. S.
Published: March 2008

Summary Clonal haemopoiesis has previously been demonstrated in some 30% of patients in remission of acute myeloid leukaemia (AML). Whilst a ‘clonal remission’ in many such patients may represent a skewed X‐chromosome inactivation pattern in haemopoietic cells, its relationship to an underlying preleukaemic state remains uncertain. We therefore analysed the clonal status of 48 female patients in remission of AML using X‐chromosome linked restriction fragment length polymorphisms (RFLPs) within the X‐linked PGK and HPRT genes and the DXSZ55 (M27β) locus, and carried out in conjunction a detailed study of the morphological and karyotypic features of the patients' bone marrows. During remission, 35 patients (73%) with AML demonstrated non‐clonal haemopoiesis, and their bone marrows were morphologically normal. Remission haemopoietic tissue in nine cases (19%) showed a skewed X‐chromosome inactivation pattern and remission bone marrows in these patients had features of trilineage myelodysplasia (TMDS), with seven having similar features at presentation. Analysis of constitutional DNA showed a non‐clonal pattern in seven of these patients, but was unsuccessful in two cases. These nine patients with post‐chemotherapy TMDS were considered to have true clonal haemopoiesis. Four patients (8%) with a skewed X‐chromosome inactivation pattern had normal remission bone marrows. Analysis of constitutional DNA showed a skewed pattern in two of these patients, but was unsuccessful in two cases. Cytogenetic investigation during remission in the nine patients with TMDS showed a normal karyotype in four cases and the acquisition of new karyotypic abnormalities in three cases. In contrast, 10 female patients in remission of de novo acute lymphoblastic leukaemia (ALL) were shown to have non‐clonal haemopoiesis. We conclude that the majority of patients with AML who achieve remission after cytoreductive chemotherapy have non‐clonal haemopoiesis, and when clonal remissions are observed these are commonly associated with the development of trilineage myelodysplasia in the bone marrow, with or without karyotypic abnormalities. True clonal remission in association with morphologically normal haemopoiesis is a rare entity. the significance and frequency of which remain uncertain.
Allogeneic bone marrow transplantation following busulfan‐cyclophosphamide with or without etoposide conditioning regimen for patients with acute lymphoblastic leukaemia
Authors: Bueltzingsloewen Anne von , Bélanger Robert , Perreault Claude , Bonny Yvette , Roy Denis‐Claude , Boileau Jacques , Kassis Jeannine , Lavallée René , Lacombe Michel , Gyger Martin
Published: March 2008

Summary We have investigated the feasibility and efficacy of administering a radiation‐free preparative regimen in the setting of allogeneic bone marrow transplantation (BMT) in 40 consecutive patients with acute lymphoblastic leukaemia (ALL). Busulfan (4 mg/kg/d × 4 d) and cyclophosphamide (50 mg/kg/d × 4 d) (BuCy4) were given in 29 patients and 11 received busulfan (4 mg/kg/d × 4 d), etoposide (60 mg/kg) and cyclophosphamide (60 mg/kg/d × 2 d) (BuCy + VP‐16). Median age was 22 years (range 1–50); 11 patients were children ≤ 15 years of age. All children and 20 adults were at high risk of relapse pretransplant. Nine adults and one child died from transplant‐related toxicity. 11 patients relapsed at a median of 11 months post‐transplant (range 2–27). The 3‐year Kaplan‐Meier estimated probability of relapse was 42.1% and found to he significantly lower in patients with chronic GVHD (P=0.03). 19 patients are leukaemia‐free survivors with a median follow‐up of 33 months (range 7–59). The Kaplan‐Meier actuarial probability of disease‐free survival at 3 years was 43% for all patients, 63.6% for children versus 30.2% for adults (P = 0.24) and 51.6% for patients transplanted in first remission versus 30.2% for those transplanted in subsequent remissions (P = 0.20).
Elevated platelet‐associated IgG in SLE patients due to anti‐platelet autoantibody: differentiation between autoantibodies and immune complexes by ether elution
Authors: Kurata Yoshiyuki , Hayashi Satoru , Kosugi Satoru , Kashiwagi Hirokazu , Tomiyama Yoshiaki , Kanayama Yoshio , Matsuzawa Yuji
Published: March 2008

Summary The level of platelet‐associated IgG (PAIgG) is reported to be elevated in patients with systemic lupus erythematosus (SLE). However. the nature of PAIgG is unclear. We have investigated whether the PAIgG of SLE consists of anti‐platelet autoantibodies or immune complexes (IC). The PAIgG values measured by flow cytonietry were elevated in 11/25 patients with SLE. 3/6 SLE patients with thrombocytopenia had a high level of PAIgG (the mean fluorescence intensity >10). We used an ether elution technique to determine whether elevated PAIgG consists of anti‐platelet antibodies or IC. Preliminary experiments showed that the eluates prepared from platelets sensitized with anti‐HPA‐4a antibody reacted with normal platelets. while the eluates prepared from platelets sensitized with heat‐aggregated IgG or model IC failed to react with normal platelets. These results indicate that the reactivity of eluates can distinguish between platelet‐bound antibody and IC. We applied this technique to analysis of the PAIgG of SLE platelets. The eluates from SLE platelets (the mean fluorescence intensity > 10) reacted with normal platelets. indicating that the PAIgG of SLE platelets has the nature of anti‐platelet autoantibodies. Furthermore, we investigated the target antigens which bind PAIgGs of SLE, using the direct immunoprecipitation procedure and modified antigen capture ELISA (MACE). Both methods identified GPIIb/IIIa as the target antigens. We conclude that the ether elution technique can distinguish between anti‐platelet antibodies and TC. and that the PAIgGs of SLE with a high PAIgG value and thrombocytopenia have the nature of anti‐platelet autoantibodies.
A favourable effect of long‐term α‐interferon therapy in refractory idiopathic thrombocytopenic purpura
Authors: Kumakura Shunichi , Ishikura Hiroto , Tsumura Hiroto , Endo Jiro , Tsunematsu Tokugoro
Published: March 2008

Summary We successfully treated two patients with refractory idiopathic thrombocytopenic purpura (ITP) with a weekly or monthly administration of α‐interferon followed by short‐course treatment with α‐interferon. In both cases an increase in platelet count was brought about within 1 week after administration of α‐interferon and the favourable response in platelet count was sustained over a few years with the weekly or monthly administration of α‐interferon in the absence of any noticeable side‐effects except for fever. So. α‐interferon therapy seems to be beneficial in some cases of refractory ITP in terms of both remission induction and maintenance therapy.
Analysis of a Pst I polymorphism of the human erythrocyte band 3 gene (EPB3)
Authors: Jenkins Patricia B. , Gallagher Patrick G. , Forget Bernard G.
Published: March 2008

Summary The genomic structure of a 1. 5 kb region near the 5′ end of the human erythrocyte band 3 gene (EPB3) was determined and the location of a two‐allele PstI polymor phism in this region identified. Using this information, a polymerase chain reaction (PCR)‐based assay was designed for rapid, simple identification of the PstI polymorphism.
A novel 203 kD aberrant BCR‐ABL product in a girl with Philadelphia chromosome positive acute lymphoblastic leultaemia
Authors: Lnukai Takeshi , Sugita Kanji , Suzuki Toshio , Ijima Kiyomu , Goi Kumiko , Tezuka Toru , Kojika Satoru , Hatakeyama Kazuo , Kagami Keiko , Mori Taijiro , Okazaki Toshiko , Mizutani Shuki , Nakazawa Shinpei
Published: March 2008

Summary We report a girl with Ph1‐positive ALL with the aberrant BCR‐ABL product. In this case. bcr exon 3 jointed not to ordinal abl exon 2 but to exon 3 resulting in the production of a 20 3 kD BCR‐ABL fusion protein with marked tyrosine kinase activity. To our knowledge, this is the first report of an aberrant BCR‐ABL product in childhood. This case was characterized with younger age and low leucocyte count at the onset, but relapsed early like the typical Phl‐positive ALL, suggesting the diversity in the clinicopathogenesis of Ph1‐positive ALL.
Response of aplastic anaemia and scleroderma to cyclosporin
Authors: Tooze J. A. , Marsh J. C. W. , Wickham N. , Duke O. L. , Behrens J. , Gordon‐Smith E. C.
Published: March 2008

Summary Scleroderma and aplastic anaemia (AA) occurred simultaneously in a patient. Treatment with antilymphocyte globulin (ALG) resulted in some improvement of the scleroderma and a partial, temporary response of the AA. Both the scleroderma and AA then responded dramatically to cyclosporin (CSA) therapy. Subsequently, a positive Ham's test, together with a reduction in the phosphatidyl‐inositolglycan (PIG) anchored membrane proteins decay accelerating factor (DAF, CD5 5) and membrane inhibitor of reactive lysis (MIRL. CD59). confirmed a diagnosis of paroxysmal nocturnal haenioglobinuria (PNH) affecting erythroid, myeloid and lymphoid cell lineages. We hypothesize that the pathogenesis of the bone marrow failure in this patient was a stem cell defect with a secondary immune response involving T‐lymphocytes that may have simultaneously triggered the pancytopenia and scleroderma.
Proliferating normal bone marrow cells do stain for Ki‐67 antigen
Authors: Cattoretti Giorgio , Orazi Attilio , Gerdes Johannes
Published: March 2008

Summary The normal human bone marrow has been investigated in the past for the presence of the proliferation associated antigen detected by the Ki‐67 antibody and aberrantly low scores for this antigen have been reported. We used a new Ki‐67 equivalent antibody (MIU 1) and a formalin fixation of smears and trephine biopsies, and we report 52.8%±9.2 SD of the normal BM cells to stain for MIB 1.Late forms of the maturing blood cells compartment such as metamyelocytes and megakaryocytes were stained in cytokine‐treated marrows.
Cytokine production by peripheral blood monocytes and T cells during haemopoietic recovery after intensive chemotherapy
Authors: Takamatsu Yasushi , Akashi Koichi , Harada Mine , Teshima Takanori , Inaba Shoichi , Shimoda Kazuya , Eto Tetsuya , Shibuya Tsunefumi , Okamura Seiichi , Niho Yoshiyuki
Published: March 2008

Summary We studied the production of cytokines by peripheral blood monocytes and T cells during the period of haematological recovery following intensive chemotherapy. Twelve adults with haematological malignancies received consolidation chemotherapy of complete remission. Monocytes and T cells were collected during the phase of recovery from intensive chemotherapy, and were incubated for 24 h in a culture medium with 10% FCS. Concentrations of cytokines in the culture supernatant were measured with an enzymelinked immunosorbent assay. During the recovery phase, concentrations of IL‐6, G‐CSF and IL‐1β in the culture supernatant of the collected monocytes significantly exceeded those of the monocytes obtained from normal healthy subjects. Similarly, the concentrations of GM‐CSF and IFN‐γ in the supernatant of recovery phase T cells significantly exceeded those of normal T cells. Plasma levels of these cytokines were also elevated. These data suggest that the monocytes and T cells may be activated in vivo to produce haemopoietic cytolines during haematological recovery, and that, during haematological recovery, the monocytes and T cells may be actively involved in the induction of haematopoiesis following the myelosuppression induced by chemotherapy.
Resolution of transfusion dependence by recombinant human erythropoietin (rHuEPO) in acquired pure red cell aplasia (PRCA) associated with myeloid metaplasia
Authors: Zeigler Z. R. , Rosenfeld C. S. , Shadduck R. K.
Published: March 2008

Summary A patient with acquired pure red cell aplasia and agnogenic myeloid metaplasia (AMM) was treated with rHuEPO. She became transfusion independent. Weekly injections of rHuEPO have maintained the response without side‐effects or disease progression. In addition, marrow fibrosis and splenomegaly have decreased. Therefore, rHuEPO may be effective therapy for some patients with acquired PRCA and/or AMM.
Somatostatin analogue scintigraphy of malignant lymphomas
Authors: Vanhagen P. M. , Krenning E. P. , Reubi J. C. , Mulder A. H. , Bakker W. H. , Oei H. Y. , Löwenberg B. , Lamberts S. W. J.
Published: March 2008

Summary Normal as well as activated lymphocytes, macrophages and leukaemic cells have previously been shown by radio receptor analysis to express receptors for somatostatin. An 111In labelled somatostatin analogue was already used successfully in the visualization of a variety of neuro‐endocrine tumours.

We investigated 10 consecutive patients with malignant lymphomas (Hodgkin's disease and non‐Hodgkin's lymphomas). In all patients the lymphoma deposits could be visualized with somatostatin receptor imaging.

In four patients additional tumour localizations were observed as compared to the results of combined physical and radiological (CT and ultrasound) examinations. In four cases tissue biopsies were taken and confirmed by autoradiography to be somatostatin receptor positive.

These data indicate that malignant lymphomas may express somatostatin receptors in sufficient numbers and density to allow in vivo tumour visualization with a radiolabelled somatostatin analogue.
Substitution of cysteine for phenylalanine 751 in mature von Willebrand factor is a novel candidate mutation in a family with type IIA von Willebrand disease
Authors: Gaucher Christine , Hanss Michel , Dechavanne Marc , Mazurier Claudine
Published: March 2008

Summary Type IIA is a variant form of von Willebrand disease (vWD) characterized by the absence of von Willebrand factor (vWF) high molecular weight multimers in plasma. Most of the candidate missense mutations potentially responsible for type IIA vWD have been found clustered within a short segment of vWF, lying between Gly742 and Glu875 of the mature subunit. The present work reports a single heterozygous T → G transversion in eight patients from a large type IIA vWD family, resulting in the substitution Phe751→Cys. The absence of this mutation in 100 normal vWF genes as well as the lack, in these patients, of any other abnormality within the whole exon 28 encoding amino acids 463–921 of mature vWF, provide a strong support that this non‐conservative mutation may be at the origin of the disease in this family. The presence of an additional cysteine at position 751 may induce a conformational change of the vWF subunit affecting either its ‘in vivo’ sensitivity to proteolytic cleavage or, more likely, its intracellular transport as suggested by the abnormal multimeric pattern of platelet vWF observed in these patients.
Effects of vitamin K and oral anticoagulants on urinary calcium excretion
Authors: Jie Kon‐Siong G. , Gijsbers Birgit L. M. G. , Knapen Marjo H. J. , Hamulyák Karly , Frank Herman L. , Vermeer Cees
Published: March 2008

Summary In a subgroup of postmenopausal women vitamin K induced a decrease of the urnary calcium loss. This effect was significant (P < 0.0001) in the so‐called fast losers of calcium (calcium/creatinine ratio > 0.5). To find out whether vitamin K antagonists would have an opposite effect, a study was started among 141 persons on long‐term oral anticoagulant therapy. In this population the number of fast losers was recorded, and compared to that in a group of age‐ and sex‐matched non‐treated controls. Notably in young men the fraction of fast losers was significantly higher in the anticoagulant‐treated group than in the control group (25 v 0%, P < 0.02). Differences between treated and non‐treated groups may also be found in other markers for calcium and bone metabolism, notably in serum osteocalcin concentration and in urinary hydroxyproline excretion. The conclusion of our study is that oral anticoagulant treatment must be regarded as a potential risk factor for a high loss of urinary calcium.
Rapid analysis of ‐α3.7 thalassaemia and αααanti 3.7 triplication by enzymatic amplification analysis
Authors: Dodé Catherine , Krishnamoorthy Rajagopal , Lamb Janette , Rochette Jacques
Published: March 2008

Summary In this report we describe a PCR‐based method for the diagnosis of the most common form of α thalassaemia, the –α3.7 deletion which occurs throughout all tropical and subtropical regions of the world. The same procedure also identifies the reciprocal recombinant chromosome (αααanti 3.7). Restriction mapping of the PCR products has enabled us to distinguish between the type I (–α3.7I), type II (–α3.7II) and type III (–α3.7III) deletions. This strategy will be very useful in screening programmes of α thalassaemia occurring on its own or in association with β thalassaemia and sickle cell disease.
Age dependency of red blood cell deformability and density: studies in transient erythroblastopenia of childhood
Authors: Linderkamp O. , Friederichs E. , Boehler T. , Ludwig A.
Published: March 2008

Summary Several investigators have demonstrated that red blood cell (RBC) deformability decreases progressively with increasing cell density and proposed that reduction in deformability plays a role in the senescence process of normal RBCs. Transient erythroblastopenia of childhood (TEC) results from temporary cessation of erythropoiesis. Since no new RBCs are produced for some time, the circulating RBCs are relatively old. RBS density (phthalate‐oil method) and RBC deformability (RBC elongation in a counter‐rotating rheoscope) were studied in seven children with TEC and in 10 control children. The mean values of MCHC, RBC density and RBC deformation were not significantly different between TEC and control children. Compared to controls, the frequency distribution of RBC density in TEC was slightly shifted to higher values. The percentage of RBCs with extremely low densities (< 1.090 g/ml) was 0.9 ± 1.2% in the patients and 5.6 ± 2.3% in the controls (P < 0.001). The percentage of RBCs with high density (> 1.106 g/ml) was 6.4 ± 2.1% in the patients and 4.9 ± 1.8% in the controls (P > 0.10). The reduction of RBCs with low density in TEC suggests that RBCs with low density are relatievly young. Since the percentage of RBCs with high density increased only slightly in TEC, we conclude that only a fraction of dense RBCs is old. In TEC, the frequency distribution of RBC elongation was slightly shifted to lower values. 5% of the RBCs studied in the control children had RBC elongation values above 0.39 (TEC 1.2%) and 5% had elongation values below 0.16 (TEC 6.7%). Thus, only a small fraction of highly deformable RBCs was diminished in TEC. These data suggest that a decrease in deformability is not a significant part of the normal ageing process of human RBCs.
The effect of endotoxin and tumour necrosis factor on erythrocyte and leucocyte deformability in vitro
Authors: Betticher Daniel C. , Keller Hansuli , Maly Friedrich E. , Reinhart Walter H.
Published: March 2008

Summary Microcirculatory disorders are a common finding in sepsis. We have analysed the influence of two factors released in sepsis, endotoxin and tumour necrosis factor (TNF), on rheological properties of blood cells. The deformability of mixed cell suspensions, isolated erythrocytes, mononuclear cells, or polymorphonuclear leucocytes exposed to endotoxin and TNF in vitro was assessed by filtration through pores of different sizes. Mixed blood cell suspensions showed an increase in cell rigidity when incubated with 100 ng/ml endotoxin. The filtration resistance of isolated erythrocytes, mononuclear or polymorphonuclear leucocytes was not affected by endotoxin. Incubation with TNF in physiological concentrations increased the rigidity of mixed blood cells and of isolated polymorphonuclear leucocytes in a dose‐ and time‐dependent manner, while erythrocytes and mononuclear leucocytes remained unaffected. Polymorphonuclear cells showed decreased deformability associated with shape changes (polarized and non‐polar cells with surface protrusions and a shift of F‐actin into protrusions). The decrease in deformability was reversed by cytochalasin B or xanthin derivatives such as pentoxifylline. We conclude that TNF decreases the passive deformability of polymorphonuclear leucocytes, which may affect the microcirculation in sepsis. The reversibility with xanthin derivatives may represent a new therapeutic approach for the high morbidity and mortality in sepsis.
Proliferation and differentiation of myelodysplastic CD34+ cells in serum‐free medium: response to individual colony‐stimulating factors
Authors: Sawada K. , Sato N. , Tarumi T. , Sakai N. , Koizumi K. , Sakurama S. , Ieko M. , Yasukouchi T. , Koyanagawa Y. , Yamaguchi M. , Ohmoto A. , Kohno M. , Koike T.
Published: March 2008

Summary. The presence of serum or contaminant cells may alter clonal development of haematopoietic progenitor cells in vitro. To investigate the pathogenesis of myelodysplastic syndromes (MDS), marrow progenitor cells from 13 MDS patients were highly purified using monoclonal antibodies including CD34 and immunomagnetic microspheres. The cells positive for CD34 in the purified cells were in the range from 87% to 98%. These purified cells were cultured in serum‐free medium with individual colony stimulating factors (CSFs) to investigate whether CD34+ cells from MDS patients have abnormal responses to individual CSFs. Dose response experiments with the purified CD34+ cells and recombinant human macrophage‐CSF (rM‐CSF), granulocyte‐CSF (rG‐CSF), granulocyte/macrophage‐CSF (rGM‐CSF), interleukin‐3 (rIL‐3) or erythropoietin (rEP) were performed in serum‐free fibrin clots in 11 patients. Five patients showed a diminished response to rG‐CSF and one patient to rEP. In the remaining six patients the purified CD34+ cells did not respond to a stimulation of any individual CSFs. The results indicate that the progenitor cell growth abnormalities in these disorders involve a defect in the capacity of progenitor cells to respond to stimulation with G‐CSF, and present direct evidence for the manner in which myelodysplastic CD34+ cells are impaired.
Role of granulocyte‐macrophage colony‐stimulating factor, Interleukin‐3 and Interleukin‐5 in the eosinophilia associated with T cell lymphoma
Authors: Fermand Jean‐Paul , Mitjavila Maria‐Teresa , Couedic Jean‐Pierre Le , Tsapis Andréas , Berger Roland , Modigliani Robert , Seligmann Maxime , Brouet Jean‐Claude , Vainchenker William
Published: March 2008

Summary. We studied two patients with a leukaemic T cell lymphoma who presented with a marked increase in blood eosinophilia. To investigate the mechanism of the eosinophilia, supernatants of peripheral blood cells containing more than 80% lymphoma cells were tested by biological assays for the presence of colony stimulating factors (CSF). In one case supernatants stimulated the growth of granulocyte‐macrophage (GM), erythroid and eosinophil colonies. These effects were neutralized by anti‐GM‐CSF antibodies; anti‐IL5 antibodies slightly decreased eosinophil colony formation. Supernatants derived from the second patient cells stimulated the same lineages. Neutralizing experiments demonstrated that in addition to GM‐CSF it contained interleukin 3 (IL‐3) and interleukin 5 (IL‐5). In agreement with the biological data. RNA studies using the polymerase chain reaction showed that cells from the first patient expressed GM‐CSF transcripts; IL‐5 transcripts were also detected in very low amounts. GM‐CSF, IL‐3 and IL‐5 transcripts were detected in cells from the second patient. Thus eosinophilia associated with some T cell lymphoma is likely due to secretion of different combinations of cytokines by malignant cells.
The management of recurrent acute myelogenous leukaemia at a single centre over a fifteen‐year period
Authors: Davis C. L. , Rohatiner A. Z. S. , Lim J. , Whelan J. S. , Oza A. M. , Amess J. , Love S. , Stead E. , Lister T. A.
Published: March 2008

Summary. One hundred and sixty‐two patients initially treated at St Bartholomew's Hospital between 1974 and 1988 developed recurrent acute myelogenous leukaemia (AML). In the majority, the intention was to administer intensive chemotherapy again; 22/162 were re‐treated palliatively. A second complete remission (CR) was achieved in 50/126 (40%) evaluable patients. Several different regimens were employed over this time period; the treatment used was the only factor that correlated with achievement of second CR (P=0.04). Ten of the 50 patients received myeloablative therapy with either allogeneic or autologous bone marrow transplantation in second CR. The median duration of second CR for those patients who did not proceed to intensive consolidation with bone marrow transplantation was 7 months. There was no correlation between second remission duration and age, or with the administration of either intensive or conventional consolidation therapy.

The median survival from first recurrence for all patients was 4 months, increasing to only 5 1/2 months for those re‐treated with chemotherapy and to 12 months for those patients in whom a second CR was achieved, confirming the very poor prognosis of these patients.