bluuebunny/crossref_metadata_2025 huggingface dataset:
A case of peroxidase‐positive acute leukaemia expressing only T lineage lymphoid markers
Authors: Lelong F. , Chretien P. , Jouault H. , Bayani N. , Bernaudin F. , Lemerle S.
Published: March 2008

We report the clinical presentation and the morphological, immunophenotypic, cytogenetic and molecular genetic characteristics of a 14·1/2‐year‐old boy who had French–American–British (FAB) type M1 acute non‐lymphocytic (ANLL) leukaemia with a common T‐ALL immunological phenotype, with no myeloid associated antigen, either on the membrane or in the cytoplasm. ALL‐directed induction therapy induced complete remission.
Ploidy in bone marrow cells from healthy donors: a MAC (morphology antibody chromosomes) study
Authors: Larramendy Marcelo L. , Nylund Stella J. , Wessman Maija , Ruutu Tapani , Knuutila Sakari
Published: March 2008

The ploidy of human bone marrow cells belonging to the megakaryocytic, granulocytic‐monocytic and erythrocytic lineages was studied by in situ hybridization using the biotin‐labelled Y chromosome‐specific DNA probe pY431 and the chromosome 1‐specific probe pUC1.77 on cells identified morphologically and immunologically by the MAC procedure. Cells of the granulocytic‐monocytic and erythrocytic lineages were seen to be 2N in ploidy, whereas the ploidy of the megakaryocytic lineage ranged from 2N to 32N, with the ploidy classes 4N and 8N being predominant. The frequency of megakaryocytes with 2N chromosomes was also high.
Spontaneous remission in acute myeloid leukaemia
Authors: Paul R. , Remes K. , Lakkala T. , Pelliniemi T. T.
Published: March 2008

A summary of the literature of adult patients with acute myeloid leukaemia (AML) who have undergone spontaneous remission (SR) is presented together with a report of a patient whose SR was accompanied by cytogenetic remission. There are less than 20 reports of SR since the 1980s. SR is by no means synonymous with cure, since the average duration of the remission is only 7·1±9·2 months. Neither infections nor transfusions are absolute requirements of SR.
Successful re‐treatment of an anti‐interferon resistant polycythaemia vera patient with lymphoblastoid interferon‐αN1 and in vitro studies on the specificity of the antibodies
Authors: Brand C. M. , Leadbeater L. , Budiman R. , Lechner K. , Gisslinger H.
Published: March 2008

A polycythaemia vera patient who initially responded to recombinant IFN‐α2 (rIFN‐α2) treatment developed neutralizing antibodies (NA) against it and lost response. Despite raising the dose, clinical resistance persisted and NA increased when two alternative rIFN‐α2 preparations were used. When treatment was switched to lymphoblastoid IFN‐α (lyIFN‐αN1), clinical response was restored and maintained. During re‐treatment, NA specific for the earlier rIFN‐α2 preparations redeveloped and cross‐reacted extensively with each other but not with ‘whole’lyIFN‐αN1 and only minimally with the lyIFN‐α2 subtype within it. These findings demonstrate the relevance of NA specificities in the re‐treatment of antibody‐compromised patients.
Control of polycythaemia vera with photochemotherapy in a patient with cutaneous T‐cell lymphoma
Authors: Cottrill Carolyn , Geller Alex , DiSpaltro Francis X. , Weissglass Bennett , Klainer Albert S. , Bisaccia Emil
Published: March 2008

We report the use of photochemotherapy in a patient with polycythaemia vera (PV) and cutaneous T‐cell lymphoma (CTCL). The patient was diagnosed with PV and required monthly phlebotomy. Subsequently he was diagnosed with CTCL and was started on photochemotherapy. A clinical improvement in both conditions, with haematocrit stabilizing at a level of 50%, eliminated the need for further phlebotomy. It is conceiveable that photochemotherapy induced the clinical and laboratory improvement in PV by targeting the aberrant clonal haemopoietic cell, similar to the mechanism thought to be operative in CTCL, a clonal helper T cell malignancy.
Therapy‐related acute promyelocytic leukaemia
Authors: Bhavnani M. , Azzawi S. Al , Liu Yin J. A. , Lucas G. S.
Published: March 2008

We report three patients with acute promyelocytic leukaemia (APL) occurring after treatment for other malignant disorders. One patient had had razoxane (a drug affecting DNA topoisomerase II) for cancer of the colon, and the other two had had treatment for cancer of the breast. Two out of the three patients went into complete remission. We review the published literature on therapy‐related acute promyelocytic leukaemia (t‐APL) and suggest that it is a genuine clinical entity which may be caused by drugs affecting DNA topoisomerase II, and has a prognosis similar to de novo APL.
The very‐long‐term course of polycythaemia: a complement to the previously published data of the Polycythaemia Vera Study Group
Authors: Najean Yves , Dresch Catherine , Rain Jean‐Didier
Published: March 2008

The very‐long‐term follow‐up of patients initially included in the PVSG protocols provides useful information. The excess risk of cancer after chlorambucil appears to persist for 5 years after stopping this treatment. The risk of leukaemia induced by marrow suppression (32P or chemotherapy) was marked before the 10th year, but low thereafter. Phlebotamy is unacceptable as permanent treatment because of the poor clinical tolerance and the frequency of vascular complications. This treatment is also associated with a risk of early progression towards myelofibrosis with myeloid splenomegaly.

In the very long term, 15 years or more after the diagnosis, this complication is the major clinical risk, affecting almost 50% of our patients surviving at this time. The prevention of this type of complication could constitute one of the objectives of future protocols dealing with this disease.
Patients with severe von Willebrand disease are insensitive to the releasing effect of DDAVP: evidence that the DDAVP‐induced increase in plasma factor VIII is not secondary to the increase in plasma von Willebrand factor
Authors: Cattaneo Marco , Simoni Loredana , Gringeri Alessandro , Mannucci Pier Mannuccio
Published: March 2008

Summary It is generally held that factor VIII (FVIII) does not increase in the plasma of severe von Willebrand disease (vWD) patients treated with DDAVP because they lack von Willebrand factor (vWF), which is the plasma carrier for FVIII. To test this hypothesis, FVIII plasma levels were monitored in severe vWD patients treated with DDAVP after normalization of vWF plasma levels with infusions of cryoprecipitate. Each of four severe vWD patients underwent four different treatments at intervals of at least 15 d: (1) cryoprecipitate plus DDAVP; (2) cryoprecipitate plus saline; (3) cryoprecipitate plus recombinant FVIII (rFVIII); (4) saline plus rFVIII. Cryoprecipitate increased the plasma levels of FVIII and vWF. The infusions of saline or DDAVP after cryoprecipitate did not further increase FVIII and vWF plasma levels and had no effect on the plasma levels of tissue plasminogen activator (tPA), which are raised by DDAVP in normal subjects and in patients with vWD of other types. The infusion of rFVIII further increased by 182 ± 32 U/dl (mean ± SEM) the plasma levels attained after cryoprecipitate, which disappeared from the circulation with a half‐life of 11.95 ± 0.86 h. In contrast, the infusion of rFVIII after saline increased by only 107 ± 18 U/dl the plasma levels of FVIII, which disappeared from the circulation with a half‐life of 2.68 ± 0.14 h, indicating that the vWF infused with cryoprecipitate is able to bind additional FVIII. These studies indicate that DDAVP does not increase the plasma levels of FVIII in patients with severe vWD even after normalization of plasma vWF. The possibility is discussed that severe vWD patients may be insensitive to the releasing effect of DDAVP.
The contribution of the haematocrit to thrombocytopenic bleeding in experimental animals
Authors: Blajchman Morris A. , Bordin José O. , Bardossy Leslie , Heddle Nancy M.
Published: March 2008

Summary Clinical studies in anaemic uraemic patients have shown that increasing the haematocrit with either red blood cell (RBC) transfusions or erythropoietin corrects the prolonged bleeding time (BT) often seen in such individuals. In this present study we evaluated experimentally the effect of the haematocrit on the BT using a microvascular BT technique in New Zealand White rabbits. The correlation between haematocrit and BT was studied in both normal and thrombocytopenic rabbits. In non‐thrombocytopenic animals the microvascular BT varied inversely with the haematocrit (r=‐0.799); animals with haematocrit levels above 35% having significantly shorter BTs than animals with haematocrit values lower than 35% (P < 0.001). To assess the role of the haematocrit on the BT in thrombocytopenic animals, thrombocytopenia was induced by a combination of γ‐irradiation and heterologous platelet antiserum. Such experiments showed that anaemic rabbits had significantly longer BTs than non‐anaemic animals with a similar degree of thrombocytopenia (P=0.0001). These data thus provide evidence that anaemia contributes significantly to the prolonged BT in both thrombocytopenic and non‐thrombocytopenic rabbits, and that RBC transfusions are capable of shortening the BT in thrombocytopenic anaemic animals. While results obtained from animal models cannot necessarily be extrapolated to the clinical situation, the fact that haematocrit influences the BT must be taken into account in the assessment of anaemic patients, particularly those who may have an associated haemostatic disorder.
α and β thalassaemia among Chinese children in Guangxi Province, P.R. China: molecular and haematological characterization
Authors: Liang R. , Liang S. , Jiang N. H. , Wen X‐J. , Zhao J‐B. , Nechtman J. F. , Stoming T. A. , Huisman T. H. J.
Published: March 2008

Summary We have studied nearly 100 patients with β thalassaemia major and 60 patients with Hb H disease who were attending the Haematology Clinic of Guangxi Medical College. Treatment of the patients was limited and only a few patients with β‐thalassaemia major received blood transfusion(s). As a result, the severe anaemia has led to early death at 3–4 years for β+‐thalassaemia homozygotes, and 8–12 years for β+‐thalassaemia homozygotes. Four β‐thalassaemia alleles are responsible for nearly 90% of all β‐thalassaemia chromosomes. This information has resulted in the initiation of a prenatal testing programme at the local level. The patients with Hb H disease maintained a haemoglobin level of 6–10 g/dl and early death was infrequently observed. The SEA deletion was the major type of α‐thalassemia‐1, while three smaller deletions (−2.7, −3.7 and −4.2 kb) and two nondeletional α‐thalassaemia determinants (Hbs Constant Spring and Quong Sze) were the α‐thalassaemia‐2 types.
Stereological studies on red corpuscle size produce values different from those obtained using haematocrit‐ and model‐based methods
Authors: Mayhew T. M. , Mwamengele G. L. M. , Self T. J. , Travers J. P.
Published: March 2008

Summary The routine methods for estimating erythrocyte size (volume and surface area) are not generally unbiased. Using geometric models introduces a bias determined by the ways in which real red blood corpuscles depart in size and shape from the simplistic ideal. Employing haematocrit and red corpuscle count to estimate mean corpuscular volume relies on an implicit assumption that erythrocytes fill three‐dimensional space. However, a biconcave circular disc cannot be space‐filling. The bias is positive and its magnitude depends on erythrocyte geometry and packing density within the centrifuge pellet.

We have estimated the volumes, surface areas and shape factors of red blood corpuscles in normal adults (three males, three females) using transmission electron microscopy and stereology. We confirm the lack of sexual dimorphism and find that the mean corpuscular volume is 44 fl (between‐subject coefficient of variation 5%), mean membrane surface area is 87 μm2 (6%) and the shape factor is 18.4 fl fl (11%). The surface is 44% greater than that of a sphere of equal volume. The possible reasons for discrepancies with standard haematological values are discussed. On the basis of observed volumes, we predict that red blood corpuscles account for only 51% of the volume of the red column in standard haematocrit pellets.
Plasma soluble interleukin‐2 receptor in patients with primary myelofibrosis
Authors: Wang Jen C. , Wang Anne
Published: March 2008

Summary Using an enzyme‐linked immunosorbant assay (ELISA) test, the level of soluble Tac peptide, one chain of the human interleukin‐2 receptor, was measured in the plasma of 26 patients with primary myelofibrosis (MF), seven patients with polycythaemia vera and 11 normal controls. The plasma soluble interleukin‐2 receptor (sIL‐2R) was found to be significantly elevated in patients with primary MF compared to polycythaemia vera or controls (P<0·01), while the plasma sIL‐2R of patients with polycythaemia vera also was found to be significantly elevated compared to controls (P<0·01). The significantly elevated value of sIL‐2R seen in primary MF may be secondary to T cell activation resulting from autoimmune phenomena, and myeloblast activation with release of sIL‐2R may also be a contributing factor. In primary MF, plasma sIL‐2R levels were also found to be correlated to survival, circulating blast cell counts, and thrombocytopenia, but not to white blood cell counts, LDH levels, degree of marrow fibrosis, or degree of splenomegaly. Patients with primary MF with higher titre of plasma sIL‐2R had a shorter survival. Further studies involving more patients and longer follow‐up may substantiate that plasma sIL‐2R is an important prognostic indicator in primary MF.
Adult T‐cell leukaemia/lymphoma featuring a large granular lymphocyte leukaemia morphologically
Authors: Sakamoto Yukihiro , Kawachi Yasunori , Uchida Tatsumi , Abe Takako , Mori Masaharu , Setsu Kojun , Indo Naohito
Published: March 2008

Summary A 70‐year‐old man from an endemic area of human T‐cell lymphotropic virus type I (HTLV‐I) developed rapid generalized lymphadenopathy and abdominal tumours. The white blood cell count was 198·3%× 109/l with 93% lymphocytes, 66·3% of which expressed large granular lymphocytes (LGLs). Bone marrow and lymph nodes were also infiltrated by LGLs. Surface markers were positive for CD4, CD25 and HLA‐DR, and negative for CD3, CD8, CD16, CD56 and CD57. A monoclonal integration of HTLV‐I proviral DNA was demonstrated on these LGLs by Southern blot hybridization analysis. This fact indicates that some adult T‐cell leukaemia/lymphoma may morphologically present LGL leukaemia.
Interleukin‐1α as an autocrine growth factor for acute lymphoblastic leukaemia cells
Authors: Mori Naoki , Shirakawa Fumihiko , Murakami Shuichi , Oda Susumu , Eto Sumiya
Published: March 2008

Summary We describe a case of B‐lineage acute lymphoblastic leukaemia (ALL) with proliferation of leukaemic cells through an interleukin‐1α (IL‐lα) autocrine mechanism. Flow cytometric analysis using the IL‐1 receptor type II (IL‐1Rt II) monoclonal antibody (mAb) demonstrated the expression of the IL‐1Rt II on leukaemic cells; this is the first report in IL‐1RtII‐positive freshly isolated ALL cells from a patient. In accordance with the expression of IL‐1RtII, the leukaemic cells proliferated in response to exogenous IL‐1α. In addition, anti‐IL‐1α mAb markedly inhibited the spontaneous cell proliferation. Furthermore, Northern blot analysis detected IL‐1α mRNA without any stimulation. These observations suggest that IL‐1α may play an important role as an autocrine growth factor in some cases of ALL.
Cytogenetic conversion in a case of polycythaemia vera treated with interferon‐alpha
Authors: Messora Carlo , Bensi Laura , Vecchi Angela , Longo Rosina , Giacobbi Francesca , Temperani Paola , Bevini Mirco , Emilia Giovanni , Sacchi Stefano
Published: March 2008

Summary Polycythaemia vera is a clonal disorder of the haemopoietic stem cell causing a pathologic expansion of the erythroid and sometimes the megakaryocytic and myeloid elements. In order to avoid the possible mutagenic effects of radioactive phosphorus, alkylating agents and hydroxyurea, since 1988 α‐IFN has been used for the treatment of PV and has been shown to induce and maintain haematological remission. We describe a 24‐year‐old PV patient with chromosomal abnormalities who achieved not only a reduction of the proliferation of erythroid elements and reticulin content in the bone marrow, but also a complete cytogenetic remission after IFN treatment.
GATA‐1 is expressed in acute erythroblastic leukaemia
Authors: Ekert Henry , Sievers Eric L. , Tan Andrew , Martin David I. K. , Smith Franklin O. , Barnard Dorothy , Bernstein Irwin D.
Published: March 2008

Summary Previous studies have demonstrated the expression of GATA‐1 (a DNA‐binding nuclear protein) in erythrocytes, megakaryocytes, eosinophils, basophils, mast cells, early marrow progenitor cells and in mouse and human erythroid leukaemia cell lines. We studied 31 bone marrow specimens from patients with acute myeloid leukaemia (AML) for GATA‐1 expression by reverse transcriptase/polymerase chain reaction (RT/PCR) analysis. GATA‐1 expression was detected in all of the patients with erythroleukaemia, and in one of nine patients with megakaryoblastic leukaemia, but absent from 17 patients with French‐American‐British (FAB) M1–5 leukaemia. In AML, GATA‐1 expression is indicative of differentiation to the erythroid and possibly megakaryocytic lineages, analogous to its expression in normal haemopoiesis.
Parvovirus B19‐associated haemophagocytic syndrome with prominent neutrophilia
Authors: Tsuda Hiroyuki , Maeda Yosuke , Nakagawa Kazumi , Nakayama Masafumi , Nishimura Hachiro , Ishihara Akira , Miyayama Haruhiko
Published: March 2008

Summary We describe a patient with both haemophagocytic syndrome and acute myocarditis probably associated with parvovirus B19 infection. The patient had a marked neutrophilia instead of neutropenia more usually observed in virus‐associated haemophagocytic syndrome (VAHS). Endogenous serum concentrations of macrophage colony‐stimulating factor (M‐CSF), granulocyte colony‐stimulating factor (G‐CSF), and tumour necrosis factor‐α (TNF‐α) were higher than normal, suggesting that these cytokines may be involved in the genesis of the observed syndrome.
Platelet aggregation in response to four low molecular weight heparins and the heparinoid ORG 10172 in patients with heparin‐induced thrombocytopenia
Authors: Keeling D. M. , Richards E. M. , Baglin T. P.
Published: March 2008

Summary A simple rapid platelet aggregation test was used to evaluate cross‐reactivity of four low molecular weight heparins and the heparinoid ORG 10172 in three patients with heparin‐induced thrombocytopenia. The low molecular weight heparins cross‐reacted in 11 out of 12 tests. The heparinoid ORG 10172 did not cross‐react in any of the patients. One patient was treated with ORG 10172 and thrombocytopenia resolved.
Haemopoietic progenitor cells are reduced in aplastic anaemia
Authors: Scopes John , Bagnara Maree , Gordon‐Smith Edward C. , Ball Sarah E. , Gibson Frances M.
Published: March 2008

Summary We investigated the frequencies of early populations of progenitors in aplastic anaemia (AA) bone marrow, from patients with a range of disease severity, compared with normal. Double‐colour immunofluorescent staining for CD34 and CD33 was carried out on bone marrow mononuclear cells (BMMC) and analysed using fluorescence activated cell sorting (FACS), AA CD34+ cells were reduced by 68% compared to normal. In addition, AA CD33+ cells and the three progenitor subsets (CD34+/CD33–, CD34+/CD33+ and CD34–/CD33+) were reduced by 44–80%. Our data lend further support for an early stem cell deficiency in AA.
Evaluation of mixed chimaerism and origin of bone marrow derived fibroblastoid cells after allogeneic bone marrow transplantation
Authors: Tanaka Junji , Kasai Masaharu , Imamura Masahiro , Masauzi Nobuo , Ohizumi Hiroko , Matsuura Atsushi , Morii Ken , Kiyama Yoshio , Naohara Tohru , Saitoh Masao , Higa Toshio , Sakurada Keisuke , Miyazaki Tamotsu
Published: March 2008

Summary We assessed the origin of bone marrow derived fibroblastoid cells (BMF) in long‐term cultures of 13 samples obtained from nine patients after allo‐BMT by polymerase chain reaction (PCR) amplification of MCT118, one of the variable number of tandem repeats regions (VNTR). BMF showed a complete recipient pattern in nine samples obtained from seven patients; however, a recipient‐predominant mixed chimaeric pattern was detected in BMF from four patients. Also, two of the four patients died with bone marrow hypoplasia. These data suggest that mixed chimaeric pattern of BMF may be correlated with bone marrow hypoplasia.
The in vitro effects of stem cell factor, interleukin 3 and granulocyte‐macrophage colony stimulating factor on haemopoietic progenitor cells from premature infants
Authors: Westwood N. B. , Chung R. , Emmerson A. J. B. , Pearson T. C.
Published: March 2008

Summary. Circulating haemopoietic progenitor cells from premature infants were assessed for their ability to respond to interleukin 3, granulocyte‐macrophage colony stimulating factor and stem cell factor (SCF) in vitro. All three cytokines increased the number of colonies derived from burst forming units erythroid (BFU‐E), colony forming units granulocyte‐macrophage (CFU‐GM) and multi‐lineage progenitors (CFU‐Mix) grown in the presence of erythropoietin (Epo). The size and haemoglobin content of BFU‐E derived colonies also increased in the presence of the cytokines. Of those tested, SCF was found to be the most potent additive to Epo for the enhanced growth of BFU‐E and CFU‐Mix. In short‐term liquid cultures without Epo, SCF alone induced globin synthesizing cells. Progenitors from premature infants were at least as responsive to all three cytokines as those from healthy adults. The use of SCF in combination with Epo in the prevention or treatment of anaemia in premature infants warrants further investigation.
Immunodeficiency and IL‐6 production by peripheral blood monocytes in multicentric Castleman's disease
Authors: Ishiyama Taijiro , Nakamura Shinji , Akimoto Yoshihisa , Koike Michiaki , Tomoyasu Shigeru , Tsuruoka Nobuyoshi , Murata Yaeko , Sato Tadatsugu , Wakabayashi Yoshihisa , Chiba Shyozo
Published: March 2008

Summary. To study the pathogenesis of multicentric Castleman's disease (MCD), IL‐6 producing cells and immune function were investigated in four MCD patients. The expression of IL‐6 mRNA in one MCD lymph node was analysed by in situ hybridization. IL‐6 mRNA expressing cells were scattered in the interfollicular areas and did not resemble plasma cells. Spontaneous IL‐6 production was detected in the culture supernatants of peripheral blood mononuclear cells (PBMNC) from four patients. The IL‐6 producing cells among the PBMNC were found to be monocytes by both in situ hybridization and immunohistochemistry. We evaluated immune function in four MCD patients. These studies show: (1) a negative PPD skin test in 3/4 patients. (2) decreased IL‐2 production in 3/4 patients, (3) decreased T cell colony formation in 3/4 patients, (4) decreased NK activity and NK cell number in 2/4 patients, (5) increased soluble IL‐2 receptor in 4/4 patients, and (6) decreased CD4/CD8 ratio in 3/4 patients. These results show that MCD resembles, in several ways, acquired immunodeficiency syndrome (AIDS).
Regulation of transcription factors NFκB and AP‐1 following tumour necrosis factor‐α treatment of cells from chronic B cell leukaemia patients
Authors: Jabbar Shireen A. B. , Hoffbrand A. Victor , Wickremasinghe R. Gitendra
Published: March 2008

Summary. Malignant B lymphocytes from patients with B‐chronic lymphocytic leukaemia (B‐CLL) or hairy cell leukaemia (HCL) are refractory in vitro to mitogenic stimulation by several agents which trigger proliferation of normal B cells. Tumour necrosis factor (TNF) is a growth factor for these malignant cells, although the proliferative response is usually small. TNF regulates some of its cellular responses via induction of the transcription factors NFκB and AP‐1 (jun/fos). The induction of NFκB by TNF is mediated via a novel signalling pathway involving the generation of reactive oxidative intermediates. Induction of jun and fos proteins (polypeptide components of AP‐1) are mediated via pathways involving protein kinase C and the protein kinase encoded by the raf proto‐oncogene. Here we have used an electrophoretic mobility shift assay to show that TNF induced NF/B in malignant cells isolated from 3/3 HCL and 15/15 B‐CLL patients. By contrast. phorbol myristate acetate (PMA), a direct activator of protein kinase C. failed to activate this transcription factor in 1/1 HCL and 5/5 B‐CLL isolates. The induction of jun and fos proteins (as detected by Western blot analysis) showed greater heterogeneity. Nuclear jun was induced by TNF in 5/12 chronic B cell leukaemia isolates. PMA induced this protein in 4/5 samples. Nuclear fos was induced by TNF in only 2/12 isolates and by PMA in 2/5. The data suggest that the pathways for the activation of jun and fos by TNF are defective in some B‐CLL and HCL cells and that these defects may be heterogeneous. The induction of AP‐1 is crucial in securing the mitogenic response to TNF. It is therefore plausible that these lesions may contribute to the refractory nature of B‐CLL and HCL cells to proliferative stimuli in vitro.
N‐desmethylclozapine: a clozapine metabolite that suppresses haemopoiesis
Authors: Gerson Stanton L. , Arce Cheryl , Meltzer Herbert Y.
Published: March 2008

Summary. Clozapine, a novel antipsychotic drug that is particularly effective in treatment‐resistant schizophrenia, causes severe agranulocytosis of unknown aetiology in approximately 0·8% of U.S. patients. We evaluated potential toxic mechanisms of drug‐induced agranulocytosis. Clozapine, the two major metabolites N‐desmethylclozapine and N‐oxide clozapine, and five other clozapine derivatives were screened for toxicity to normal haemopoietic precursors. For all compounds except N‐des‐methylclozapine, toxicity to CFU‐GM, BFU‐E and CFU‐GEMM occurred at concentrations at least 10 times the normal serum levels reported in unaffected patients. In contrast, the LD50 for N‐desmethylclozapine was 2·5 μg/ml for CFU‐GM, 3·2 μg/ml for BFU‐E, and 2·4 μg/ml for CFU‐GEMM, only 3–6 times the normal serum concentration. Bone marrow from patients with acute clozapine‐induced agranulocytosis was not more sensitive to clozapine or N‐desmethylclozapine than bone marrow from normal donors. These studies suggest that N‐desmethylclozapine, the major metabolite of clozapine, is itself toxic or is further metabolized to an unstable compound which is toxic to haemopoietic precursors of both myeloid and erythroid lineages.
A novel haemophilia B defect due to partial duplication of the factor IX gene
Authors: Chan Vivian , Au P. , Lau P. , Chan T. K.
Published: March 2008

Summary. A patient with mild FIX deficiency was found to have partial duplication of the 3′ region of the gene, giving, in addition to the a normal gene, another piece of DNA containing exons 5′, 6′, 7′ and 8′ and the intervening sequences. Cloning and sequencing of the junction region revealed that crossover occurred at nt 31927 in the 3° untranslated region of one chromosome/chromatid and nt 10640 in intron 4 of the other. No homology or topoisomerase specific sequences were observed in the crossover region. PCR and sequencing of illegitimate FIX transcripts from the patient's lymphocytes showed at least three different species of mRNAs. Translation of two of these ‘novel’mRNAs should result in truncated proteins. Possibilities for the splicing of the mature mRNA are offered to explain the translation of a normal‐size FIX protein, which was the only product demonstrated on Western blot analysis.
Binding and uptake of exogenous isoferritins by cultured human erythroid precursor cells
Authors: Meyron‐Holtz E. G. , Fibach E. , Gelvan D. , Konijn A. M.
Published: March 2008

Summary. The interaction of extracellular human isoferritins with normal erythroid precursors developing in a two‐phase liquid culture was studied. Cells at the stage of polychromatic normoblasts exhibited substantial specific binding of radio‐iodinated placental isoferritins. Considerably more acidic isoferritin was bound than basic isoferritin. The binding of ferritin was significantly higher at 37°C than at 4°C. All of the 125I‐acidic isoferritin bound at 4°C, but only part of that bound at 37°C, could be dislodged by the addition of a 500‐fold excess of non‐labelled acidic isoferritin. Acidic isoferritin displaced radio‐iodinated acidic isoferritin from the erythroid cells more efficiently than intermediate or basic isoferritins. Kinetic analysis suggests a dissociation constant (Kd) of 3·9 × 10‐8m for acidic ferritin and 3·7 × 10‐7m for basic isoferritin. The average number of binding sites for acidic isoferritin was 1·3 × 105 per cell. The results point to specific binding and receptor‐mediated internalization for predominantly acidic isoferritin by developing human erythroid cells.
Successful use of all‐trans retinoic acid in acute promyelocytic leukaemia presenting during the second trimester of pregnancy
Authors: Harrison P. , Chipping P. , Fothergill G. A.
Published: March 2008

Summary. The development of acute leukaemia in pregnancy is a rare event, with an estimated incidence of 0·9–1·2 cases per 100 000 per year and with the majority of cases occurring in the second and third trimesters. We report a case of acute promyelocytic leukaemia (APL) developing during the second trimester of pregnancy which was successfully treated with all‐trans retinoic acid (ATRA) resulting in a complete morphological remission.
Changes in the phenotypic characteristics of eosinophils from patients receiving recombinant human interleukin‐2 (rhIL‐2) therapy
Authors: Rodgers Sheila , Rees Robert C. , Hancock Barry W.
Published: March 2008

Summary.We have studied the properties of eosinophils from 26 patients with malignant melanoma and 16 patients with renal cell carcinoma (RCC) who were entered into a phase II clinical trial using various schedules of low‐dose rhIL‐2 immunotherapy. Eosinophilia was observed in 65% of melanoma patients and 100% of renal patients when receiving rhIL‐2 therapy.

The eosinophil count increased up to 20‐fold approximately 5 d after the appearance of lymphocyte activation markers. This would be consistent with eosinophil kinetics and the release of soluble mediators, for example IL‐5, from lymphocytes. Eosinophils from eosinophilic patients became hypodense compared to their pre‐treatment levels as demonstrated by sedimentation through a discontinuous metrizamide density gradient; they also showed an increased expression of CD4, CD25 and CD11b cell surface activation markers. Eosinophil count could not be correlated to either patient survival or response to therapy in melanoma patients: however, patients with renal cell carcinoma demonstrated a significant correlation (P0·05) between eosinophil count and survival but not with clinical response. Therefore the maximum eosinophil count achieved during rhIL‐2 therapy is of prognostic significance in patients with renal cell carcinoma.
Successful treatment of transfusion‐associated graft‐versus‐host disease
Authors: Yasukawa Masaki , Shinozaki Fumiki , Hato Takaaki , Takada Kiyonori , Ishii Yuko , Tamai Tomonori , Takamatsu Hideyuki , Shiobara Shintaro , Yoshizaki Kenichi , Fujita Shigeru
Published: March 2008

Summary. We present an immunocompetent patient with transfusion‐associated graft‐versus‐host disease (GVHD), in which chimaerism of peripheral blood lymphocytes was demonstrated by analysis of a highly polymorphic genome. The patient was treated successfully with anti‐CD 3 monoclonal antibody, OKT3 and cyclosporin A. Although it is undoubtedly important to prevent transfusion‐associated GVHD by irradiation of cellular blood components, intensive therapy with OKT3 and cyclosporin A in the early phase of onset may be effective for treatment of this potentially fatal condition. The mechanism of the effectiveness of this treatment for transfusion‐associated GVHD is discussed.
Marked basophilia in acute promyelocytic leukaemia treated with all‐trans retinoic acid: molecular analysis of the cell origin of the basophils
Authors: Iwakiri Rika , Inokuchi Koiti , Dan Kazuo , Nomura Takeo
Published: March 2008

Summary. We report a patient with acute promyelocytic leukaemia who developed marked basophilia during all‐trans retinoic acid treatment. We studied genomic DNA and RNA extracted from the patient's peripheral leucocytes in order to determine the origin of the basophils. The RARα rearranged band in the Southern blot analysis and a chimaeric product of PML‐RARα by polymerase chain reaction were strongly visible before ATRA treatment, but at the time of maximal basophilia both of them were markedly diminished. These findings suggest that the basophils which appeared during the ATRA treatment are reactive in nature rather than a leukaemic clone.
Establishment of a human T‐cell line with deficient surface expression of glycosylphosphatidylinositol (GPI)‐anchored proteins from a patient with paroxysmal nocturnal haemoglobinuria
Authors: Masuda Tetsuya , Yonemura Yuji , Fujimoto Koji , Hidaka Michihiro , Nagakura Shoichi , Nakakuma Hideki , Hata Hiroyuki , Sanada Isao , Kawakita Makoto , Takatsuki Kiyoshi
Published: March 2008

A novel interleukin‐2 dependent T‐cell line, PMT‐2Y, was established from the peripheral blood of a patient with paroxysmal nocturnal haemoglobinuria (PNH) by human T lymphotropic virus type I (HTLV‐1)‐mediated transformation. PMT‐2Y cells are positive for CD2, CD3, CD4, CD25, T cell receptor αβ and HLA‐DR, but negative for CD1, CD7, CD8, CD19 and CD20, indicating that the clone belongs to a helper/inducer subset of T cells. PMT‐2Y cells have the monoclonal integration of HTLV‐I proviral DNA, suggesting that they derived from a single clone. Moreover, they lack surface expression of complement regulatory proteins such as DAF (CD55) and CD59, that are the most important glycosylphosphatidylinositol (GPI)‐anchored membrane proteins defective in haemopoietic cells of patients with PNH. Northern blot analysis, however, revealed the production of normal levels of DAF mRNAs. Thus, PMT‐2Y is derived from a PNH T cell clone and may be a useful model to study PNH.
An autoantibody to human plasma prekallikrein blocks activation of the contact system
Authors: Page J. D. , Cadena R. A. DeLa , Humphries J. E. , Colman R. W.
Published: March 2008

. A patient without a history of bleeding or thromboembolism presented with an activated partial thromboplastin time (aPTT) of 55.1 s (normal 24‐38 s). Incubation of the patient plasma with an equal volume of normal plasma failed to correct the aPTT. suggesting the presence of an inhibitor. The MRVVT (modified Russell Viper venom time) was normal, and the anti‐cardiolipin antibody titres were not elevated, indicating that the presence of a lupus anticoagulant was unlikely. Plasma prekallikrein (PK) measured by a coagulant assay (2 U/dl) was very low, but PK was in the low normal range (65%) when measured by an enzymatic assay (amidolytic) or by an antigenic assay (ELISA). The purified patient IgG reacted with purified PK. the heavy chain, and the 28 kD fragment of the heavy chain, indicating that it contained an autoantibody to PK. The purified IgG did not directly inhibit the amidolytic activity of kallikrein, but it did inhibit the activation of PK to kallikrein by activated factor XII. Activation of the contact system by dextran sulphate, as reflected by the cleavage of HK on a Western blot, was inhibited when the patient IgG was added to pooled normal plasma. The antibody appears to be oligoclonal with IgG1 being most abundant. followed by IgG4. This report appears to be the first of a spontaneously occurring antibody to prekallikrein.
Fine epitope mapping of monoclonal antibodies to the NH2‐terminal part of von Willebrand factor (vWF) by using recombinant and synthetic peptides: interest for the localization of the factor VIII binding domain
Authors: Jorieux Sylvie , Gaucher Christine , Piéatu Geneviégve , Chéarel Ghislaine , Meyer Dominique , Mazurier Claudine
Published: March 2008

. Two different approaches were used in order to define the epitope of three monoclonal antibodies (MoAbs) against the NH2‐terminal part of the mature subunit of von Willebrand factor (vWF) which contains its factor VIII (FVIII) binding site. First, a vWF cDNA fragment library using the bacteriophage λgt11 expression vector was screened with radiolabelled MoAbs. The epitope of each MoAb was defined, following sequence analysis, by the overlapping DNA sequence of immunoreactive clones. MoAb 32B12, a potent inhibitor of FVIII/vWF interaction, binds within the Glu35‐Ile81 sequence of vWF subunit. MoAb 14A12, a non‐inhibitory antibody, recognizes a sequence within Thr141‐Val220. MoAb 31H3, a partial inhibitory antibody, gives no positive clone. In the second method, a panel of 24 synthetic pentadecapeptides corresponding to the first NH2‐terminal 105 amino acid residues was used to block the binding of inhibitor MoAbs to immobilized vWF in an ELISA system. The localization of MoAb 32B12 epitope was confirmed and restricted to the Met51‐Ala60 sequence, The MoAb 31H3 binding to vWF is inhibited by two synthetic peptides with the overlapping sequence Cys66‐Gly76. All these data confirm that the FVIII binding site of vWF is not limited to the binding area (Thr78‐Thr96) of the previously described MoAbs inhibiting FVIII/vWF interaction but is composed of several key sequences.
Demonstration of a marked reduction in the amount of GPIIb in most type II patients with Glanzmann's thrombasthenia
Authors: Tomiyama Yoshiaki , Kashiwagi Hirokazu , Kosugi Satoru , Shiraga Masamichi , Kinoshita Seiji , Kanayama Yoshio , Kurata Yoshiyuki , Matsuzawa Yuji
Published: March 2008

Summary. In this study employing a sensitive immunoblot assay, we have characterized GPIIb and GPIIIa in thrombasthenic platelets from seven type II and four type I patients from 10 unrelated families. The amounts of GPIIb and GPIIIa were both markedly reduced in all these patients, and abnormal molecular weight GPIIb or GPIIIa was not detected. In all of four type I patients the amount of GPIIb was much lower than that of GPIIIa. In this study, however, we found that the amount of GPIIb was also lower even in six out of seven type II patients. Immunodepletion of patients’platelets with AP2 (a monoclonal antibody specific for the GPIIb‐IIIa complex), AP3 (specific for GPIIIa) or AMF7 (specific for αv) further confirmed that GPIIIa existed in excess, and demonstrated that excess GPIIIa were mostly in free form and not associated with GPIIb or αv. The reduction of GPIIb may represent an abnormality in GPIIb processing in these type II and type I thrombasthenic platelets. It remains unclear whether these two subgroups represent distinct categories.
A novel polyadenylation signal mutation in the α2‐globin gene causing α thalassaemia
Authors: Harteveld C. L. , Losekoot M. , Haak H. , Heister J. G. A. M. , Giordano P. C. , Bernini L. F.
Published: March 2008

Summary. In a family of Indian origin we have identified a deletion of two bases at the polyadenylation signal sequence of the α2‐globin gene (AATAAA AATA). Three individuals heterozygous for this mutation display an αo‐thalassaemialike phenotype. Single‐stranded conformation analysis and automatic sequencing showed no additional mutations in either α1‐ or α2‐globin genes. A previously described polyadenylation sequence mutation (AATAAA AATAAG), αTSaudiα, causes HbH disease in homozygotes. In this study the patients heterozygous for the AATA(‐AA) mutation show a similar phenotype observed in the αTSaudiα heterozygotes. This confirms the observation that the inefficient transcriptional termination due to mutations of the polyadenylation sequence of the α2‐gene might interfere with the α1‐gene expression.
Recombinant human erythropoietin for the treatment of the anaemia associated with autologous bone marrow transplantation
Authors: Ayash Lois J. , Elias Anthony , Hunt Myla , Demetri George , Wheeler Catherine , Tepler Isidore , Schwartz Gary , Mazanet Rosemary , Reich Elaine , McCauley Mary , Antman Karen , Anderson Kenneth C.
Published: March 2008

Summary. Patients with solid tumours undergoing highdose chemotherapy with autologous bone marrow transplantation use an average of 10 units of packed red blood cells (PRBC) while awaiting haemopoietic reconstitution. They are also known to have inappropriately low endogenous erythropoietin levels for their degree of anaemia. This pilot study was designed to determine the effects of recombinant human erythropoietin (rHuEPO) on erythroid recovery and PRBC transfusion requirements.

Ten patients received high‐dose chemotherapy (days ‐ 7 to ‐3), bone marrow reinfusion (day 0), and then rHuEPO (day 1 onward). RHuEPO (200 units/kg intravenous bolus daily), along with iron supplementation, was administered for 28 d or until a haematocrit (Hct) of 35% (independent of transfusions) was reached, whichever occurred first. PRBCs were routinely given for Hct 25% and platelets for counts < 20000/μl.

Eight (80%) patients developed a brisk reticulocytosis (median peak reticulocyte count 0.32 × 109/l) and a haematocrit 30% independent of red blood cell transfusions within 32 d of receiving marrow, as compared to 20/37 (54%) similarly treated controls. An unexpected finding was the more rapid engraftment in myeloid and platelet lineages in a subset of rHuEPO‐treated patients. Quick return of red blood cells (17 v 33d) (P = 0.0001), platelets (14 v 19d) (P = 0.04), and neutrophils (13 v 25d) (P = 0.01) (with circulating myeloblasts and early myeloid forms) characterized recovery from an ifosfamide‐based intensification with rHuEPO support. Similar trilineage enhancement of haemopoiesis did not occur with the possibly more myeloablative cyclophosphamide‐based regimens. Despite the enhancement by rHuEPO on reticulocytosis, there was no significant decrease in PRBC transfusion requirements.

RHuEPO proved to be a well‐tolerated agent in enhancing reticulocytosis following high‐dose chemotherapy. further study to elucidate the activity of erythropoietin on both erythroid and non‐erythroid growth and maturation appears warranted.
Pure red cell aplasia: further evidence of T cell clonal disorder
Authors: Maung Zor T. , Norden Jean , Middleton Peter G. , Jack Fergus R. , Chandler John E.
Published: March 2008

Summary. We report a patient with pure red cell aplasia and type I autoimmune polyglandular syndrome who also had an expansion of suppressor T lymphocytes in the peripheral blood. Southern blotting of DNA from these cells suggested T cell receptor (TCR) γ gene rearrangement. We confirmed true clonality of this by amplification of the gene rearrangement using the polymerase chain reaction and subsequent analysis of the product by gene cloning and DNA sequencing.
Familial occurrence of LGL leukaemia
Authors: Loughran Thomas P. , Kidd Jr. Pamela , Poiesz Bernard J.
Published: March 2008

Summary. The aetiology of LGL leukaemia is now known: however, we recently detected HTLV‐II from such a patient. We describe here the occurrence of LGL leukaemia in a mother and her son. Serum from the son reacted to HTLV‐I/II gag proteins, but not to recombinant HTLV‐I env protein p21e; serum from the mother was negative. PCR analyses in both patients were negative for pX and pol sequences shared by HTLV‐I/II and also for specific gag sequences of HTLV‐I and HTLV‐II. These data show that familial cases of LGL leukaemia are not associated with prototypical HTLV‐I or HTLV‐II infection.
Reversal of diabetes associated with escape of myeloma: evidence for inappropriate IGF‐II secretion
Authors: Snowden J. A. , Greaves M. , Page K.
Published: March 2008

Summary. We report the sudden and dramatic reversal of maturity onset diabetes in a 57‐year‐old woman in association with relapse of IgA myeloma diagnosed 3 years earlier. Prior to the relapse of the myeloma, twice daily insulin had been administered at a dose which had been stable for 3 years. However, the same dose produced hypoglycaemic coma at the time of relapse and, subsequently, blood glucose was controlled by diet alone. There had been no signifficant change in weight or renal function prior to the hypoglycaemic episode. Investigations showed a suppressed fasting serum insulin level in association with an inappropriately high serum level of IGF‐II compared with IGF‐I and a ‘big’IGF‐II concentration at the upper end of the normal range. Pituitary, adrenal and liver disease, as well as the autoimmune insulin syndrome, were excluded. The findings are consistent with the hypothesis that the plasma cell tumour was associated with excessive production of insulin‐like peptides with consequent reduction in the blood glucose level.
Peripheral mononuclear cells of haemophiliacs with chronic liver disease are infected with replicating hepatitis C virus
Authors: Henin Christine , Makris Mike , Brown Julie , Peake Ian R. , Preston Eric F.
Published: March 2008

Most haemophiliacs treated with non‐virally inactivated concentrates have been infected with the hepatitis C virus (HCV). 19/21 (90%) patients with HCV antibody were found to have HCV RNA in their serum and in 16 of these it was also detected in peripheral blood mononuclear cells (PBMC). We used specific primers to detect the negative (replicative) strand of this single positively stranded virus. The negative strand was detected in the serum or PBMC of 16 patients indicating active replication outside the liver. The detection of replicating HCV RNA in the cells of HIV negative haemophiliacs could explain at least some of the immunological abnormalities previously reported in these patients and attributed to clotting factor concentrates.
Long‐term survivors in chronic granulocytic leukaemia: a study by the International CGL Prognosis Study Group
Authors: Cervantes Francisco , Robertson Judith E. , Rozman Ciril , Baccarani Michele , Tura Sante , Gómez Germán A. , Braun Thomas J. , Clarkson Bayard D. , Pereira Arturo
Published: March 2008

SUMMARY. The purposes of the present work were to identify the initial characteristics associated with long‐term survival in chronic granulocytic leukaemia (CGL) and to analyse the accuracy of prognostic models in identifying long‐term survivors. 813 Philadelphia (Ph) chromosome‐positive, nonblastic CGL patients from six American and European institutions, the majority treated conventionally, with a minimum follow‐up < 10 years, were studied. Stepwise logistic regression was performed to ascertain the association between the initial clinicohaematological variables and survival ≤8 years, and a prognostic index was derived. The usefulness of both Sokal's and the new prognostic index to identify long‐term survivors was assessed by calculating their positive and negative predictive accuracies, sensitivity and specificity. Median survival of the series was 45 months (range 1‐255), with 784 patients (96.4%) having died and 109 (13.4%) surviving 8 years or longer. Younger age, smaller spleen, platelets ≤600 × 109/I, and lower blood blast percentage were associated with survival ≤8 years; platelets ≤ 600 × 109/I and lower blood blast percentage were the predictive factors in patients 50 years old or younger. Two‐thirds of long survivors belonged to Sokal's low‐risk group, but the positive predictive accuracy and specificity for prolonged survival of Sokal's index were very low. This was also the case for the new predictive index. In 366 patients with information on the presence or not of additional karyotypic abnormalities at diagnosis, younger age, platelets ≤ 600 × 109/I, lower blood blast percentage, and absence of additional cytogenetic abnormalities were associated with survival ≤8 years. The addition of the cytogenetic data improved the positive predictive accuracy and specificity of the model, albeit modestly. We conclude that, although most CGL long survivors show favourable prognostic features at presentation, clinicohaematological data do not allow a precise definition of such a subpopulation.