bluuebunny/crossref_metadata_2025 huggingface dataset:
Haemophilia B mutations in a complete Swedish population sample: a test of new strategy for the genetic counselling of diseases with high mutational heterogeneity
Authors: Green P. M. , Montandon A. J. , Ljung R. , Bentley D. R. , Nilsson I.‐M. , Kling S. , Giannelli F.
Published: March 2008

Summary. Carrier and prenatal diagnosis based on the identification of the gene defect (direct diagnosis) increases the proportion of haemophilia B families that can be offered precise genetic counselling from the 50–60% attainable by DNA markers, to 100%, and they also provide information on the molecular biology of the disease. We propose that in order to maximize the practical and scientific benefits of direct diagnosis the gene defect of complete (possibly national) populations of patients should be characterized and the information stored in appropriate confidential databases.

We demonstrate the feasibility of such a strategy by characterizing the mutations of all the patients registered with the Malmö haemophilia centre.

These patients (44♂ and 1♀) are from 45 unrelated families and 24 (53%) have negative family history. The 25 patients with similar reduction of factor IX:C and factor IX:Ag (24♀+1♂) have: two gross deletions, three frame‐shifts, four translation stops, six mutations expected to affect pre‐mRNA splicing and 10 amino acid substitutions. The six patients with greater reduction of factor IX:C than factor IX:Ag and the seven with reduced IX:C and normal IX:Ag have only amino acid substitutions. Patients with inhibitors have: one complete deletion, one frameshift and three translation stops. One patient has both a translation stop and a functionally neutral amino acid substitution (His257→Tyr). Characterization of the factor IX mutation was successful in every case, usually entailed 4 person‐days work, and has led to the identification of 12 amino acid residues essential for the factor IX structure and function.
An HphI‐polymorphism in exon 28 of the von Willebrand factor gene, and its frequency among patients with various forms of von Willebrand's disease
Authors: Donnér Mikael , Holmberg Lars , Kristoffersson Ann‐Charlotte , Nilsson Inga Marie
Published: March 2008

Summary. Besides having a large number of restriction fragment length polymorphisms (RFLP) the von Willebrand factor (vWF) gene contains several sequence polymorphisms in the coding regions. Eight nucleotide substitutions have been reported in two or more independent cDNA clones. Four of them give rise to amino acid substitutions, two of which are in the mature vWF subunit (at positions 26 and 709). We have investigated a previously suggested putative alaninethreonine polymorphism at position 618 of the mature subunit in normal subjects and patients with various types of von Willebrand's disease (vWD). The codon for amino acid 618 is located in exon 28, which encodes several important vWF functional domains. We amplified the whole exon 28 and parts of it by polymerase chain reaction (PCR) and distinguished gene from pseudogene sequences. The alanine→lthreonine (G→A) substitution was studied with restriction enzyme cleavage of the products, since it creates a new HphI site. Moreover, in two individuals we confirmed the polymorphism by cDNA sequencing. In 23 normals the frequencies of the h‐ (Ala) and the h+ (Thr) alleles were 0·50/0·50. In eight patients with type III vWD from seven different families, the h– allele was present in 13 of 16 genes, but whether this signifies a common mutation in some of the patients is not known. In types I and II. both alleles were present in roughly similar proportions. Owing to the high frequency of heterozygosity, the polymorphism should prove useful as an aid in genetic counselling.
A novel δº mutation in cis with Hb Knossos: a study of different genetic interactions in three Egyptian families
Authors: Olds R. J. , Sura T. , Jackson B. , Wonke B. , Hoffbrand A. V. , Thein S. L.
Published: March 2008

Summary. We have defined the molecular basis of normal HbA2β‐thalassaemia associated with Hb Knossos. DNA sequence analysis of the δ globin gene in cis with βKnossos showed deletion of a single A in codon 59 leading to a premature termination at codon 60. This δº/βKnossos allele has been observed in three unrelated Egyptian families and associated with a single β haplotype (+ ‐ ‐ ‐ ‐ ++). One individual who was homozygous for the δº/βKnossos allele as well as heterozygous for a non‐deletional α thalassaemia, was completely clinically asymptomatic, while others have coinherited the δº/βKnossos allele with different β and α thalassaemia determinants. A study of the different genetic interactions giving rise to a spectrum of clinical phenotypes is reported.
Human T‐cell lymphotropic virus type I associated adult T‐cell leukaemia/lymphoma in Taiwan Chinese
Authors: Shih Lee‐Yung , Kuo Tseng‐Tong , Dunn Po , Liaw Shiumn‐Jen
Published: March 2008

Summary Twenty‐five Chinese patients with human T‐cell lymphotropic virus type I (HTLV‐I) associated adult T‐cell leukaemia/lymphoma (ATLL) were identified in Taiwan. No patients had been outside Taiwan and none were descendants of Japanese heritage. Their ages ranged from 28 to 71 years. There were 17 men and eight women. Main clinical and laboratory features at presentation were lymphadenopathy (16), skin lesions (11), hepatosplenomegaly (11), pulmonary lesions (11), hypercalcaemia (10) and bone marrow infiltration (14). Peripheral blood was characterized by leucocytosis with presence of pleomorphic abnormal lymphocytes but rare anaemia or thrombocytopenia. The clinical subtypes were acute in 15, chronic in three, smouldering in one, and lymphoma type in six. The immunopheno‐types of the ATLL cells were characterized by the expression of CD2+, CD4+, CD7‐, CD8‐ and CD25+. The overall prognosis was poor with a median survival of 5 months. The acute form had a significantly shorter survival (2 months) than lymphoma type (13 months). Susceptibility to various infections was common. Pulmonary complications accounted for 73% of the causes of death. The clinicopathologic features of ATLL in Taiwan are indistinguishable from those in HTLV‐I endemic areas. The present series adds to the knowledge of the worldwide pattern of the disease.
In vivo administration of granulocyte‐macrophage colony stimulating factor enhances neutrophil function in patients with myelodysplastic syndromes
Authors: Verhoef Gregor , Boogaerts Marc
Published: March 2008

Summary We studied the long‐term in vivo effects of recombinant granulocyte‐macrophage colony stimulating factor (rhGM‐CSF) on granulocyte functions in nine patients with myelodysplastic syndrome (MDS). The treatment schedule consisted of a 14 d course of rhGM‐CSF (250 μg/m2/d s.c.) for patients with refractory anaemia (RA) and refractory anaemia with ringed sideroblasts (RARS), while patients with refractory anaemia with excess of blasts (RAEB) and refractory anaemia with excess blasts in transformation (RAEBt) received a 14 d combination course of rhGM‐CSF (250 μg/m2 s.c.) and low dose cytosine arabinoside (20 mg/m2 s.c.). rhGM‐CSF increased the mean neutrophil count from 3.9 × 109/l to 44 × 109/l. Significant increases of myeloperoxidase content in granulocytes occurred during treatment (P=0.003). Phagocytosis and killing of Staph. aureus by granulocytes was markedly enhanced during treatment. Microbicidal capacity normalized in four out of six patients during GM‐CSF therapy. However, chemotaxis in response to zymosan‐activated serum (ZAS) and f‐Met‐Leu‐Phe (f‐MLP), was further impaired on the last day of treatment, which was associated with a marked increase in the expression of the granulocyte adhesion receptors CD11a (P=0.01), CD11b (P=0.002), CD11c (P=0.00015) and CD18 (P=0.0014). GM‐CSF therapy did not cause significant changes in hexose monophosphate (HMP)‐shunt activity, chemiluminescence, nor superoxide production. The present results show that in vivo administration of GM‐CSF is able to repair at least in part the neutrophil anomalies in patients with myelodysplastic syndrome (MDS), which might be useful in modulating host response to infections. However, increased adherence and impaired chemotaxis may explain some toxicities observed during treatment with GM‐CSF.
Thioguanine used in maintenance therapy of chronic myeloid leukaemia causes non‐cirrhotic portal hypertension. RESULTS FROM MRC CML II TRIAL COMPARING BUSULPHAN WITH BUSULPHAN AND THIOGUANINE
Authors: Shepherd P. C. A. , Fooks J. , Gray R. , Allan N. C.
Published: March 2008

Summary Portal hypertension with varices developed in 18/675 patients with chronic myeloid leukaemia (CML) in a randomized trial comparing busulphan with busulphan and thioguanine. All 18 had received the drug combination and none busulphan alone (P<0.0001). Ascites was also seen significantly more often in the combination arm (P>0.05). These results strongly suggest that the addition of thioguanine was responsible for the development of portal hypertension. The histological features were predominantly those of non‐cirrhotic portal hypertension—either idiopathic portal hypertension with minimal morphological abnormalities, nodular regenerative hyperplasia or in two cases leukaemic infiltration only was noted. Cirrhosis was present in 3/16 cases studied.

Both treatment groups developed abnormal liver function tests during the chronic phase, but particularly with progression of the disease. During chronic phase abnormalities were significantly more frequent in those receiving busulphan and thioguanine‐alkaline phosphatase (P<0.02), transaminases (P<0.04), bilirubin (P<0.05), multiple abnormalities (P<0.01). The development of portal hypertension was often associated with abnormalities of these tests; however, lack of specificity precludes their use as a predictor of subsequent clinical problems. Thioguanine confers no survival advantage in this disease. In view of its hepatotoxicity it should not be used routinely for maintenance of control in chronic phase CML.
Characterization of a human plasmacytoma line
Authors: Weinreich Stephanie S. , Kr.von dem Borne Albert E. G. , Van Lier René A. W. , Feltkamp Constance A. , Slater Rosalyn M. , Wester Mieke R. , Zeijlemaker Wim P.
Published: March 2008

Summary The TH line was established by bringing tumour cells from a multiple myeloma patient into suspension culture and subsequently cloning them by limiting dilution. The cultured cells show marked heterogeneity: there are ultrastructural differences between small and large TH cells, particularly with respect to the rough endoplasmatic reticulum (RER). Karyotyping revealed chromosome numbers in the triploid range, with many structural abnormalities, at the 14q32 region among others. A t(14:18) could not be demonstrated. TH was shown to have germline and a rearranged allele for kappa light chain, and only a single rearranged gene for heavy chain immunoglobulin. TH expressed PCA‐1, CD9, CD28 and CD38 antigens, HLA class II, RER and kappa light chain, but few or no other antigens associated with the B‐cell lineage. Light chain kappa and trace amounts of IgG3 were found intracellularly as well as in culture supernatant. The addition of IL‐6 to cultures of TH increased proliferation, as well as the secretion of kappa light chain and the membrane expression of CD28 and CD38 antigens. Because TH has relatively few B cell markers on its membrane, it may be useful for the induction of monoclonal antibodies specific for human plasma cells. It also provides a model for the demonstration that IL‐6 can act as a paracrine growth and differentiation factor for cells of myelomal origin.
Inherited platelet‐storage pool deficiency associated with a high incidence of acute myeloid leukaemia
Authors: Gerrard Jon M. , Israels Esther D. , Bishop Agnes J. , Schroeder Marlis L. , Beattie Laura L. , McNicol Archibald , Israels Sara J. , Walz Daniel , Greenberg Arnold H. , Ray Mano , Israels Lyonel G.
Published: March 2008

Summary A family with an inherited bleeding disorder extending over four generations, and multiple cases of myeloblastic and myelomonoblastic leukaemia was studied. Ten members of the family had, by history, a haemorrhagic diathesis. There were three documented cases of myeloblastic leukaemia, two documented cases of myelomonoblastic leukaemia and two more cases of leukaemia by history. In four of the cases the bleeding diathesis clearly antedated the leukaemia, in two by many years. The bleeding disorder is characterized by a long bleeding time, abnormal platelet aggregation, low platelet ADP and decreased numbers of platelet dense bodies consistent with a dense granule storage pool deficiency. The number of dense granules was decreased by immunofluorescence employing quinacrine or using an antibody to the dense granule membrane protein, granulophysin, confirming an absolute decrease in dense granule numbers rather than the presence of empty granule sacs. This congenital storage pool deficiency is associated with a high incidence of acute myeloid leukaemia in this family.
Activation of human platelets by exposure to a monoclonal antibody, PM6/248, to glycoprotein IIb‐IIIa
Authors: Hornby E. J. , Brown S. , Wilkinson J. M. , Mattock C. , Authi K. S.
Published: March 2008

Summary Monoclonal antibody PM6/248, which recognizes the GPIIb‐IIIa complex on human platelets, causes platelet aggregation in platelet‐rich plasma or in gel‐filtered platelet suspensions. Aggregation follows a concentration‐dependent lag phase and reaches a maximum at 8 μg/ml. High concentrations of antibody (< 30 μg/ml) produce complete inhibition of the aggregation response. Aggregation is accompanied by serotonin secretion and thromboxane A2 synthesis, neither of which are inhibited by high concentrations of antibody, and by the mobilization of intracellular Ca2+. The F(ab')2 fragment of PM6/248 does not cause platelet activation and pre‐incubation of platelets with this fragment inhibits all platelet responses stimulated by the whole antibody. Pre‐incubation with the F(ab')2 fragment of the anti‐FcγRII Mab, IV. 3, also inhibits all responses to PM6/248. These data indicate that platelet activation stimulated by PM6/248 is caused by cross‐linking of GPIIb‐IIIa to the FcγRII which stimulates signal transduction across the plasma membrane through a conformational change in the FcγRII.
Familial clustering of defective release of t‐PA
Authors: Petäjäu Jari , Rasi Vesa , Vahtera Elina , Myllylä Gunnar
Published: March 2008

Summary 91 unrelated patients with idiopathic or familial deep vein thrombosis (DVT) and 72 (34 with DVT) relatives from 26 families were screened for hypofibrinolysis by measuring tissue plasminogen activator antigen (t‐PA: Ag) after venous occlusion (VO) for 10 and 20 min and by measuring t‐PA inhibitor activity (PAI) at rest. 21 healthy subjects served as controls. Defective release of t‐PA: Ag was found in eight out of the 91 patients (9%). A partial family study of six of these eight patients was performed. This study included 10 family members with and 21 without DVT. A defective release of t‐PA: Ag was found in 50% (5/10) of the family members with DVT, which is significantly more frequent than the 9% (8/91) prevalence in the unrelated patients (P<0.001). Furthermore, 24% (5/21) of asymptomatic members of these families also had defective release of t‐PA: Ag. In the 18 families where the propositus had a normal level of t‐PA: Ag, none of the 24 studied family members with DVT had defective release of t‐PA : Ag. In contrast to the defective release of t‐PA: Ag, increased basal level of PAI did not show familial clustering. In conclusion, low release of t‐PA during VO shows familial clustering in a proportion of the cases.
The Corfu δβ thalassaemia mutation in Greece: haematological phenotype and prevalence
Authors: Traeger‐Synodinos Joanne , Tzetis Maria , Kanavakis Emmanuel , Metaxotou‐Mavromati Anna , Kattamis Christos
Published: March 2008

Summary The Corfu δβ thalassaemia mutation, a 7.2 kb deletion partially removing the (δ‐globin gene and a single nucleotide mutation (GA) at intervening sequence I (IVSI‐n5) in the β‐globin gene in cis, was first described in a family from Corfu; the carriers for this mutation had the unusual haematological phenotype of heterozygous β‐thalassaemia with normal levels of HbA2.

To investigate the frequency and haematological characteristics of Corfu δβ thalassaemia in Greece we analysed 25 unrelated normal HbA2 type 2 β‐thalassaemia heterozygotes and their 23 clinically affected offspring. Gene mapping demonstrated that nine (36%) of the 25 normal HbA2β‐thalassaemia heterozygotes were in fact Corfu δβ thalassaemia heterozygotes and of the 23 patients, two were Corfu δβ thalassaemia homozygotes and five compound heterozygotes for Corfu δβ thalassaemia and another β‐thalassaemia defect.

Detailed haematological analysis demonstrated that: the Corfu δβ thalassaemia mutation does not completely abolish the expression of the β‐globin gene; the HbA2 levels are slightly lower (P<0.01) and the HbF levels slightly higher (P<0.01) in Corfu δβ thalassaemia heterozygotes compared to β‐thalassaemia heterozygotes with the normal HbA2‐type 2 phenotype who do not have the Corfu δβ chromosome.
Combined flow cytometric assessment of cell surface antigens and nuclear TdT for the detection of minimal residual disease in acute leukaemia
Authors: Drach J. , And C. Gattringer , Huber H.
Published: March 2008

Summary. To define more precisely the immunophenotype of lymphoid blast cells, a new flow cytometric technique for the simultaneous detection of surface antigens and nuclear terminal deoxynucleotidyl transferase (TdT) was established. After staining for the cell surface marker, mononuclear cells were treated with paraformaldehyde (1 %) and methanol to permeabilize the cell membrane. Then the cells were stained by indirect immunofluorescence using a rabbit anti‐human TdT antibody. Dilution experiments were performed to reveal the sensitivity of the described flow cytometric assay: 0·02% leukaemic cells could reliably be detected above background level among normal peripheral blood lymphocytes. It is concluded that the double‐staining procedure described here is a sensitive tool that contributes to the detection of minimal residual disease in a substantial proportion of acute leukaemias.
Refractory myelomatosis treated with mitoxantrone in combination with vincristine and prednisone (NOP‐regimen): a phase II study
Authors: Gimsing Peter , Bjerrum Ole W. , Brandt Eystein , Ellegaard Josrgen , Evensen Stein A. , Hansen Mogens Mosrk , Hedenus Michael , Hippe Erik , Keldsen Nina , Palva Ilmari , Roudjer Stig , Talstad Ingebrigt , And Jan Westin , Wislosff Finn
Published: March 2008

Summary. In a phase II study, patients with refractory myelomatosis were treated with a combination chemotherapy (NOP regimen): mitoxantrone (bolus injection of 4 mg/m2 on days 1‐4), vincristine (continuous infusion of 0·4 mg/24h on days 1‐4) and prednisone (250 mg/d on days 1‐4 and 17‐20). The treatment was repeated every 4 weeks. Ninety‐two patients were treated after they were found refractory to treatment with melphalan and prednisone (and occasionally vincristine) (n= 50) or more intensive treatment regimens (n= 42) including anthracyclines (n= 18). Response (≥ 50% reduction of M protein) was obtained in 23 patients and minor response (clinical improvement but less than 50% reduction in M protein) in 22 patients. The median duration of the response was 7·5 months. Equal response rates were observed irrespective of the type of previous treatment. The major toxicity was myelosuppression with severe granulocytopenia and infections. However, the frequency decreased throughout the cycles. The NOP treatment is recommended in refractory myelomatosis, especially in patients refractory to other intensive regimens. Patients in a poor clinical condition or with thrombocytopenia before treatment should have a reduced mitoxantrone dose in the first treatment cycles.
Increased intraplatelet levels of platelet‐derived growth factor and transforming growth factor‐β in patients with myelofibrosis with myeloid metaplasia
Authors: Martyré Marie‐Claire , Magdelenat Henri , Bryckaert Marie‐Claude , Laine‐Bidron Christine , Calvo Fabien
Published: March 2008

Summary. Platelet‐derived growth factor (PDGF) is thought to play some role in the genesis of fibrosis associated with myeloproliferative disorders. In addition, transforming growth factor‐β (TGF‐β) has been confirmed to promote fibrotic process. Both PDGF and TGF‐β have been shown to cooperate with epidermal growth factor (EGF) in regulating the growth of human marrow fibroblasts. All three are contained in platelet α‐granules.

We report the results of a study in patients with myelofibrosis with myeloid metaplasia (MMM). We evaluated PDGF, TGF‐β and EGF‐like activities in circulating platelets from patients compared to healthy subjects. In contrast to EGF‐like intraplatelet levels which were similar in patients and in normal donors (1‐4 ng/109 platelets), we found constantly higher values for both PDGF and TGF‐β in MMM patients. In both radioimmunoassay (RIA) and assay for mitogenic activity on human bone marrow fibroblasts, PDGF levels were increased on the average 2‐3·5‐fold over the levels found in normal donors (P<0·01 and P<0·001, respectively). PDGF serum levels in patients were consistent with those found in platelets. In platelet‐poor plasma (PPP), PDGF concentrations were undetectable or ≃ 2 ng/ml in patients and in control donors as well. The total TGF‐β activity in platelet lysates, determined using a competitive radioreceptor binding assay on Swiss 3T3 mouse cells and an inhibition growth assay on CCL64 cells, was found 2‐3‐fold increased in patients with MMM as compared to control subjects (P<0·003).

These results emphasize that, not only PDGF, but also TGF‐β are implicated in the myelofibrosis with myeloid metaplasia.
Pharmacokinetics and biodistribution of technetium 99m labelled standard heparin and a low molecular weight heparin (enoxaparin) after intravenous injection in normal volunteers
Authors: Laforest M. D. , Colas‐Linhart N. , Guiraud‐Vitaux F. , Bok B. , Bara L. , Samama M. , Marin J. , Imbault F. , Uzan A.
Published: March 2008

For a better understanding of low molecular weight heparin pharmacokinetics, 99m technetium labelled heparin and enoxaparin were injected intravenously to four normal volunteers, after approval by the Ethics Committee and preliminary animals studies.

In vitro and in vivo, the labelled products proved to be stable and identical to the non‐labelled drugs.

Radioactivity curves in blood, organs and urines were similar for both products. Anti Xa plasma half‐life was 3 times longer for enoxaparin than for heparin. Anti Ha plasma half‐lives were similar. However, radioactivity persisted much longer than biological activities for both products. After chromatography, most of the radioactivity was bound to AT III, where an anti Xa activity peak was also detected. The anti Xa activity peak seen after adding AT III to plasma was much higher with heparin than with enoxaparin. In urine, biological activities, measured with AT III supplementation, were higher with enoxaparin than with heparin.

These results suggest that phenomena other than biodistribution are responsible for the differences in pharmacokinetics observed between these two products. The two most likely explanations are differences in metabolism and/or a release of an endogenous factor.
The role of recombinant haematopoietic growth factors in human long‐term bone marrow culture in serum‐free medium
Authors: Douay Luc , Giarratana Marie‐Catherine , Drouet Xavier , Bardinet Dominique , Gorin Norbert‐Claude
Published: March 2008

We have previously reported prolonged in vitro maintenance of human bone marrow progenitor cells using a serum‐free (SF) liquid culture system. The present study was undertaken to determine recombinant growth factor (rGF) requirement of long‐term marrow culture (LTMC) in absence of exogenous serum, to avoid interference of any undefined components. Our data clearly show that the presence of serum is a major obstacle to the correct evaluation of rGF activity. However, in SF conditions the sequential analysis of these rGFs, alone or in combination, clearly showed a stimulating activity of interleukin 3 (IL3), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and erythropoietin (EPO) on granulopoiesis and erythropoiesis. Indeed, the cumulative number of progenitors recovered during an 8‐week period exceeded the initial input by a fractor of 1·7 for granulocyte‐macrophage (CFU‐GM), of 3 for erythroid blast‐forming units (BFU‐E) and of 5·45 for CFU‐E when EPO, GM‐CSF and IL3 were combined. This study has confirmed that the system is able to sustain haematopoiesis for 8 weeks in a way similar to that in serum‐dependant LTMC, despite diminished stromal adherent layer development which never covered more than 50% of the flask surface. We conclude that this defined SF‐LTMC system provides a reproducible technique for studying human haematopoiesis.
Primary thrombocythaemia associated with systemic mastocytosis: a report of five cases
Authors: Tourneau A. Le , Gaulard P. , D'Agay M. F. , Vainchencker W. , Cadiou M. , Devidas A. , Haioun C. , Clauvel J. P. , Audouin J. , Diebold J.
Published: March 2008

Mastocytosis is an uncommon disease characterized by a proliferation of tissue mast cells involving various organs, particularly the bone marrow. Though rare, the association of mastocytosis and haematological malignancies is well established. However, the frequency of mastocytosis reported in patients with essential thrombocythaemia is relatively low. We report five cases of such an association. Our five patients were undergoing evaluation for thrombocy‐when the bone marrow biopsy revealed the presence of mastocytosis. The pathogenetic significance of this association is poorly understood. The different hypotheses are discussed.
A new β‐thalassaemia frameshift mutation detected by PCR after selective hybridization to immobilized oligonucleotides
Authors: Deidda G. , Novelletto A. , Hafez† M. , El‐Ziny M. , Terrenato L. , Felicetti L.
Published: March 2008

A previously undescribed mutation (−1, +3, codon 24) causing β‐thalassaemia was identified in an Egyptian patient. It consists in the concomitant deletion of a G in codon 24 and its replacement with the new trinucleotide CAC, thus resulting in the shift of the β‐globin reading frame. The sequence of the chromosome of interest was isolated from the homologous one by means of selective hybridization to an immobilized oligonucleotide. The presence of this mutation in the proband's family was confirmed by dot blot hybridization with an oligonucleotide probe.
Immunophenotypic, genomic and clinical characteristics of blast crisis of chronic myelogenous leukaemia
Authors: Hernández J. M. , González‐Sarmiento R. , Martin C. , González M. , Sánchez* I. , Corral* J. , Orfao A. , Canizo M. C. , san Miguel J. F. , López‐borrasca A.
Published: March 2008

We have studied phenotypic and clinical features in a consecutive series of 45 patients with chronic myelogenous leukaemia (CML) in blast crisis (BC). In addition, in 22 of these patients we have analysed the genotypic characteristics including immunoglobulin, T‐cell receptor (TCR) and major breakpoint cluster region (M‐bcr) gene organization. The granulomonocytic and megakaryoblastic lineages are the most commonly involved in these BC of CML (33% and 33% of cases, respectively); only 18% of our cases displayed a lymphoid phenotype. Moreover, both morphological and immunophenotypic studies revealed the frequent coexistence of two or three cell populations, especially when the megakaryoblast component is involved. The lymphoid BC displayed the highest incidence of complete remissions although this was not associated with a longer survival. Only minor differences between the different myeloid subgroups were observed. Immunoglobulin heavy chain (IgH) gene rearrangement was found in five of the six lymphoid BC and in one myeloid BC. Only one case showed k light chain gene rearrangement. In all but one myeloid BC the TCR‐β gene was in germline configuration. The TCR‐γ gene was rearranged in all lymphoid and one myeloid BC, while TCR‐δ gene rearrangement was detected in 67% and 16% of the lymphoid and myeloid BC, respectively. Most of the lymphoid BC (4/5) had the M‐bcr breakpoint in subregion 3, while the myeloid BC had the breakpoint either in subregion 2 or 3. No differences between the different myeloid phenotypic subgroups were observed in relation to breakpoint.
Mitozantrone and cytosine arabinoside as first‐line therapy in elderly patients with acute myeloid leukaemia
Authors: Yin J. A. Liu , Johnson P. R. E. , Davies J. M. , Flanagan N. G. , Gorst D. W. , Lewis M. J.
Published: March 2008

We have prospectively evaluated a regimen of mitozantrone and cytosine arabinoside (Ara‐C) as first‐line therapy in elderly patients with acute myeloid leukaemia (AML). One hundred and four patients with a median age of 68 (range 60–81) were studied, in whom 86 hadde‐novoAML, and 18 had preceding myelodysplasia or secondary AML. Complete remission was achieved in 64% ofde‐novocases, in 28% of MDS/secondary cases, and in 58% overall. The incidence of early death within 28 d of chemotherapy was 11%. The median disease‐free survival (DFS) was 11 months with an actuarial DFS of 15% at 43 months. The median overall survival was 9 months with an actuarial survival of 10% at 44 months. The incidence of non‐haematological toxicity was acceptably low, and usually of mild to moderate severity. Quality of life was improved, or unchanged, in 90% of responders. We conclude that mitozantrone and ara‐C is an effective and well‐tolerated regimen which produces high remission rates in elderly patients with AML.
Diagnostic criteria and classification of clinical subtypes of adult T‐cell leukaemia‐lymphoma
Authors: Shimoyama M. ,   members of The Lymphoma Study Group (1984–87)
Published: March 2008

The following diagnostic criteria are proposed to classify four clinical subtypes of HTLV‐1 associated adult T‐cell leukaemia‐lymphoma (ATL): (1) Smouldering type, 5% or more abnormal lymphocytes of T‐cell nature in PB, normal lymphocyte level (< 4 × 109/1), no hypercalcaemia (corrected calcium level < 2·74 mmol/1), lactate dehydrogenase (LDH) value of up to 1·5 × the normal upper limit, no lymphadenopathy, no involvement of liver, spleen, central nervous system (CNS), bone and gastrointestinal tract, and neither ascites nor pleural effusion. Skin and pulmonary lesion(s) may be present. In case of less than 5% abnormal T‐lymphocytes in PB, at least one of histologically‐proven skin and pulmonary lesions should be present. (2) Chronic type, absolute lymphocytosis (4 × 109/1 or more) with T‐lymphocytosis more than 3·5 × 109/1. LDH value up to twice the normal upper limit, no hypercalcaemia, no involvement of CNS, bone and gastrointestinal tract, and neither ascites nor pleural effusion. Lymphadenopathy and involvement of liver, spleen, skin, and lung may be present, and 5% or more abnormal T‐lymphocytes are seen in PB in most cases. (3) Lymphoma type, no lymphocytosis, 1% or less abnormal T‐lymphocytes, and histologically‐proven lymphadenopathy with or without extranodal lesions. (4) Acute type, remaining ATL patients who have usually leukaemic manifestation and tumour lesions, but are not classified as any of the three other types. A total of 818 ATL patients with a mean age of 57 years, newly diagnosed from 1983 to 1987, were analysed by this criteria. There were 448 males and 370 females, and 253 were still alive with a median follow‐up time of 13·3 months from diagnosis, while 565 were dead with a median survival time (MST) of 5·4 months. MST was 6·2 months for acute type, 10·2 months for lymphoma type, 24·3 months for chronic type, and not yet reached for smouldering type. Projected 2‐ and 4‐year survival rates were 16·7% and 5·0% for acute type, 21·3% and 5·7% for lymphoma type, 52·4% and 26 × 9% for chronic type, 77·7% and 62·8% for smouldering type, respectively. Distinct clinical features and laboratory findings of each clinical subtype are described.
Consistently normal CD4+, CD8+ levels in haemophilic boys only treated with a virally safe factor VIII concentrate (BPL 8Y)
Authors: Evans J. A. , Pasi K. J. , Williams M. D. , Hill F. G. H.
Published: March 2008

Prospective biochemical, virological and selected immunological follow up has been done for up to 32 months on 15 previously untreated haemophilic boys following treatment with an intermediate purity dry heated factor VIII concentrate (BPL 8Y). Tests for liver function and antibodies to blood‐borne viruses have been assessed monthly for the first year after starting treatment and thereafter every 2 months. All patients were immunized against hepatitis B and have not developed hepatitis B core antibodies and no boy has shown any rise in alanine transaminase level nor has anyone developed antibodies to hepatitis C (HCV). All patients have remained anti‐HIV seronegative. T lymphocyte subsets have been measured approximately every 4 months and in no patient has there been a significant rise in CD8+ cells: one patient showed a significant decrease in CD4+ cells but these and all CD4+ values for the other boys remained within normal age related limits. Changes in CD4+ levels in this one boy were not related to the total amount of treatment received. This group of patients who appear not to have contracted HIV, hepatitis B or non A non B hepatitis following treatment with this intermediate purity factor VIII concentrate have also not shown any consistent changes in CD4+ or CD8+ cells, which have been recorded previously in frequently treated haemophiliacs.
Hepatitis C antibody profile and viraemia prevalence in adults with severe haemophilia
Authors: Tedder R. S. , Briggs M. , Ring C. , Tuke P. W. , Jones P. , Savidge G. F. , Rodgers B. , Garson J. A.
Published: March 2008

Sera from 21 patients who had Received large amounts of unheated factor VIII concentrate were tested for antibodies to the hepatitis C virus (HCV) by both commercial (Ortho C100) and‘in house’ ELISAs. ‘In house’ assays utilized recombinant structural (core) or non‐structural (replicase) HCV proteins generated by a baculovirus expression system. Antibodies to HCV were detected in 100% of the sera by the core protein based ELISA but in only 62% and 19% by the C100 and replicase based ELISAs, respectively. Hepatitis C viraemia was demonstrated in 90% of the patients by in vitro amplification of the 5’non‐coding region of the HCV genome. Amplification with primer sets from two other regions of the genome proved less efficient at detecting viraemia. We conclude that the prevalance of hepatitis C infection in haemophiliacs may have been underestimated previously and that almost all HCV‐infected patients have evidence of on‐going viral replication.
Detection of endothelial cell‐reactive immunoglobulin in patients with anti‐phospholipid antibodies
Authors: McCrae Keith R. , DeMichele Angela , Samuels Philip , Roth Denise , Kuo Alice , Meng Qiang‐hua , Rauch Joyce , Cines Douglas B.
Published: March 2008

Individuals with anti‐phospholipid antibodies are at increased risk for the development of thrombosis and fetal loss. The pathogenesis of these syndromes is unknown, but may involve antibody‐mediated alterations in endothelial cell coagulant activity. To address this possibility, we determined the incidence of endothelial cell‐reactive antibodies in 76 patients whose plasma contained anti‐phospholipid antibodies, but who had no clinically‐evident immune disorder. Plasma from 47 patients deposited significantly more immunoglobulin on cultured endothelial cells than control plasma. Positive tests were more frequent in patients with a history of thrombosis than in those without (17/19 v 23/48; P=0.004). However, we observed no correlation between immunoglobulin deposition on cardiolipin and endothelial cells by individual plasmas. Furthermore, endothelial cell reactivity was not diminished by adsorption of anti‐cardiolipin antibodies from patient sera using liposomes. Immunoglobulin fractions prepared from 5/6 patient sera immuno‐precipitated a ≈ 70 kDa endothelial cell surface protein; 4/5 of these fractions also induced the release of von Willebrand factor from endothelial cells. These results demonstrate that plasma from many patients with anti‐phospholipid antibodies, but no clinically‐evident autoimmune disease, also contains endothelial cell‐reactive antibodies. Detection of such antibodies might help identify individuals in this patient population at greatest risk for thrombosis.
Rare β‐thalassaemia mutations in Asian Indians
Authors: Varawalla N. Y. , Old J. M. , Weatherall D. J.
Published: March 2008

Five β‐thalassaemia mutations hitherto undescribed in Asian Indians were identified in β‐thalassaemia carriers originating from the Indian subcontinent by direct sequencing of their β‐globin genes which were amplified by the polymerase chain reaction (PCR). A T‐G substitution at IVS 2 position 837, which probably creates an alternative acceptor splice site and a T insertion in codon 88, resulting in a shift in the reading frame with a premature stop codon, are new β‐thalassaemia mutations. The others were framshift codon 5 (‐CT), IVS 1 position 110 (G‐A) and IVS‐1 minus 1 (G‐A) which have been described previously in other populations. These results complete the characterization of the β‐thalassaemia mutations in 708 carriers of Asian Indian origin and will enable a comprehensive programme of carrier screening and prenatal diagnosis of β‐thalassaemia in this population.
Clinical experience with the combined use of recombinant interleukin‐2 (IL2) and interferon alfa‐2a (IFNα) in metastatic melanoma
Authors: Kruit W. H. J. , Goey S. H. , Monson J. R. T. , Stahel R. A. , Calabresi F. , Mertelsmann R. , Holdener E. E. , Eggermont A. M. M. , Bolhuis R. L. H. , de Mulder P. H. M. , Stoter G.
Published: March 2008

A multicentre study of IL2 and IFNα has been performed in 58 patients with metastatic melanoma. The scheme consisted of IL2 30 BRMP MU/m2/d as a continuous infusion for 4 d combined with subcutaneous administration of IFNα 6 MU/m2/d, day 1 + 4. The cycle was repeated every 2 weeks for a maximum duration of 26 weeks. 54 patients were evaluable for response. One (2%) achieved a complete and 10 (19%) a partial response. 19 (35%) patients were stable and 24 (44%) showed progressive disease. Common side‐effects included fever, chills, fatigue, skin rash, anorexia, nausea and diarrhoea. Hypothyroidism was noted in 10% of the patients. These results show that this regimen of IL2 and IFNα is active but, in contrast to what could be expected, not superior to IL2 alone possibly due to suboptimal dosing.

In an ongoing study in Rotterdam and Nijmegen, a more intense schedule was chosen, consisting of three daily i. v. doses of IL2 45 BRMP MU/m2 and IFNα 30 MU/m2 for 5 d. This regimen is repeated at intervals of 3 weeks for a total of three cycles. Presently, nine patients have been entered. One patient achieved a complete response, four a partial response (overall 56%), three had stable disease and one progressed. Toxicity was severe and treatment was prematurely stopped in five patients: myocardial infarction (one patient), atrial fibrillation (one patient), negative T waves and myocardial hypokinesia (one patient) and psychosis (two patients). This regimen can only be justified if the therapeutic results are superb, which has yet to be awaited.
Maintenance therapy in the myeloproliferative disorders: the current options
Authors: Giles Francis J.
Published: March 2008

Recombinant interferon alfa‐2a has been shown to be an effective induction agent in essential thrombocythaemia and thrombocythaemia associated with other myeloproliferative disorders, including chronic granulocytic leukaemia, polycythaemia rubra vera and myelofibrosis. Few data exist on the use of the recombinant interferons as maintenance agents in patients with thrombocythaemia. A cohort of 22 previously untreated patients, with essential thrombocythaemia, were treated with recombinant interferon alfa‐2a maintenance therapy for a minimum period of 6 months. Effective long‐term control of platelet counts, without evidence of haematological toxicity, was achieved in 19/22 patients. No objective haemorrhagic or thrombotic event occurred in 298 patient‐months of interferon therapy. Three patients discontinued interferon alpha therapy due to adverse side‐effects. Interferon alfa‐2a is an effective maintenance agent in essential thrombocythaemia and is discussed in this context.
Effects of low doses of recombinant human granulocyte‐macrophage colony stimulating factor (GM‐CSF) in patients with myelodysplastic syndromes
Authors: Estey Elihu H. , Kurzrock Razelle , Talpaz Moshe , McCredie Kenneth B. , O'Brien Susan , Kantarjian Hagop M. , Keating Michael J. , Deisseroth Albert B. , Gutterman Jordan U.
Published: March 2008

There has been no previously published experience with granulocyte‐macrophage colony stimulating factor (GM‐CSF) doses < 12 μg/m2 daily in patients with myelodysplastic syndromes, and most observations have been made at doses ≥ 120 μg/m2 daily. We administered 5 μg/m2 daily by subcutaneous injection to 29 such patients increasing the dose in patients who did not show a haemato‐logic response. Doses of 5 or 10 μg/m2 (‘low‐dose GM‐CSF”) produced an increase in neutrophils in 14/29 patients. Response was significantly (P=0·03) more frequent in patients who had a higher pre‐treatment neutrophil count (e.g. 11/16 in patients with ≥0·5 × 109/1). A rise in blasts followed administration of low‐dose GM‐CSF in five patients, all with either refractory anaemia with excess blasts (RAEB) or refractory anaemia with excess blasts in transformation (RAEBT). Platelets decreased in five patients, four of whom had no change in blasts, reverting to baseline when GM‐CSF was discontinued. We and others have previously observed simliar rises in blasts or decreases in platelets at doses of 120 μg/m2 daily. Low‐dose GM‐CSF produced no constitutional side effects. Our results suggest that low doses of GM‐CSF might be initially employed in neutropenic patients with myelodysplastic syndromes who present with pre‐treatment neutrophil counts > 0·5 × 109/1, increasing the dose, and hence the risk of extramedullary toxicity, only in patients who do not respond to the low dose. Patients who present with lower pre‐treatment neutrophil counts might begin treatment at doses above 10 μg/m2, but below the 120 μg/m2 commonly employed, which may be necessary in relatively few patients.
Leukaemia as a manifestation of large cell lymphoma
Authors: Bain Barbara , Matutes Estella , Robinson Danièle , Lampert I. A. , Brito‐Babapulle Vasanta , Morilla K. , Catovsky D.
Published: March 2008

A leukaemic phase is uncommon in large cell lymphoma, particularly at presentation of the disease, and very few cases have been reported since immunological markers became available. We have studied 24 cases presenting over a period of 1 5 years. I h were H‐lineage and eight T‐lineage. In five patients the large cell lymphoma represented a transformation of a preceding low grade, small cell lymphoproliferative disease. Three other patients with T‐lineage large cell lymphoma presented with skin infiltration (two cases) or lymphoma‐associated nephrotic syndrome (one case) 3‐9 months before leukaemia occurred. The other 16 patients (12 B‐lineage and four T‐lineage) presented with de novo large cell leukaemia/lymphoma. With the exception of skin infiltration, clinical features did not differ between T and B‐lineage cases. Prognosis was generally poor although a minority of patients lived 1‐2 years. Median survival was 7 months. In the B‐cell lymphomas immunological markers were those expected but among the T‐cell cases there were many unusual immunophenotypes with frequent failure to express markers which are usually positive in peripheral T cells. Without the availability of immunological markers diagnosis would have been difficult, with acute myeloid leukaemia being the most important differential diagnosis. T‐lineage and B‐lineage cases could not be distinguished on cytological features. In the majority of cases the cells were very pleomorphic with nuclear lobulation, prominent nucleoli and marked cytoplasmic basophilia being commonly observed. Marked nuclear lobulation was not confined to T‐lineage cases but was seen also in seven of 12 cases of de novo B‐lineage leukaemia/lymphoma; it was readily apparent in histological sections and on ultrastructural examination.
The effect of human T cell leukaemia virus type I infection on a herpes simplex virus‐specific CD8+ cytotoxic T cell clone
Authors: Inatsuki Akira , Yasukawa Masaki , Kobayashi Yuzuru
Published: March 2008

In an effort to clarify the effect of human T cell leukaemia virus type I (HTLV‐I) infection on virus‐specific CD8+ cytotoxic T cells, a herpes simplex virus‐specific CD8+ cytotoxic T cell clone was infected with HTLV‐I in vitro. The cytotoxic activity of the clone was found to have declined early after HTLV‐I infection when the expression of T cell receptor‐CD 3 complex on the cell surface still showed no difference in comparison with that of uninfected parent cells. After 16 weeks of HTLV‐I infection, expression of T cell receptor‐CD3 complex on HTLV‐I‐infected clone cells became decreased. This phenomenon is similar to the effect of HTLV‐I infection on CD4+ cytotoxic T cells as we previously reported, and suggests that there are common mechanisms of declined cytotoxic activity mediated by both CD4+ and CD8+ cytotoxic T cells following infection with HTLV‐I. Such functional alterations of cytotoxic effector cells might be one of the mechanisms underlying immunodeficiency caused by HTLV‐I infection.
Non‐Hodgkin's lymphomas with t(11;14)(q13;q32): a subset of mantle zone/intermediate lymphocytic lymphoma?
Authors: Leroux Dominique , Marc'hadour F. Le , Gressin R. , Jacob Marie‐Christine , Keddari E. , Monteil Michèle , Caillot P. , Jalbert P. , Sotto J. J.
Published: March 2008

We here describe 13 patients with non‐Hodgkin's lymphoma (NHL) and a translocation t(11;14)(q13;q32). They were part of a series of 163 patients with NHL and an abnormal karyotype. serially referred to our institution between January 1984 and 1990. Patients with t(11;14) seem to present several common and interesting features. Males are more frequently affected than females, and old people more than young. They present at diagnosis with advanced disease and usually show involvement of epithelium and bone marrow. With respect to histologic diagnoses, these patients are usually considered to be of low‐grade malignancies. However, most of them do very poorly, have short complete remission and frequent relapses whatever the treatment. As a whole, the median survival rate is rather low. The cytologic, histologic as well as the immunologic patterns tend to be uniform: tumours are composed of small cells and display features of mantle zone/intermediate lymphocytic lymphoma. They express high IgM and low IgD levels and more commonly bear Ig lambda light chains. They also express all pan‐B antigens (except CD23) as well as the CD 5 antigen, but usually lack the CD10. According to these characteristics, these tumours could be placed in between lymphocytic lymphomas (which usually express CD23) and follicular lymphomas (which commonly lack IgD and CD5 and bear CD 10 as well as a t(14:18)).