bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel mutation in the erythrocyte protein 4.2 gene of Japanese patients with hereditary spherocytosis (protein 4.2Fukuoka)
Authors: Takaoka Yutaka , Ideguchi Hiroshi , Matsuda Miho , Sakamoto Norihiro , Takeuchi Takayuki , Fukumaki Yasuyuki
Published: March 2008

Summary. Human erythrocyte protein 4.2 (band 4.2; pallidin) is a major membrane protein that comprises 5% of the total weight of the human erythrocyte membrane. Deficiencies of this protein have been observed in hereditary spherocytosis with anaemia, suggesting a role of protein 4.2 in erythrocyte stability and integrity. The molecular basis of this disorder remains unknown. As a first step in elucidating the pathogenesis of hereditary spherocytosis associated with protein 4.2 deficiency, we cloned and sequenced the erythrocyte protein 4.2 gene from a normal Japanese person. We prepared sets of oligonucleotide primers for polymerase chain reaction (PCR) and determined nucleotide sequences of exons and exon‐intron boundaries of the protein 4.2 gene from three unrelated Japanese patients with hereditary spherocytosis due to a complete defect of protein 4.2, using PCR‐related techniques. Two patients were homozygous for a missense mutation in codon 142 with the Ala (GCT) → Thr (ACT) amino acid substitution that has been reported previously (protein 4.2NIPPON), whereas one patient was compound heterozygous for the same missense mutation in codon 142 and a guanine‐adenine transition in codon 119 that changes the codon for Trp (TGG) to the termination codon (TGA) (protein 4.2Fukuoka). No additional mutation was identified in other exons of the protein 4.2 genes. Dot‐blot hybridization with allele‐specific oligonucleotide probes showed that homozygosity for the missense mutation in codon 142 and compound heterozygosity for the codon 142 and the codon 119 mutations were related to protein 4.2 deficiency in the families. Although two alleles of missense mutation of the codon 142 were also detected in 100 alleles of healthy Japanese, results obtained in this study indicate that the two mutations described above are closely related to the pathogenesis of hereditary spherocytosis due to protein 4.2 defect.
Karyotypic and ras gene mutational analysis in idiopathic myelofibrosis
Authors: Reilly J. T. , Wilson G. , Barnett D. , Watmore A. , Potter A.
Published: March 2008

Summary. Karyotypic analysis was performed in a total of 69 patients with well‐characterized idiopathic myelofibrosis. Karyotypic abnormalities were detected in 46% of cases examined during the chronic phase (29/63); with three abnormalities, del (13q), del(20q) and partial trisomy 1q, accounting for 75% of all abnormalities at diagnosis. The absence of del(5q), trisomy 8 and 21, as well as the rarity of monosomy 7, contrasts with pooled published data and may reflect our exclusion of closely related disorders, in particular MDS with fibrosis. Chromosomal aberrations increased to approximately 90% (8/9) in patients analysed during acute transformation. Mutational activation of codons 12, 13 and 61 of N‐, Ha‐ and Ki‐ras genes were assessed by polymerase chain reaction and hydridization with synthetic non‐radioactive digoxigenin‐labelled probes. Three mutations were detected in samples of peripheral blood DNA taken from 50 patients during the chronic phase of their disease: one N12 Asp (GGT GAT) and two N12 Ser (GGT AGT) mutations. The results from this study indicate that karyotypic abnormalities are present in at least 29% of cases at diagnosis and that del(13q), del(20q) and partial trisomy 1q are the most frequent findings. Ras mutations were relatively infrequent (6%) and appeared restricted to the N‐ras gene. Karyotypic analysis at diagnosis was found to be of prognostic significance.
Purification and characterization of human platelet von Willebrand factor
Authors: Williams Sybil B. , McKeown Laurie P. , Krutzsch Henry , Hansmann Kristin , Gralnick Harvey R.
Published: March 2008

Summary. Platelet von Willebrand factor (vWf) was purified from human platelet concentrates. The multimeric structure of the purified platelet vWf was similar to that observed in the initial platelet lysate, and, like the platelet lysate, the purified platelet vWf contained higher molecular weight multimers than plasma vWf. The apparent molecular weight of the reduced platelet vWf subunit was similar to the plasma vWf subunit. The N‐terminal amino acid of the purified platelet and plasma vWf was blocked. In concentration dependent binding to botrocetin‐ or ristocetin‐stimulated platelets, 125I‐plasma vWf bound with a higher affinity than platelet. The ristocetin cofactor activity per mg of purified plasma vWf was 5‐fold greater than the platelet vWf activity. Platelet and plasma vWf bound to collagen with similar affinities; however, platelet vWf bound to thrombin‐stimulated platelets and to heparin with a higher affinity than plasma vWf. The differences in the binding affinity(s) of plasma and platelet vWf to platelet GPIb and GPIIb/IIIa and extracellular matrix proteins may reflect different roles for plasma and platelet vWf in the initial stages of haemostasis and thrombosis.
Fraxiparin, a low‐molecular‐weight heparin, stimulates megakaryocytopoiesis in vitro and in vivo in mice
Authors: Shen Z. X. , Basara N. , Xi X. D. , Caen J. , Maffrand J. P. , Pascal M. , Petitou M. , Lormeau J. C. , Han Z. C.
Published: March 2008

Summary. The effect of a low‐molecular‐weight heparin, faxiparin (Nadroparinŕ;), on murine megakaryocytopoiesis in vitro and in vivo was studied in comparison with unfractionated heparin. The addition of fraxiparin at 1–20 IU/ml into plasma clot cultures but not serum‐free agar culture significantly enhanced MK colony growth. Furthermore, fraxiparin was found to potentiate the stimulating activity of aplastic anaemia serum (AAS) but not stem cell factor (SCF), interleukin‐3 (IL‐3), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and erythropoietin (Epo), on MK colony growth in vitro, and to neutralize the inhibitory effect of platelet factor 4 (PF4) in vitro and in vivo. Fraxiparin also acted synergistically with heparin confactor II and antithrombin III to promote megakaryocyte colony formation. Intraperitoneal administration of fraxiparin twice daily for 4d at 0.1–25IU/injection increased in mice the level of blood platelet counts and the number of single MKs and CFU‐MK in bone marrow. These data demonstrate that fraxiparin is able to positively regulate megakaryocytopoiesis.
Ultrasound examination of pathological cervical lymph nodes in patients with non‐Hodgkin's lymphoma and Hodgkin's disease
Authors: Gerrits C. J. H. , Overhagen H. van , Lom K. van , Adriaansen H. J. , Löwenberg B.
Published: March 2008

Summary. The choice of treatment of malignant lymphoma may vary considerably and is mainly determined by the pathology and clinical stage. Ultrasonography is currently one of the most sensitive techniques for evaluation of the cervical area. We set out to assess the value of ultrasound imaging when added to conventional staging (physical examination, laryngoscopy, computer tomography of thorax and abdomen, bone marrow cytology and histology) of maliganant lymphoma in 47 patients with untreated lymphoma. Hodgkin's disease was present in 14 patients and non‐Hodgkin's lymphoma in 33 individuals. The ultrasound results were independently compared with physical examination of the neck. Ultrasonography revealed additional pathological lymph nodes in 6/47 cases (13%). Furthermore, the diagnosis non‐Hodgkin's lymphoma could be established in one patient merely as a result of ultrasonography. Ultrasonography of the neck may reveal more pathological lymph nodes in a significant number of patients and may be of value in the initial staging of patients with malignant lymphoma.
A biclonal large granular lymphocyte (LGL)/NK‐associated (NKa) disorder of CD4+ and CD8+ lymphocyte subpopulations characterized by the simultaneous presence of distinct TCR rearrangements
Authors: Richards Stephen J. , Short Michael , Steed Andrew J. , Scott Colin Stephen
Published: March 2008

Summary. This communication reports a patient with concomitant expansions of CD4+ and CD8+ large granular lymphocytes. Immunological analyses revealed that the abnomally increased CD4+ LGL fraction was phenotypically similar to other reported persistent CD4+ LGL expansions, whereas the phenotypic profile for the CD8+ LGL component was unusual. of particular note was the finding that both the CD4+ and CD8+ LGL fractions showed high membrane the CD45RO isoform expression, thus suggesting their ‘primed’ status. Molecular biology studies of immunomagnetically fractionated cells using a Tγ9 TCR gamma gene primer further revealed that the CD4+ and CD8+ components were both clonal but showed different patterns of rearrangement It is suggested that the simultaneous presence of CD4+ and CD8+ clonal populations are unlikely to have been derived from a common progenitor and that they reflect expansions of functionally restricted subpopulations
Human platelet antigen4 (Br) genotyping by ASPA: allele‐specific primer amplification (PCR‐SSP)
Authors: Simsek S. , Bleeker P. M. M. , Heeremans J. , von Dem Borne A. E. G. Kr.
Published: March 2008

Summary. We have developed a PCR assay named ASPA (allele‐specific primer amplification) to determine the HPA‐5a and −5b genotypes. It consists of two PCR‐reactions. One primer of each priomer set has a 3’‐end nucleotide which is specific for A or G at position 505 of the nHPA‐5b or −5a allele respectively. The HPA‐5 genotypes determined in this way were strictly concordant with the genotypes established by the PCR‐ASRA and with the phenotypes established using MAIPA. The ASPA is a rapid and reliable technique and can be used for the determination of alleles which code for platelet antigen allotypes.
Metaphase fluorescence in situ hybridization (FISH) in the follow‐up of 60 patients with haemkopietic malignancies
Authors: Nylund Stella J. , Ruutu Tapani , Saarinen Ulla , Knuutila Sakari
Published: March 2008

Summary. Metaphase DNA fluorescence in situ hybridization (metaphase‐FISH) was performed on follow‐up samples from 60 patients suffering from haemopoietic malignancies (acute and chronic myeloid leukaemia, acute lymphoblastic leukaemia, non‐Hodgkin's lymphoma and myelodysplastic syndorme). All patients had clonal chromosomal trisomies or translocations at diagnosis, and were treated by bone marrow transplantation (BMT), chemotherapy (CT) or interferon‐α therapy. Metaphase‐FISH was performed during therapy‐induced complete haematological remission (CR) (BMT and CT patients) using biotin‐labelled whole chromosme paint probes. 28% of all patients in CR were shown by FISH to have abnormal metaphase cells, and 62% of this group suffered a clinical relapse. Of those with negative FISH results (72%), 12% relapsed. In three CML patients treated with BMT a small population of t(9;22)‐positive cells was demonstrated. These cells disappeared during follow‐up without causing a relapse. One ALL patient had abnormal cells a short time after start of therapy but was also later found FISH‐negative. Furthermore, we demonstrated that metaphase‐FISH is a suitable method for quantifying the proportion of abnormal cells in CML patients during interferon‐α therapy. Metaphase‐FISH was also employed to detect a local replase in an ALL patient. Thus, metaphase‐FISH was found reliable and sensitive for detection of minimal residual disease in patients with haemopoietic malignancies.
Pentosan‐induced thrombocytopenia: support for an immune complex mechanism
Authors: Goad Kathryn E. , Horne McDonald K. , Gralnick Harvey R.
Published: March 2008

Summary Pentosan polysulphate is a low molecular weight heparinoid that is used as an anticoagulant. Because the drug also has antineoplastic properties, it has been used experimentally at the National Institutes of Health to treat metastatic malignancies. We present the case of a patient who developed thrombocytopenia resembling Type II heparin‐induced thrombocytopenia (HIT) during the course of pentosan therapy. The patient's plasma demonstrated platelet reactivity both by aggregometry and 14Cserotonin release in the presence of pentosan. Heparin and other polyanions could substitute for pentosan in aggregation studies. The aggregating activity co‐purified with the patient's IgG and was inhibited by pre‐incubation with monoclonal antibody (MoAb) to the platelet Fc receptor. To elucidate the relationship between the platelet, the polyanion and the antibody, we measured the binding of 3H‐heparin to platelets in the presence of the patient's IgG and found that it was increased 6‐fold over binding in the presence of control IgG. Heparin binding was not reduced by MoAb against the Fc receptor. Taken together, these data support a model in which polyanion‐antibody complexes attach to the platelet surface by the polyanion and secondarily stimulate the platelet via their Fc termini.
The development of antiphospholipid antibodies in haemophilia is linked to infection with hepatitis C
Authors: Al‐Saeed A. , Makris M. , Malia R. G. , Preston F. B. , Greaves M.
Published: March 2008

Summary In haemophilia the presence of antibodies to antiphospholipid has been linked with HIV infection, but other possibilities have not been fully explored and the specificity for various phospholipids not established. In order to investigate further the pathogenesis and clinical significance of these antibodies, we have determined IgG and IgM antibodies to a variety of phospholipids, including cardiolipin, in the serum of 52 haemophiliacs, and related our findings to the presence of antibodies to HIV and hepatitis C virus (HCV), as well as to clotting factor concentrate usage and blood markers of liver biochemistry. Our results demonstrate that the presence of infection with hepatitis C virus is strongly associated with raised serum levels of antiphospholipid antibody even in the absence of HIV infection. They suggest that earlier conclusions on the relationships of antiphospholipid to HIV infection in haemophilia should be revised and that chronic infection with the hepatitis C virus should be added to the list of infective cause for the development of angtiphospholipid antibody.
Correlation between disease activity and T‐cell CD3 chain expression in a B‐cell lymphoma
Authors: Massaia Massimo , Attisano Carmela , Beggiato Eloise , Bianchi Alberto , Pileri Alessandro
Published: March 2008

Summary. The T‐cell receptor (TCR)/CD3 complex is the surface molecule responsible for the highly specific response to foreign antigens, including tumour‐associated antigens. T‐cell responses are frequently defective in cancer patients, and correlations have been observed with disease status. Recently, low amounts of CD3γ and CD3 chains in the CD3/TCR complex have been reported in long‐term tumour‐bearing mice as a possible mechanism for the impaired T‐cell responses. In this study we report a B‐cell lymphoma in which T‐cell CD3 expression correlated with disease activity.
Intragenic dinucleotide repeats in factor VIII gene for the diagnosis of haemophilia A
Authors: Yip B. , Chan Vivian , Chan T. K.
Published: March 2008

Summary . Two multi‐allelic microsatellite polymorphisms within the factor VIII gene were studied in 138 Chinese subjects. The allele sizes detected were higher than those found in Caucasian populations, whereas the heterozygosity rates were lower, being 0.5370 for intron 13 and 0.4444 for intron 22 repeats respectively. Their usefulness in diagnosis was compared to other intragenic and extragenic RFLPs, using previous data on the same 31 unrelated haemophilia A families. These intragenic microsatellite repeat polymorphisms were only informative for 18/31 families (58%); however, with the combined use of all existing RFLPs, the cumulative informativeness would be 100%.
A distinct coagulopathy associated with Interleukin‐2 therapy
Authors: Oleksowicz Leslie , Strack Margery , Dutcher Janice P. , Sussman Ira , Caliendo Geralyn , Sparano Joseph , Wiernik Peter H.
Published: March 2008

Summary We studied the coagulation profiles of 14 patients with advanced malignancies treated with Interleukin‐2 (IL‐2). A 43% prolongation of the PTT (P<0.001) and a significant decrease in functional levels of factors II, IX, X, XI, and XII were observed 6 h post IL‐2 treatment in comparison to pretreatment values. These parameters normalized within 2–3 d following IL‐2 administration. The PT, factors V. VII, VIII, fibrinogen and D‐dimer levels were unchanged with IL‐2 treatment. This pattern of coagulopathy has not previously been reported.
Effect of recombinant human interleukin‐3 on haematological recovery from chemotherapy‐induced myelosuppression
Authors: Tepler I. , Elias A. , Kalish L. , Shulman L. , Strauss G. , Skarin A. , Lynch T. , Levitt D. , Resta D. , Demetri G. , Gaynes L. , Schnipper L.
Published: March 2008

Summary. Patients with non‐small‐cell lung cancer (NSCLC) were treated with ICE chemotherapy (ifosfamide 2000 mg/m2, days 1‐3; carboplatin 300 mg/m2, day 1; etoposide 75 mg/m2, days 1‐3) intravenously (i.v.) during the first 3d of a maximum of four 28 d treatment cycles. Interleukin‐3 (IL‐3) was administered in cycles 2 and 4 as a daily subcutaneous (s.c.) injection on days 5‐18. Cohorts of three patients were treated at dosage levels of 0.5, 1.25, 2.5, 5.0, 10.0 and 15.0 μg/kg/d. At 15.0 μg/kg/d a ‘flu‐like’ syndrome of myalgias, arthralgias and fatigue was considered dose‐limiting. Other toxicities were headache, fever, urticaria, arrhythmia, chills and flushing. Subsequently, nine patients were added to the group receiving 10 μg/kg/d. 27 patients received IL‐3 after their second course of ICE. At 10 and 15 μg/kg/d, IL‐3 in cycle 2 was associated with enhanced haematological recovery. Depth of neutrophil nadir and days of neutropenia (ANC < 0.5 × 109/1) were reduced in 9/13 patients and in 8/11 patients, respectively. No effect was seen on platelet nadir or days of thrombocytopenia. IL‐3 was well tolerated up to 10 μg/kg/d when given as a daily s.c. injection. Results suggest IL‐3 as a potential adjunct to chemotherapy, and further studies to explore administration of IL‐3 in combination with other cytokines in this setting are warranted.
Accessory cells mediate hairy‐cell proliferation by mechanism (s) involving both adhesion and TNFα secretion
Authors: Till K. J. , Cawley J. C.
Published: March 2008

Summary. The mechanisms responsible for hairy‐cell (HC) growth both in vitro and in vivo are still unclear. In a recent study we showed that monocytes/macrophages induce HC proliferation in vitro. The purpose of the present paper is to examine the specificity of this accessory cell effect and to establish the mechanism(s) involved.

We demonstrate that the effect is not confined to monocytes/macrophages but is also potentially seen with a range of other cell types. However, at low accessory cell:HC ratios (< 1:20) only human umbilical vein endothelial cells (HUVEC) and macrophages induce HC proliferation. We suggest that these observations are of pathophysiological significance in relation to the close association frequently observed between HCs and endothelial cells/macrophages in the liver and spleen of patients with hairy‐cell leukaemia (HCL).

Regarding the mechanisms of the accessory cell effect, we show that both soluble factors and cell contact are important. A blocking anti‐TNFα antibody abrogated the HC proliferation induced by HUVEC supernatant. indicating the involvement of this cytokine. Interaction of HCs with HUVEC via CD11b and 11c leucocyte integrins was shown to be important in the contact effect.

Out demonstration of the involvement of both cytokines and cell contact in HC proliferation is in accord with what is already known about the control of B‐cell growth and differentiation. More specifically, our results suggest that TNFα and interaction with endothelial cells/macrophages via leucocyte integrins are involved in the proliferation of late B‐cells of the maturational stage represented by HCs.
Establishing the incidence of myelodysplastic syndrome
Authors: Williamson P. J. , Kruger A. R. , Reynolds P. J. , Hamblin T. J. , Oscier D. G.
Published: March 2008

Summary. We studied the incidence of myelodysplastic syndrome (MDS) in a population served by one district general hospital over a 10‐year period (1981‐90). Cases were sought through investigation of individuals presenting with suggestive peripheral blood findings. 279 new cases were diagnosed in a population rising from 203000 to 226 000 over the study period. The observed incidence rate did not change over the study period. The crude incidence rate was 12.6/100000/year. Age‐specific incidence rates per 100000 per year were 0.5 for age < 50, 5.3 for age 50‐59, 15 for age 60‐69, 49 for age 70‐79, and 89 for age 80+. A cohort study of one large general practice revealed several previously unknown and asymptomatic cases, indicating that the observed figures for the population overall may still underestimate the real incidence.

We conclude that MDS is more common than reported in previous studies and that incidence continues to rise into very old age. Difficulties in case ascertainment hamper reliable estimates of true incidence.
Human melanoma cell lines differ in their capacity to release ADP and aggregate platelets
Authors: Boukerche Habib , Berthier‐Vergnes Odile , Penin Franclois , Tabone Eric , Lizard Gearard , Bailly Mireille , McGregor John L.
Published: March 2008

Summary. In this study we have investigated, using three different human melanoma cell lines (M1Do., M3Da., M4Be.), the varying capacity of melanoma cells to induce platelet aggregation in the presence or absence of inhibitors of ADP or thrombin. The expression levels of different integrins (αv, β3, αvβ3, αIIb, αvβ3) were evaluated by immunoprecipitation, binding and flow cytometry studies. The level of ADP in supernatants of melanoma cells were quantified by ADP bioassay and HPLC. Platelets were irreversibly aggregated by M3Da. as shown by electron microscopy, in contrast to M1Do, which induced a slow reversible aggregation. M4Be. did not induce platelet aggregation. In both cases, with M3Da. or M1Do., apyrase but not PPACK inhibited platelet induced aggregation. An anti‐αvβ3 monoclonal antibody (LYP18) or polyclonal antibody inhibited platelet aggregation. A similar number of LYP18 molecules bound to the surface of M1Do., M3Da. and M4Be. cell lines. Biological HPLC assays of ADP present in the supernatant of tumour cell lines showed the highest concentration of ADP to be secreted by M3Da., followed by M1Do., and none detected for M4Be. These results show that differences in in vitro aggregating potential of the three human melanoma cell lines are not related to low integrin expression levels but to their ability to generate ADP. Generation of ADP by human melanoma cells may act as important modulator of melanoma‐platelet interactions.
A cross‐over pharmacokinetic and thrombogenicity study of a prothrombin complex concentrate and a purified factor IX concentrate
Authors: Thomas D. P. , Hampton K. K. , Dasani H. , Lee C. A. , Giangrande P. L. F. , Harman C. , Lee M. L. , Preston F. E.
Published: March 2008

Summary. A prospective cross‐over study was carried out on 19 patients with haemophilia B, comparing the pharmacokinetics of a purified factor IX concentrate prepared by metal chelate affinity chromatography (9MC) with a conventional three‐factor prothrombin complex concentrate (9A). The highly purified factor IX concentrate was shown to have a half‐life comparable to the PCC; the in vivo recovery of the purified concentrate was significantly greater than that of the complex (P < 0.01). The 20% change in the value of the International Standard for Factor IX Concentrate, introduced in 1988, might have been expected to lower the recovery values. However, the in vivo recovery for both concentrates was somewhat higher than reported previously, particularly in the older literature.

In nine patients, serial assays for fibrinopeptide A, prothrombin fragment FI+2 and thrombin‐antithrombin complexes (TAT) were performed to assess the potential thrombogenicity of the two concentrates. Evidence was obtained that there was significantly less activation of coagulation following administration of purified factor IX (9MC), as compared to the activation that occurred after the PCC.
Factor IX gene haplotypes in Brazilian Blacks and characterization of unusual Ddel alleles
Authors: Figueiredo M. S. , Bowen D. J , Silva W. A. , Zago M. A.
Published: March 2008

Analysis of factor IX gene polymorphisms is considered the best approach for prenatal diagnosis and carrier detection of haemophilia B when the identification of the gene mutation is not possible. Studies involving factor IX gene polymorphisms in Black populations are scarce and essentially restricted to the North‐American Black population whose composition is substantially different from that of the Brazilian and presumably other Black populations of South America. In this paper we report the analysis of eight factor IX gene polymorphisms in Brazilian Blacks: 5’ BamHI, DdeI, intron 2 BamHI, XmnI, TaqI, MspI, MnII and HhaI. Characterization of the VNTR‐like Ddel polymorphism revealed six different alleles: B, AB, A2B, A2B2, A3B and A5, the last being described here for the first time. The 5’ BamHI, DdeI, MspI and Hhal polymorphisms showed the highest heterozygosities (0.40‐0.50) and are in linkage equilibrium with one another. 19 complete haplotypes could be identified in this population. Based on the results we propose a systematic strategy for carrier detection and prenatal diagnosis of haemophilia B in this population. The combined analysis of four polymorphisms (5’ BamHI, HhaI, MspI and DdeI) provided an informative genetic marker in 85% of the females. The use of all eight polymorphisms allows information in 95% of females. Additionally, differences in gene frequencies and haplotype distribution suggest dissimilarities in factor IX gene polymorphisms between the Brazilian and the North‐American Black populations.
Ten‐year survival and prognostic factors in multiple myeloma
Authors: Tsuchiya Jun , Murakami Hirokazu , Kanoh Tadashi , Kosaka Masaaki , Sezaki Tatsuo , Mikuni Chikara , Kawato Masafumi , Takagi Toshiyuki , Togawa Atsushi , Isobe Takashi , Suzuki Kenshi , Imamura Yukio , Takatsuki Kiyoshi
Published: March 2008

Summary. Among 1119 Japanese patients with symptomatic multiple myeloma diagnosed between 1965 and 1981, 38 (3.4%) survived more than 10 years. Younger age, low tumour mass (absence of severe anaemia, hypercalcaemia, renal failure, and multiple bone lesions), low plasma cell percentage in bone marrow, mature and intermediate myeloma according to Greipp's criteria, and a positive response to subsequent treatment, were related to long‐term survival according to univariate analysis. Multivariate logistic regression analysis indicated younger age and low tumour mass as pretreatment characteristics to be related to long‐term survival. Prognostic factors proposed applicable to myeloma were also related to 10‐year survival.
Detection of trisomy 12 in chronic lymphocytic leukaemia: comparison of a polymerase chain reaction based technique with fluorescence in situ hybridization
Authors: Reining G. , Clodi K. , König M. , Geissler K. , Haas O. A. , Mannhalter C.
Published: March 2008

Trisomy 12 is the most frequent chromosomal aberration in chronic lymphocytic leukaemia (CLL) and seems to indicate a poor prognosis. To detect this abnormality we tested the applicability of the polymerase chain reaction (PCR) and compared it to the current standard fluorescence in situ hybridization (FISH). Two DNA regions containing variable numbers of tandem repeats (VNTR) located on (a) the long and (b) the short arm of chromosome 12 were chosen for PCR analysis. 8/72 patients (11%) were trisomy 12 positive compared to 16% by FISH. Chromosomal imbalances were only detected by PCR if at least 20% of the cells carried the numerical aberration.
Familial discrepancy between the one‐stage and two‐stage factor VIII methods in a subgroup of patients with haemophilia A
Authors: Duncan Elizabeth M. , Duncan Bruce M. , Tunbridge Lynn J. , Lloyd John V.
Published: March 2008

A higher result for plasma factor VIII:C measured by the one‐stage as compared with the twostage method has been described in some patients with haemophilia A or with von Willebrand's disorder. We used both methods to measure FVIII:C in 95 patients with haemophilia A. The results were equivalent in all 21 patients with severe haemophilia (16 families) and in 45 of the patients with mild or moderate haemophilia (18 families). However, the results were discrepant (FVIII: C by one‐stage assay 2‐7‐fold higher than by two‐stage assay) in the other 29 patients with mild or moderate haemophilia (12 other families). For each patient with discrepant FVIII: C results the classification was the same for all other affected members of his family. In some families with haemophilia A the gene defect leads to a discrepancy between the one‐stage and twostage FVIII: C results and may be more widespread than previously recognized.
Hypercalcaemia in B cell chronic lymphocytic leukaemia
Authors: Fain Olivier , M'Selmi Abdellatif El , Dosquet Christine , Meseure Didier , Lejeune Francloise , Garel Jean‐Michel , Thomas Michel
Published: March 2008

Hypercalcaemia is common in some lymphoproliferative disorders such as myeloma or T‐cell leukaemialymphoma. but is rarely described in B cell chronic lymphocytic leukaemia (BCLL). We report the case of a patient with BCLL, hypercalcaemia and osteolytic bone lesions. Parathyroid hromone‐related protein (PTHrP) mRNA was identified by Northern blot analysis of liver. spleen and lymph node tumour samples. Serum levels of tumour necrosis factor alpha (TNFα) were increased.
Mutations in the PIG‐A gene causing partial deficiency of GPI‐linked surface proteins (PNH II) in patients with paroxysmal nocturnal haemoglobinuria
Authors: Bessler Monica , Mason Philip J. , Hillmen Peter , Luzzatto Lucio
Published: March 2008

Paroxysmal nocturnal haemoglobinuria (PNH) is due to the absence or marked reduction of glycan phosphatidylinositol (GPI)‐anchored proteins on the surface of blood cells. Affected patients may have a population of red blood cells that are completely deficient (PNH III) or partially deficient (PNH II) in these proteins, or they may have both. PNH III has recently been shown to be due, in all cases examined, to a somatic mutation in the PIG‐A gene, whose product is required for an early step in GPI anchor synthesis. We now show that two patients with PNH II cells also have somatic mutations of the same gene: these produce a partial rather than a total loss of PIG‐A function.
Burkitt type leukaemia associated with multiple myeloma
Authors: Kawachi Yasunori , Sakamoto Yukihiro , Shidahara Toshiki , Abe Takako , Uchida Tatsumi , Sasaki Kazunori , Ikeda Kazuma
Published: March 2008

We describe a 77‐year‐old man with multiple myeloma (MM) who developed Burkitt type leukaemia (BTL) 16 months after the initial diagnosis of MM. MM was positive for CD10 with immunoglobulin (Ig) κ, κ, Bence Jones protein (BJP) and a normal karyotype. BTL was positive for CD10, CD19, CD20 and HLA‐DR with Ig μ and γ, γBJP and a clonal abnormal karyotype of 46, XY, ‐13, t(8;14)(q24;q32), 11q+,14q+, +mar. The patient died 9 d after diagnosis of BTL despite treatment with multiple agents. This is, to our knowledge, the first such case to be reported.
The in vitro and in vivo effect of recombinant interferon α‐2a on circulating haemopoietic progenitors in polycythaemia vera
Authors: Castello G. , Lerza R. , Cerruti A. , Cavallini D. , Bogliolo G. , Pannacciulli I.
Published: March 2008

In four patients with polycythaemia vera (PV) who received interferon alpha (IFN‐α) (3 MU subcutaneously three times a week) for 5 months, peripheral blood levels of granulocyte‐macrophage colony‐forming units and erythroid burst‐forming units were assessed monthly. Circulating progenitors significantly decreased throughout the treatment period. Moreover, we observed an inhibitory activity of IFN‐α on haemopoietic progenitor cells (HPC) from patients with PV in vitro.

The data presented confirm previous research which has shown an inhibitory effect of IFN‐α on HPC from patients with myeloproliferative disorders.

Significant improvement in the patients’clinical and haematological conditions should encourage larger studies on IFN‐α administration in PV patients.
Elevated levels of soluble factors in the cerebrospinal fluid in patients with adult T‐cell leukaemia complicated with meningeal infiltration
Authors: Uozumi Kimiharu , And Shuichi Hanada , Arima Terukatsu
Published: March 2008

We measured some soluble factors in the cerebrospinal fluid (CSF) of patients with adult T‐cell leukaemia (ATL) complicated with meningeal infiltration. Interleukin‐4 (IL‐4) was not detectable in the CSF of all cases with meningeal infiltration. Interleukin‐6 (IL‐6) was detected in a few patients with ATL. Measurement of IL‐4 and IL‐6 in CSF had no diagnostic value for meningeal infiltration of ATL cells. Soluble CD4 (sCD4) was highly elevated in all ATL patients with meningeal infiltration. Soluble interleukin‐2 receptor (sIL‐2R; sCD25) in CSF was markedly elevated in 13/18 ATL patients (72.2%) with meningeal infiltration. Levels of sCD4 and sCD25 in the CSF of ATL patients with central nervous system (CNS) symptoms were significantly higher than those of non‐ATL patients with CNS symptoms. These observations indicate that sCD4 and sCD25 in the CSF are probably associated with meningeal infiltration of leukaemia cells that expressed CD4 and CD25 on the surface membrane and new markers for the meningeal infiltration of ATL cells.
Splenectomy for treatment of immunoblastic lymphadenopathy
Authors: Hirose Masao , Nakagawa Ryuji , Okada Akira , Sano Toshiaki , Kuroda Yasuhiro
Published: March 2008

Summary. A 5‐month‐old girl was diagnosed with immunoblastic lymphadenopathy (IBL) on the basis of clinical symptoms and pathological findings. α‐interferon. immunosuppressive agents such as cyclosporin A, deoxyspagarine, and azathioprine, and an alternating regimen of low‐dose 6‐ mercaptopurine, cyclophosphamide and methotrexate were tried with partial success. Finally, splenectomy was carried out at 58 months of age, inducing prolonged complete remission without any medication.
Long‐term treatment with interferon‐α2b for severe pruritus in patients with polycythaemia vera
Authors: Muller E. W. , Wolf J. Th. M. de , Egger R. , Wijermans P. W. , Huijgens P. C. , Halie M. R. , Vellenga E.
Published: March 2008

Summary. Pruritus is a major clinical problem in patients with polycythaemia vera (PV). Conventional symptomatic treatment is unsatisfactory. Recently, a favourable effect of interferon‐α on pruritus in patients with PV has been reported. Also, interferon‐α suppresses the increased haemopoiesis in PV. However, long‐term treatment with interferon‐α may be hampered by side‐effects and the inconvenience of chronic subcutaneous injection therapy.

We conducted a long‐term study (median follow‐up 13 months) of the efficacy and tolerability of interferon‐α in 15 patients (mean age 68 years) with PV and severe pruritus. Six patients were evaluable after 1 year. Pruritus significantly improved in 12/15 patients. Haematological control improved, as evidenced by a decreased number of phlebotomies from a mean of 4.3 in the year before the study to 1.8 while on interferon‐α. Leucocyte and platelet numbers also decreased significantly. Five patients (33%) did not tolerate interferon‐α. The effects of interferon‐α could not be ascribed to an inhibitive effect on histamine production or to the disappearance of the abnormal erythroid progenitor clone, because erythropoietin‐independent erythroid colony formation persisted during interferon‐α treatment. We conclude that long‐term interferon‐α treatment is feasible and effectively relieves pruritus in patients with PV, but side‐effects are an important concern. The optimal dose regimen that is well tolerated, relieves pruritus, and offers satisfactory haematological control at the same time remains to be established.
Detection of BCR‐ABL and E2A‐PBX1 fusion genes by RT‐PCR in acute lymphoblastic leukaemia with failed or normal cytogenetics
Authors: Devaraj Prema E. , Foroni Letizia , Kitra‐Roussos Vasiliki , Secker‐Walker Lorna M.
Published: March 2008

Summary. To evaluate the use of molecular analysis as a complement to karyotypic analysis in the detection of specific chromosomal abnormalities, the occurrence of t(1;19)(q23;p13) and t(9;22)(q34;q11) was investigated by RT‐PCR in 43 diagnostic acute lymphoblastic leukaemia cases in whom cytogenetic investigations had failed (32 cases) or showed only a normal karyotype (20 normal metaphases, 11 cases). One child (aged 14 years) and five adults (aged 18–60 years) were BCR‐ABL positive on first round for M‐BCR‐ABL (one case) or m‐BCR‐ABL (one case), or on nested PCR for m‐BCR‐ABL (three cases). Coexpression of M‐BCR‐ABL (first‐round PCR) and m‐BCR‐ABL (nested PCR was seen in one case. One m‐BCR‐ABL‐positive case also expressed the E2A‐PBX1 fusion transcript. Patients positive for the transcript(s) were older, had higher white blood cell counts and a significantly poorer event‐free survival (P<0.001) than those negative for the transcript.
The two CD23 isoforms display differential regulation in chronic lymphocytic leukaemia
Authors: Fournier S. , Yang L. P. , Delespesse G. , Rubio M. , Biron G. , Sarfati M.
Published: March 2008

Summary. B lymphocytes from chronic lymphocytic leukaemia (B‐CLL) patients express the two CD23 isoforms (type A and B), which differ only in their intracytoplasmic domain. The abnormal regulation of the CD23 antigen in response to IL‐4, IFNs α and γ results in CD23 over‐expression on B‐CLL cells. Our present study shows that the two CD23 isoforms are differentially and abnormally regulated on B‐CLL cells. IL‐4 selectively up‐regulates CD23 type A mRNA in five different B‐CLL patients, whereas in normal B cells it enhances CD23 type A and is the most potent inducer of type B. In contrast, phorbol esters (PMA) up‐regulate both CD23 isoforms in the malignant B cells and specifically increases type B in normal B cells. We next postulated that cytokines other than IL‐4 regulate CD23 B isoform in B‐CLL cells and therefore examined the effect of IL‐2, IFN‐γ and IFN‐α. We found that the ability of a given cytokine to induce B‐CLL growth (i.e. IL‐2 and IFNα) is concurrent with a selective up‐regulation of CD23 type B mRNA, whereas lymphokines that have no B cell growth activity (i.e. IL‐4 and IFNγ) specifically increase CD23 type A mRNA. We next showed that IL‐4 and IFNγ prevent hydrocortisone‐induced programmed cell death and that the rescued malignant B cells mainly express CD23 type A. Given that CD23 molecule has been reported to play a role in normal B cell proliferation and survival, it is therefore proposed that in B‐CLL cells the expression of CD23 type A may be related to cell viability and that of type B to cell proliferation. These data suggest that the CD23 molecule may contribute to the physiopathology of the disease which is characterized by the accumulation of long‐lived and slow‐dividing monoclonal B cells.
Haptoglobin therapy for acute favism: a Japanese boy with glucose‐6‐phosphate dehydrogenase Guadalajara
Authors: Ohga Shouichi , Higashi Eriko , Nomura Akihiko , Matsuzaki Akinobu , Hirono Akira , Miwa Shiro , Fujii Hisaichi , Ueda Kohji
Published: March 2008

Summary. We report the case of a 2‐year‐old Japanese boy with acute favism who was treated with human haptoglobin products. He had been exhibiting chronic nonspherocytic haemolytic anaemia until the diagnosis of glucose‐6‐phosphate dehydrogenase (G6PD) deficiency when 14 months old. He suffered a favic crisis at 24 months of age, when the administration of haptoglobin was effective for relieving bilirubinaemia and haemoglobinuria. Serum‐free Hb rapidly decreased to normal levels despite the sustained level of serum lactate dehydrogenase. His G6PD gene was G6, Guadalajara. This is the first application of haptoglobin therapy for acute favism and the first reported case of Japanese G6PD deficiency with typical favic crisis. Haptoglobin treatment might be helpful for managing the haemolytic crisis in the disease.
Leukaemic non‐Hodgkin's lymphomas with hyperdiploid cells and t(ll;14)(ql3;q32): a subtype of mantle cell lymphoma?
Authors: DANIEL MARIE‐THERESE , TIGAUD ISABELLE , FLEXOR MARIE‐ANGE , NOGUEIRA MARIA‐ELENA , BERGER ROLAND , JONVEAUX PHILIPPE
Published: March 2008

The present study describes five patients with leukaemic non‐Hodgkin's lymphoma (NHL) detected on the basis of particular morphology and cytogenetic findings. With respect to histological, immunological and cytogenetic features these NHL are closely related to mantle cell Iymphoma/intermediate differentiated lymphocytic lymphoma. However, the presence of unusual large cells associated with the t(ll;14)(ql3;q32) translocation and numerical chromosome changes, in the near triploid or near tetraploid range, could delineate a particular subtype of mantle cell lymphoma.
Prognostic implications of DNA aneuploidy in 156 untreated multiple myeloma patients
Authors: GARCIA‐SANZ R. , ORFAO A. , GONZALEZ M. , MORO MA J. , HERNANDEZ J. M. , ORTEGA F. , BORREGO D. , CARNERO M. , CASANOVA F. , JIMENEZ R. , PORTERO J. A. , MIGUEL J. F. SAN
Published: March 2008

In this study the incidence of DNA aneuploidy in a large series of untreated multiple myeloma (MM) patients was assessed in order to determine its clinical and prognostic significance. A total of 156 MM patients were included in the study. DNA measurements were performed in all cases at diagnosis using two different flow cytometry methods: (1) propidium iodide (PI) staining on isolated nuclei, and (2) CD38/PI double staining on whole cells. The DNA ploidy status was correlated with the most relevant clinical and haematological disease characteristics. From the 156 cases analysed, 91 (58%) were aneuploid (56% hyperdiploid and 2% hypodiploid). The correlation between the two techniques on the detection of DNA aneuploidy was excellent, although CD38/PI double staining would be preferable in cases with <5% of DNA aneuploid plasma cells (PC). Upon comparing the clinical and haematological disease characteristics of hyperdiploid versus diploid cases, the former group was characterized by a lower age, reduced incidence of anaemia, lower 02M levels, higher proliferative activity within the residual normal haemopoietic cells, increased expression of CD 5 6 antigen in PC, and higher proportion of PB CD4+ T cells. In contrast, diploid cases had a higher expression of the CD10, CD20 and CD15 antigens and greater numbers of PB CD56+CD3 NK cells (P < 0–05). Circulating PC were identified in six cases, all being diploid. Overall survival was significantly longer in hyperdiploid compared to diploid MM (P = 0–02).
Der(16)t(l;16)(qll;qll) in myelodysplastic syndromes: a new non‐random abnormality characterized by cytogenic and fluorescence in situ hybridization studies
Authors: MUGNERET FRANCINE , DASTUGUE NICOLE , FAVRE BERNARDINE , SIDANER ISABELLE , SALLES BRUNO , HUGUET‐RIGAL FRANCINE , SOLARY ERIC
Published: March 2008

The der(16)t(l;16)(qll;qll) is a frequent recurrent rearrangement in solid tumours such as breast carcinomas and Ewings sarcomas. Recently, this abnormality was described also in multiple myeloma. We identified a der(16)t(l;16)(qll;qll) in three patients with myelodysplastic syndrome, either during preleukaemic phase (n = 2) or at the time of blastic transformation (n= 1). Breakpoints were ascertained by fluorescence in situ hybridization (FISH) using specific centromeric a‐satellite probes and whole chromosome painting for chromosome 1 and chromosome 16. These observations, combined with isolated cases of the literature, suggest that der(16)t(l;16)(qll; qll) is a non‐random abnormality associated with myelodysplastic syndromes.
Characterization of genomic BCR‐ABL breakpoints in chronic myeloid leukaemia by PCR
Authors: ZHANG J. G. , GOLDMAN J. M. , CROSS N. C. P.
Published: March 2008

In order to understand better the mechanism of translocation between the BCR and ABL genes in CML, we have exploited a‘bubble PCR’technique to clone genomic breakpoints. BCR‐ABL junction fragments were successfully amplified and sequenced in 14/32 (43%) patients tested. Breakpoints were dispersed throughout the major breakpoint cluster region without any clustering or hot spots. In three cases Alu sequences were found at or near the breakpoint on the ABL side of the translocation but no other obvious sequence homologies were found either in BCR or ABL. The translocation event was characterized further in three other patients by amplifying the reciprocal ABL‐BCR junction on the 9q+ chromosome and also normal ABL around breakpoints. In two of these patients a few nucleotides of BCR and ABL were either duplicated or deleted on translocation, suggesting that staggered cuts had been made in the DNA strands prior to recombination. In the third patient 50 bp of ABL was deleted and 159 bp of M‐BCR including exon b3 was duplicated, indicating either that the single‐stranded cuts may span a larger distance than previously thought or that another mechanism, perhaps involving gene conversion, may be involved in this instance.
Analysis of heat‐shock protein expression in myeloid leukaemia cells by flow cytometry
Authors: CHANT IAN D. , ROSE PETER E. , MORRIS ALAN G.
Published: March 2008

Expression of heat‐shock proteins (hsp) was analysed in the leukaemic cells of 12 patients with acute myeloid leukaemia (AML) and nine patients with chronic myeloid leukaemia (CML). Using monoclonal antibodies to hsp70, hsp90 and hsp60 (ML30, a mycobacterial antigen with homology to human hsp60), we measured hsp levels by flow cytometry of permeabilized cells. Mononuclear cells from 10 healthy volunteers were also examined. The results demonstrate that hsp expression is significantly increased (P<0′01) in the circulating cells of patients with AML compared with cells from CML patients, and compared with normal peripheral blood mononuclear cells. This increased pattern of expression was found for all three heat‐shock protein families included in this study. Mononuclear cells from leukaemic patients showed a heterogenous pattern of hsp expression, between different patients, between cells from individual patients, and between the different hsp proteins examined. It is possible that hsp expression relates to the differentiation state or proliferative potential of these leukaemic cells.
Attenuated‐dose idarubicin in acute myeloid leukaemia of the elderly: pharmacokinetic study and clinical results
Authors: LEONI F. , CIOIXI S. , GIULIANI G. , PASCARELLA A. , CAPORALE R. , SALTI F. , CERVI L. , FERRINI P. Rossi
Published: March 2008

AML in the elderly is characterized by intrinsic biological features implying an enhanced chemoresistance. Intensive chemotherapy should be the treatment of choice, but the standard doses could induce unacceptable rates of aplastic deaths. We evaluated the efficacy of an induction protocol with attenuated‐dose idarubicin (IDA) 8 mg/m2 for 3 d plus cytarabine and etoposide in 26 AML patients aged >60. 18 patients (69%) achieved CR, five (19%) were non‐responders and three (12%) died during induction. To compare the pharmacokinetics of IDA between elderly and young patients, we assayed daily the serum level of the drug and of its metabolite (idarubicinol, IDAol) in a group of eight elderly patients who received a dose of 8 mg/m2 (group A) and in a group of nine younger AML patients treated with 12 mg/m2 (group B). The apparent terminal half‐life of IDAol was significantly longer in the elderly than in the younger patients (mean half‐life 59–7h versus 41–4h, P< 0–05). The values of the area under the serum concentration curve of IDAol indicated that the two patient groups received a very similar exposure to the drug despite the different doses.

In conclusion, this protocol, based on attenuated doses of IDA, compares well with the results obtained previously in similar age‐matched patient series.