bluuebunny/crossref_metadata_2025 huggingface dataset:
Polycythaemia vera. III. Burst‐forming units‐erythroid (BFU‐E) response to stem cell factor and c‐kit receptor expression
Authors: Dai Chun Hua , Krantz Sanford B. , Green Wayne F. , Gilbert Harriet S.
Published: March 2008

We previously demonstrated that highly purified normal human blood burst‐forming units‐erythroid (BFU‐E) need the direct action of recombinant human stem cell factor (rSCF) in the presence of recombinant human erythropoietin (rEP) and recombinant human interleukin‐3 (rIL‐3) for further development in a serum‐free medium. To study the response of polycythaemia vera (PV) BFU‐E to rSCF, we performed dose–response experiments in a serum‐free medium using highly purified BFU‐E from PV patients. A marked increase in the number of PV bursts occurred with increasing concentrations of rSCF, compared to normal burst formation, when the cells were cultured in the presence of rIL‐3 at 1 U/ml. The percentage of maximum growth for normal BFU‐E was 31±11% while for PV it was 64·9% at the highest concentration of rSCF (P<0.01). Without rIL‐3, only 11% of maximum normal BFU‐E growth occurred as the rSCF concentration was increased and the size of the colonies was very small, but PV BFU‐E still expressed 48% of the maximum number of large erythroid bursts (P<0.001). This demonstrated an enhanced sensitivity of PV BFU‐E to rSCF, compared to normal BFU‐E. The pattern of 59Fe incorporation into haem after 8 d of cell culture indicated that PV BFU‐E had a time course of maturation and a degree of cellular maturity similar to normal BFU‐E. The percentage positivity and intensity of c‐kit receptors on PV erythroid cells were examined using immunofluorescence flow cytometry. When BFU‐E, CFU‐E, or erythroblasts were incubated with phycoerythrin‐conjugated SR‐1 anti‐c‐kit receptor monoclonal antibody, 90% of the PV and normal BFU‐E displayed c‐kit receptor at comparable intensities, as well as 80% of the PV and normal CFU‐E. A distinct loss of c‐kit expression occurred with erythroid differentiation beyond the CFU‐E stage, but at all stages no difference of c‐kit receptor expression was evident for PV erythroid precursors compared to normal precursors. These results indicate that the hypersensitivity to rSCF did not appear to be related to the number of c‐kit receptors. Since we have previously shown that highly purified PV BFU‐E are hypersensitive to rIL‐3 and rGM‐CSF, as well as rEP, it is now evident that PV BFU‐E are hypersensitive to each of the cytokines that have a prominent role in guiding their normal proliferation and differentiation. While the precise mechanism for the hypersensitivity is not known, it may be a key factor in the pathogenesis of PV.
Interleukin 8 in serum in granulocytopenic patients with infections
Authors: Waage Anders , Remick Daniel , Steinshamn Sigurd , Deforge Laura , Lamvik Jon
Published: March 2008

Serum levels of interleukin 8 (IL‐8) were examined in eight patients with acute myeloid leukaemia during 16 courses of chemotherapy. The patients experienced 14 episodes of fever which occurred in periods with granulocyte counts <0·5 × 109/l. Febrile episodes were classified as bacteriologically defined infection (n= 6), clinically defined infection (n= 2), and unexplained fever (n= 6). IL‐8 was detected in 18/25 (72%), 2/3 (67%) and 3/7 (43%) of the serum samples in the respective groups. In contrast, IL‐8 was detected in 22/90 (24%) of the samples taken when no fever was present (P<0·00003 versus bacteriologically defined infection). The median concentration of IL‐8 in samples taken during febrile episodes was 194 ng/ml (range 0–6358 ng/ml) and 0 (range 0–5392 ng/ml) on days without fever (not significant). In three patients with infections caused by, respectively, Streptococcus sanguis, Acinetobacter calcoanitratus and Candida albicans, IL‐8 rose to a peak levels and declined during recovery. We conclude that IL‐I is released systemically during infections with gram‐positive and gram‐negative bacteria and Candida albicans in patients with acute myeloid leukaemia and peripheral granulocytopenia due to chemotherapy. However, IL‐8 can also be detected when no sign of infection is present.
Allogeneic bone marrow transplantation for acute lymphoblastic leukaemia: risk factors and clinical outcome
Authors: Weisdorf Daniel J. , Woods William G. , Nesbit Mark E. , Uckun Fatih , Dusenbery Kathryn , Kim Tae , Haake Robert , Thomas William , Kersey John H. , Ramsay Norma K. C.
Published: March 2008

We report 12 years’experience with histocompatible, related donor marrow transplantation for 123 patients with acute lymphoblastic leukaemia; 104 second remission. Four regimens were studied: cyclophosphamide (Cy)+total body irradiation (TBI) (n= 35); Cy + fractionated TBI (n= 45); TBI + high‐dose cytarabine (n= 15); and hyperfractionated TBI + Cy (n= 28). 45 patients survive (34 ± 9%: 95% confidence interval) between 1 and 12·7 years (median 7·8 years) following BMT and 29 ± 8% survive leukaemia‐free. Significantly improved disease‐free survival was observed in patients with an initial WBC < 50 x 109/l (P= 0·02). Conditioning regimens tested yielded similar outcomes, though TBI/cytarabine led to greater treatment associated mortality. Leukaemia relapse was the most frequent cause of failure in 56 ± 11%; median time of relapse 8 months following BMT, none beyond 2·2 years. Relapse was more frequent with higher WBC, shorter initial remission and previous CNS leukaemia. Acute and/or chronic GVHD was associated with a strong trend (P= 0·06) towards less relapse. Allogeneic BMT may be curative for a substantial fraction of patients with ALL, but additional anti‐leukaemic measures beyond these conditioning modifications tested will be required to prevent post‐transplant leukaemia recurrence.
Haemopoietic reconstitution after autologous blood stem cell transplantation in patients with malignancies: a multicentre retrospective study
Authors: Pierelli Luca , Iacone Antonio , Quaglietta Anna Maria , Nicolucci Antonio , Menichella Giacomo , Panici Pierluigi Benedetti , D'Antonio Domenico , de Laurenzi Antonio , de Rosa Luca , Fioritoni Giuseppe , Indovina Alessandro , Leone Giuseppe , Majolino Ignazio , Montuoro Aldo , Scimé Rosanna , Torlontano Glauco
Published: March 2008

A retrospective study was undertaken to evaluate the efficacy of autologous blood stem cell transplantation (ABSCT) in terms of haemopoietic reconstitution after ablative chemotherapy or chemo‐radiotherapy. 55 patients with malignancies, observed in four Italian institutions from January 1987 to June 1991, were eligible for evaluation. This series included 19 non‐Hodgkin's lymphoma, 11 multiple myeloma, nine ovarian cancer, seven Hodgkin's disease, seven non‐lymphocytic leukaemia, one acute lymphoblastic leukaemia, one neuroblastoma. 522 PBSC collections were performed on 55 patients. Following ABSCT, the rate of engraftment was positively related to the dose of CFU‐GM infused and negatively to the presence of bone marrow involvement at conditioning. 48 patients out of 55 transplanted (87%) had rapid, complete and sustained engraftment. Three patients (5%) died of transplant‐related complications. Considering that 60% of the patients in this series were in partial remission or in progressive disease at the time of ABSCT, we conclude that ABSCT is a safe approach for the use of ablative conditioning therapy in patients with a wide scope of malignancies, provided that a large number of CFU‐GM have been collected after mobilizing treatment.
Severe cytopenias associated with the sequential use of busulphan and interferon‐alpha in chronic myeloid leukaemia
Authors: Shepherd Patricia C. A. , Richards Susan , Allan N. C.
Published: March 2008

We describe four patients who developed severe thrombocytopenia which progressed to aplasia after the use of α‐interferon in maintenance therapy of chronic phase CML after busulphan induction. On reviewing over 400 patients in the MRC CML III trial we found that there is a risk of cytopenia developing after busulphan therapy and a lesser risk of cytopenias developing after α‐interferon therapy. If the therapies are given in a sequential fashion the risk of cytopenia developing appears to be additive, may be pronounced, and may lead to clinically significant problems. Hydroxyurea alone does not lead to sustained cytopenia. Care should be taken to ensure that counts are stable after the use of busulphan before starting α‐interferon as maintenance therapy.
The correction of coagulometer effects on international normalized ratios: a multicentre evaluation
Authors: Poller L. , Thomson J. M. , Taberner D. A. , Clarke D. K.
Published: March 2008

One hundred and twenty‐nine centres in the U.K. participated in a study to test the reliability of the three methods of correction for coagulometer effects on international normalized ratios (INR). Results from 37 centres which tested three warfarinized plasmas by the manual method were taken as the ‘true’INR for the assessment of coagulometers. 63 centres (11 manual and 52 using coagulometers) determined their local International Sensitivity Index (ISI) in a calibration exercise. This was performed with a set of 20 lyophilized plasma calibrants with certified manual prothrombin times for the thromboplastin used in the study.

The following methods of INR derivation were compared by assessing the percentage deviation from the three INR values established by 37 manual users:

I. No coagulometer correction, i.e. (local PT/reference manual normal pT)manual ISI.

II. Coagulometer ratio correction, i.e. (local coagulometer PT/local coagulometer MNPT)manual ISI.

III. Local system ISI, i.e. (local coagulometer PT/local coagulometer MNPT)local system ISI.

IV. System ISI, i.e. (local coagulometer PT/local coagulometer MNPT)system ISI.

The local system ISI with the plasma calibrants (method III) gave the most reliable correction (mean deviation from ‘true’INR 4·87%). The method which gave the least was with the coagulometer ratio correction, i.e. the manual ISI and local coagulometer MNPT (mean 11·25%). The system ISI tested with ACL coagulometers gave less correction than the local ISI calibration.

The local system calibration with lyophilized plasmas also avoids some of the constraints on conventional thromboplastin calibrations.
Association of a novel high oxygen affinity haemoglobin variant with δβ thalassaemia
Authors: Rochette J. , Barnetson R. , Kiger L. , Kister J. , Littlewood T. J. , Webster R. , Poyart C. , Thein S. L.
Published: March 2008

We report an uncommon association of δβ thalassaemia and a haemoglobin (Hb) variant with high oxygen affinity in an Asian Indian family. Minimal polycythaemia was seen in a heterozygote for this novel Hb variant, Hb Headington (β72 (E16) Ser→Arg), while compound heterozygosity for Hb Headington and the Indian Gγ (Aγδβ) thalassaemia produces a marked increase in erythrocytosis with a concomitant increase in the level of the variant Hb. The HbF in such compound heterozygotes remains at a level consistent with that usually observed in individuals heterozygous for the Gγ (Aγδβ)° thalassaemia alone. The purified Hb variant showed an increased oxygen affinity, moderately decreased co‐operativity and a normal Bohr effect. Results of functional studies suggest that the high oxygen affinity of Hb Headington is due to the Ser→Arg substitution which disrupts the normal and tight interaction between A. B and E helices leading to a destabilization of the T deoxy‐structure of the abnormal haemoglobin.
Two cases of congenital dyserythropoietic anaemia type I associated with unusual skeletal abnormalities of the limbs
Authors: Brichard B. , Vermylen C. , Scheiff J. M. , Michaux J. L. , Ninane J. , Cornu G.
Published: March 2008

This report describes the cases of two young female patients with congenital dyserythropoietic anaemia (CDA) type I who presented similar hand and foot skeletal abnormalities: lack of distal phalanges and nails, and syndactyly. Up to now, some morphological malformations have been described in association with CDA type I but there is no report of skeletal defects.

These two cases are suggestive of a possible association between this hereditary anaemia and specific skeletal abnormalities of the distal extremities.
Avascular necrosis of bone following intensified steroid therapy for acute lymphoblastic leukaemia and high‐grade malignant lymphoma
Authors: Chan‐Lam D. , Prentice A. G. , Copplestone J. A. , Weston M. , Williams M. , Hutton C. W.
Published: March 2008

Five out of nine adults (55%) with lymphoblastic disease developed severe avascular necrosis of bone (AVN) when treated with a Berlin‐Frankfurt‐Munster (BFM) ALL protocol similar to the current joint MRC–ECOG ALL trial (UKALL XII). The principal purpose of these intensified regimens is to improve long‐term disease‐free survival without necessarily increasing toxicity and secondary morbidity. The presentation of all five was non‐specific bone pain occurring after the re‐intensification block of chemotherapy containing high doses of dexamethasone. Three types of diagnostic imaging were performed and magnetic resonance imaging (MRI) proved superior in demonstrating AVN and showed it at an earlier stage than plain radiographs or isotopic scans. We believe that the dose of corticosteroids was the major factor in the development of AVN. The five men in our series all remain in first remission with a median disease‐free survival of 3·5 years (range 2–8 years) but with varying degrees of disability due to AVN. Clinicians involved in UKALL XII and similar trials should be aware of this debilitating and potentially crippling complication when using high‐dose steroid‐containing regimens, perform MRI scan early and modify treatment if necessary.
Differential regulation of M‐CSF and IL‐6 gene expression in monocytic cells
Authors: de Wit Harry , Esselink Mariet T. , Halie M. Ruud , Vellenga Edo
Published: March 2008

Summary Using the human monocytic cell line Mono Mac 6 we studied the involvement of Ca2+, protein kinase A (PKA), and protein kinase C (PKC) dependent pathways in the regulation of M‐CSF and IL‐6 gene expression. The results demonstrate that on activation with the calcium ionophore A23187 both M‐CSF and IL‐6 mRNA are induced after 3 and 6 h respectively. Co‐stimulation with A23187 plus PMA resulted in an up‐regulation of M‐CSF mRNA and a downregulation of IL‐6 mRNA. Conversely co‐stimulation with A23187 plus DBcAMP resulted in a down‐regulation of M‐CSF mRNA and an up‐regulation of IL‐6 mRNA.

Nuclear run‐on and mRNA half‐life studies showed that the effects on the M‐CSF expression were related to changes at transcriptional and post‐transcriptional level. In contrast, the effects on the IL‐6 gene expression seems to be mediated at post‐transcriptional level. With regard to the secretion of the IL‐6 protein it was shown that it closely follows the accumulation of IL‐6 mRNA. Taken together, the data show that several intracellular signalling pathways control strictly the cytokine expression in monocytic cells which gives the cells the opportunity to respond variably to external activation signals.
Beta‐2 microglobulin as a predictor of prognosis in HIV‐infected men with haemophilia: a proposed strategy for use in clinical care
Authors: Sabin Caroline A. , Phillips Andrew N. , Lee Christine A. , Elford Jonathan , Timms Anthony , Bofill Margarita , Janossy George
Published: March 2008

Summary Whilst the prognostic value of serum beta‐2 microglobulin (s‐β2m) is well documented, the lack of a simple strategy for its use means that it is rarely ever measured in clinical practice. The prognosis associated with s‐β2m at two different points in HIV infection, as defined by the CD4 count, was studied in a cohort of 111 men with haemophilia registered at the Royal Free Hospital School of Medicine, London. At CD4 counts of 0·5 and 0·2 × 109/1, a raised s‐β2m level was significantly associated with an increased risk of developing AIDS (P= 0·002 and 0·022 respectively, adjusted for the patient's age). Kaplan‐Meier progression rates to AIDS by 4·5 years after a CD4 count of 0·5 × 109/1 were 57% (95% CI 32–82%) in those with s‐β2m levels of 3 mg/1 or more, but 20% (95% CI 4–36%) in those with s‐β2m levels of less than 3 mg/1. By 3·5 years after a CD4 count of 0·2 × 109/1, Kaplan‐Meier progression rates to AIDS were 75% (95% CI 52–98%) in those with s‐β2m levels of 3·8 mg/1 or more, and 47% (95% CI 29–66%) in those with s‐β2m levels of less than 3·8 mg/1. In the absence of acute viral infections, a raised s‐β2m indicates those who will tend to progress to AIDS more rapidly than those with lower s‐β2m levels and the same CD4 count. S‐β2m levels in general are likely to be higher in haemophilia patients than in other, nonhaemophilic risk groups. Whilst care should be taken, therefore, when applying our chosen cut‐off values to nonhaemophilic patients, our findings support the introduction of prophylaxis and antiviral therapies at a higher CD4 count in those with raised s‐β2m levels relative to other patients in the same risk group whilst delaying treatment in those with lower CD4 counts, but relatively normal s‐β2m levels.
Severe thalassaemia intermedia caused by interaction of homozygosity for α‐globin gene triplication with heterozygosity for β thalassaemia
Authors: Oron Varda , Filon Dvora , Oppenheim Ariella , Rund Deborah
Published: March 2008

Summary A 3‐year‐old child was evaluated for β‐thalassaemia intermedia. Molecular characterization including β‐globin gene sequence analysis revealed heterozygosity for a single β‐thalassaemia mutation, IVSI nt1 (GA). In addition the patient was found to be homozygous for α‐globin gene triplication (αααanti3,7/αααanti3,7). The propositus has a significantly more severe phenotype than has been previously reported with this combination of genetic defects. In contrast, four individuals heterozygous for both triplicated α and for β‐thalassaemia had a phenotype of thalassaemia minor, and a fifth had very mild thalassaemia intermedia.
Possible involvement of protein kinase C activation in down‐regulation of CD3 antigen on adult T cell leukaemia cells
Authors: Matsuda Mitsuhiro , Maeda Yasuhiro , Shirakawa Chikashi , Morita Satoshi , Koyama Atsuko , Horiuchi Fusanari , Hamazaki Hiroyuki , Irimajiri Kiyohiro , Horiuchi Atsushi
Published: March 2008

Summary The role of protein kinase C (PKC) system on CD3 expression on adult T‐cell leukaemia (ATL) was examined. The down‐regulation of CD3 on ATL cells is reportedly induced by CD3 down‐regulating factor (CD3DF) contained in serum and culture supernatants of leukaemia cells from acute type ATL patients. After we cultured normal PBMC with a PKC inhibitor, H‐7, CD3DF activity for PBMC was reduced significantly. Culture with H‐7 of HTLV‐1 transformed T cells, ATL‐2 cells whose CD3 expression had been decreased, led to enhancement of CD3 expression in a time‐dependent manner. These findings suggest that CD3DF may play an important role as a PKC system activator, resulting in CD3 down‐regulation.
Infection by Epstein‐Barr virus in Hodgkin's disease is not restricted to the Reed‐Sternberg cells
Authors: Valente Guido , Negro Francesco , Pacchioni Donatella , Palestro Giorgio
Published: March 2008

Summary The presence of Epstein‐Barr virus (EBV) DNA was analysed by a non‐radioisotopic in situ hybridization procedure within eight Hodgkin's disease tissues. Seven specimens contained EBV DNA, that was found not only within the Reed‐Sternberg tumour cells, but also, to a varying degree, within the surrounding lymphocytes. The staining was more diffuse in the nodular sclerosis and interfollicular types, whereas it involved only a minority of lymphocytes in cases with mixed cellularity. Appropriate controls established the specificity of these findings. The widespread expression of EBV DNA within Hodgkin's disease tissue heavily underscores the involvement of EBV in the pathogenesis of this illness.
Lack of MDM2 amplification in human leukaemia
Authors: Ridge Susan A. , Dyer Martin , Greaves Mel F. , Wiedemann Leanne M.
Published: March 2008

Summary While deletion or mutation of the p53 gene is one of the most common molecular alterations detected in a wide variety of tumours, it has been shown to occur in only a relatively small percentage of the leukaemia cases examined. However, it may be that other components of the p53 pathway are involved. Amplification of the MDM2 gene has recently been demonstrated in human sarcomas resulting in an increase in MDM2 protein levels. This protein can bind to p53 preventing the transactivation of p53 responsive genes, thus mimicking mutation or deletion of p53. We have investigated the prevalence of MDM2 amplification in human leukaemias. 101 leukaemia or lymphoma samples and nine cell lines were studied using Southern blotting. In no case was MDM2 amplification present. We conclude that MDM2 amplification is not a common event in human leukaemias.
Sweet's syndrome in myeloid malignancy: a report of two cases
Authors: van Kamp Harmen , van den Berg Eva , Timens Wim , Kraaijenbrink René A. , Halie M. Ruud , Daenen Simon M. G. J.
Published: March 2008

Summary Two patients with a myeloid malignancy in whom Sweet's syndrome (acute febrile neutrophilic dermatosis) was diagnosed, are described. They suffered from fever and showed cutaneous lesions, with infiltration of the skin by mature neutrophils without signs of vasculitis. In one of them the clonal origin of the infiltrating neutrophils could be demonstrated by in situ hybridization. In this patient an association with the use of recombinant human granulocyte‐colony stimulating factor was suspected. In the other patient. Sweet's syndrome was the initial symptom of haematological disease. Inadequate wound healing after surgical procedures led to the diagnosis.
Domiciliary desensitization therapy for young boys with haemophilia and factor VIII inhibitors
Authors: Lilleyman J. S.
Published: March 2008

Summary Immune tolerance was induced in five consecutive young boys with haemophilia and factor VIII inhibitors using frequent conventional doses of human factor VIII.

All therapy was given at home by parents and the four youngest boys had surgically implanted central venous catheters (Port‐a‐Cath). Two catheters eventually became infected after 2 and 4 years respectively, otherwise the devices were trouble‐free and liked by the families concerned.

Regular domiciliary factor VIII therapy in pre‐school children is not difficult with an aid to venous access and immune tolerance can be achieved in those developing factor VIII inhibitors.
Interleukin‐6 serum levels in patients with multiple myeloma
Authors: Emile Carole , Fermand Jean‐Paul , Danon Françoise
Published: March 2008

Summary Studying the prognostic value of serum interleukin‐6 (IL‐6) levels in multiple myeloma, we observed important daily variations in some patients. Therefore a unique serum IL‐6 measurement should be interpreted with caution and requires confirmation by multiple determinations performed over a period of several days.
Assessment of therapeutic potential of interleukin 2 for myelodysplastic syndromes
Authors: Ogata K. , Yokose N. , Ito T. , An E. , Tamura H. , Dan K. , Hamaguchi H. , Sakamaki H. , Onozawa Y. , Nomura T.
Published: March 2008

Summary. The therapeutic potential of interleukin 2 (IL‐2) for myelodsplastic syndromes (MDS) was evaluated in vitro, IL‐2‐induced lymphokine‐activated killer (LAK) cells were prepared from 38 MDS patients and 20 normal subjects. The cytotoxicity of LAK cells against K562 and Raji cell lines and MDS blasts was significantly reduced in high‐risk MDS (refractory anaemia with excess blasts (RAEB). RAEB in transformation, and leukaemic transformation of MDS), but was relatively well‐preserved in low‐risk MDS (refractory anaemia (RA) and RA with ringed sideroblasts). Examination of the immunophenotypes of freshly‐isolated lymphocytes showed that the percentage of CD4+ cells in low‐risk MDS and the percentage of CD3+, CD4+ and CD8+ cell populations in high‐risk MDS was significantly reduced compared with these populations in normal subjects. After cultivation with IL‐2, these three cell populations were still reduced in the corresponding MDS groups and the percentage of CD3‐CD56+ cells were significantly reduced in high‐risk MDS. There was a positive correlation between the percentage of K562 cells lysed by MDS LAK cells and the percentage of CD3‐CD56+ lymphocytes in MDS LAK cells. These aberrant lymphocyte subpopulations appeared to explain, at least in part, the reduced LAK cell cytotoxicity in MDS. These results present a possibility that IL‐2 and LAK therapies are ineffective for most high‐risk MDS patients, whereas they have potential value for low‐risk MDS patients whose lymphocyte cytotoxicity is usually preserved.
Sweet's syndrome during therapy with granulocyte colony‐stimulating factor in a patient with aplastic anaemia
Authors: Fukutoku Masaaki , Shimizu Shiro , Ogawa Yoshimi , Takeshita Shouichi , Masaki Yasuhumi , Arai Toshihide , Hirose Yuko , Sugai Susumu , Konda Susumu , Takiguchi Tomoo
Published: March 2008

Summary. A patient with aplastic anaemia developed Sweet's syndrome (a febrile neutrophilic dermatosis) during granulocyte colony‐stimulating factor (G‐CSF) therapy. Three repeated episodes of appearance and disappearance of erythematous nodules after administration and withdrawal of G‐CSF confirmed that G‐CSF induced Sweet's syndrome in the patient. Sweet's syndrome has been reported in patients with myelodysplastic syndrome and acute leukaemia, but not in patients with aplastic anaemia. This is the first report of a patient with aplastic anaemia who developed G‐CSF‐induced Sweet's syndrome.
Graft‐derived anti‐HPA‐2b production after allogeneic bone‐marrow transplantation
Authors: Taaning Ellen , Jacobsen Niels , Morling Niels
Published: March 2008

Summary. We report on a male who received a bone‐marrow allograft from his HLA identical sister for acute myelogenous leukaemia. After transplantation, the patient suffered from refractoriness to the transfusions of HLA‐matched platelets and a strong platelet‐specific antibody, anti‐HPA‐2b, of IgG1 subclass was demonstrated in the patient's serum. In the serum of the bone‐marrow donor a weak IgG1 anti‐HPA‐2b was demonstrated. IgG allotyping of the patient and donor showed identical results. We could not determine the origin of the anti‐HPA‐2b, although we hypothesize that the anti‐HPA‐2b was produced by immunocompetent donor lymphocytes infused with the suspension of bone‐marrow cells.
The human platelet alloantigens, HPA‐5(a+ b‐) and HPA‐5(a‐ b+), are associated with a Glu505/Lys505 polymorphism of glycoprotein Ia (the α2 subunit of VLA‐2)
Authors: Simsek S. , Gallardo D. , Ribera A. , von dem Borne A. E. G. Kr.
Published: March 2008

Summary. GP Ia/IIa (also called VLA‐2 or α2β1) is the primary receptor for collagen on platelets. The human platelet alloantigens HPA‐5a(Brb) and HPA‐5b(Bra) have been found to reside on the platelet GP Ia/IIa complex. In order to establish the molecular basis of the HPA‐5 system, platelet RNA was isolated from HPA‐5 (a+ b‐) and HPA‐5(a b+) individuals. After reverse transcription, cDNA coding for glycoprotein Ia (GP Ia) was amplified by the polymerase chain reaction (PCR). Nucleotide sequence analysis of the PCR products revealed an A·G polymorphism at base pair 1648 of the coding region of the mature protein, resulting in a substitution of lysine (AAG) in HPA‐5b(Bra) by glutamic acid (GAG) in HPA‐5a (Brb) at amino acid 505. Subsequent PCR‐ASRA (allele‐specific restriction enzyme analysis) with Mnl I using cDNA derived from three HPA‐5 (a+b‐), one HPA‐5 (a+b+) individuals demonstrated that HPA‐5a and −5b alleles are distinguishable by DNA typing.

In addition to the A→G substitution at base pair 1648, three silent mutations were identified, G→C (195 bp), C→T (837 bp), G→A (1041 bp).
Elevation of serum M‐CSF concentrations during pregnancy and ovarian hyperstimulation
Authors: Praloran Vincent , Coupey Lionel , Donnard Magali , Berrada Lamya , Naud Marie Françoise
Published: March 2008

Summary. Macrophage colony stimulating factor (CSF‐1 or M‐CSF) is involved in haemopoiesis and probably in mouse gestation. Sexual steroids induce its production by uterine glandular epithelial cells and its receptor (product of the protooncogene C‐FMS) is expressed on placental trophoblastic cells. We measured M‐CSF serum levels in 119 pregnant women and in eight women undergoing ovarian hyperstimulation for in vitro fertilization. M‐CSF increased early (4–8 weeks) and progressively during gestation. Its rapid elevation during the course of ovarian hyperstimulation suggests that its synthesis is probably induced by sexual steroids. This locally produced M‐CSF could play a role in human pregnancy and in the pathogenesis of thrombocytopenias observed during pregnancy.
PRAD1 gene over‐expression in mantle‐cell lymphoma but not in other low‐grade B‐cell lymphomas, including extranodal lymphoma
Authors: Oka Kouji , Ohno Toshiyuki , Kita Kenkichi , Yamaguchi Motoko , Takakura Nobuyuki , Nishii Kazuhiro , Miwa Hiroshi , Shirakawa Shigeru
Published: March 2008

Summary. Employing Northern blot analysis and the polymerase chain reaction, we investigated PRAD1 gene over‐expression in the tumour tissues of 58 patients with B‐cell lymphoma. These findings were then examined in relation to the patients' clinical and immunohistological characteristics. The over‐expression of this gene was detected in 6/8 patients with mantle cell lymphoma (MCL) and in only 1/50 other lymphomas, indicating its close association with MCL. The patients with MCL had common clinical findings of advanced disease with generalized lymphadenopathy on admission, and they had a CD5+CD10–IgD+ phenotype. The patients with chronic lymphocytic leukaemia (CLL) also showed findings indicating a distinctive disease entity: a CD5+CD10–IgD+ phenotype and lack of PRAD1 over‐expression. In contrast, most patients with diffuse low‐grade lymphoma other than MCL and CLL had localized extranodal disease, expressed a CD5‐CD10–IgD– phenotype, and lacked PRAD1 over‐expression. These findings suggest that extranodal low‐grade lymphomas differ from nodal MCL and are not part of the spectrum of CLL.
Rapid clearance of hepatitis C virus RNA in peripheral blood mononuclear cells of patients with clotting disorders and chronic hepatitis C treated with alpha‐2b interferon is not a predictor for sustained response to treatment
Authors: Peerlinck K. , Willems M. , Sheng L. , Nevens F. , Fevery J. , Yap S. H. , Vermylen J.
Published: March 2008

Summary. Thirteen patients with congenital coagulation disorders and chronic hepatitis C were treated with alpha‐interferon (IFN). Serum transaminases (ALT) and hepatitis C virus (HCV)‐RNA in plasma and peripheral blood mononuclear cells (PBMC) were followed during and after treatment. During IFN treatment ALT levels normalized in 8/13 patients. In all patients HCV‐RNA disappeared from PBMC. In 9/13 HCV‐RNA also became undetectable from plasma; in six of these nine this occurred earlier in PBMC than in plasma; in the remaining three the disappearance of HCV‐RNA occurred simultaneously in plasma and PBMC. All but two patients relapsed within 3 weeks after cessation of IFN, one relapsed after 6 weeks and one patient remains in remission after 6 months. HCV‐RNA reappeared either earlier in plasma (3/12) or simultaneously in plasma and PBMC (9/12). Viral replication in PBMC thus can easily be suppressed by IFN, whereas longer treatment is needed for the eradication of HCV‐RNA in plasma. The detection of HCV‐RNA in PBMC cannot be used as a prognostic marker for the identification of patients with chronic hepatitis C who will have a sustained response to IFN treatment.
t(8;13) (p11;q12) translocation in a myeloproliferative disorder associated with a T‐cell non‐Hodgkin lymphoma
Authors: Leslie J. , Barker T. , Glancy M. , Jennings B. , Pearson J.
Published: March 2008

Summary. An unusual cytogenetic translocation, t(8;13) (p11;q12), is described in a patient presenting with a CML‐like myeloproliferative disorder associated with a high‐grade T‐cell lymphoma. Evidence is presented suggesting that the breakpoint region in the translocation involves a site implicated in both the T cell malignancy and the abnormal granulocyte proliferation.
Inhibition of erythropoietin production in vitro by human interferon gamma
Authors: Vannucchi A. M. , Grossi A. , Rafanelli D. , Statello M. , Cinotti S. , Rossi‐Ferrini P.
Published: March 2008

The effects of interferon‐gamma (IFN‐γ). alone and in combination with IL‐1, IL‐6 and tumour necrosis factor‐alpha (TNF‐α), on in vitro erythropoietin (Epo) production by the human hepatoma Hep 3B cell line were evaluated. The addition of IFN‐γ to either unstimulated or cobalt chloride (CoCl2)‐treated Hep 3B cells resulted in a dosedependent inhibition of Epo release in the medium by as much as 70% at 1000 U/ml. Half‐maximal inhibition was observed at around 50 U/ml. According to previous observations, IL‐6 had a stimulatory effect on Epo production by CoCl2‐treated Hep3B cells: however, the simultaneous addition of IFN‐γ and IL‐6 resulted in a reversal of the stimulatory effects due to IL‐6. IFN‐γ and IL‐1 had an additive inhibitory effect, whereas IFN‐γ and TNF‐α acted in a synergistic fashion in inhibiting Epo production by Hep 3B cells. The inhibitory effect of IFN‐γ appeared to be due to a down‐modulation of Epo mRNA levels in CoCl2,‐treated Hep 3B cells, as shown by Northern blot analysis.

These data indicate that Epo production by hepatoma cells in vitro is inhibited by IFN‐γ, and that a complex network of interacting cytokines may regulate Epo production in response to an hypoxic stimulus. Overall, these results also suggest that IFN‐γ might have a role in the defective Epo production observed in several inflammatory and immunemediated disorders characterized by relatively high IFN‐γ plasma levels.
Evidence that the clonogenic cell in multiple myeloma originates from a pre‐switched but somatically mutated B cell
Authors: Bakkus Marleen H. C. , Riet Ivan Van , Camp Ben Van , Thielemans Kris
Published: March 2008

Summary. There is still much controversy about the precursor cell type in multiple myeloma (MM). Some authors claim that it is a pre‐B cell, others state that it is a memory B cell or plasmablast. We have recently shown that the VDJ region of the MM immunoglobulin heavy chain gene is somatically hypermutated and antigen selected, without intraclonal variation or evolution in time. By using a patient‐specific PCR approach we have now obtained evidence that the premyeloma cell can be situated in the pre‐switched B‐cell compartment and that heavy chain switching can occur without further somatic mutation. Based on the MM immunoglobulin sequences derived from the bone marrow, patient‐specific CDR2 and CDR3 oligonucleotides were designed. B lymphocytes were separated from plasma cells based on the expression of CD19 and HLA class II or surface bound IgM using immunomagnetic beads. The expressed Ig sequences were amplified by RT‐PCR using patient specific CDR2 primers and isotype specific primers (Cμ, Cγ and Cα). Myeloma‐specific Ig sequences were detected by a myeloma‐specific CDR 3 probe and sequenced. In one out of five cases we found in the peripheral blood clonally related IgM and IgA sequences with the same somatic mutations as the MM‐IgG sequence. In another case of an IgG MM we found in the bone marrow clonally related IgA sequences with the same somatic mutations. These findings, together with the fact that myeloma‐Ig genes contain somatic mutations without intraclonal variation, suggest that the clonogenic cell in multiple myeloma can originate from a pre‐switched but somatically mutated B cell.
Familial aggregation of haematological neoplasms: a controlled study
Authors: Shpilberg Ofer , Modan Michaela , Modan Baruch , Chetrit Angela , Fuchs Zahava , Ramot Bracha
Published: March 2008

Summary. Advances in molecular biology techniques suggest that many haematological neoplasms originate from a transformation process at the level of the haemopoietic pluripotential stem cell. While familial aggregation has been reported for many haematological neoplasms, most studies were uncontrolled and examined the presence of the same haematological neoplasm as the index case. We assessed the familial aggregation of all haematological neoplasms in 4061 family members of 189 patients with various haematological neoplasms and two control groups: 955 relatives of 36 patients with non‐malignant haematological disorders and 508 relatives of 33 patients with type II diabetes mellitus. Data collection included self‐administered questionnaires. The odds ratio for haematological neoplasms among relatives of the index cases adjusted for age, sex, ethnicity, number of relatives in the family, and degree of familial linkage in the study group versus the two control groups was 3.62 (95% confidence interval, 1.44‐9.07; P<0.01). The vast majority of the haematological neoplasms among family members did not belong to the same histopathological category as the index cases. The data support the hypothesis of a genetic predisposition to haematological neoplasms. The fact that the aggregation is not disease specific is consistent with a defect in the pluripotent haemopoietic stem cell.
Mercury compounds induce a rapid increase in procoagulant activity of monocyte‐like U937 cells
Authors: Kaneko Hiroshi , Kakkar Vijay V. , Scully Michael F.
Published: March 2008

. When monocytic leukaemia line U937 cells were incubated in the presence of HgCl2there was a rapid increase in tissue factor (TF)‐dependent procoagulant activity, reaching a maximum (equivalent to the total TF activity observed when cells had been subjected to a freeze/thaw cycle) after 15 min at 50 μmHgCl2and after 30 min at 10 μmHgCl2. Two other heavy metal compounds, AgNO3and phenylmercuric acetate, caused a similar increase in TF activity. The increase was independent of protein synthesis. Other reagents tested, CdCl2, ZnCl2, NiCl2, ADP, FMLP and monocyte chemotactic factor (MCF‐1), did not cause a rapid increase in functional activity, when tested under the same experimental conditions. The addition of HgCl2to the cells causes, in a concentration‐dependent manner, a 10‐12‐fold increase in intracellular calcium (Cai) which coinicides with increase in TF activity. Calcium ionophore also caused an increase in TF activity of the U937 cells. Upon treatment with HgCl2the cells surface of U937 cells showed a large increase in the level of phosphatidylserine (PS) on the cell surface (as measured by potentiation of the rate of activation of prothrombin by factor Xa‐factor Va) but with no change in the level of TF antigen on the cell surface. We consider that the TF is present on the cell surface of the monocyte but relatively inactive towards the physiological substrate, factor X (FX), until HgCl2causes a change in the polarity of the cell membrane exposing PS on the outer leaflet by a mechanism likely to be enhanced by the increase in intracellular calcium.
Activation of the coagulation cascade in severe falciparum malaria through the intrinsic pathway
Authors: Clemens R. , Pramoolsinsap C. , Lorenz R. , Pukrittayakamee S. , Bock H. L. , White N. J.
Published: March 2008

. The mechanisms involved in the activation of the coagulation cascade in severe falciparum malaria were studied in 22 adult patients (19 male, three female) aged 18‐45 (mean ± SD 31 ± 11) years. Of these, nine had multiple vital organ dysfunction, and bleeding occurred in four patients, two of whom died. During acute illness the reduction in plasma antithrombin III (AT III) concentrations and elevation in thrombin‐AT III complexes were associated with significant reductions in factor XII and prekallikrein activities, and an increase in the C1 inhibitor antigen/activity ratio. Serial plasminogen activity remained within the normal range in all patients while protein C activity was significantly reduced. All patients had markedly elevated plasma polymorphonuclear leucocyte elastase (PMN‐elastase) levels with mild depletion of alpha‐2 macroglobulin but normal concentrations of alpha‐1 antitrypsin. There was no correlation between PMN‐elastase concentrations and any of the coagulation parameters or concentrations of proteinase inhibitors. These results suggest that the intrinsic pathway of the clotting cascade is activated in severe malaria. This may cause activation of the complement system and release of bradykinin and PMN‐elastase and could contribute to the pathogenesis of severe malaria.
Pancytopenia in allogeneic marrow transplant recipients: role of cytomegalovirus
Authors: Bilgrami S. , Almeida G. D. , Quinn J. J. , Tuck D. , Bergstrom S. , Dainiak N. , Poliquin C. , Ascensao J. L.
Published: March 2008

SUMMARY. We describe the clinical course of three cytomegalovirus‐antibody‐positive allogeneic marrow graft recipients who developed progressive pancytopenia during the third month post‐transplant. Bone marrow biopsy cores were hypocellular without evidence of disease recurrence. Haemopoietic progenitor assays demonstrated a decrease of all assayable progenitors. Cytomegalovirus was identified from the blood and urine of all three patients. However, none of the patients developed other manifestations of cytomegalovirus infection such as pneumonitis, hepatitis and enteritis. The therapeutic use of ganciclovir and intravenous immunoglobulins resulted in prompt resolution of both viraemia and viruria in all three patients, and of pancytopenia in two patients.
Red cell dimorphism in a young man with a constitutional chromosomal translocation t(11;22)(p15.5;q11.21)
Authors: Rees Mark I. , Worwood Mark , Thompson Peter W. , Gilbertson Clive , May Alison
Published: March 2008

SUMMARY. A constitutional, balanced chromosomal translocation t(11;22)(p15.5;q11.21) was discovered in a tall young man during investigation of a red cell dimorphism. The red cells are predominantly normochromic and normocytic with a small population of hypochromic, microcytic cells. Contained within the regions involved in the translocation are determinants of height (IGF2:11p15.5), red cell haemoglobinization (non‐α globin gene complex: 11p15.5) and oncogenesis (cHa‐Ras‐1, Beckwith‐Wiedemann syndrome: 11p15.5; BCR, Burkitts lymphoma, Ewings sarcoma: 22q11.21). To map these regions in the patient, somatic cell hybrids were generated and cell lines that segregated the chromosomes 11, 22 and 22q‐ were obtained. All 11p15.5 sequences investigated, in particular the whole of the non‐α globin gene complex including its 5’and 3’regulatory sequences, were found to be translocated to 22q‐. All chromosome 22 sequences studied were missing from the 22q‐ cell lines, including the proximal anonymous marker D22S24, and therefore assumed to be translocated to 11p+. These results suggest that the non‐α globin gene complex has been moved close to the centromeric region of chromosome 22q‐. It is postulated that such a positioning subjects the complex to a variegated position‐effect bringing about a clonal exclusion of the complex and thus producing a β‐thalassaemia trait mosaic.
Myelodysplasia following paroxysmal nocturnal haemoglobinuria: evidence for the emergence of a separate clone
Authors: Kamp Harmen van , Smit Jan W. , van den Berg Eva , Halie M. Ruud , Vellenga Edo
Published: March 2008

SUMMARY. A patient with paroxysmal nocturnal haemoglobinuria (PNH) who developed a myelodysplastic syndrome (MDS) is described. After the onset of myelodysplasia the neutrophils of the patient fully expressed GPI‐linked proteins. It is concluded that the myelodysplasia does not originate from transformed PNH stem cells, but represents the emergence of a separate clone arising from an injured marrow.
Increased soluble IL‐2 receptor levels in HCV‐infected haemophiliacs: a possible indicator of liver disease severity
Authors: Makris M. , Preston F. E. , Ralph S.
Published: March 2008

SUMMARY. We have measured sIL‐2R in 60 haemophiliacs and 20 male control subjects. Haemophiliacs were grouped according to their HIV/HCV antibody status. Group 1 (n= 20) comprised HIV + ve/HCV + ve, group 2 (n= 27) HIV ‐ ve/HCV + ve and group 3 (n= 13) HIV ‐ ve/HCV ‐ ve. Group 4 comprised the normal control subjects. We also examined, retrospectively, the relationship between the severity of chronic liver disease, assessed histologically, and sIL‐2R levels in selected patients. There was no significant difference between sIL‐2R levels of the group 1 and group 2 patients, and the levels for both were significantly greater than those of either the group 3 patients or the control subjects. sIL‐2 levels were also higher in selected patients with cirrhosis than in those with chronic active hepatitis (CAH) or chronic persistent hepatitis (CPH). We conclude that in haemophiliacs, chronic HCV‐related liver disease is associated with increased plasma levels of sIL‐2R and that the degree of elevation may reflect the severity of the associated chronic liver disease.
Relapse of acute lymphoblastic leukaemia 14 years after presentation: use of molecular techniques to confirm true re‐emergence
Authors: Levasseur Mark , Maung Zor T. , Jackson Graham H. , Kernahan Jennifer , Proctor Stephen J. , Mddleton Peter G.
Published: March 2008

SUMMARY. Late relapse after successful treatment of acute lymphoblastic leukaemia (ALL) in children is well recognized but rare. It is often uncertain whether this represents a true relapse of the original disease or a second malignancy. We present the case of a patient who relapsed 14 years after the original diagnosis of childhood ALL in whom both the orignal leukaemic cells and those taken at relapse had an identical T cell receptor gamma (TCRG) gene rearrangement. This analysis confirms that this relapse is a true re‐emergence of the patient's original disease. The term ‘cure’ should be used with caution in childhood ALL, even after long periods in continuous remission.
Myeloid haemopoietic cells of patients with chronic granulomatous disease are relatively resistant to TNF
Authors: Sakashita Akiko , Curnutte John T. , Koeffler H. Phillip
Published: March 2008

Summary. Generation of superoxide may be a key step in the cytotoxicity mediated by tumour necrosis factor (TNF); cells that cannot produce oxygen radicals might be resistant to TNF. Myeloid haemopoietic cells from patients with chronic granulomatous disease (CGD) cannot produce a large burst of oxygen radicals; therefore we examined the ability of TNF to inhibit clonal growth of myeloid haemopoietic cells from patients and carriers with several types of CGD. Mononuclear light‐density cells from the peripheral blood of 13 CGD patients (11 patients with defects of gp91‐phox and two with p47‐phox), five gp91‐phox carriers and 10 normal volunteers were cultured with the appropriate growth factor and TNF in methylcellulose. As expected, TNF (0.001–100 ng/ml) inhibited colony formation of myeloid cells of normal volunteers in a dose‐dependent manner. In contrast, clonal growth of myeloid cells of CGD patients was resistant to inhibition by TNF ≤ 100 ng/ml. As expected, the effects of TNF on erythroid clonogenic cells, which are not capable of producing an oxygen burst, and the action of TGF‐β on clonal growth of myeloid cells, were similar in both the individuals with CGD and the normal volunteers. In X chromosome‐linked female carriers of CGD (gp91‐phox deficiency), TNF showed an intermediate cytotoxicity on clonal growth of myeloid cells, and analysis of NBT reduction demonstrated that the colonies derived from myeloid cells deficient in gp91‐phox were resistant to TNF and those derived from the myeloid cells expressing gp91‐phox were inhibited in their proliferation by TNF. This study shows for the first time that myeloid haemopoietic cells from patients with CGD are relatively resistant to the growth‐inhibiting effects of high concentrations of TNF.