bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitation of human cytomegalovirus DNA in bone marrow transplant recipients
Authors: Gerna Giuseppe , Furione Milena , Baldanti Fausto , Percivalle Elena , Comoli Patrizia , Locatelli Franco
Published: March 2008

Summary. HCMV DNA was retrospectively quantitated in the early post‐transplant period in 36 paediatric bone marrow transplant (BMT) recipients prospectively monitored for human cytomegalovirus (HCMV) infection on the basis of antigenaemia and viraemia assays. Viral DNA was quantitated in peripheral blood leucocytes (PBL) by PCR using an internal control of amplification and a series of external standards. Densitometric analysis of hybridization results obtained on PCR products enabled construction of a standard curve from which DNA amounts of clinical samples, expressed in terms of genome equivalents (GE), were interpolated. Of the 36 BMT recipients, three had clinically symptomatic HCMV infection with mean peak levels of viral DNA > 5000 GE (antigenaemia and viraemia mean peak levels were 873 and 35, respectively), whereas 19 with HCMV reactivation were asymptomatic (five of them had abortive HCMV infection) showing mean peak DNA levels of 131 GE (and of 6‐8 and 13 for antigenaemia and viraemia, respectively) (P≤0.01). Single or multiple courses of pre‐emptive therapy with ganciclovir or foscarnet were given to 14/19 asymptomatic children in whom antigenaemia levels were > 2 or lower yet persisting. Overall, in the 14 asymptomatic treated patients the mean antigenaemia level was 9‐3 (range 1‐22), and the mean DNA level was 184‐6 (range 20‐710) GE. Antiviral drugs were also administered to the three symptomatic patients who, due to late diagnosis of HCMV infection, escaped preemptive therapy. Antiviral treatment caused marked decrease or disappearance of viral DNA, antigenaemia and viraemia in both symptomatic and asymptomatic patients. In conclusion, our study suggests that: (i) starting therapy in the presence of a mean antigenaemia level of 9′3 (range 1‐22) corresponding to a mean DNA level of 184′6 (range 20‐710) GE avoided occurrence of any major HCMV‐related clinical complication; (ii) clinical symptoms were associated with antigenaemia levels > 100 and DNA levels > 1000 GE; (iii) the effect of antiviral treatment could be more carefully monitored by quantitation of viral DNA.
Autoimmune‐prone (NZW × BXSB)F1 (W/BF1) mice escape severe thrombocytopenia after treatment with deoxyspergualin, an immunosuppressant
Authors: Nemoto Kyuichi , Mae Takako , Saiga Kan , Matsuura Eiji , Koike Takao
Published: March 2008

Summary. Male (NZW × BXSB)F1 mice spontaneously develop a disease which closely resembles human systemic autoimmune disease, involving idiopathic thrombocytopenic purpura and glomerulonephritis. We investigated whether autoimmune thrombocytopenia in the mice responded to deoxyspergualin, as immunosuppressant. Deoxyspergualin completely prevented the development of thrombocytopenia and suppressed the increase in circulating autoantibodies against platelets. This agent also ameliorated lupus nephritis. These findings suggest that deoxyspergualin may be effective in the prevention of idiopathic thrombocytopenic purpura.
Activated platelets in paroxysmal nocturnal haemoglobinuria
Authors: Gralnick H. R. , Vail M. , McKeown L. P. , Merryman P. , Wilson O. , Chu I. , Kimball J.
Published: March 2008

Summary. One of the major causes of morbidity and mortality in paroxysmal nocturnal haemoglobinuria (PNH) is venous thrombosis. We have studied fibrinolysis, coagulation and platelets in 11 patients with PNH in an attempt to identify the possible mechanism(s) of thrombosis in PNH. In this study we did not identify any fibrinolytic defects, evidence of coagulation activation, nor reduction in coagulation inhibitors. In contrast, in this cohort of 11 PNH patients we have identified varying degrees of platelet activation as defined by the surface expression of activation dependent proteins and the binding of adhesive proteins to the platelet surface. The thrombotic events in PNH usually occur in the venous system. Our studies and previous experimental studies suggest that anti‐platelet therapy may be efficacious in reducing the incidence and severity of venous thrombosis in PNH.
Prediction of iron deficiency in chronic inflammatory rheumatic disease anaemia
Authors: Kurer Sylvia Baumann , Seifert Burkhardt , Michel Beat , Ruegg Regula , Fehr Jörg
Published: March 2008

Summary. We prospectively studied 45 anaemic patients (3 7 women, 8 men) with chronic inflammatory rheumatic diseases. The combination of serum ferritin and CRP (as well as ESR) in its predictive capacity for bone marrow iron stores was examined. The relationship between other iron‐related measurements (transferrin, transferrin saturation, soluble transferrin receptor, erythrocyte porphyrins and percentage of hypochromic/microcytic erythrocytes) and bone marrow iron stores was also investigated. Stainable bone marrow iron was taken as the most suitable standard to separate iron‐deficient from iron‐replete patients. 14 patients (31%) were lacking bone marrow iron. Regression analysis showed a good correlation between ferritin and bone marrow iron (adjusted R2=0.721, P<00001). The combination of ferritin and CRP (ESR) did not improve the predictive power for bone marrow iron (adjusted R2=0.715) in this cohort of patients with low systemic inflammatory activity. With respect to the bone marrow iron content the best predictive cut‐off value of ferritin was 30μg/l (86% sensitivity, 90% specificity). The other iron‐related parameters both individually and when combined were less powerful in predicting bone marrow iron than ferritin alone. Only zinc bound erythrocyte protoporphyrin in combination with ferritin slightly improved prediction (adjusted R2=0.731). A cut‐off point of 11% hypochromic erythrocytes reached a high specificity (90%), but was less sensitive (77%).
Clinical significance of serum cytokine patterns during start of fever in patients with neutropenia
Authors: Engervall Per , Andersson Birger , Björkholm Magnus
Published: March 2008

Summary. Serum concentrations of tumour necrosis factor alpha (TNF‐α), interleukin‐1 receptor antagonist (IL‐1ra), interferon gamma (IFN‐γ), interleukin‐6 (IL‐6) and interleukin‐10 (IL‐10) were studied in 31 patients with haematological malignancies during febrile neutropenia. Samples were obtained when blood cultures were performed (time 0) and, when possible, after 2, 4, 6, 12 and 24 h. Increased levels of all cytokines were detected after start of fever with peak values in gram‐negative (Gr−) bacteraemias after 2 h (TNF‐α, IL‐lra and IFN‐γ), 4h (IL‐6) and 6 h (IL‐10), respectively. At time 0 the median TNF‐α value was higher in the Gr− group (80 pg/ml; range 54‐516 pg/ml) as compared to both gram‐positive bacteraemias (Gr+, 14pg/ ml; range 7‐60 pg/ml; P < 0 05) and blood culture negative episodes (BCN, 8pg/ml; range 0‐87pg/ml; P<0 05).

Furthermore, the peak values of TNF‐α, IL‐lra, IL‐6 and IL‐10 during the 24 h study period were significantly and/or numerically higher in the Gr‐ group in comparison to the Gr+ and BCN groups, respectively. It may be concluded that neutropenic patients have increased levels of both pro‐ and anti‐inflammatory cytokines at start of fever, with the highest values recorded during the first hours in Gr‐ bacteraemias. Prospective studies will show whether monitoring of serum cytokines may be used as an early diagnostic tool before results of blood cultures are available, which may have important therapeutic implications.
Platelets prime PMN via released PF4: mechanism of priming and synergy with GM‐CSF
Authors: Aziz Khalil A. , Cawley John C. , Zdzel Mirko
Published: March 2008

Summary. Platelet‐PMN interactions have been extensively studied and a spectrum of possible effects has been demonstrated. However, the physiological relevance of many of the observed in vitro phenomena remains obscure. Here we report a novel, and potentially pathophysiologi‐cally important, mechanism by which platelets can enhance PMN reactivity. We first observed that addition of platelets to PMN suspensions enhanced the chemilumines‐cence response of PMN to FMLP. This enhancement occurred without augmentation of superoxide generation and did not involve mutual platelet‐PMN adhesion. The soluble material responsible was biochemically and immunologically identified as PF4 derived from platelet α‐granules. The α‐granule release was shown to be selective and required minimal platelet stimulation. Since the PF4 effect did not influence NADPH oxidase activation, it differed markedly from that of other priming agents such as GM‐CSF. Further studies showed that the PF4 effect was attributable entirely to the surface translocation and secretion of primary granule myeloperoxidase. There was marked synergy between PF4 and GM‐CSF and both were required for maximal potentiation of PMN reactivity. These results demonstrate that PF4 and GM‐CSF employ different pathways in PMN priming. The ease with which platelets could release PF4 at sites of vessel‐wall damage and inflammation suggests that platelet‐PMN interaction via PF4 is likely to be of major pathophysiological importance.
Expression of the leucocyte common antigen (LCA, CD45) isoforms RA and RO in acute haematological malignancies: possible relevance in the definition of new overlap points between normal and leukaemic haemopoiesis
Authors: Schiavone Ettore Mariano , Pardo Catia Lo , Noto Rosa Di , Manzo Ciro , Ferrara Felicetto , Vacca Clemente , Vecchio Luigi Del
Published: March 2008

Summary. The membrane expression of CD45RA and CD45RO on fresh leukaemic cells taken from 529 cases of acute haemopoietic malignancies, including 117 B‐origin acute lymphoblastic leukaemia (B‐origin ALL), 3 7 T‐origin acute lymphoblastic leukaemia (T‐origin ALL), 297 de novo acute myeloid leukaemia (AML), 42 refractory anaemia with excess of blasts in transformation (RAEB‐T) and 36 myeloid blastic phase of chronic myelogenous leukaemia (CML‐BP‐my), was analysed. B‐origin ALLs were characterized by the lack of the RO isoform along with the consistent presence of RA. Conversely, a differential expression of the two isoforms was detected in different subsets of T‐origin ALL, in that T‐stem cell leukaemias (T‐SCL: CD7+, CD4−, CD8−, CD1−) preferentially expressed CD45RA whereas conventional T‐acute lymphoblastic leukaemias (T‐ALL: CD7+, CD4+ and/or CD8+ and/or CD1+) were consistently marked by CD45RO. Within myeloid malignancies, most of AMLs displayed CD45RA, while a substantial group of CML‐BP‐my preferentially exhibited CD45RO. As a general rule, a reciprocal exclusion of the two isoforms was observed in AML as well as in ALL. Nevertheless, a frequent coexpression of CD45RA and CD45RO was observed in CD14+ AML.

In vitro treatment with all‐trans retinoic acid (ATRA) was able to promote a switch from CD45RA to CD45RO expression in 2 7 de novo AML, independently from morphological subtyping.

To our knowledge, this is the first report on CD45 isoform expression in a large series of patients with acute leukaemia. The knowledge of the differential expression of CD45RA and CD45RO can ameliorate our classificative approach to haematological malignancies, as well as disclose new multiple overlap points between normal and leukaemic cell differentiation.
Expression of the human OX40 (hOX40) antigen in normal and neoplastic tissues
Authors: Dürkop Horst , Latza Ute , Himmelreich Petra , Stein Harald
Published: March 2008

Summary. The Ber‐ACT35 mAb was raised against the human T‐cell lymphotrophic virus (HTLV) 1‐transformed, CD4+ HUT 102 cell line and recognizes the human homologue of the 0X40 (hOX40) antigen. The analysis of the expression of hOX40 by immunohistochernical techniques in malignant lymphomas, carcinomas and non‐malignant tissues of different organs shows that hOX40 expression was almost completely restricted to T lymphocytes. Besides T cells only a small subpopulation of macrophages in Langerhans’cell histiocytosis and a few blasts in B‐cell non‐Hodgkin's lymphoma (B‐NHL) revealed a faint immunostaining with the Ber‐ACT35 mAb. Furthermore, most of the hOX40+ T‐cells are CD4+.
Expression of lymphoid‐associated antigens in mast cells: report of a case of systemic mast cell disease
Authors: Escribano Luis , Orfao Alberto , Villarrubia Jesús , Cerveró Carlos , Velasco José L. , Martín Flor , Miguel Jesus F. San , Navarro Jose L.
Published: March 2008

Summary. In this study the expression of‘classically’considered lymphoid‐associated antigens (CD2, CD3, CD4, CD5, CD7, CD8, CD10, CD19, CD20, and CD22) was explored both in peripheral blood (PB) and bone marrow (BM) mast cells (MC) in a case of systemic mast cell disease (SMCD) by means of using multiple stainings and a direct immunofluorescence technique. CD2 and CD22 were expressed in both PB and BM MC, all the remaining lymphoid‐associated markers were negative. Our results suggest that the reactivity for both CD2 and CD22 in PB and BM MC would be aberrant.
Propagation of large numbers of cells of a human mixed‐lineage T‐lymphoid/myeloid
Authors: SCHMIDT‐WOLF INGO G. H. , GRIMM BEATE , LEFTEROVA PETJA , JOHNSTON VALERIE , SCHEFFOLD CHRISTIAN , HUHN DIETER , SERKE STEFAN
Published: March 2008

Summary. Previously, a subset of T cells co‐expressing the myeloid antigen CD33 has been described in patients with acute myelogenous leukaemia. However, normal lymphocytes have been viewed as not expressing the CD33 antigen. We have developed culture conditions which allow for the rapid expansion of CD3+CD33+ cells from patients with myeloid leukaemia as well as normal individuals. The protocol for cellular expansion includes the addition of interferon‐gamma on day 0, interleukin‐1, interleukin‐2 and a monoclonal antibody against CD 3 on day 1 to peripheral blood lymphocytes. Using this protocol, total cell number increased more than 600‐fold within 16 d of culture. Cells could be kept in culture for more than 6 months. Cells of the CD3+CD33+ phenotype increased to 15‐2 –4‐6% using this protocol after 16 d in culture. These cells have been characterized by flow cytometry and have been found to express the alpha, beta T‐cell receptor, co‐express the CD2, CD5, CD7 and HLA‐DR antigens and did not express CD14 or CD15 antigens. Cells of the CD3+CD33+ phenotype were unable to lyse tumour cells as determined in a 51Cr release assay. In patients with chronic myeloid leukaemia, CD3+CD33+ cells seem to be negative for expression of bcr/abl transcript in contrast to CD33 cells. Our data suggest that CD3+CD33+ cells do exist in peripheral blood from normal individuals.
Influence of white blood cells on the fibrinolytic response to sepsis: studies of septic patients with or without severe leucopenia
Authors: HAJ MONTASER A. , NEILLY IAN J. , ROBBIE LINDA A. , ADEY GILLIAN D. , BENNETT BRUCE
Published: March 2008

Summary. In septic patients capable of normal white cell responses, high plasma levels of PAI‐I, t‐PA antigen and t‐PA‐PAI‐I complex were observed. The ratios of t‐PA and PAI‐I were such that free PA activity was almost never observed. In patients severely leucopenic prior to becoming septic the changes were significantly less marked, so presence of leucocytes enhances the fibrinolytic inhibition occurring in sepsis. The non‐leucopenic septic group showed greater evidence of thrombin generation in that FPA levels were higher but fibrinogen levels were only slightly less and antithrombin levels not different from those in the leucopenic group. A greater tendency to fibrin deposition and the striking fibrinolytic inhibition noted in patients with normal white cell responses may contribute to the development of some of the complications of sepsis in which fibrin deposition participates and may explain their relative rarity in leucopenic patients. When shock supervened, levels of PAI‐I were high in both leucopenic and non‐leucopenic groups, indicating that a source of PAI‐I outwith the leucocytes themselves contributes to the phenomena observed.
Differential sensitivity of CD30+ neoplastic cells to gelonin delivered by anti‐CD30/anti‐gelonin bispecific antibodies
Authors: SFORZINI SABRINA , BOLOGNESI ANDREA , MEAZZA RAFFAFXLA , MARCIANO SABRINA , CASALINI PATRIZIA , DiiRKOp HORST , TAZZARI PIER LUIGI , STEIN HARALD , STIRPE FIORENZO , FERRINI SILVANO
Published: March 2008

Summary. Lymphocyte activation antigens, such as CD30, represent suitable target molecules for antibody‐driven drug delivery in haemopoietic malignancies. A ribosome‐inactivating protein (RIP) type 1 of potential interest for mAb targeting is gelonin, which displays a lower toxicity, as compared to other RIPs. In this study, two anti‐CD 3 0/anti‐gelonin bispecific monoclonal antibodies (bimAbs), secreted by hybrid hybridomas, were used to deliver this RIP to CD30+ tumour cells. The two bimAbs, termed D4 and A18, were produced using the same anti‐CD30 mAb and two anti‐gelonin mAbs, directed to unrelated epitopes of the gelonin molecule. These bimAbs enhanced gelonin toxicity (IC50 5 × 10−8 M, in the absence of mAbs) against the CD30+ L540 Hodgkin's lymphoma cell line in a protein synthesis inhibition assay. Thus, in the presence of 10−9 M D4 bimAb, protein synthesis was inhibited with an ICs0 of 5 × 1CT10M as gelonin, whereas with A18 bimAb the ICS0 was 8 × 1CT11 M. More interestingly, the combined use of the two bimAbs had a synergistic effect, since the IC50 of gelonin reached 6 × 10_12M. Among CD30 tumour cell lines, the Hodgkin's lymphoma L428 was also sensitive to gelonin delivered by bimAbs (IC50 6 × 1CTU M), whereas the COLE Hodgkin's cell line and the T‐ALL Jurkat were completely resistant to the toxic effect of gelonin and bimAbs. COLE and Jurkat cells were also resistant to a gelonin/anti‐CD30 conventional immunotoxin, whereas they were sensitive to a saporin/anti‐CD30 immunotoxin. This suggests that the resistance to gelonin is not related to a lack of internalization through the CD30 molecule but is associated with some property of the RIP.
Allogeneic marrow grafts from donors with congenital chromosomal abnormalities in marrow cells
Authors: BARQUINERO JORDI , WITHERSPOON ROBERT , SANDERS JEAN , HOROWITZ MARY M. , MONTUORO ALDO , PATTON DONNA F. , BACIGALUPO ANDREA , ABECASIS M. M. , MIALE THOMAS , ROZMAN CIRIL , BRYANT EILEEN , STORB RAINER
Published: March 2008

Summary. To determine whether siblings with chromosomal abnormalities in marrow cells which are associated with cellular defects (e.g. Down syndrome or heterozygosity for Fanconi syndrome) are suitable donors for allogeneic bone marrow transplants, we have reviewed the patient files at the Fred Hutchinson Cancer Research Center (FHCRC) and carried out a survey among member centres of the International Bone Marrow Transplant Registry (IBMTR). The 57 of 253 (23%) member centres which responded to the survey reported seven transplants from donors with the following conditions: Down syndrome (n = 2), suspected heterozygotes for Fanconi syndrome (n = 3), and 47.XXX syndrome (n = 2), among a total of 5561 allogeneic transplants from HLA‐identical siblings. Adding the three cases seen at the Fed Hutchinson Cancer Research Center among 2927 HLA‐identical sibling transplants during 1992 resulted in 10 transplants among 8488 cases transplanted overall: four with Down syndrome, four suspected of being heterozygous for Fanconi syndrome, and two trisomy X. Three out of four grafts from siblings with Down syndrome had complications, including poor graft function (n = 2) and graft failure (n= 1). Two of four recipients of marrow from presumed Fanconi syndrome heterozygotes presented with poor graft function and a third recipient developed graft failure after initial evidence of engraftment. The two patients given marrow from siblings with 47.XXX syndrome engrafted uneventfully. The experience reported here shows a low frequency of encountering an HLA‐identical sibling donor who has chromosomal abnormalities in marrow cells consistent with Down syndrome or heterozygosity for Fanconi syndrome, about one case among 1000 transplants. The much higher than expected incidence of graft problems with marrow from such a donor would make it reasonable to look either for an alternative marrow donor or consider an autologous transplant, in case a sibling marrow donor with Down syndrome or heterozygosity for Fanconi syndrome is encountered, although a donor with trisomy X seems acceptable.
Redistribution of GPIb/IX and GPIIb/IIIa during spreading of discoid platelets
Authors: WHITE JAMES G. , KRUMWIEDE MARLYS D. , JOHNSON DEBRA K. , ESCOLAR GINES
Published: March 2008

Summary. The purpose of the present study was to prelabel mobile receptors on discoid platelets with specific ligands identifiable in the electron microscope and follow their redistribution during spreading. Platelets were incubated in suspension with cytochalasin E (CE) to preserve discoid form, chilled and mounted on cold formvar grids or glass slide fragments to inhibit receptor movement, covered with cold bovine or ristocetin‐activated human plasmas as sources of vWF to bind GPIb/IX, fibrinogen‐coated gold particles (Fgn/ Au) to couple GPIIb/IIIa, or both probes simultaneously, washed to remove CE and rewarmed to 37°C for intervals up to 30min to stimulate spreading. After brief fixation grids and glass fragments were incubated with anti‐vWF antibody and, subsequently, staphylococcal protein A coupled to 5 nm and 10 nm gold particles to detect vWF multimers. Virtually all of the CE‐treated chilled platelets retained their discoid shape. Half of the discs (53‐3%) bound Fgn/Au, and all bound vWF. Receptors for both ligands were randomly dispersed on discoid cells from edge to edge. During re warming discoid platelets expanded into spread forms. Fgn/Au‐GPIIb/IIIa complexes moved into caps over cell centres and into residual channels of the open canalicular system (OCS). vWF bound to GPIb/IX moved with the cell membrane as the surface expanded during spreading. Discoid platelets prelabelled with both ligands demonstrated similar findings. During rewarming Fgn/Au‐GPIIb/IIIa complexes moved to cell centres and into OCS channels. vWF multimers bound to GPIb/IX moved apart from each other toward peripheral margins of the spread cells. Thus, surface activation resulting in conversion of discoid platelets to spread forms does not cause clearance of GPIb/IX receptors to cell centres and channels of the OCS in the manner that GPIIb/IIIa receptors coupled to Fgn/Au are simultaneously translocated and concentrated in OCS channels.
Monoclonal gammopathy of undetermined significance: chromosome changes are a common finding within bone marrow plasma cells
Authors: ZANDECKI MARC , OBEIN VALERIE , BERNARDI FRANCK , SOENEN VALERIE , FLACTIF MARTIAL , LAI JEAN LUC , FRANCOIS MICHEL , FACON THIERRY
Published: March 2008

Summary. We used two indirect approaches [image analysis (Feulgen staining) and fluorescence in situ hybridization (FISH)] to study bone marrow plasma cells (BMPC) in 28 patients fulfilling criteria for MGUS. 61% of patients were found to be aneuploid after image analysis: three were hypodiploid and 14 were hyperdiploid. 12/14 hyperdiploid patients also revealed abnormalities after FISH: 12‐72% of BMPC exhibited trisomy for at least one of chromosomes 3, 7, 9 and 11. These latter chromosomes are the four chromosomes most frequently implicated (in the shape of trisomy) in MM, confirming the tight relationship between both conditions. After a median follow‐up of 19 months (12‐41 months) no patient developed overt MM. Also, we failed to find any relationship between currently available biological parameters and DNA findings. As literature data give a transformation rate of 20‐30% after a follow‐up of 20‐35 years, it is worth presuming that some aneuploid patients will evolve to MM, whereas others (also with aneuploid bone marrow plasma cells) will never develop cancer. Our findings indicate that numeric abnormalities, as they are shared both by MGUS and MM patients, are certainly an additional or a prerequisite event, but are not related to an overt disease. They also emphasize the importance of cytogenetic study in the pathophysiology of MGUS.
Combined immunophenotyping and in situ hybridization (FICTION): a rapid method to study cell lineage involvement in myelodysplastic syndromes
Authors: SOENEN VALERIE , FENAUX PIERRE , FLACTIF MARTIAL , LEPELLEY PASCALE , LAI JEAN LUC , COSSON ALAIN , PREUDHOMME CLAUDE
Published: March 2008

Summary. We present a study in which we used a recently described method combining fluorescence in situ hybridization (FISH) and immunophenotyping, i.e. FICTION, to assess the involvement of different cell lineages in myelodysplastic syndrome (MDS) with monosomy 7 (–7), trisomy 8 (+8) or loss of Y chromosome (–Y). Blood or marrow smears or cytocentrifuge preparations were stained both by antibodies to granulocytes (CD15), monocytes (CD14), T lymphocytes (CD3), B lymphocytes (CD2o) and by probes specific for chromosomes 7, 8 or Y. Of nine cases of MDS with –7, four with +8 and two with – Y studied, none showed lymphocytic involvement by the chromosome abnormality. In contrast, ‐7,‐1‐8 and – Y were found in granulocytes and monocytes in all patients studied, but they involved a variable proportion of those cells. The partial involvement by –7 and +8 seen in some cases suggests that myelopoi'esis was only partially clonal in those cases, or that the chromosome abnormality was a secondary event in the MDS process. FICTION therefore appears to be a simple and easily reproducible method that can be used for the assessment of lineage involvement in MDS and other haematological malignancies.
The gp 130 family cytokines IL‐6, LIF and OSM but not IL‐11 can reverse the anti‐proliferative effect of dexamethasone on human myeloma cells
Authors: JUGE‐MORINEAU NADINE , FRANCOIS SYLVIE , PUTHIER DENIS , GODARD ANNE , BATAILLE REGIS , AMIOT MARTINE
Published: March 2008

Summary. In order to understand the mechanisms supporting steroid escape in patients with multiple myeloma (MM), three IL‐6 autocrine human myeloma cell lines, LP1, OPM2 and L363, have been treated with dexamethasone in the presence or absence of cytokines belonging to the gp 130 family: IL‐6, LIF, OSM and IL‐11. With pharmacological doses of dexamethasone, a dramatic growth arrest was observed in all the cell lines. IL‐6 completely reversed this inhibition. Of note, this IL‐6 induced reversion was still seen with very low amounts of IL‐6 (12 pg/ml). Finally, whereas LIF and OSM had clear growth‐promoting effects on OPM2 only, both cytokines (but not IL‐11) reversed the dexa‐methasone‐induced growth arrest in all the cell lines. Therefore the high levels of IL‐6 (ng/ml) observed in the MM intermediate milieu and the putative presence of LIF and OSM can easily counteract the effects of dexamethasone in vivo.
Flow cytometric analysis of decay‐accelerating factor (CD55) on neutrophils from aplastic anaemia patients
Authors: FORES RAFAEL , ALCOCER MARIA , DIEZ‐MARTIN JOSE L. , FERNANDEZ MANUEL N.
Published: March 2008

Summary. Using a flow cytometric analysis, CD55 (decay‐accelerating factor), CD59 and CD58 have been measured on neutrophils from 12 aplastic anaemia (AA) patients who were long‐term survivors after immunosuppressive therapy (IS), 17 healthy individuals, four patients with PNH, and six patients with other haematological disorders.

The neutrophils from normal control patients and the six patients with other haematological disorders showed 98 ±2% (mean ±SD) positive granulocytes for CD55. Corresponding values were low (12%, 26%, 51% and 58%) on the primarily PNH patients. Among the 12 AA patients examined, seven had normal and five low values (59% in two, 70%, 71% and 82%).

Among the five A A patients who showed CD 5 5 neutrophil deficiency, four had showed an incomplete response after the initial IS treatment and the other relapsed following an initial haematological complete response; three cases had a positive Ham's test and two were negative.

Our data suggest that the development of PNH clones is a frequent finding in A A long‐term survivors, mainly in those who had shown an incomplete response following IS. Neutrophil CD55 expression analysis by flow cytometry could be useful to detect clonal evolution in these patients.
Granulocyte colony‐stimulating factor administration modulates the surface expression of effector cell molecules on human monocytes
Authors: Ohsaka Akimichi , Saionji Katsu , Kuwaki Tomoaki , Takeshima Toshisuke , Igari Jun
Published: March 2008

Granulocyte colony‐stimulating factor (G‐CSF) has been shown to stimulate human neutrophil functions, both in vitro and in vivo. We examined the effects of G‐CSF administration on the surface expression of effector cell molecules on human neutrophils and monocytes. G‐CSF (50 μg/m2/d) was administered subcutaneously to five healthy volunteers once a day for 7 d. Venous blood was obtained immediately before and after the completion of G‐CSF administration and 1 week after the last G‐CSF administration. The surface expression of complement receptors (CR), Fc receptors for IgG (FcR) and cellular adhesion molecules on human neutrophils and monocytes were determined by indirect immunofluorescence using flow cytometry and monoclonal antibodies. The expression of CR1, CR3, FcRI and FcRII on neutrophils increased significantly after G‐CSF administration and then decreased after the last G‐CSF administration. The expression of human leucocyte adhesion molecule‐1 (LAM‐1) on neutrophils reflected the above expression. On the other hand, the administration of G‐CSF increased the expression of CR1, CR3, FcRI and FcRIII on monocytes. The expression of CR1, CR3 and FcRI on monocytes then decreased after the last G‐CSF administration, whereas the expression of FcRIII remained at an increased level. These findings indicate that G‐CSF administration modulates the expression of effector cell molecules on circulating monocytes as well as on neutrophils, resulting in enhanced defence against selected infections or in potentiation of the tumouricidal capacity of phagocytes in cancer patients.
Relationship between transfusion regimen and suppression of erythropoiesis in β‐thalassaemia major
Authors: Cazzola Mario , Stefano Piero De , Ponchio Luisa , Locatelli Franco , Beguin Yves , Dessì Carlo , Barella Susanna , Cao Antonio , Galanello Renzo
Published: March 2008

In the management of β‐thalassaemia major, different transfusion schemes are employed with baseline haemoglobin levels ranging from 8 to over 12 g/dl. We studied the relationship between transfusion regimen and suppression of erythropoiesis in 52 patients with β‐thalassaemia major whose mean pretransfusion haemoglobin levels ranged from 8.6 to 10.9 g/dl. Multiple, regression analysis showed that serum transferrin receptor was the parameter more closely related to mean pretransfusion haemoglobin (r = ‐0.77, P < 0.001). As measured through serum transferrin receptor, erythroid activity was 1‐2 times normal for pretransfusion haemoglobin levels between 10 and 11 g/dl, 1‐4 times normal for levels from 9 to 10 g/dl, and 2‐6 times normal for levels from 8.6 to 9 g/dl. Mean pretransfusion haemoglobin was also inversely related to serum erythropoietin (r = ‐0.72, P < 0.001), whereas it showed no or a weak relationship with Hb F, reticulocyte count, or circulating nucleated red cell count. This study suggests that serum transferrin receptor is a reliable indicator of suppression of erythropoiesis in β‐thalassaemia major. On the basis of our findings, pretransfusion haemoglobin values of ≦ 9 g/dl should be adopted with caution, because these levels can be associated with an insufficient inhibition of erythroid marrow expansion. However, a transfusion programme, with a baseline haemoglobin of 9‐10 g/dl, may provide enough suppression of erythropoiesis and allow a reduction in blood consumption as compared with the classic hyper‐ or supertransfusion schemes. Since fixed haemoglobin levels may not be the best target for transfusion treatment in all thalassaemic patients, assay of serum transferrin receptor may be helpful for individualizing the transfusion regimens.
Thalassaemia in Vanuatu, South‐West Pacific: frequency and haematological phenotypes of young children
Authors: Ganczakowski M. , Bowden D. K. , Maitland K. , Williams T. N. , O'Shaughnessy D. , δ J. Viji , Lucassen A. , Clegg J. B. , Weatherall D. J.
Published: March 2008

The archipelago of Vanuatu situated in the South‐West Pacific has a high frequency of α+ thalassaemia and additionally on some of the islands there is a high frequency of β thalassaemia. As part of a large cohort study to investigate the clinical effect of thalassaemia on malaria on the islands of Espiritu Santo and Maewo in Vanuatu, the gene frequencies of the thalassaemias were determined and blood counts were performed on a cohort of infants from birth to 3 years. The haematological phenotypes of the different thalassaemic genotypes are compared, providing a detailed description of the clinical manifestations of α+ thalassaemia during early development. In addition, crosssectional surveys of the population of the two islands were performed to establish the frequency of thalassaemia and other red cell polymorphisms and their geographical distribution.
α‐Thalassaemia in the population of Cyprus
Authors: Baysal E. , Kleanthous M. , Bozkurt G. , Kyrri A. , Kalogirou E. , Angastiniotis M. , Ioannou P. , Huisman T. H. J.
Published: March 2008

We have determined the α‐thalassaemia (α‐thal) determinants in 78 patients with Hb H disease from Cyprus; 25 were Turkish Cypriots and 53 were Greek Cypriots. Four deletional and three non‐deletional α‐thal alleles were present; the ‐α(3.7 kb) α‐thal‐2 and the —MED‐1α‐thal‐1 were most frequently seen; —MED‐II and ‐(α)20.5 deletions occurred at considerably lower frequencies. About 15% of all chromosomes carried a non‐deletional α‐thal‐2 allele; of these the 5 nucleotide (nt) deletion at the first intervening sequence (IVS‐I) donor splice site was present in ˜ 8% of all chromosomes. Two types of polyadenylation signal (poly A) mutations were observed. No striking frequency differences were seen between Greek and Turkish Cypriot patients. Combinations of the various types of α‐thal resulted in eight different forms of Hb H disease. The phenotypes were comparable except for great variations in the level of Hb H which was highest (average ˜ 22%) in the 12 patients with the α5ntα/—MED‐I combination. One patient with the same form of Hb H disease but with an additional β‐thal (IVS‐I‐110, G → A) heterozygosity had a most severe microcytosis and hypochromia with < 1% Hb H. Variations in the level of Hb H might correlate with the severity of the disease, although this was not evident from the haematological data.
Frequent association of t(3;14) or variant with other lymphoma‐specific translocations
Authors: Horsman Douglas E. , McNeil B. Kelly , Anderson Michelle , Shenkier Tamara , Gascoyne Randy D.
Published: March 2008

Malignant lymphomas (ML) with t(3;14) or variant t(2;3) and t(3;22) have recently been recognized. These translocations have been shown to associate predominantly with B‐cell diffuse large cell lymphoma (DLCL) and less frequently with follicular lymphoma (FL). The molecular alterations associated with these translocations involve one of the immunoglobulin gene (Ig) loci and a recently cloned gene, bcl‐6 located at 3q27 which codes for a zinc‐finger protein that may function as a transcription factor.

We have identified by cytogenetic analysis 22 cases of ML with a 3q27/Ig translocation. The pathologic diagnoses of these cases include DLCL, FL, small non‐cleaved non‐Burkitt lymphoma and chronic lymphocytic leukaemia. Molecular analysis confirmed a bcl‐6 rearrangement in 10/12 cases tested.

The karyotype in 5/22 cases revealed the t(3;14) or variant in association with another lymphoma‐specific translocation, t(14;18) in three cases and t(8;14) in two cases. ML with dual translocations that implicate Ig genes in the deregulation of proto‐oncogenes are being increasingly recognized and may represent distinct subtypes or ‘hybrid’forms of malignant lymphoma.
Attenuated platelet sensitivity to collagen in patients with neurofibromatosis type 1
Authors: Rasko John E. J. , North Kathryn N. , Favaloro Emmanuel J. , Grispo Linda , Berndt Michael C.
Published: March 2008

Haemostatis has not previously been studied in patients with neurofibromatosis 1 (NF‐1), despite case reports of an association with von Willebrand disease and reported excessive bleeding in those undergoing surgery for neurofibromas. Platelets from NF‐1 patients (n= 28) were tested for aggregation and ATP release with agonists including ADP, arachidonic acid, thrombin and collagen. Mepacrine staining of platelets and three different assays for von Willebrand factor (VWF) were also performed.

In response to collagen as the platelet agonist, tested at both 2 and 1 μg/ml, NF‐1 patients had an attenuated rate of aggregation (P < 0.007), aggregation lag phase (P < 0.005) and ATP release (P < 0.045), as well as requiring higher collagen concentrations to attain threshold aggregation response (P= 0.041). Normal platelets resuspended in selected NF‐1 plasma exhibited significantly reduced platelet aggregation and release compared to controls, which was not corrected by mixing 1:1 with normal plasma. Collagen binding activity was reduced in NF‐1 patients compared with controls (127% v 161%, P= 0.05).

As a group, patients with NF‐1 display defective platelet function characterized by in vitro evidence of impaired responsiveness to collagen. It is suggested that a plasma factor, present in a significant proportion of NF‐1 patients, may interfere with the ability of collagen to interact with other proteins such as von Willebrand factor and the platelet collagen receptor.
Antithrombins Southport (Leu 99 to Val) and Vienna (Cln 118 to Pro): two novel antithrombin variants with abnormal heparin binding
Authors: Chowdhury V. , Mille B. , Olds R. J. , Lane D. A. , Watton J. , Barrowcliffe T. W. , Pabinger I. , Woodcock B. E. , Thein S. L.
Published: March 2008

We report the characterization of three variant antithrombins with reduced heparin binding as the primary abnormality. Two of these variants, antithrombin Southport (Leu 99 to Val, 2759 C to G) and antithrombin Vienna (Gln 118 to Pro, 5349 A to C) were novel, whereas the third, Pro 41 to Leu, has been previously described as antithrombin Basel. All three variants exhibited reduced binding for heparin on crossed immunoelectrophoresis and in a quantitative monoclonal antibody‐based assay. The mutations were characterized by direct. sequence analysis of enzymatically amplified genomic DNA and all affected individuals were heterozygous for the mutations. These three mutations do not occur at the sites of the basic amino acids directly involved in heparin binding nor do they result in a change in charge of the affected residue. It seems probable that they reduce heparin affinity either by perturbing the initial contact site involved in the heparin‐binding domain (Arg 47, Arg 129 and possibly Arg 24), or by preventing the subsequent heparin‐induced conformational change.
Effective treatment of disease‐related anaemia in B‐chronic lymphocytic leukaemia patients with recombinant human erythropoietin
Authors: Pangalis Gerassimos A. , Poziopoulos Christos , Angelopoulou Maria K. , Siakantaris Marina P. , Panayiotidis Panos
Published: March 2008

Nine B‐chronic lymphocytic leukaemia (B‐CLL) patients suffering from anaemia, due to no obvious cause except their disease, were treated with recombinant human erythropoietin (r‐HuEPO). The treatment protocol provided a closed label phase of 3 months duration, during which the patients received r‐HuEPO or placebo in a ratio of 2:1, followed by an open label phase, also of 3 months duration, during which r‐HuEPO was administered to all patients three times a week s.c. r‐HuEPO was given at a dose of 150 U/kg of body weight with an escalation of 50 U/kg up to a maximum of 300 U/kg three times a week. Complete response was achieved in 5/9 (55%) patients and partial response in 3/9 (33%). The response obtained was independent of the pretreatment serum EPO levels, the duration of anaemia, the concomitant administration of chemotherapy, the presence of splenomegaly, or the degree of bone marrow infiltration by lymphocytes. It appears that r‐HuEPO is very effective in reversing the disease‐related anaemia of B‐CLL patients.
Epinephrine test and plasma elastase as diagnostic tools in a patient with CD3+ large granular lymphocyte proliferation
Authors: Seebach Jörg , Schleiffenbaum Boris , Rüegg Regula , Fehr Jörg
Published: March 2008

Large granular lymphocytes (LGL) proliferation is characterized by expansion of cytotoxic lymphocytes and associated with neutropenia. In a case of CD3+ LGL‐proliferation the epinephrine stimulation test (EST) induced a striking elevation of CD3+, CD8+, CD57+ LGL in peripheral blood from 2.7 x 109/1 to 20 x 109/1 and might be an additional diagnostic tool in patients with normal or low absolute numbers of circulating LGL. After treatment with steroids, plasma elastase ‐ a marker of neutrophil destruction ‐ decreased from 162 to 40μg/1 (normal < 47μg/1) which correlated well with a simultaneous increase in peripheral neutrophil counts from 0.14 to 1.0 x 109/1. This finding supports the hypothesis that neutropenia in CD3+ LGL proliferation is due to neutrophil destruction, possibly mediated by LGL.
Non‐radioactive SSCP for genotyping human platelet alloantigens
Authors: Peyruchaud Olivier , Nurden Alan , Bourre Franclois
Published: March 2008

Human platelet alloantigen systems are responsible for neonatal and post‐transfusional thrombocytopenias. The determination of the different allotypes can be performed using immunological or DNA‐based methods. The most used DNA‐based procedure requires the digestion by specific restriction enzymes of PCR products containing the genetic determinants of these alloantigens. We now report a rapid method of genotyping which does not use restriction enzymes and is less prone to misinterpretation. This is non‐radioactive PCR‐SSCP (single strand conformation polymorphism), which we illustrate for two different HPA system, one on GPIIIa (HPA‐1) and the other on GPIIb (HPA‐3).
Low‐dose cladribine for symptomatic hairy cell leukaemia
Authors: Juliusson Gunnar , Lenkei Rodica , Tjønnfjord Geir , Heldal Dag , Liliemark Jan
Published: March 2008

Cladribine is an effective therapy for hairy cell leukaemia (HCL), but the standard regime is frequently complicated by neutropenic fever and prolonged T‐cell depression. We studied 102 patients with active HCL following treatment with various doses of cladribine given for 7 d. Two patients received 1 mg cladribine/m2/d without toxicity or effect. Eight subsequent patients received 2 mg cladribine/m2/d, and normalized cytopenia as quickly as 94 control patients receiving a standard dose (3.4mg/m2 or 0.085 mg/kg), with significantly less lymphopenia and a similar complete remission rate.
Microscopic haematuria as a relative contraindication for tranexamic acid
Authors: Schultz Marc , Van Der Lelie Hans
Published: March 2008

Tranexamic acid has been advocated for patients with severe bleeding tendency due to thrombocytopenia not responding to platelet transfusions. Macroscopic haematuria is a well‐known contraindication for its use in such patients. We present three clinical cases with microscopic haematuria, in whom tranexamic acid caused problems of clot formation in the urinary tract, indicating that microscopic haematuria should also be considered as a contraindication for tranexamic acid.
Summary of findings from a participant country pollen beetle questionnaire
Authors: Richardson D. M.
Published: March 2008

A questionnaire was sent to all participating countries to gain some background information on rape growing and on pollen beetle (Meligethes aeneus). Information was sought on the use of thresholds, the availability of active substances and on the year in which resistance, if present, had been observed. A summary of the main findings of the questionnaire are presented. In total 20 countries responded and resistance in pollen beetle was reported in 12 of those countries. Information presented shows the range of active substances and doses recommended, frequency of use of insecticides on rape, and year when resistance was reported.
Experiences in controlling resistant pollen beetle by type I ether pyrethroid Trebon 30 EC in Germany
Authors: Glattkowski H. , Saggau B. , Goebel G.
Published: March 2008

Oil seed rape (OSR) is an important crop in Germany with a long growing history. Due to the prevalence of various springtime pests an average of two pyrethroid applications are made. Pyrethroid resistance in Germany was found locally in 2003, widespread distribution of resistance was observed in 2006, a year of very strong pest pressure. Field trials in 2005 and the following years and laboratory analysis showed good control of resistant pollen beetle (Meligethes aeneus) by type I ether pyrethroid etofenprox, the active substance of Trebon 30 EC. Etofenprox differs significantly from common pyrethroids, which are usually ester pyrethroids including an α‐amino‐group and halogen substitutions. Trebon 30 EC has comparable efficacy on all important OSR pests but it has superior efficacy on resistant pollen beetle compared to common pyrethroids. To cope with pyrethroid resistance of pollen beetle, three different modes of action (MOA)s are postulated, which may be type I pyrethroids, neonicotinoids and organophosphates (OPs). Because type I pyrethroids, such as Trebon 30 EC, perform well at low temperatures, have a broad spectrum of activity, are considered as not to be toxic to bees, and give a high level of control of resistant pollen beetle, they will have an important role in the insecticide spraying regime of OSR.
Occurrence of insecticide resistant pollen beetles (Meligethes aeneus F.) in Danish oilseed rape (Brassica napus L.) crops
Authors: Hansen L. M.
Published: March 2008

The pollen beetle is the most important pest in Danish oilseed rape crops and it is essential that farmers are able to control this pest, especially in years when the economic damage threshold will be exceeded. About seven years ago, Danish oilseed rape growers began reporting that the pyrethroids seemed to be losing effectiveness towards pollen beetles. In 2001 18 populations collected from winter and spring oilseed rape fields were tested and very high level of insecticide resistance was found. In 2003 100 populations of pollen beetles were tested, using a FAO dip‐test, for resistance to fenitrothion, lambda‐cyhalothrin and tau‐fluvalinate. It can be concluded that many Danish populations of pollen beetles are resistant to the pyrethroid lambda‐cyhalothrin and to a lesser extent to the pyrethroid tau‐fluvalinate. No resistance to the organophosphate fenitrothion was found.
Les insectes entomophages d’intérêt agricole acclimatés en France métropolitaine depuis le début du 20ème siècle
Authors: Malausa J.‐C. , Rabasse J.‐M. , Kreiter P.
Published: March 2008

Au moment où les introductions de nouveaux phytophages ravageurs augmentent de façon préoccupante dans la zone paléarctique, un bilan de la lutte biologique classique en France depuis l’origine est présenté. Les références‐clefs concernant les expériences d’acclimatations réussies sont complétées par des informations inédites et des commentaires d’ensemble. Cet acquis devrait servir de base à un renforcement de la mise en œuvre de cette stratégie de lutte qui représente toujours une méthode efficace, économique et durable pour répondre à l’afflux de nouvelles espèces invasives.