bluuebunny/crossref_metadata_2025 huggingface dataset:
Erythroid response to treatment with G‐CSF plus erythropoietin for the anaemia of patients with myelodysplastic syndromes: proposal for a predictive model
Authors: Hellström‐Lindberg Eva , Negrin Robert , Stein Richard , Krantz Sanford , Lindberg Greger , Vardiman James , ÖSt ÅKe , Greenberg Peter
Published: November 2003

Previous studies have shown that approximately 40% of patients with myelodysplastic syndrome (MDS) and anaemia respond to treatment with human recombinant granulocyte‐CSF (G‐CSF) plus erythropoietin (epo). The present study was designed to investigate pre‐treatment variables for their ability to predict erythroid responses to this treatment. 98 patients with MDS (30 RA, 31 RARS, 32 RAEB, five RAEB‐t) were treated with a combination of G‐CSF (0.3–3.0 μg/kg/d, s.c.) and epo (60–300 U/kg/d, s.c.) for at least 10 weeks. Minimum criteria for erythroid response was a 100% reduction of red blood cell (RBC) transfusion need or an increase in haemoglobin level of  1.5 g/dl. 35 patients (36%) showed responses to treatment. Medium duration of response was 11–24 months. In multivariate analysis, serum erythropoietin levels and initial RBC‐transfusion need retained high statistical significance (P < 0.01). Using pre‐treatment serum epo levels as a ternary variable (< 100, 100–500 or > 500 U/l) and RBC transfusion need as a binary variable (< 2 or  2 units per month), the analysis provided a predictive score for erythroid response. This score divided patients into three groups: one group with a high probability of erythroid responses (74%), one intermediate group (23%) and one group with poor responses to treatment (7%). This predictive scoring system could be used in decisions regarding use of these cytokines for treating the anaemia of MDS, both for defining patients who should not be given the treatment and for selecting patients for inclusion in prospective trials.
Ala601‐Thr type dysplasminogenaemia genetically diagnosed in patients with retinochoroidal vascular disorders
Authors: Murata Masatoshi , Ooe Asako , Izumi Tomonori , Nakagawa Michihiro , Takahashi Shigeki , Ishikawa Masaaki , Mori Kazuo , Ichinose Akitada
Published: November 2003

Dysplasminogenaemia has been reported in patients with retinochoroidal vascular disorders. The precise genetic defects of these cases, however, remain unclear because of the limitations of conventional diagnostic techniques. In this study, three patients with these diseases were investigated at the DNA level for the first time to define the molecular bases of these disorders. Polymerase chain reaction–restriction fragment length polymorphism analysis revealed that all three cases carried the same Ala601‐Thr mutation. This defect may also play a role in the pathogenesis of circulation disorders in small local vessels because of reduced fibrinolytic activity due to decreased functional plasminogen levels.
Sickle cell disease: relation between procoagulant activity of red blood cells from different phenotypes and in vivo blood coagulation activation
Authors: Helley Dominique , Girot Robert , Guillin Marie‐Claude , Bezeaud Annie
Published: November 2003

In the present study we examined if, among other mechanisms, the abnormal exposure of phosphatidylserine at the surface of sickle red blood cells (RBCs) contributes to the hypercoagulability which characterizes homozygous sickle cell disease (SCD). The question was addressed by comparison of the procoagulant properties of RBCs from subjects with various phenotypes (SS, SC and AS) that differ in clinical presentation. As previously reported, SS‐RBCs accelerated the prothrombin activation by factor Xa, by decreasing the Km of the reaction compared to normal RBCs. SC‐RBCs and AS‐RBCs also promoted prothrombin activation although their procoagulant properties were milder compared to SS‐RBCs. A significant increase of the thrombin–antithrombin complexes was observed in SS subjects. Prothrombin fragment 1+2 (F1+2) was elevated in half of the SS subjects, but the difference with controls did not reach significance. Elevated levels of thrombin–antithrombin complexes were observed in a number of SC (4/11) and AS (3/12) subjects, but the difference with controls was not significant. A significant correlation was observed between the plasma levels of thrombin–antithrombin complexes in the subjects with SS, AS and AA phenotypes, and the procoagulant properties of RBCs. Our results strongly suggest that the procoagulant properties which characterize SS‐RBCs also affect SC‐RBCs and AS‐RBCs, and that exposure of phosphatidylserine by RBCs contributes to the hypercoagulable state observed in SCD.
PIG‐A gene mutations in Four Taiwanese Patients with Paroxysmal Nocturnal Haemoglobinuria Following Aplastic Anaemia
Authors: Lin Liang‐In , Liu Chun‐Hao , Chen Yao‐Chang , Shen Ming‐Chin , Wang Chiu‐Hwa , Huang Yi‐Ling , Lin Jen‐Kun
Published: November 2003

Paroxysmal nocturnal haemoglobinuria (PNH) is an acquired haemolytic disorder caused by deficient biosynthesis of the glycosyl phosphatidylinositol (GPI) anchor in haemopoietic stem cells. PIG‐A, an X‐linked gene that participates in the first step of GPI‐anchor synthesis, is responsible for PNH. Various abnormalities of the PIG‐A gene have been demonstrated in all patients with PNH so far examined. In this study we characterized the somatic mutations in PIG‐A gene in four Taiwanese patients with PNH. We identified five novel mutations in the PIG‐A gene, three single nucleotide substitution mutations (−342, C → G, codon 335, GGT → AGT and codon 405, GCT → GTT) and two frameshift mutations (codon 22, GGA → G‐A and codon 356, TGT → TGTT) in the PIG‐A gene. The −342 mutation was judged to be a polymorphism. Furthermore, three patients had previous clinicopathologic evidence which suggested aplastic anaemia (AA), before the development of PNH. One of these was found to have thrombocytopenia during follow‐up. We suggest that the somatic PIG‐A gene mutations highlight a subgroup of AA having a pathogenetic link with PNH.
The rate and kinetics of molecular response to donor leucocyte transfusions in chronic myeloid leukaemia patients treated for relapse after allogeneic bone marrow transplantation
Authors: Raanani Pia , Dazzi Francesco , Sohal Jastinder , Szydlo Richard M. , Van Rhee Frits , Reiter Andreas , Lin Feng , Goldman John M. , Cross Nicholas C. P.
Published: November 2003

We have assessed the molecular response of 30 consecutive patients with chronic myeloid leukaemia (CML) treated for relapse after allogeneic bone marrow transplantation (BMT) by donor leucocyte transfusions (DLT). Response was evaluated by qualitative nested and quantitative competitive RT‐PCR for BCR‐ABL mRNA at various time intervals before and after DLT. The probability of attaining molecular remission at 2 years was 61% (95% CI 42–78%). Disease state at the time of DLT was significantly associated with response: molecular remission was achieved for 9/10 (90%) patients treated early (cytogenetic or molecular relapse) compared to only 8/20 (40%) patients treated late (haematological relapse; P=0.009). The Kaplan–Meier estimates of molecular remission at 2 years post DLT for patients treated in early or late relapse were 86.6% and 47.3% respectively (P=0.004). The median time interval from DLT to molecular remission was 11.0 months (range 2.5–32). Molecular remissions were durable for most (15/17) patients (median follow‐up 21.2 months; range 0–55). Two patterns of molecular response were found: a very rapid decline after an initial lag phase or a more gradual decline over a period of several months. We conclude that molecular monitoring is a sensitive indicator of response to DLT; different kinetics of molecular response may reflect disease heterogeneity or differences in the mode of action of DLT.
p53 abnormalities in CLL are associated with excess of prolymphocytes and poor prognosis
Authors: Lens Daniela , Dyer Martin J. S. , Garcia‐Marco Jose M. , De Schouwer Pierre J. J. C. , Hamoudi Rifat A. , Jones David , Farahat Nahla , Matutes Estella , Catovsky Daniel
Published: November 2003

To determine the role of the p53 gene in chronic lymphocytic leukaemia (CLL) and its possible involvement in the pathogenesis of a progressive form of CLL characterized by > 10% prolymphocytes (CLL/PL), we selected 32 cases, 17 with typical morphology and 15 CLL/PL. The extent of inactivation of p53 was examined by assessing loss of heterozygosity (LOH) at 17p13.3, by sequencing the highly conserved region (exons 5–9) of the p53 gene and by analysing p53 protein expression. LOH was detected in 8/28 (29%) cases, p53 mutations in 5/32 (16%) cases and p53 expression in 5/27 (19%) cases. Overall 11 cases (30%) had p53 abnormalities of which eight cases had CLL/PL. There was a significant association between CLL/PL and p53 abnormalities (P = 0.05); 75% of cases with LOH, 80% of p53 mutations and 80% of cases positive for p53 protein had CLL/PL. Thus, p53 inactivation is the first gene abnormality identified so far to be involved in the development of CLL/PL. All the cases with typical CLL and p53 abnormalities had only one allele affected whereas 4/6 CLL/PL had both alleles inactivated. This difference in the extent of p53 inactivation suggests that accumulation of p53 abnormalities may be associated with progression of CLL to CLL/PL.

CLL cases with p53 abnormalities were characterized by a higher incidence of stage C (P < 0.025), a higher proliferative rate (P = 0.05), short survival (P < 0.005) and resistance to first‐line therapy (P < 0.02) but not to nucleoside analogues. Analysis of the correlation between p53 status and incidence of trisomy 12 by fluorescence in situ hybridization (FISH) showed that trisomy 12 was more frequent in cases without p53 abnormalities, suggesting that trisomy 12 and p53 may represent different pathways of transformation in CLL.
Fludarabine, cytarabine, G‐CSF and idarubicin (FLAG‐IDA) for the treatment of poor‐risk myelodysplastic syndromes and acute myeloid leukaemia
Authors: Parker J. E. , Pagliuca A. , Mijovic A. , Cullis J. O. , Czepulkowski B. , Rassam S. M. B. , Samaratunga I. R. , Grace R. , Gover P. A. , Mufti G. J.
Published: November 2003

Nineteen patients with high‐risk myelodysplastic syndrome (MDS)/acute myeloid leukaemia (AML) received fludarabine, cytarabine, granulocyte‐colony stimulating factor (G‐CSF), and idarubicin chemotherapy (de novo MDS/MDS‐AML, nine; relapsed/refractory MDS/AML, seven; therapy‐related MDS, three). Median age was 44 years and median disease duration 10 months. 16/19 (84%) patients had abnormal cytogenetics with seven (37%) harbouring abnormalities of chromosome 7. 18/19 (94.7%) patients responded to FLAG‐idarubicin with 12 (63%) achieving complete remission (CR) (<5% blasts and normal cytogenetics). 7/9 (78%) patients with de novo MDS/MDS‐AML achieved CR compared to 5/10 (50%) with alternative diagnoses. Response was associated with age < 50 years, disease duration < 3 months, and cytogenetics other than abnormalities of chromosome 7. Haemopoietic regeneration was rapid in most patients and there were no toxic deaths. Nine patients received a second course of chemotherapy, three have proceeded to allogeneic bone marrow transplant and three to autologous blood stem cell/bone marrow transplantation. Follow‐up is short (median 10 months). 12/19 (63%) patients remain alive and 5/12 (42%) have relapsed at a median 5 months following CR achievement. FLAG‐idarubicin was well tolerated. High rates of morphological and cytogenetic remission, especially in de novo MDS, offer a window of opportunity for assessment of autologous BMT in this group of diseases where no treatment except alloBMT has led to prolongation of survival.
Virus inactivation and prevalence of GBV‐C in haemophiliacs
Authors: Uhle Christian , Zimmermann Rainer , Goeser Tobias , Seelig Renate
Published: November 2003

A new putative hepatitis virus has recently been discovered and termed GB virus C (GBV‐C). We investigated the prevalence of this virus among 50 haemophiliacs treated with non‐virus‐inactivated clotting factor concentrates prior to 1985 and 21 haemophiliacs treated exclusively with virus‐inactivated clotting factor concentrates. In the first group the prevalence of GBV‐C based on PCR and ELISA was 46%. In the second group the prevalence of GBV‐C was similar to that of healthy blood donors (5%). We therefore conclude that GBV‐C is reliably inactivated by modern virucidal methods such as vapour heating, pasteurization and treatment with solvent/detergent mixtures.
Interferon increases serum thrombopoietin in patients with chronic hepatitis C
Authors: Shiota Goshi , Okubo Michiko , Kawasaki Hironaka , Tahara Tomoyuki
Published: November 2003

We measured serum thrombopoietin (TPO) in chronic hepatitis C treated with interferon (IFN). The platelet count before the therapy was 161.9 ×109 ± 64.1 × 109/l, which decreased to 116.3 × 109 ±  48.4 × 109/l 1 week after IFN therapy (P <0.01). On the other hand, serum TPO increased from 1.96 ± 0.60 fmol/ml to 2.68 ± 0.69 fmol/ml (P < 0.02). Contrary to a recent report that serum TPO was not altered in liver cirrhosis, these data indicate that serum TPO was increased in chronic hepatitis C in response to thrombocytopenia by IFN therapy.
Pilot study of HLA alloimmunization after transfusion with pre‐storage leucodepleted blood products in aplastic anaemia
Authors: KILLICK S. B. , WIN N. , MARSH J. C. W. , KAYE T. , YANDLE A. , HUMPHRIES C. , KNOWLES S. M. , GORDON‐SMITH E. C.
Published: November 2003

We have performed a pilot study to examine the incidence of alloimmunization using pre‐storage leucocyte‐depleted blood products (PLDP) in 16 previously transfused aplastic anaemia (AA) patients with no detectable HLA antibodies. A further eight AA patients with HLA antibodies received HLA‐matched PLDP. Leucodepleted apheresed platelets were obtained using either Cobe spectra or Haemonetics system with an integral pall filter. Pall BPF4 filters were used for red cell preparation. Patients’ sera were tested for HLA antibodies using lymphocytotoxicity (LCT). Patients who were HLA antibody negative by LCT at study entry were further tested with enzyme‐linked immunoassay (ELISA). Out of 16 patients, two (12%) formed anti‐HLA antibodies with a median follow‐up of 9 months (range 1–15), but did not display platelet refractoriness to random donor platelets. Two patients were inadvertently transfused with non‐leucodepleted blood products when later referred back to their local hospital. Both subsequently demonstrated HLA antibodies by LCT and became platelet refractory. These results contrast with a 50% incidence of HLA alloimmunization in a control group of AA patients transfused prior to this study with non‐PLDP. HLA antibodies could no longer be detected by LCT in follow‐up of three out of eight patients with HLA antibodies at study entry. Only one patient experienced non‐haemolytic febrile transfusion reactions (NHFTR). We conclude that PLDP reduce the risk of alloimmunization even in previously transfused AA patients, PLDP are associated with a low incidence of NHFTR, and all new AA patients should receive PLDP from diagnosis.
Reversible suppression of bone marrow response to erythropoietin in Plasmodium falciparum malaria
Authors: Kurtzhals J. A. L. , Rodrigues O. , Addae M. , Commey J. O. O. , Nkrumah F. K. , Hviid L.
Published: November 2003

To study the importance of bone marrow inhibition in the pathogenesis of malarial anaemia, haematological and parasitological parameters were followed in patients with acute malaria. Three patient categories were studied, severe malarial anaemia (SA), cerebral malaria (CM) and uncomplicated malaria (UM). Red cell distribution width (RDW) was used as a surrogate marker of release of young erythrocytes and reticulocytes. Initially RDW was low in all patients in spite of markedly increased concentrations of erythropoietin (EPO). 3 d after institution of treatment and coinciding with parasite clearance RDW increased dramatically, reaching the highest levels 1–2 weeks later. Although severe anaemia was corrected by blood transfusion during the first 3 d of treatment, the peak RDW correlated significantly with the initial EPO levels. This suggests that Plasmodium falciparum infection causes a rapidly reversible suppression of the bone marrow response to EPO. Furthermore, the inhibition of bone marrow response was a general finding irrespective of initial haemoglobin levels suggesting that the severity of anaemia depends upon the degree of peripheral erythrocyte destruction in patients with suppressed bone marrow response to EPO.
Multiple bcl‐2/Ig gene rearrangements in persistent polyclonal B‐cell lymphocytosis
Authors: Delage Robert , Roy Jean , Jacques Louise , Bernier Vincent , Delâge Jean‐Marie , Darveau André
Published: November 2003

Persistent polyclonal B‐cell lymphocytosis is a benign lymphoproliferative disorder of unknown aetiology occurring exclusively in women, characterized by typical binucleated lymphocytes, polyclonal expansion of B cells and elevated serum IgM. Owing to the role of Bcl‐2 oncogene in inhibition of apoptosis, we have investigated the presence of the bcl‐2/Ig gene rearrangement. Bcl‐2/Ig gene rearrangement was determined by polymerase chain reaction targeting the usual breakpoint regions of the t(14;18). Bcl‐2/Ig gene rearrangement was identified in all six patients and, more importantly, multiple rearrangements were present in five patients. The frequency of the bcl‐2/Ig gene rearrangement is estimated to be of one translocation in 1 × 102 to 1 × 103 peripheral blood mononuclear cells. We conclude that persistent polyclonal B‐cell lymphocytosis is associated with bcl‐2/Ig gene rearrangement. These findings are of clinical importance because these patients may be misdiagnosed as having a leukaemic expression of non‐Hodgkin's lymphoma.
Blood viscosity and risk of cardiovascular events: the Edinburgh Artery Study
Authors: Lowe G. D. O. , Lee A. J. , Rumley A. , Price J. F. , Fowkes F. G. R.
Published: November 2003

We examined the relationships of whole blood viscosity and its major determinants to incident cardiovascular events (ischaemic heart disease and stroke) in a prospective study of a random population sample of 1592 men and women aged 55–74 years (the Edinburgh Artery Study). 272 fatal and non‐fatal cardiovascular events occurred during 5 years of follow‐up (cumulative incidence 17.1%). Age and sex adjusted mean levels of blood viscosity (3.70 v 3.55 mPa.s), haematocrit (46.2 v 45.7%), haematocrit‐corrected blood viscosity (3.57 v 3.48 mPa.s), plasma viscosity (1.35 v 1.33 mPa.s) and fibrinogen (2.88 v 2.67 g/l) were significantly higher in subjects who experienced events than in subjects who did not. The relationships of these rheological variables to cardiovascular events were at least as strong as those of conventional risk factors (smoking habit, diastolic blood pressure, and low‐density lipoprotein cholesterol). After adjustment for these conventional risk factors, the associations of blood viscosity and haematocrit remained significant for stroke, but not for total events; whereas the associations of plasma viscosity and fibrinogen remained significant for total events and for stroke.

These findings suggest that increased blood viscosity may be one plausible biological mechanism through which increases in haematocrit and fibrinogen may promote ischaemic heart disease and stroke. Randomized controlled trials of viscosity reduction in the prevention of cardiovascular events (e.g. by lowering high levels of haematocrit or plasma fibrinogen) are suggested.
The human thrombin receptor and proteinase activated receptor‐2 genes are tightly linked on chromosome 5q13
Authors: Schmidt Valentina A. , Nierman William C. , Feldblyum Tamara V. , Maglott Donna R. , Bahou Wadie F.
Published: November 2003

The thrombin receptor (TR) and proteinase activated receptor‐2 (PAR‐2) may represent the prototypes of an emerging family of cell‐surface receptors that effect cell activation events mediated by serine proteases generated during inflammatory, fibrinolytic or haemostatic‐regulated pathways. To further characterize the molecular genetics of these receptors, we have refined the genetic and physical mapping of both PAR‐2 and TR. Utilization of two distinct radiation hybrid mapping panels with different levels of resolution demonstrated that both genes are tightly linked to the microsatellite markers D5S424, D5S1977, D5S2529 and D5S2596 (in order of decreasing LOD scores, from 13.7 for D5S424 to 7.7 for D5S2596). Physical mapping using yeast artificial chromosomes (YACs) and inversion field gel electrophoresis demonstrated that they are maximally separate by 90 kb. If the association of TR and PAR‐2 genes resulted from a relatively recent gene duplication event from a common ancestral gene, these observations provide a general framework for the identification of gene transcripts representing alternative proteolytically activated receptors which may be clustered within this region of the human genome. These observations are especially relevant given recent evidence that murine and human platelets express alternative signalling mechanisms or receptors for thrombin.
Thymic function in adults: evidence derived from immune recovery patterns following myeloablative chemotherapy and stem cell infusion
Authors: Cavenagh J. D. , Milne T. M. , Macey M. G. , Newland A. C.
Published: November 2003

We studied 45 patients aged 14–66 years who had undergone stem cell transplantation for a variety of malignant conditions at least 12 months previously. Compared to normal controls, they had significantly reduced absolute numbers of CD4+, CD4+CD45RA+ and CD4+CD45RO+ T cells and a reduced CD4+CD45RA+: CD4+CD45RO+ ratio. In all subsets T‐cell numbers were significantly greater 24 months, compared to 12–24 months, after transplantation and there was a non‐significant trend towards lower T‐cell numbers with increasing age. We conclude that the thymus, or putative thymic‐equivalent tissue, remains functional in older adults.
Body iron stores in relation to growth and pubertal maturation in healthy boys
Authors: Anttila Raija , Cook James D. , Siimes Martti A.
Published: November 2003

During male puberty, erythropoiesis is exceptionally active. Pubertal development and iron status were followed in 60 healthy boys at 3‐month intervals for 24 months to evaluate changes in body iron stores with the serum transferrin receptor–ferritin ratio. The estimated amount of stored iron declined by about 50% over a 2‐year period. Remarkable changes in iron stores were found even after as short an interval as 3 months and pubertal development was closely linked with a decrease in stored iron. The annual increments of estimated red blood cell (RBC) iron showed strong positive correlations with velocities in testicular volume and certainly in body height and weight. In contrast, the estimated changes in individual iron stores were not associated with any of those parameters. The lacking associations between iron stores and growth parameters are probably indicative of increasing intestinal absorption. Despite the relatively small quantitative role of iron stores in supplying iron for growth, falling iron stores probably play a more important regulatory role by stimulating iron absorption.
Genetic typing of human platelet antigen 1 (HPA‐1) by oligonucleotide ligation assay in a specific and reliable semi‐automated system
Authors: Zotz R. B. , Giers G. , Maruhn‐Debowski B. , Scharf R. E.
Published: November 2003

Genotyping of platelet alloantigens with the possibility of using any type of cellular material as a source of DNA has become a preferred procedure, particularly in thrombocytopenic patients when platelet counts are too low for phenotyping. Recently human platelet antigen 1 (HPA‐1) has been identified as an inherited risk factor for coronary thrombosis. The different detection methods currently used have disadvantages for large‐scale DNA diagnosis, including the need for electrophoresis (allele‐specific restriction enzyme analysis, amplification with sequence‐specific primers) or the potential risk of reduced specificity (allele‐specific oligonucleotide hybridization). In this report we describe the adaptation of an automated oligonucleotide ligation assay to genotype HPA‐1 in polymerase chain reaction (PCR)‐amplified DNA samples. HPA‐1a and HPA‐1b phenotypes corresponded to the results of the different genotyping assays. The genotypes determined with the ELISA‐based PCR‐oligonucleotide ligation assay were in 100% concordance with the results obtained by conventional allele‐specific restriction enzyme site analysis and PCR amplification with sequence‐specific primers. The automated oligonucleotide ligation assay provides a rapid, reliable, nonisotopic method to genotype human platelet antigens that can rapidly be applied to large population screening.
Analysis of T‐cell clonality in bone marrow of patients with acquired aplastic anaemia
Authors: Melenhorst J. Joseph , Fibbe Willem E. , Struyk Linda , Van DerElsen Peter J. , Willemze Roel , Landegent James E.
Published: November 2003

Acquired aplastic anaemia (AA) represents a state of bone marrow (BM) failure which is characterized by BM hypocellularity and pancytopenia. It has been hypothesized that in some AA patients, bone marrow failure is secondary to the targeted destruction of haemopoietic stem cells by autoreactive T cells. The response of T cells to antigenic stimulation has been shown, in a number of animal models and in autoimmune diseases, to result in the (oligo)clonal expansion of positively reacting T cells. For this reason, we studied the utilization of 24 T‐cell receptor‐β variable gene segments (TCRBV) and the clonality in BM aspirates and peripheral blood (PB) of seven AA patients. BM from transplant donors served as controls. Determination of TCRBV gene segment usage revealed no significant differences between patients and controls.

Clonality within each family was analysed by single‐strand conformation polymorphism (SSCP) analysis. Clonal and clonally predominant bands were seen in BM of three AA patients in five to eight TCRBV families. Clonal rearrangements were encountered less often in BM of control subjects. In conclusion, our results suggest an antigen‐driven T‐cell response in the BM of predominantly AA patients resulting in oligoclonal T‐cell outgrowth.
Interferon regulatory factor 1 induces the expression of the interferon‐stimulated genes
Authors: Henderson Ying C. , Chou Marian , Deisseroth Albert B.
Published: November 2003

The interferon regulatory factor 1 (IRF‐1) is a positive transcriptional regulatory protein which acts in the interferon signal transduction pathway to activate the transcription of the type I interferon genes by binding to the PRDI response element. The aim of this study was to explore the role of IRF‐1 in regulating the expression of other interferon‐stimulated genes in the interferon signal transduction pathway. A transient transfection assay was used to show that IRF‐1 induced the expression of interferon‐stimulated genes. The induction was a direct result of IRF‐1 binding to the promoters of the interferon‐stimulated response element (ISRE). The levels of endogenous mRNA of two interferon‐stimulated genes, 6‐16 and 9‐27, were increased in cells containing increased levels of IRF‐1. In addition, IRF‐1 activates the expression of IRF‐2, a negative regulator of the type I interferon genes themselves. Two sequences were found in the IRF‐2 promoter which were the binding sites for IRF‐1. Mutations in the oligonucleotide sequences of these sites could abolish the binding of the IRF‐1. These data suggested that IRF‐1 not only plays an important role in the induction of type I interferon genes, but also in the activation of interferon‐stimulated genes.
Treatment of chronic myeloid leukaemia in first chronic phase with idarubicin and cytarabine: mobilization of Philadelphia‐negative peripheral blood stem cells
Authors: CHALMERS E. A. , Franklin I. M. , KELSEY S. M. , NEWLAND A. C. , CLARK R. E. , SPROUL A. M. , CROTTY G. , MCCANN S. R. , FIELDING A. , GOLDSTONE A. H. , HEPPLESTONE A. , WATSON W. , SHARP R. A. , TANSEY P.
Published: November 2003

Peripheral blood stem cell (PBSC) mobilization using idarubicin and cytarabine was investigated in 40 patients with chronic myeloid leukaemia in first chronic phase (CML CP1). Disease contamination was evaluated in harvests from 41/44 (93%) mobilization episodes. Using cytogenetics, 22/37 (59%) showed a complete or major response; Southern blot analysis demonstrated a complete or major response in 9/17 (53%). No harvests were RT‐PCR negative. In the 41 evaluable episodes, more complete or major responses were seen when PBSC mobilization occurred within 24 months [17/23 (74%) versus 6/18 (33%); P =0.02] and within 12 months of diagnosis [10/11 (91%) versus 13/30 (43%); P = 0.018]. 20 patients underwent PBSC transplantation and 18/20 successfully engrafted. Post‐transplant cytogenetic analysis was available on 15 cases, of whom five achieved a major cytogenetic response at 1–3 months, with five partial cytogenetic remissions. Two of 40 patients died during mobilization therapy (5%) and three of 20 after the transplant (15%). Overall mortality was high at five of 40 patients, and the procedural mortality was 20%. This study demonstrates that Ph‐negative PBSCs can be mobilized in a significant proportion of patients with CML CP1, with the best results observed within a year of diagnosis. These cells can subsequently be used for autologous transplantation, however, the impact on long‐term survival requires longer follow‐up, and potential benefits may be compromised by the high mortality.
Presence of immunoglobulins, C3 and cytolytic C5b‐9 complement components on the surface of erythrocytes from patients with β‐thalassaemia/HbE disease
Authors: Malasit Prida , Mahasorn Weerawuth , Mongkolsapaya Juthathip , Singhathong Benjawan , Fucharoen Suthat , Wasi Prawase , Bhakdi Sucharit
Published: November 2003

The occurrence of IgG, IgM, IgA, C3 and C5b‐9 complement complexes on erythrocytes from 43 patients with β‐thalassaemia HbE disease was investigated. Indirect immunoradiometric assays using radioiodinated protein A were employed to quantify the individual components. We confirmed that circulating erythrocytes from thalassaemic patients contained elevated amounts of IgG, and small but significant amounts of C3. In addition, small but significant amounts of C5b‐9 were detected. Levels of cell‐bound IgG, C3 and C5b‐9 were higher in splenectomized versus non‐splenectomized patients. The presence of C5b‐9 on circulating cells from five splenectomized patients was confirmed by an ELISA employing a monoclonal antibody specific for a C5b‐9 neoantigen. When C5b‐9 positive cells from two patients were solubilized with detergent and subjected to sucrose density gradient centrifugation, the terminal complexes sedimented as 25–40S macromolecules, thus behaving as membrane C5b‐9 complexes. The presence of C8 and C9 in these high molecular weight fractions was directly demonstrated by Western blotting. These results constitute the first demonstration that circulating diseased erythrocytes may carry low numbers of potentially cytolytic C5b‐9 complement complexes which may be partly responsible for the known ionic disturbances found in thalassaemic cells. Both bound C3 and C5b‐9 could promote removal of diseased cells in the reticuloendothelial system.
A comparison of vincristine and doxorubicin infusional chemotherapy with methylprednisolone (VAMP) with the addition of weekly cyclophosphamide (C‐VAMP) as induction treatment followed by autografting in previously untreated myeloma
Authors: RAJE NOOPUR , POWLES RAY , KULKARNI SAMAR , MILAN SARAH , MIDDLETON GARY , SINGHAL SEEMA , MEHTA JAYESH , MILLAR BARBARA , VINER CHRISTINE , RAYMOND JULIAN , TRELEAVEN JENNIFER , CUNNINGHAM DAVID , GORE MARTIN
Published: November 2003

In a sequential nonrandomized study, 204 consecutive unselected patients aged < 70 years received induction chemotherapy with infusional vincristine and adriamycin with oral methyl prednisolone (VAMP; n=75) or with additional cyclophosphamide, C‐VAMP (n=129). 38/129 C‐VAMP patients also received verapamil during induction as part of a controlled trial with the aim to overcome drug resistance. A median of five courses (range 1–11) of chemotherapy were required before maximal response was attained and this was similar in both groups. An over‐all response rate of 71% was noted at the end of induction. The complete remission (CR) rate with C‐VAMP was 24%, which was significantly higher (P=0.04) than the CR rate with VAMP alone (8%). The addition of verapamil did not alter the response rate of C‐VAMP. Compliance to VAMP was overall 83% and not affected by the addition of cyclophosphamide. The proportion of patients going on to receive high‐dose chemotherapy and an autograft was the same for VAMP and C‐VAMP treated patients (71%). The median overall survival (OS) and progression‐free survival (PFS) for the whole group were 4.4 years and 2.0 years and no difference in outcome was observed between the different treatment groups. Therefore the addition of weekly cyclophosphamide to VAMP induction therapy has significantly improved the response rates of previously untreated myeloma patients. C‐VAMP was not more toxic and did not compromise the chances of receiving an autograft. Verapamil was without influence on any parameters in this study.
Effect of clopidogrel treatment on ADP‐induced phosphorylations in rat platelets
Authors: SAVI P. , ARTÇANUTHURRY V. , BORNIA J. , GRELAC F. , MACLOUF J. , LEVY‐TOLEDANO S. , HERBERT J. M.
Published: November 2003

Phosphorylations induced by 2‐MeS‐ADP, a potent agonist of platelet ADP receptors, have been studied in rat platelets, and the effect of clopidogrel, a compound which inhibits platelet aggregation by selectively reducing the binding of ADP to its low affinity receptors on platelets, has been determined. 2‐MeS‐ADP induced platelet activation (shape change and aggregation) simultaneously with the phosphorylation of myosin light chain (P20) and plekstrin (P47). Phosphorylation of P20 and P47 was transient, a maximum being observed 10 s after addition of the agonist when shape change reached its maximum. P20 and P47 phosphorylations were not strongly affected by clopidogrel treatment. Following stimulation of platelets with 2‐MeS‐ADP, several proteins were phosphorylated at tyrosine residues. Clopidogrel treatment inhibited the increase in phosphorylation of P140 , P100 , P80/85 , P66 and P55 concomitantly with the inhibition of platelet aggregation. However, clopidogrel did not interfere with the early phosphorylation of the P80/85 kD doublet which occurs at the time of the shape change. P80/85 , identified by immunodetection as cortactin, could be involved in the reorganization of the cytoskeleton necessary for morphological changes.  Thus, by using clopidogrel‐treated rat platelets, we were able to determine some of the phosphorylations coupled either to clopidogrel‐resistant high‐affinity ADP receptors leading to shape change or to clopidogrel sensitive low‐affinity ADP receptors coupled to the aggregation process.
Flow cytometric method for the routine follow‐up of red cell populations after bone marrow transplantation
Authors: Hendriks E. C. M. , De Man A. J. M. , Van Berkel Y. C. M. , Stienstra S. , De Witte T.
Published: November 2003

A simple and sensitive flow cytometric method is presented for the quantitation of erythrocyte subpopulations. Cells were indirectly labelled using human antisera (mostly crude) and fluorescein isothiocyanate (FITC)‐conjugated anti‐human IgG Fab. The method was evaluated by analysing artificial mixtures of blood group antigens A, B, D, E, c, K and Fya. Intra‐assay coefficients of variation were found to be 7.2% for 1% D‐positive mixtures and 2.2% for 10% D‐positive mixtures; the inter‐assay coefficients of variation were 9.5% and 6.0% respectively. The sensitivity of the method was found to be 0.31%. The method has shown to be suitable for the routine follow‐up of patients after allogeneic bone marrow transplantation (BMT).
Comparative genomic hybridization detects genomic abnormalities in 80% of follicular lymphomas
Authors: AVET‐LOISEAU Hervé , VIGIER Magda , MOREAU Anne , MELLERIN Marie‐Paule , GAILLARD Fanny , HAROUSSEAU Jean‐Luc , Bataille Régis , MILPIED Noël
Published: November 2003

Comparative genomic hybridization (CGH) was used to analyse 34 follicular lymphoma (FL) samples. 27 samples showed DNA sequence copy number changes of at least one genomic region (26 samples with at least one gain and nine with at least one loss). Some chromosomes or chromosomal regions were preferentially involved. The most frequently gained regions were chromosome 18q (29% of samples), chromosome X (21%), chromosome 7 (18%), chromosomes 2, 6p and 8q (12%). Two regions were preferentially lost: 6q (12%) and 17p (9%). All these gained and lost regions have been previously reported in cytogenetic studies, confirming the accuracy of CGH in detecting genetic abnormalities in FL. 21% of samples displayed normal profiles, probably reflecting the absence of unbalanced abnormality, which is also in agreement with the cytogenetic data. In conclusion, we showed that CGH is an accurate, reliable and rapid method and we propose the inclusion of CGH in the evaluation of FL at diagnosis.
Cytokine and chemokine production by CD34+ haemopoietic progenitor cells: detection in single cells
Authors: Behringer Dirk , Kresin Vera , Henschler Reinhardt , Mertelsmann Roland , Lindemann Albrecht
Published: November 2003

In vitro expansion of haemopoietic progenitor cells (HPC), lineage‐specific differentiation, and gene transfer are all based on in vitro culture systems using haemopoietic growth factors (HGF). A close control of the actual culture conditions, however, is difficult due to secondary mediators secreted by the heterogenous population of mature and immature cells in culture. Although monocytes and granulocytes have already been identified as active producers, this study specifically addressed the role of CD34+ progenitor cells in this respect. Using an immunostaining method that enables simultaneous detection of cytokines and phenotype, 56±6% CD34+ peripheral blood progenitor cells (PBPC) were found to contain cytoplasmic IL‐8 after stimulation with phorbol myristate acetate+ionomycin for 90 min, 19±4% stained positive after TNF‐α induction (20 h), and 7±1% expressed IL‐8 in the presence of culture medium alone. Intra‐cytoplasmic TNF‐α and IL‐1β were detected at lower frequency, and <1% of CD34+ cells expressed IL‐1ra or IL‐6, whereas IL‐1α, IL‐10 and G‐CSF were not detected. Thus, CD34+ HPC are able to synthesize chemo‐ and cytokines that may operate in an auto‐ or paracrine manner to modulate in vivo as well as in vitro growth and differentation of haemopoietic cells.
Identification of a Monoclonal Thrombin Inhibitor Associated with Multiple Myeloma and a Severe Bleeding Disorder
Authors: Colwell Niall S. , Tollefsen Douglas M. , Blinder Morey A.
Published: November 2003

We investigated a patient with a long‐standing IgGκ monoclonal gammopathy who developed severe haemorrhagic complications. At IgG concentrations of ∼50 g/l the patient had severe bleeding associated with prolongation of the thrombin time, activated partial thromboplastin time, and reptilase time. Plasmapheresis resulted in improvement in the thrombin time and resolution of bleeding. Depletion of the IgG by absorption of plasma with protein G–Sepharose in vitro resulted in normalization of the thrombin time and reptilase time. The purified IgG bound to immobilized thrombin and immunoprecipitated human α‐, β‐ and γ‐thrombin but not prothrombin, other vitamin K‐dependent coagulation factors, or fibrinogen. Purified IgG at concentrations >1×10−2 g/l decreased (∼50%) the rate of hydrolysis of a chromogenic substrate by thrombin. Addition of purified IgG to normal pooled plasma at concentrations >1×10−2 g/l prolonged the thrombin time and activated partial thromboplastin time, but the reptilase time was prolonged only at IgG concentrations >1 g/l. This finding suggests that at low concentrations the IgG produces a specific antithrombin effect, but at higher concentrations it also affects fibrin polymerization; the combination of these effects probably produced clinical bleeding. This is the first report of a monoclonal antithrombin antibody associated with bleeding in a patient with multiple myeloma.
Issues in multidisciplinary teamwork for children with disabilities
Authors: Yerbury M
Published: November 2003

This paper presents a review of key issues surrounding multidisciplinary work in relation to the care of children with disabilities and their families. The discussion is based on experience of working in a child development team and research visits to teams around Britain at the beginning of the 1990s. An outstanding feature of child disability services is their organizational diversity. The research found that district interagency planning groups were a rarity, as were multi‐agency management groups for teams. Thus it was rare for teams to have clear lines of external accountability. Similarly leadership and management structures within the teams had a profound influence on their effectiveness. Issues are highlighted for identifying good practice and for further development of services.
Motor difficulties in children with developmental disorders of speech and language
Authors: Owen S E , McKinlay I A
Published: November 2003

The motor hand function of 16 children, aged between 4 and 7 years, with developmental speech and language disorders, was compared with that of 16 control children. The children with developmental speech and language disorders were significantly slower than controls on three out of four motor tasks. They were also more likely than controls to have mixed hand preference although this results was not significant. Children with developmental speech and language disorders should be assessed to ensure that motor deficits are diagnosed and that appropriate support is given.
Child Health Surveillance in England and Wales: the good news
Authors: Butler J R
Published: November 2003

Postal surveys were conducted in 1993 among all, or samples of, six groups of providers and managers of pre‐school child health surveillance (CHS) in England and Wales. Content analyses were also carried out of strategic policy statements for CHS produced by 54 district health authorities in England and Wales. The surveys aimed to document the views and experiences of CHS providers and managers about the impact of recent changes affecting the structure and operation of CHS, including the publication of Health for All Children, the 1990 Contract for General Practitioners (GPs), the implementation of the National Health Service and Community Care Act 1990, and the changing roles of community doctors and health visitors. Five positive findings from the surveys are discussed: the impact of the first edition of Health for All Children; improvements in the development and use of child health information systems; the beneficial effects of the growing involvement of GPs in CHS; the developing understanding of, and commitment to, the principle of clinical audit in CHS; and the growing collaboration between providers in the NHS internal market. A separate paper reports the negative findings from the study.
Prompt epithelial damage and restitution processes in allergen challenged guinea‐pig trachea in vivo
Authors: ERJEFÄLT J. S. , KORSGREN M. , NTLSSON M. C. , SUNDLER F. , PERSSON C. G. A.
Published: November 2003

Background
Little is known about the induction and the morphology of epithelial damage, and of the ensuing epithelial restitution processes in allergic airways.

Objective
To examine epithelial damage and restitution in allergen challenged guinea‐pig trachea.

Methods
Whole‐mount techniques, transmission and scanning electron microscopy, cryosectioning, and histochemical staining were used. Cell proliferation was monitored by BrdU‐immunohistochemistry.

Results
Allergen challenge produced patchy, crater‐like, and leucocyte‐rich epithelial damage sites. At 1, 5, and 24 h damage was associated with poorly differentiated epithelial restitution cells. Already at 1 h the epithelial craters had a floor of flattened restitution cells and the damaged areas comprised < 1% of the mucosal surface area (whole‐mount preparations). In contrast, cryo sections displayed large areas (∼ 20%, 1 h) of denudation. Epithelial, and subepithelial (fibroblasts, smooth muscle) proliferation was increased 5 and 24 h after challenge (P < 0.01).

Conclusion
Within 1 h allergen challenge has induced patchy damage sites where epithelial restitution is already advanced; although easily produced by cryosectioning frank denudation was not evident in whole‐mount preparations. The present findings may explain the well maintained, functional tightness of allergic airways displaying epithehal damage, shedding, and even denudation. The present data also suggest the possibility that epithelial damage‐restitution may be causative to allergic airway remodelling.
Dysfunction of guinea‐pig pulmonary surfactant and type II pneumocytes after repetitive challenge with aerosolized ovalbumin
Authors: LIU M. , WANG L. , HOLM B. A. , ENHORNING G.
Published: November 2003

Background
Asthma symptoms may partially be caused by a surfactant dysfunction. The inflammatory reaction, so characteristic of asthma, involves a protein invasion of airways which harmfully affects the surfactant function. However, mild asthma attacks might also impede the surfactant synthesis in alveolar type II cells.

Objective
The present study evaluates the hypothesis that type 11 pneumocyte metabolic function might be disturbed in a model of mild asthma.

Methods
Immunized, as well as not immunized control guinea‐pigs, were challenged three times at two‐day intervals with 0.04% ovalbumin aerosol. Bronchoalveolar lavage (BAL) was performed one day after the last challenge and the fluid was evaluated for surface activity, and content of phospholipids and proteins. Alveolar type II cells were isolated and their ability to incorporate a 3H labeled surfactant precursor was evaluated.

Results
BAL fluid from immunized and challenged animals showed less surface activity (P < 0.01) when compared with BAL fluid from controls, not immunized but challenged. Most likely the reduced surface activity was caused by a 14% increase in the protein concentration (P < 0.05). Isolated type II cells from immunized and challenged animals had 33% less phospholipids than cells from controls (P < 0.05), and phosphatidylcholine synthesis was reduced 35% (P < 0.05).

Conclusion
Conclusion These results suggest that the synthesis, intra‐cellular storage, and biophysical activity of surfactant are decreased in an intermittent and mild form of asthma.