bluuebunny/crossref_metadata_2025_split huggingface dataset:
The use of two substrates to improve the sensitivity of indirect immunofluorescence in the diagnosis of pemphigus
Authors: Harman K.E. , Gratian M.J. , Bhogal B.S. , Challacombe S.J. , Black M.M. 
Published: March 2003

Abstract
The aim of this study was to re-evaluate indirect immunofluorescence (IIF) comparing two substrates, normal human skin (HS) and monkey oesophagus (MO) using serum from 29 pemphigus patients classified according to the presence of serum autoantibodies to either desmoglein (Dsg) 1 or Dsg3 detected by enzyme-linked immunosorbent assay (ELISA). Overall, the sensitivity of IIF was 83% on HS and 90% on MO. When data from both substrates were combined, the sensitivity increased to 100%. When sera from pemphigus foliaceus (PF) patients were studied, which contained Dsg1 antibodies only, the sensitivity of IIF was greatest on HS and titres were on average 4·8 doubling dilutions higher than on MO. In contrast, when sera containing autoantibodies only to Dsg3 from pemphigus vulgaris (PV) patients was studied, the sensitivity was greatest on MO and titres were on average 4·4 doubling dilutions higher than on HS. There was a significant correlation between Dsg1 antibody levels and IIF titres on HS and between Dsg3 antibody levels and IIF titres on MO. The investigation of immunobullous disorders in the future is likely to move towards antigen-specific techniques such as the Dsg ELISAs used in this study. However, in laboratories which currently rely on IIF for detecting pemphigus autoantibodies, the data presented in this study strongly suggest that two substrates should be used for IIF screening: one rich in Dsg1, such as HS, and the other rich in Dsg3, such as MO. This combination of substrates should not only increase the sensitivity of detecting pemphigus antibodies, but will aid in the differentiation of PV from PF. It is also possible that the data might be more useful for disease monitoring.
B‐lymphocyte suppression in multiple myeloma is a reversible phenomenon specific to normal B‐cell progenitors and plasma cell precursors
Authors: Rawstron Andrew C. , Davies Faith E. , Owen Roger G. , English Anne , Pratt Guy , Child J. Anthony , Jack Andrew S. , Morgan Gareth J. 
Published: March 2003

The reduced levels of normal immunoglobulin in patients with myeloma may be due to suppression of normal B‐cell differentiation. However, reports on the numbers of B cells vary, with some finding decreases consistent with immunoparesis, and others reporting expansions of phenotypically aberrant cells. We have therefore assessed the phenotype and levels of B lymphocytes in patients at presentation (n = 23), in plateau or complete remission (PB n = 42, BM n = 18), and in relapse (PB n = 17, BM n = 14), in comparison to normal individuals (n = 10). Phenotypic analysis was performed using five‐parameter flow cytometry, with CD14 used to exclude monocytes where necessary.We found no evidence of a phenotypically distinctive blood or marrow B‐cell population in patients with myeloma, nor of an increase in the levels of any B‐cell subset. Numbers of blood CD19+38+ normal plasma cell precursors were significantly reduced in presentation/relapse patients, but not in patients in plateau/remission. Total CD19+ cells were significantly reduced only in patients with circulating myeloma cells, detected by IgH‐PCR. In the marrow, CD19+ B cells expressing CD5, CD10, CD34, CD38, CD45low and Syndecan‐1 were significantly decreased at presentation/relapse, but not in patients in plateau/remission. The majority of these antigens are expressed by normal B‐cell progenitors, indicating that myeloma also affects the early stages of B‐cell development. The suppression of progenitor cells was not restricted to B‐lymphoid differentiation, as total CD34+ cells were also significantly reduced in the marrow of myeloma patients at presentation.These results indicate that, if neoplastic B cells are present in myeloma, they are low in number and have a phenotype similar to their normal counterparts. Furthermore, there is a reversible suppression of CD19+ B lymphocytes that correlates inversely with disease stage, and specifically affects the early and late stages of normal B‐cell differentiation.
Unrelated donor marrow transplantation for chronic myelogenous leukaemia
Authors: Dini  , Lamparelli  , Rondelli  , Lanino  , Barbanti  , Costa  , Manfredini  , Guidi  , Rosti  , Alessandrino  , Locatelli  , Marenco  , Soligo  , DI Bartolomeo  , Aversa  , LA Nasa  , Busca  , Majolino  , DE Laurenzi  , Bacigalupo  
Published: March 2003

Between January 1989 and July 1995 the search for an unrelated donor (UD) was started for 379 consecutive Italian patients with Philadelphia positive (Ph+) chronic myelogenous leukaemia (CML). 89 (23%) were transplanted. The overall probability of transplant before and after December 1991 was 16% and 49%, respectively (P = 0.0001), and average interval between search activation and graft was 23 months and 13 months, respectively (P = 0.0001). Disease‐free survival (DFS) following 60 consecutive transplants performed before February 1996 was 41.5% at 48 months and was 64% for patients grafted after January 1993. In univariate analysis, five variables had a favourable effect on DFS: year of bone marrow transplantation (BMT) after 1993 (P = 0.0002), HLA‐DRB1 donor/recipient (D/R) match (P = 0.0006), total body irradiation (TBI) containing regimen (P = 0.0006), graft‐versus‐host disease (GvHD) prophylaxis including ‘early’ cyclosporin before the transplant, and a marrow cell dose > 3 × 108/kg of recipient body weight (P = 0.04). Multivariate analysis confirmed that HLA identity (P = 0.006), TBI‐containing regimen (P = 0.0001) and ‘early cyclosporin’ (P = 0.04) were associated with higher DFS. Transplant‐related mortality (TRM) was 67% in patients grafted before January 1993 and 30% in patients grafted subsequently (P = 0.002). Multivariate analysis confirmed DRB1 identity (P = 0.03) and TBI‐containing regimen (P = 0.0005) to be independent factors predictive of low TRM.This suggests that the outcome of patients transplanted from an HLA DRB1 matched donor, after a TBI‐containing preparative regimen, is similar to results recently reported in patients transplanted from geno‐identical siblings.These results indicate that the search should be initiated at diagnosis for patients < 45 years of age and UD BMT should be considered early in the disease course for those with an available DRB1‐matched unrelated donor.
Prevention of EBV‐induced B‐lymphoproliferative disorder by ex vivo marrow B‐cell depletion in HLA‐phenoidentical or non‐identical T‐depleted bone marrow transplantation
Authors: Cavazzana‐Calvo  , Bensoussan  , Jabado  , Haddad  , Yvon  , Moskwa  , Tachet Des Combes  , Buisson  , Morand  , Virion  , LE Deist  , Fischer  
Published: March 2003

HLA‐mismatched bone marrow transplantation (BMT) is hampered by three major complications: graft rejection, acute graft‐versus‐host disease (aGVHD) and delayed immune reconstitution. Infusion of anti‐LFA1 plus anti‐CD2 monoclonal antibodies (MAb), combined with ex‐vivo T‐cell depletion of the graft, was efficient in preventing graft rejection and aGVHD. Nevertheless, disease‐free survival was limited by the high frequency of lethal infections, including EBV‐induced lymphoproliferative disease (BLPD), which originates mostly from donor B cells, with an incidence of 5–30%. To decrease the rate of this complication, ex‐vivo B‐cell depletion was attempted.This study compares a group of 19 patients who received a T‐ and B‐cell‐depleted marrow from an HLA‐mismatched related donor with a retrospective control group of 19 patients, who had received T‐cell‐depleted marrow by the same method. The level of T‐cell depletion was similar in the two groups. For B‐cell depletion, two different methods were compared. The median number of B cells infused in the study group was 0.46/kg. Engraftment and aGVHD incidence were similar in the two groups. No EBV donor‐derived BPLD occurred in the study group, compared with seven in the control group, four of whom died because of EBV‐BPLD. Event‐free survival was significantly different between the two groups. We conclude that ex‐vivo B‐cell depletion of the graft may be a useful means of preventing EBV‐BPLD, and warrants further study on a larger group of patients.
Heterogeneity of blood group RhE variants revealed by serological analysis and molecular alteration of the RHCE gene and transcript
Authors: Noizat‐Pirenne France , Mouro Isabelle , Gane Pierre , Okubo Yasuto , Hori Yasuto , Rouger Philippe , LE Pennec Pierre‐Yves , Cartron Jean‐Pierre 
Published: March 2003

After testing red cells from 12 RhE variants with a panel of anti‐E monoclonal antibodies (MoAbs), four patterns of reactivity were detected indicating that the MoAbs may recognize four distinct E epitopes designated epE1, epE2, epE3 and epE4. The variants were classified into four categories (cat EI to EIV) which carried epE1 and epE2, epE1 and epE4, epE1, epE3 and epE4, and all four epitopes, respectively. Molecular analysis of the transcripts and genomic DNA of the variants from cat EI, EII and EIII displayed three distinct genetic alterations. Cat EI variants exhibited a point mutation (T500A) in exon 4 of the RHCE gene that resulted in a Met167Lys substitution in the third extracellular loop of the RhcE protein. Cat EII variant carried a hybrid gene structure characterized by replacement of exons 1–3 (or 2–3) of the RHCE gene by their specific counterparts in the RHD gene. This latter variant was also associated with a weak expression of the RhC antigen. In cat EIII variants there was a partial DNA exchange of exon 5 sequences (nt 697 and 712) between the RHCE and the RHD genes, generating a hybrid Rh cE‐D‐cE protein carrying the Glu233 and Val238 substitutions.The serological and molecular studies of the RhE variants indicated that: (i) the RhE antigen is a mosaic composed of at least four epitopes and proline at position 226 is necessary but not sufficient for the full expression of the E antigen, (ii) the lack of RhE epitope(s) is associated with heterogenous molecular alterations of the RHCE gene, and (iii) amino‐acids located on the third and fourth extracellular loops of the RhCE polypeptide are critical for some RhE epitopes expression.
P‐glycoprotein, lung resistance‐related protein and multidrug resistance associated protein in de novo acute non‐lymphocytic leukaemias: biological and clinical implications
Authors: Michieli Mariagrazia , Damiani Daniela , Ermacora Anna , Masolini Paola , Raspadori Donatella , Visani Giuseppe , Scheper R. J. , Baccarani Michele 
Published: March 2003

P‐glycoprotein (PGP), lung resistance‐related protein (LRP) and multidrug resistance associated protein (MRP) expression and the blast cells' intracellular daunorubicin accumulation (IDA) were evaluated in 96 previously untreated cases of de novo acute non‐lymphocytic leukaemia (ANLL). 47/96 patients (49%) were classified as PGP+, 44/96 (46%) as LRP+, and 8/96 (8%) as MRP+. The more frequent MDR clusters were PGP−/LRP−/MRP− (32/96 cases, 33%) and the PGP+/LRP+/MRP− (27/96 cases, 28%) followed by PGP+/LRP−/MRP− (15/96 cases, 16%) and PGP−/LRP+/MRP− (14/96 cases, 14%). A favourable karyotype was observed more frequently in PGP− and LRP− cases. A highly significant correlation was found between either PGP or LRP overexpression and leukaemic blast cell IDA.All the patients received standard induction and consolidation treatments containing MDR‐related (idarubicin, mitoxantrone, etoposide) and other (arabinosyl cytosine) drugs. Multivariate analysis showed that PGP overexpression was significantly associated with a poor response to treatment, both in terms of primary resistance or shorter survival. Other independent prognostic factors were age and cytogenetics. LRP overexpression did not reach statistical significance, although for LRP+ cases the trend was unfavourable. Due to small numbers, no conclusion could be made regarding MRP overexpression, but 5/8 cases showed unfavourable karyotypic abnormalities, 8/8 had a defective IDA and 6/8 failed to achieve remission. This study showed that both PGP and LRP overexpression are common features in de novo ANLL at onset whereas MRP overexpression is more rare. It suggested that overexpression of one of the MDR related proteins was associated with a defective IDA, and confirmed that, in addition to age and cytogenetics, PGP retains an independent prognostic value. It also suggested that LRP did not affect clinical outcome when patients were treated with idarubicin or mitoxantrone and arabinosyl cytosine.
Primary prophylaxis in severe haemophilia should be started at an early age but can be individualized
Authors: Astermark Jan , Petrini Pia , Tengborn Lilian , Schulman Sam , Ljung Rolf , Berntorp Erik 
Published: March 2003

The frequency of joint bleeds and orthopaedic joint scores were evaluated in 121 patients with severe haemophilia who had started prophylactic treatment with clotting factor concentrates at least once weekly before the age of 10. 75 of the patients started before the age of 3, 31 at the age of 3–5 and 15 at the age of 6–9. Each subgroup was evaluated separately. In addition, a regimen of one infusion weekly was compared with that of two (haemophilia B) or three (haemophilia A) infusions weekly in each patient.A significant decrease in the overall number of joint bleeds per year was found after shortening the infusion interval (P < 0.005), but the individual bleeding pattern varied. In survival analysis of the first pathologic joint score event, those who started prophylaxis before the age of 3 had a better outcome overall than those starting at later ages (P = 0.001). However, in subgroup analysis, no significant difference was seen in the annual number of joint bleeds and the development of arthropathy between those starting with, or shifting to, the more intensive regimen before the age of 3 and those that were put on this regimen at the age of 3–5. Age at start of prophylaxis was found to be an independent predictor for the development of arthropathy (P = 0.0002), whereas dose and infusion interval at start were not.Our data emphasize the importance of starting replacement therapy during the first years of life. However, it seems that when beginning the regimen it can be individualized and adjusted according to the bleeding pattern. In this way, the need for a venous access system may be assessed on an individual basis.
Skeletal survey in advanced multiple myeloma: radiographic versus MR imaging survey
Authors: Lecouvet Frédéric E. , Malghem Jacques , Michaux Lucienne , Maldague Baudouin , Ferrant Augustin , Michaux Jean‐Louis , Vande Berg Bruno C. 
Published: March 2003

In an attempt to compare the sensitivity of bone radiographs and bone marrow magnetic resonance (MR) imaging for bone lesion detection in patients with stage III multiple myeloma (MM) and to evaluate the possible consequences of the replacement of the conventional radiographic skeletal survey (RSS) by an MR survey of the spinal and pelvic bone marrow in these patients, we obtained MR studies of the thoracic and lumbar spine, pelvis and proximal femurs in addition to the conventional RSS (including radiographs of the skull, entire spine, pelvis, ribs, humerus and femurs) in 80 consecutive patients with newly diagnosed stage III MM according to the Durie and Salmon staging system (based on blood tests and on the RSS). The performance of MR and radiographic studies to detect bone lesions in given anatomic areas and in given patients were compared. The consequences on MM staging following the substitution of the RSS by the MR survey were assessed.MR imaging was superior to radiographs for lesion detection in the spine (76% v 42% of patients) and pelvis (75% v 46% of patients). The RSS was superior to the limited MR imaging survey for the detection of bone involvement in the patient population (87.5% v 79% of patients). If the RSS had been replaced by the MR imaging survey for patient staging, 7/80 patients would have been categorized as stage I and one as stage II MM on the basis of normal MR findings and biological findings consistent with these stages.Substitution of the RSS by a limited spinal and pelvic marrow MR survey would lead to ‘understaging’ of 10% of patients with otherwise stage III MM on the basis of blood tests and the conventional RSS.
Role of FcγRI (CD64) in erythrocyte elimination and its up‐regulation in thalassaemia
Authors: Wiener  , Allen  , Porter  , Wickramasinghe  , Porter  , Chinprasertsuk  , Siripanyaphinyo  , Pattanapanyasat  , Fucharoen  , Wanachiwanawin  
Published: March 2003

To examine any role of the high affinity Fcγ class I receptor (FcγRI) (CD64) in erythrocyte elimination by mononuclear phagocytes (MP) in thalassaemia (thal), we investigated the in vitro interaction of β‐thalassaemic erythrocytes with monocytes (Mo) whose FcγR expression had been modulated by cytokines.Treatment of Mo with interferon (IFN)‐γ or interleukin (IL)‐10 which up‐regulate FcγRI, caused a dose‐dependent increase in binding of β‐thalassaemic erythrocytes, whereas stimulation with IL‐4 which down‐regulates the receptor, reduced this interaction, in a dose‐dependent manner, to that of normal erythrocytes. Binding of thalassaemic erythrocytes by IFN‐γ or IL‐10‐treated Mo was inhibited by FcγRI‐specific reagents. In addition, Mo expression of FcγRI and HLA class II DR was determined by flow cytometry in Thai patients with HbH disease (α1/α2 or α1/Hb Constant Spring) (n = 15) or β°‐thal/HbE (n = 16). In both groups of patients FcγRI expression was increased as compared to normal controls (n = 14): mean fluorescence intensity (±SD) 124.79 ± 38.77 in HbH disease and 121.86 ± 18.23 in β°‐thal/HbE versus 91.94 ± 17.36 in normal controls (P < 0.01 and P < 0.001, respectively). In contrast, HLA class II DR expression was similar in patients and controls.The results suggest that, in thalassaemia, up‐regulated FcγRI on mononuclear phagocytes plays a role in their interaction with erythrocytes and that this process can be modified by cytokines.
An in vitro system for expression analysis of mutations of the β‐globin gene: validation and application to two mutations in the 5′ UTR
Authors: Ho P. Joy , Sloane‐Stanley Jackie , Athanassiadou Aglaia , Wood William G. , Thein Swee Lay 
Published: March 2003

We describe the setting up of an in vitro expression system for the analysis of mutations of the β‐globin gene. The system is based on the stable transfection of a normal or mutated β‐globin gene into mouse erythroleukaemia (MEL) cells. The expression construct contains an Aγ gene as an internal control and both globin genes are under the control of the HS2 element of the β LCR. The system enables analysis of transcription, RNA processing and transport, as well as mRNA stability. With non‐mutant genes, high‐level expression of both β and Aγ genes is seen and both mRNAs are stable.The system was validated by comparing the expression of the β654 thalassaemia splicing mutation in MEL cells with its well‐characterized expression in vivo. The level of the initial transcript, the proportion of abnormally spliced mRNA and its instability during erythroid cell maturation were all faithfully reproduced. The system was used to examine the mechanism by which two mutations in the β‐globin 5′ untranslated region (5′ UTR) result in β thalassaemia. Surprisingly, the mechanism appeared to differ in the two cases, with the C‐G substitution at position +33 affecting transcription, whereas the −T deletion at position +10 resulted in a translational defect. The stably transfected MEL cells, with an internal control and an endogenous enhancer, appear to be a valid and realistic experimental model, superior to transient expression studies. This system should find wide application in the analysis of the effects and mechanisms of gene inactivation in mutations affecting the β‐globin as well as other genes.
Allogeneic bone marrow transplantation in aggressive non‐Hodgkin's lymphoma (excluding Burkitt and lymphoblastic lymphoma): a series of 73 patients from the SFGM database
Authors: Dhedin  , Giraudier  , Gaulard  , Esperou  , Ifrah  , Michallet  , Milpied  , Rio  , Cahn  , Molina  , Laporte  , Guilhot  , Kuentz  ,   for the Société Française de Greffe de Moelle
Published: March 2003

The place of allogeneic bone marrow transplantation (BMT) in the treatment of aggressive non‐Hodgkin's lymphoma (NHL) remains controversial. We conducted a retrospective study of French experience in allografting NHL between 1984 and 1994. To improve the homogeneity of the study population, cases of low‐grade, Burkitt and lymphoblastic NHL were excluded. 73 patients were included in the analysis. Median age at transplantation was 35 years (range 9–61 years); 64 patients were in stage IV and 45 had bone marrow involvement at diagnosis. At the time of transplantation, 46 patients had sensitive disease (25 in complete remission; CR).The overall survival (OS) and progression‐free survival (PFS) rates were 41% and 40% respectively at 5 years (median follow‐up of survivors 90 months). The probability of disease progression was 30% at 5 years, and only one relapse occurred after 15 months. 32 patients died of transplantation‐related complications. In multivariate analysis, pretransplant complete remission was the main factor associated with longer survival (OS at 60 months of 76% among the 25 patients in CR at transplant and of 23% among the 48 patients not in CR at transplant). Neither acute nor chronic graft‐versus‐host disease (GvHD) influenced the relapse rate.In conclusion, in this high‐risk population the overall results of allogeneic BMT were encouraging, despite a high transplant‐related mortality rate. We believe this procedure should be studied further in prospective controlled trials.
Time‐course expression of polypeptides carrying blood group antigens during human erythroid differentiation
Authors: Bony Viviane , Gane Pierre , Bailly Pascal , Cartron Jean‐Pierre 
Published: March 2003

The time course expression of blood group antigens was examined by flow cytometry using a two‐phase liquid culture system that supports the proliferation and maturation of human erythroid progenitors from adult peripheral blood. The progression towards erythroid differentiation was followed by the expression changes of the transferrin receptor (CD71++) and glycophorin A (GPA+). Four main categories of blood group markers were identified: (i) those characterized by an early expression like ABO (A), Kell (K:2) and Rh50 which were detected in the Epo‐independent phase 1, (ii) those including GPC (Gerbich, Ge antigens) and Fy6 which were expressed in the late phase 1, (iii) GPA (MN antigens), Wrb (Band 3/GPA interaction), Rh(D, Cc/Ee) and LW which appeared during the Epo‐dependent phase 2 and (iv) those like Jk3 and Lub which were expressed in late phase 2.Regarding blood group molecules exhibiting adhesive properties (LW/ICAM‐4, Oka and Lu) the most significant event was a sharp decrease of Oka (neurothelin) expression with the concomitant loss of ICAMs expression during the later stage of differentiation. These studies suggest that Oka, ICAMs and LW might contribute to the adhesive interactions involved in the formation of erythroblastic islands and attachment to stroma cells and the extracellular matrix. We also noted an asynchronous expression of the proteins that compose the core of the Rh complex, since Rh50 glycoprotein was expressed earlier than Rh(D, CE) proteins.