bluuebunny/crossref_metadata_2025 huggingface dataset:
Acquired von Willebrand disease in a patient with angiodysplasia resulting from immune‐mediated clearance of von Willebrand factor
Authors: Inbal A. , Bank I. , Zivelin A. , Varon D. , Dardik R. , Shapiro R. , Rosenthal E. , Shenkman B. , Gitel S. , Seligsohn U.
Published: November 2003

A patient with a severe bleeding tendency due to acquired von Willebrand disease (VWD) is presented. Although no underlying disorder has emerged during 6 years of follow‐up, an immune‐mediated mechanism was responsible for acquired VWD in this patient as demonstrated by detection of von Willebrand factor (VWF)/anti‐VWF complexes in the patient's plasma and their removal by protein A–sepharose beads and resumption of normal haemostasis with correction of VWF antigen, VWF activity and VWF multimeric pattern after treatment of the patient with high‐dose gammaglobulin. Detection of anti‐VWF antibodies in the patient’s plasma had a significant impact on the choice of therapeutic intervention to control bleeding.
Comparison of the direct platelet immunofluorescence test (direct PIFT) with a modified direct monoclonal antibody‐specific immobilization of platelet antigens (direct MAIPA) in detection of platelet‐associated IgG
Authors: Joutsi Lotta , Kekomäki Riitta
Published: November 2003

Glycoprotein (GP)‐specific platelet‐associated IgG (PA‐IgG) may be demonstrable in autoimmune‐mediated thrombocytopenia. We studied 159 consecutive patients with histories of thrombocytopenia by a modified direct monoclonal antibody‐specific immobilization of platelet antigens (direct MAIPA) assay, which immobilizes GP IIb/IIIa, GP Ib/IX and GP Ia/IIa simultaneously. This modification requires smaller quantities of platelets than standard measurements performed separately. PA‐IgG was present in 84/159 (53%) patients, as shown by the direct platelet immunofluorescence test (PIFT) with flow cytometry as a reference. PA‐IgG against GP IIb/IIIa and/or GP Ib/IX and/or GP Ia/IIa was noted in 46 patients (29%), of whom 93% (43/46) were also PA‐IgG positive. The amount of PA‐IgG detected by PIFT correlated directly with that detected by direct MAIPA (r=0.71; P<0.0001). Only three patients 12548 with negative direct PIFT had GP‐specific PA‐IgG. GPV‐specific PA‐IgG was detected in 13 (10%) of the 125 patients, in whom further studies could be performed. In the subgroup of patients with GP‐specific PA‐IgG, the median fluorescence intensities of direct PIFT were higher than in patients with no GP‐specific PA‐IgG (P<0.001). Direct PIFT and direct MAIPA divided the patients into asymmetric subgroups. However, the relative roles of these tests in the diagnosis of autoimmune‐mediated thrombocytopenia await further studies.
Efficient RT‐PCR on platelet mRNA after long‐term storage
Authors: PEYRUCHAUD Olivier , NURDEN Alan , Bourre François
Published: November 2003

We have developed a procedure permitting RT‐PCR from mRNA even after a long‐term storage (1 year) of platelet samples in ethanol (EtOH‐platelets) at −80°C. To validate our method, we have analysed the human platelet alloantigen system (HPA‐1) which is coded by β3 mRNA. We have also demonstrated the efficiency of amplification of part of the coding region for (i) αIIb subunit mRNA, (ii) αv subunit mRNA, and (iii) the seven transmembrane domain thrombin receptor mRNA.
Glycosylphosphatidyl‐inositol (GPI)‐linked protein deficiency on the platelets of patients with aplastic anaemia and paroxysmal nocturnal haemoglobinuria: two distinct patterns correlating with expression on neutrophils
Authors: Jin Jong‐Youl , Tooze Jennifer A. , Marsh Judith C. W. , Gordon‐Smith Edward C.
Published: November 2003

Deficiencies in glycosylphosphatidyl‐inositol (GPI)‐linked proteins on erythrocytes and leucocytes in patients with paroxysmal nocturnal haemoglobinuria (PNH) are well known; however, expression on platelets in these patients is less well documented. We have studied CD55 and CD59 on the platelets of PNH and aplastic anaemia (AA) patients using flow cytometry. In all cases of PNH, CD55 and CD59 negative populations of platelets were detected with single or bimodal distribution and these results showed close correlation with the CD55 and CD59 patterns of neutrophils. Previous published studies have not demonstrated this distribution. We suggest that our findings may be due to the methodology used.
Expression and function of CD95 (FAS/APO‐1) in leukaemia‐lymphoma tumour lines
Authors: Dirks Wilhelm , Scho¨ne Stefanie , Uphoff Cord , Quentmeier Hilmar , Pradella Silke , Drexler Hans G.
Published: November 2003

Cross‐linkage of the CD95 (FAS/APO‐1) antigen is responsible for the induction of programmed cell death or apoptosis in a variety of normal and malignant cells of the haemopoietic system. In order to evaluate predominant expression of the CD95 gene in a cell lineage‐specific manner, we have determined the CD95 expression patterns in cell lines of myeloid, T‐, pre‐B‐ or B‐cell origin as well as those established from Hodgkin's disease (HD). Our results reveal constitutive transcriptional activation of the CD95 gene in all cell lines derived from the lymphoid and myeloid lineages. Despite the ubiquitous expression of CD95 transcripts in haemopoietic cells, the corresponding protein was undetectable in 2/5 cell lines derived from Burkitt lymphomas and 6/16 leukaemia cell lines of the megakaryocytic or monocytic lineage. In an effort to identify apoptosis‐resistant cell lines resulting from mutations in the death‐signalling domain of CD95 or from defects in the apoptotic pathway or in survival programmes, we applied a CD95‐mediated apoptosis assay. However, 21/38 CD95‐expressing cell lines were sensitive upon induction with an anti‐CD95 antibody whereas the remaining cell lines (predominantly of myeloid derivation) were resistant to antibody‐induced cell death. Resistance to CD95‐mediated apoptosis was not due to mutations within the CD95 open reading frame as confirmed by a combined reverse transcription PCR sequencing method. Five myeloid out of 13 tumour lines with the apoptosis‐resistance phenotype analysed showed programmed cell death, when protein synthesis was blocked by treatment with cycloheximide prior to CD95‐mediated induction. These data suggest an active cellular mechanism for the maintenance of an apoptosis‐resistant phenotype. Elucidating the steps in such an active process of resistance to apoptosis might be expected to provide new approaches for therapeutic intervention in certain tumours.
Risk of arterial thrombosis in patients with anticardiolipin antibodies and lupus anticoagulant
Authors: Nojima Junzo , Suehisa Etsuji , Akita Naoki , Toku Masayuki , Fushimi Ryo , Tada Hisato , Kuratsune Hirohiko , Machii Takashi , Kitani Teruo , Amino Nobuyuki
Published: November 2003

The relationship between arterial or venous thrombosis and the levels of anticardiolipin antibodies (aCL) and/or existence of lupus anticoagulant (LA) was studied. The 141 patients with systemic lupus erythematosus (SLE) were divided into four groups: aCL single positive (25 cases), LA single positive (11 cases), aCL and LA double positive (25 cases), aCL and LA double negative (80 cases). The prevalence of thrombosis was higher in aCL and LA double positive patients (21/25 cases, 84.0%, P < 0.01) than that in aCL single positive patients (4/25 cases, 16.0%), LA single positive patients (1/11 cases, 9.1%) and double negative patients (3/80 cases, 3.8%). Furthermore, in these double positive patients, all patients (10/10 cases) with a high positive level of aCL (>10 units/ml) had arterial thrombosis, whereas only 2/15 patients (13.3%) with a low positive level of aCL (3–10 units/ml) were affected. Venous thrombosis was frequently found in the low positive group (9/15 cases, 60.0%). On the contrary, none of 105 LA negative patients had arterial thrombosis and only seven (6.7%) had venous thrombosis. These findings indicate that a high aCL activity combined with a LA positive result might be a risk factor for arterial thrombosis.
International study to compare antigen‐specific methods used for the measurement of antiplatelet autoantibodies
Authors: Berchtold P. , Mu¨ller D. , Beardsley D. , Fujisawa K. , Kaplan C. , Kekoma¨ki R. , Lipp E. , Morell‐Kopp M. C. , Kiefel V. , McMillan R. , Kr. von dem Borne A. E. G. , Imbach P.
Published: November 2003

Platelet‐associated and plasma autoantibodies against platelet glycoproteins (GP) have been demonstrated in patients with autoimmune thrombocytopenia (AITP) using various methods. Eight laboratories in seven countries participated in this international study to evaluate the interlaboratory agreement using glycoprotein‐specific immunoassays for these autoantibodies. The participating laboratories received blind samples of frozen washed platelets and plasma from 22 normal donors and 22 AITP patients. Platelet‐associated and plasma autoantibodies against GPIIb–IIIa and GPIb–IX were measured by MAIPA, immunobead assay or modified antigen capture assay. Of the control samples, 96.0% and 97.2% of all results for platelet‐associated and plasma autoantibodies to GPIIb–IIIa/GPIb–IX, respectively, were negative. The mean variation coefficient of the control samples of platelet‐associated and plasma autoantibodies was 89.5% (range 11.1–272.9%) and 46.5% (range 21.0–78.0%), respectively. In 20/22 patient samples, platelet‐associated autoantibodies to either glycoprotein were noted by at least two laboratories. The mean degree of agreement in these samples was 74.0%. There was a significant correlation in the individual antibody measurements between all laboratories (Kendall coefficient of concordance 0.60 and 0.38, P < 0.001; Spearman rank order test, range of correlation coefficient 52.3–94.0% and 42.2–85.0%, P < 0.05, for anti‐GPIIb–IIIa and anti‐GPIb–IX, respectively). In contrast, plasma autoantibodies to either glycoprotein were noted by at least two laboratories in only 13/22 patient samples. Moreover, the degree of agreement was poor (50.1%) and a significant correlation was noted between only six pairs of laboratories. We conclude that methods used in this study yield good interlaboratory agreement in measuring platelet‐associated autoantibodies against GPIIb–IIIa and GPIb–IX. In contrast, poor agreement was found in detecting plasma autoantibodies to the same glycoproteins.
System ISI calibration: a universally applicable scheme is possible only when coumarin plasma calibrants are used
Authors: Stevenson K. J. , Craig S. , Dufty J. M. K. , Taberner D. A.
Published: November 2003

Coagulometer‐induced errors in International Normalized Ratios (INR) may be corrected by assigning an International Sensitivity Index (ISI) specific for the local combination of coagulometer and thromboplastin reagent (System ISI). In the present study, 57 laboratories determined the INR of eight frozen coumarin plasmas. There was considerable variation in INR reported at the different centres, with coefficients of variation (CV) ranging from 11.30% to 17.29%. In addition, their consensus mean INR values were consistently higher than those obtained manually. The overall mean percentage deviation was 20.06%, indicating that correction was required to reduce the over‐estimation of INR that occurred even in those Systems incorporating a thromboplastin reagent with an instrument‐specific ISI. Local correction using coumarin plasma calibrants reduced this value to ≤7% with current reference thromboplastins BCT 441, OBT 79 and RBT 90 and also lowered the CV to <5%. In contrast, when adsorbed plasma calibrants were employed, mean percentage deviations were 7.87% with BCT 441, 12.47% with OBT 79 and 77.37% with RBT 90 as reference respectively. This difference may be due, in part, to the difficulty in assigning an INR to adsorbed plasma calibrants, which contain no protein induced by vitamin K antagonists (PIVKA). This study demonstrates that coumarin plasma calibrants are superior in System ISI calibration.
A novel acute lymphoid leukaemia type BCR/ABL transcript in chronic myelogenous leukaemia
Authors: Okamoto Kiyoshi , Karasawa Masamitsu , Sakai Hirotaka , Ogura Hidemi , Morita Kimio , Naruse Takuji
Published: November 2003

Using a reverse transcription‐polymerase chain reaction (RT‐PCR), we identified a patient with typical clinical features of chronic myelogenous leukaemia (CML) in the chronic phase who showed no amplification of the CML‐type BCR/ABL transcript. RT‐PCR with primers detecting the acute lymphoid leukaemia (ALL)‐type transcript disclosed a novel fragment co‐amplified with an ALL‐type fragment. Sequencing revealed the novel transcript to be a chimaeric mRNA produced by fusion of a segment of BCR exon 2 (e2) to ABL exon 2 (a2), with a 21 base‐pair insertion of ABL intron 1b sequence between them. This transcript has not been reported previously.
Mobilization of predominantly Philadelphia chromosome‐negative blood progenitors using cyclophosphamide and rHUG‐CSF in early chronic‐phase chronic myeloid leukaemia: correlation with Sokal prognostic index and haematological control
Authors: Hughes T. P. , Grigg A. , Szer J. , Ho J. , Ma D. , Dale B. M. , Green R. M. , Norman J. E. , Sage R. E. , Herrmann R. , Cannell P. , Schwarer A. P. , Taylor K. , Atkinson K. , Arthur C.
Published: November 2003

Mobilization of Philadelphia chromosome (Ph) negative blood progenitors was attempted in 23 newly diagnosed chronic myeloid leukaemia (CML) patients using a regimen of cyclophosphamide (CY) 5 g/m2 and rHUG‐CSF 150 μg/m2 daily. This regimen was well tolerated with no major adverse events reported. More than 2 × 106/kg CD34+ cells were collected in 21 patients (91%). Predominantly Ph‐negative mobilization (0–25% Ph‐positive) was seen in 30% of cases overall and was confined to patients with a Sokal prognostic score < 1 (7/11 with Sokal score <1; 0/12 with Sokal score ≥1). Within the low Sokal index group, a low WBC count pre‐mobilization and a low WBC nadir both correlated strongly with Ph‐negative mobilization (P =0.006 and 0.02 respectively). Five of 19 patients receiving at least 6 months of Roferon A therapy post mobilization achieved a major cytogenetic response; all five patients were Ph‐negative mobilizers. Therefore CML patients can be divided into a good‐prognosis group in whom predominantly Ph‐negative progenitors can be mobilized using a regimen of moderate intensity if haematological control is achieved pre‐mobilization, and a poor‐prognosis group for whom predominantly Ph‐positive cells are mobilized with this regimen regardless of haematological control.
Coagulation abnormalities in type 1 Gaucher disease are due to low‐grade activation and can be partly restored by enzyme supplementation therapy
Authors: HLLAK Carla E. M. , Levi Marcel , Berends Fenny , Aerts Johannes M. F. G. , Van Oers Marinus H. J.
Published: November 2003

In type 1 Gaucher disease a bleeding tendency occurs which is partly caused by thrombocytopenia due to massive splenomegaly. In addition, low levels of factors IX and XI have been described. The mechanism responsible for these clotting factor abnormalities is unknown.

We performed a detailed study of parameters of coagulation and fibrinolysis in 30 type 1 Gaucher disease patients (14 splenectomized) before and after treatment with enzyme supplementation therapy. Pre‐treatment aPTT and PT were prolonged in 42% and 38% of patients, respectively. In 30–60% serious deficiencies (>50%) of coagulation factors XI, XII, VII, X, V and II were observed. The low levels of factor V correlated with platelet count and were inversely associated with splenic volume. Levels of inhibitors were mildly decreased. Markers for activation of coagulation (thrombin–antithrombin (TAT) complex) and fibrinolysis (PAP complex, fibrin cleavage product D‐dimer) were significantly elevated, especially in the splenectomized patients, indicating ongoing activation of these processes. After 12 months of enzyme supplementation therapy partial correction occurred.

Thus, severe disorders of the coagulation system occur in Gaucher disease, contributing to the bleeding tendency. The deficiencies may be the result of consumption of coagulation factors caused by ongoing low‐level coagulation activation, possibly due to mononuclear cell activation.
Acute myelogenous leukaemia with t(8;21) translocation of normal cell origin in mosaic Down's syndrome with isochromosome 21q
Authors: Sato Atsushi , Imaizumi Masue , Koizumi Yoshitsugu , Obara Yasuhiko , Nakai Hiroshi , Noro Tomoyo , Saito Toshiaki , Saisho Takako , Yoshinari Miyako , Cui Yan , Suzuki Hoshiro , Funato Tadao , Iinuma Kazuie
Published: November 2003

We report a 13‐year‐old girl with Down's syndrome (DS) having a mosaic karyotype of 46,XX/46,XX, −21,+i(21q), who developed acute myelogenous leukaemia (AML) (FAB M1). The t(8;21) translocation generating a AML1/MTG8 chimaeric gene of her blasts was demonstrated by cytogenetic analysis and reverse transcription‐polymerase chain reaction. Interestingly, the leukaemic clone with t(8;21) did not have isochromosome 21q, indicating that the blasts were of normal cell origin. These findings suggest that, in older patients with DS, 21 trisomy cells have no greater predisposition to develop AML than normal karyotypic cells.
Deficiency of p67phox, p47phox or gp91phox in chronic granulomatous disease does not impair leucocyte chemotaxis or motility
Authors: Zicha Daniel , Dunn Graham A. , Segal Anthony W.
Published: November 2003

Chronic granulomatous disease (CGD) is a syndrome characterized by failure of the NADPH oxidase of phagocytes that generates superoxide, which is central to the microbicidal process. Cytosolic components of this oxidase system include the proteins p67phox and p47phox, deficiencies of which cause the autosomal recessive form of CGD, whereas the X‐linked form of the disease is characterized by a deficiency in the plasma membrane component gp91phox. Components of the oxidase system have been reported to be associated with the cytoskeleton and neutrophils from CGD patients have been reported to have a defective chemotactic response in Boyden chambers. Using a chamber that permits the direct observation of cell behaviour in a linear gradient of a chemoattractant, we have analysed the chemotactic response of neutrophils from a patient lacking p67phox; from another lacking p47phox and from a third lacking gp91phox. The results of measuring the speeds and directions of locomotion of the cells show that their speeds are undiminished relative to cells from healthy control subjects and that their directions of migration are at least as strongly biased in the direction of the gradient as those of the control cells. We conclude that these definitive aspects of the chemotactic response are not abnormal in either the autosomal recessive or the X‐linked forms of CGD and that they are therefore not factors in the predisposition to infection that characterizes the syndrome.
Successful treatment of chronic severe neutropenia with weekly recombinant granulcyte‐colony stimulating factor
Authors: Fine Kenneth D. , Byrd Tom D. , Stone Marvin J.
Published: November 2003

Daily treatment for symptomatic chronic neutropenia with recombinant granulocyte‐colony stimulating factor (rhG‐CSF) filgrastim is costly and sometimes causes neutrophillia. We report the use of weekly filgrastim in a 40‐year‐old man with life‐long symptomatic neutropenia. Baseline neutrophil counts were <1×109/l 60% of the time, and fell below 0.5×109/l for 7 d periods every 22 d. Following 1 year of weekly filgrastim treatment, the absolute neutrophil count was maintained >1×109/l (averaging 2×109/l) and the frequency and severity of symptoms were reduced by 85%. Therefore the benefits of filgrastim for the treatment of at least one form of chronic severe neutropenia can be derived from weekly rather than daily doses.
G‐CSF‐mobilized CD34+ peripheral blood stem cells are significantly less apoptotic than unstimulated peripheral blood CD34+ cells: role of G‐CSF as survival factor
Authors: Philpott N. J. , Prue R. L. , Marsh J. C. W. , Gordon‐Smith E. C. , Gibson F. M.
Published: November 2003

The mechanism of release of CD34+ cells into the peripheral blood (PB) after mobilization treatment with chemotherapy and/or growth factors is not clearly understood. Growth factors may induce increased proliferation and self renewal within the stem cell compartment. It is possible that they alter adhesion molecule profiles or other progenitor:stroma interactions, to allow release of these cells into the periphery. However, CD34+ cells are present in the PB under steady‐state conditions, albeit in low number. Growth factors such as granulocyte colony‐stimulating factor (G‐CSF) may promote the survival of CD34+ cells in the PB by suppressing apoptosis. In order to test this hypothesis, we have quantitated apoptotic cells in the CD34+ fraction of peripheral blood stem cell (PBSC) collections, using two‐colour flow cytometry, after staining with anti‐CD34 antibody and the fluorescent DNA binding agent, 7‐amino actinomycin D (7AAD). 7AAD differentially stains live, apoptotic and dead cells, due to the altered accessibility of DNA in each subpopulation.

We have shown a significant reduction in the proportion of apoptotic cells in the CD34+ population mobilized by G‐CSF compared to CD34+ cells in unstimulated PB, consistent with the theory that G‐CSF is acting, at least in part, by suppressing apoptosis. In addition, we found that G‐CSF mobilized CD34+ cells are less apoptotic than CD34+ cells of unstimulated normal bone marrow, indicating that, at the doses used, G‐CSF is significantly altering the survival capacity of the mobilized cells.
THE EFFECTS OF CHRONIC SMOKING ON THE FIBRINOLYTIC POTENTIAL OF PLASMA AND PLATELETS
Authors: Simpson A. J. , Gray R. S. , Moore N. R. , Booth N. A.
Published: November 2003

We studied tissue‐type plasminogen activator (t‐PA) and plasminogen activator inhibitor type 1 (PAI‐1) in healthy individuals divided by smoking habit into current smokers, former smokers and non‐smokers (who had never smoked). Plasma PAI‐1 antigen was significantly higher in smokers than in non‐smokers with intermediate levels in former smokers. A similar trend was observed for plasma PAI activity but this did not reach statistical significance. Platelet PAI‐1 and plasma t‐PA were not significantly different when comparing the three groups. After venous occlusion t‐PA rose significantly in all groups; no significant change in plasma PAI‐1 was observed. The ratio of t‐PA to PAI‐1 in plasma was similar in non‐smokers and former smokers but lower in smokers, suggesting that there is at least partial restoration of plasma fibrinolytic potential after smoking cessation. Plasma PAI‐1 antigen and PAI activity correlated with estimated pack‐years of cigarettes smoked among smokers and former smokers. When all subjects were studied collectively, plasma PAI‐1 correlated strongly with plasma t‐PA and triglycerides; plasma t‐PA also correlated strongly with triglycerdes.

We conclude that chronic smoking is associated with impaired fibrinolysis in plasma and that this largely reflects elevated plasma PAI‐1 in smokers. Smoking does not appear to affect the response to venous occlusion. The postulated effect of chronic smoking on plasma PAI‐1 may be mediated by the influence of smoking on triglycerides and insulin resistance. Stopping smoking appears to return impaired fibrinolysis towards normal. Smoking does not quantitatively affect the platelet pool of PAI‐1. Smoking habit should be controlled for in clinical analyses of PAI‐1 and t‐PA.
Diverse effects of anti‐CD44 antibodies on the stromal cell‐mediated support of normal but not leukaemic (CML) haemopoiesis in vitro
Authors: Ghaffari Saghi , Dougherty Graeme J. , Eaves Allen C. , Eaves Connie J.
Published: November 2003

We have identified three non‐cross‐reacting anti‐human CD44 monoclonal antibodies that have significant positive or negative (or no) effects on normal human haemopoiesis in the long‐term culture (LTC) system. These effects manifested as increases or decreases in the number of LTC‐initiating cells (LTC‐IC), and the number of colony‐forming cells (CFC) recovered from cultures in which either unseparated or highly purified CD34+CD38− normal marrow cells were placed on pre‐established normal marrow feeder layers in the presence or absence of each antibody. The effects seen were rapid and sustained, and dependent on the presence of a preformed feeder layer. Interestingly, the same anti‐CD44 antibodies had no effect on the maintenance of leukaemic (Ph+) progenitors (from patients with chronic myeloid leukaemia) when these cells were cultured on preformed feeder layers established from normal marrow. CD44 appears to be part of a mechanism by which stromal elements can regulate primitive normal haemopoietic cells but not their leukaemic (Ph+) counterparts.
Anti‐endothelial cell antibodies to the endothelial hybridoma cell line (EAhy926) in systemic lupus erythematosus patients with antiphospholipid antibodies
Authors: Matsuda Juzo , Gotoh Moritaka , Gohchi Kengo , Kawasugi Kazuo , Tsukamoto Miyo , Saitoh Noriko
Published: November 2003

The endothelial hybridoma (EAhy926) cell line was employed to clarify whether antiphospholipid antibodies (aPA) [lupus anticoagulant (LA), antiprothrombin antibody (aPT) and/or anticardiolipin antibody (aCL)] and anti‐endothelial cell antibodies (AECA) are identical, and establish whether β2‐glycoprotein I (β2‐GPI) is needed for reactivity of aPA to endothelial cells. Ig‐G AECA was positive in 9/30 SLE patients with aPA (30.0%) and 10/22 SLE patients negative for aPA (45.5%). Ig‐M AECA was positive in one SLE patient with aPA and one SLE patient without aPA. AECA‐positivity was not significantly different among unfixed, TNF‐stimulated and fixed EAhy926. The influence of β2‐GPI on the reactivity of serum to EAhy926 was minimal, and absorption experiments of serum with cardiolipin‐liposome/β2‐GPI or phosphatidylserine‐liposome/prothrombin gave little evidence of cross‐reactivity of aPA and AECA. The results of our study suggest that aPA and AECA may have partially cross‐reacted, but were different antibodies. However, further study is needed to clarify the clinico‐pathological significance of AECA.
Morphological and cytochemical findings in 150 cases of T‐lineage acute lymphoblastic leukaemia in adults
Authors: Gassmann Winfried , Löffler Helmut , THIEL ECKHARD , Ludwig Wolf‐Dieter , Schwartz Stefan , Haferlach Torsten , Maurer Jürgen , Rieder Harald , Fonatsch Christa , Gökbuget Nicola , Hoelzer Dieter
Published: November 2003

We evaluated the morphological findings in 150 consecutive cases of T‐lineage acute lymphocytic leukaemia (T‐ALL). Cytochemistry including PAS staining and acid phosphatase reaction proved of limited value for the diagnosis of ALL.

The diagnosis of acute leukaemia was easy to establish in most instances. However, in a few cases the leukaemic cells were difficult to recognize as blasts. The nuclei of such cells showed condensed chromatin and nucleoli were lacking, and was encountered particularly in thymic ALL.

Basophilic cytoplasm combined with prominent vacuolization suggestive of mature B‐ALL (ALL‐L3 type), was observed in 16 cases. Other features, however, such as cell size, polymorphism, chromatin structure, sparse cytoplasm or focal positivity for acid phosphatase, excluded a diagnosis of ALL‐L3 in those cases.

Distinction from hybrid leukaemia was difficult in 20 cases, because of a low percentage of peroxidase‐positive blasts or other features which suggested a separate myeloid leukaemia component. In nine of these the hybrid nature of the leukaemia was considered as certain on the basis of morphology. Seven cases had been diagnosed as biphenotypic with coexpression of myeloid and lymphoid markers by immunological techniques.

In conclusion, our analysis showed some serious pitfalls of the morphology in T‐ALL, clearly indicating the need for immunological analysis of the leukaemic cells. However, morphology remains an essential component of the diagnostic repertoire, especially when the marrow is difficult to aspirate and in cases with equivocal immunological findings. Furthermore, recognition of a separate myeloid leukaemic component in addition to the lymphatic one requires a morphological analysis.
Needs and responses of parents following the diagnosis of childhood cancer
Authors: Sloper P
Published: November 2003

The diagnosis of childhood cancer can have wide‐ranging effects on the family. The study investigated parents' views of the effects on family life at 6 months post‐diagnosis, their satisfaction with services, and the support they needed and obtained. Ninety‐eight families took part in the study which involved interviews with the main carer and self‐report questionnaires completed by mothers and fathers. Levels of parental psychological distress were assessed, using the Malaise inventory: 55% of mothers and 41% of fathers scored above the cut‐off point held to indicate high levels of emotional distress. Relationships between scores on this measure and reported effects and support were investigated. Negative effects on parental employment, finance and family relationships and lack of emotional support were associated with higher levels of distress. Parental concerns regarding delay in diagnosis and unmet needs for counselling were noted.
Improving services to Asian families and children with disabilities
Authors: Shah R
Published: November 2003

There is a clear need to improve the delivery of services to Asian families and their children with disabilities. This paper describes some of the key points where communication and access may break down. It offers principles to guide assessments of children, and makes recommendations on ways of working with interpreters, monitoring uptake of services and strategies for supporting and informing Asian families.
Evaluating family‐centred service using a measure of parents’ perceptions
Authors: King G A , Rosenbaum P L , King S M
Published: November 2003

This article describes the use of a newly‐developed measure of parents’ perceptions of health care providers’ behaviours (Measure of Processes of Care — MPOC) to evaluate the family centredness of children’s rehabilitation services. The measure was developed with the participation of more than 1600 parents of children with chronic neurodevelopmental conditions throughout Ontario. It assesses five domains: enabling and partnership; providing general information; providing specific information about the child; coordinated and comprehensive care; and respectful and supportive care. By comparing the perceptions of parents receiving services from three different types of organizations or programmes, we demonstrated that the MPOC can pick up differences between parents in their experiences of caregiving. We also demonstrated that the MPOC is able to detect differences in how parents view the family‐centredness of services provided by individual centres. The data indicate that the MPOC has appreciable utility in providing programmes and services with a description of their current level of family‐centred service as perceived by parents. The strengths, limitations and potential uses of the measure in other contexts are discussed.
Mutation screening of interferon‐gamma (IFNγ) as a candidate gene for asthma
Authors: HAYDEN C. , PEREIRA E. , RYE P. , PALMER L. , GIBSON N. , PALENQUE M. , HAGEL I. , LYNCH N. , GOLDBLATT J. , LESOUËF P.
Published: November 2003

Background
Reduced levels of interferon gamma (IFNγ) mRNA and protein have been detected in the bronchoalveolar lavage fluid of atopic asthmatics. IFNγ is secreted by TH1 cells while IL‐4 and IL‐5 are secreted by TH2 cells and an imbalance in the TH1/TH2 response may be responsible for atopic asthma. The gene for IFN4gM is located on chromosome 12; a region of the genome which has been shown in linkage studies to be associated with asthma.

Objective
To determine if there are any mutations present in the coding exons and 5’flanking region of the IFNγ gene in atopic asthmatic subjects compared with controls to explain the lower levels of this cytokine as an inherited, rather than acquired, factor in the asthmatic subjects.

Methods
The four exons and 5’flanking region of the IFNγ gene were amplified by polymerase chain reaction (PCR) from genomic DNA of 265 individuals from a Western Australian and a Venezuelan population. The PCR products were examined by single strand conformational polymorphism and heteroduplex analyses to see if there were any changes in the DNA migration patterns which would suggest the presence of a sequence variation.

Results
The four exons and the 5’flanking region of the IFNγ gene were amplified from 265 individuals from two populations. Single strand conformational polymorphism and heteroduplex analyses did not reveal any mutations in the regions examined.

Conclusions
The gene for IFNγ appears to be highly conserved as no sequence variations were detected in 265 individuals. These results suggest that mutations of the IFNγ gene are unlikely to be a significant cause of an inherited asthma diathesis.
Prevalence of childhood asthma based on questionnaires and methacholine bronchial provocation test in Korea
Authors: KIM Y. Y. , CHO S. H. , KIM W. K. , PARK J. K. , SONG S. H. , KIM Y. K. , JEE Y. K. , HA M. N. , AHN Y. O. , LEE S. I. , MIN K. U.
Published: November 2003

Background
In most epidemiological survey studies, only subjective symptoms and past medical history of asthma have been used as diagnostic criteria. Even though a questionnaire survey can be performed in a large population study at low cost, limitations such as lack of objectivity and poor predictability in non‐specific bronchial hyperresponsiveness cannot be avoided.

Objectives
The purpose of this study was to elucidate the prevalence of current asthma based on questionnaires and methacholine bronchial provocation test, and the prevalence of atopy in Korea.

Methods
We performed modified ATS respiratory questionnaires and allergen skin‐prick test with 10 common inhalant allergens among 3219 subjects aged 7–19 years in Seoul and a rural part of a small city, Chungju in Korea. Methacholine bronchial provocation tests were also performed among those who had asthma symptoms according to the questionnaire. The criteria of asthma was presence of both asthma symptoms and non‐specific bronchial hyperresponsiveness. Atopy was defined as when an allergen induced weal size is same or larger than that caused by histamine.

Results
The prevalence of asthma based on questionnaires and methacholine bronchial provocation tests was 4.6%, while the prevalence of wheeze was 8.2% and 19.3% of total population complained of one or more respiratory symptoms related to asthma on the questionnaires. There was no significant difference according to age, sex and living area. The mean prevalence of atopy was 35.0% and the most common allergens were Dermatophagoides farinae (30.9%), Dermatophagoides pteronyssinus (27.5%), cat fur (20.4%) and cockroach (11.8%). The atopy prevalence in Chungju area was higher than that in Seoul and males showed a higher prevalence than females. The asthma prevalence was higher among atopies (6.8%) than among non‐atopies (2.7%). None of questionnaire items were enough to predict the presence of bronchial hyperresponsiveness in terms of sensitivity, specificity and positive predictive value.

Conclusion
The prevalence rate of current asthma in Korea was 4.6% and the prevalence rate of atopy in Korea was 35.0%. Questionnaire‐based surveys are not enough to predict the actual prevalence of asthma.
Loss of nitric oxide release in passively sensitized guinea‐pig aorta with purified immunoglobulin Gl
Authors: JONDET A. , GOGNY M. , VU A. T. , BLIN M. , BIDON J.‐C.
Published: November 2003

Background
Vascular hyperresponsiveness can be reproduced by in vitro passive sensitization of isolated aorta with immunoglobulin G1 (IgGl) taken from ovalbumen‐sensitized BFA guinea‐pig.

Objective
The aim of the present work was to investigate the role of nitric oxide in the sensitization‐induced alteration of the contractile and relaxant responses of guinea‐pig aorta to noradrenaline (NA) and acetylcholine (ACh). respectively.

Methods
Cumulative concentration‐response curves to NA or ACh were established before and after IgG1 sensitization and then after successive treatments.

Results
IgGl in vitro passive sensitization of aorta caused a significant hyperreactivity to NA and completely inhibited the relaxation to ACh. After sensitization. the addition of an intact aortic ring (with endothelium) in the organ chamber restored the maximal response to NA and ACh close to control but was ineffective in the presence of hemoglobin. The restoration of the control reactivity to NA was also inhibited in the presence of L‐NAME or when the added aortic ring was endothelium‐denuded. Moreover. L‐arginine. a nitric oxide (NO) precursor, was able to restore the control reactivity to NA.

Conclusion
The present results show that IgG| in vitro sensitization induced a loss of NO release from the vascular endothelium. This loss of NO probably plays a great role in vascular hyperreactivity by increasing the contractile response and decreasing the relaxant response to mediators and would be a component of allergic diseases pathogenesis.
Presence of antibodies to replication protein A in some patients with systemic lupus erythematosus (SLE)
Authors: Garcia-Lozano R , Wichmann I , Garcia A , Sanchez-Roman J , Gonzalez-Escribano F , Nuñez-Roldan A
Published: November 2003

Presence of antibodies directed against replication protein A (RPA), a DNA binding protein complex composed of three subunits (RPA-70, RPA-32 and RPA-14) was investigated among patients with SLE and other autoimmune diseases using immunoblot analysis to RPA-70 and RPA-32 recombinant proteins. Anti-RPA antibodies were found in two out of 108 sera from SLE patients, one of them showing reactivity against RPA-32 and RPA-70 and the other reacting only against RPA-32. Sera from 108 patients with other autoimmune disorders as well as from 42 healthy control individuals were negative. Thus, the frequency of these antibodies in SLE is estimated to be 2–3%. The study demonstrates that RPA is one target more of the wide array of autoantigens that elicit an immune response in SLE. The presence of anti-RPA autoantibodies seems to be circumscribed to a small number of patients with SLE.
Human natural killer (NK) cells present staphylococcal enterotoxin B (SEB) to T lymphocytes
Authors: D’Orazio J A , Stein-Streilein J
Published: November 2003

Superantigen-mediated T cell activation requires the participation of antigen-presenting cells (APC). Once superantigen has bound class II MHC molecules on the surface of APC, it then can interact with the T cell receptor to induce T cell activation. Superantigen-mediated T lymphocyte activation, along with its consequent cytokine production is thought to be the basis for the pathophysiology of conditions such as toxic shock syndrome, Kawasaki’s disease and possibly rheumatoid arthritis. We examined the role of CD56+ NK lymphocytes in the interaction between superantigens and T lymphocytes. First, we found that a subpopulation of CD56+ cells freshly isolated from human peripheral blood expressed class II MHC molecules. The amount of HLA-DR expression varied between individuals, ranging from 9.3% to 37.7%. CD56+ (NK) cells were purified from the peripheral blood by cell sorting and were tested for their ability to support SEB-mediated T cell activation as assessed by surface expression of IL-2 receptor α-chain (CD25) on CD3+ lymphocytes. We observed that when enriched T cells were incubated with SEB in the presence of NK cells, there was a significant up-regulation of CD25 expression on the T cells. When HLA-DR+ cells were removed from sorted CD56+ populations, the remaining HLA-DR− NK cells were unable to support SEB-mediated T cell activation. Also, SEB up-regulated the expression of HLA-DR on CD56+ cells in peripheral blood mononuclear cell (PBMC) populations after 24 h of incubation, implying that the ability of NK cells to function as superantigen-presenting cells is up-regulated by superantigens themselves. Together, these data demonstrate for the first time that human CD56+HLA-DR+ NK cells can function as superantigen-presenting cells, and imply that NK cells may be involved in the activation of non-specific T cell reactivity during early host defences against superantigen-elaborating microorganisms in vivo. Furthermore, the physical linkage of NK cells and T cells by the interaction of superantigen with HLA class II molecules and T cell receptors, respectively, may lead to NK cell activation and augmented lytic potential, helping to clear the body of superantigen-elaborating microorganisms.
Circulating IgA, IgG, and IgM class antibody against Haemophilus parainfluenzae antigens in patients with IgA nephropathy
Authors: Suzuki S , Nakatomi Y , Odani S , Sato H , Gejyo F , Arakawa M
Published: November 2003

We previously demonstrated a close relationship between the outer membranes of Haemophilus parainfluenzae (HP) antigens (OMHP) and IgA nephrophathy (IgAN). Our objective was to clarify the relationship among IgA, IgG, and IgM class antibody against OMHP in the sera of 44 patients with IgAN and 62 patients with other glomerular diseases (OGD) by ELISA. Patients with IgAN showed a significantly higher level of IgA antibodies (P < 0.0005) and IgG antibodies (P < 0.001) against OMHP, than did patients with OGD. Positive correlations were observed between IgA and IgG antibodies, between IgA and IgM antibodies, and between IgG and IgM antibodies against OMHP in the sera of patients with IgAN. Immunoblotting showed that IgA, IgG, or IgM antibodies against OHMP in the sera of all patients with IgAN bound to the components of OMHP. Amino acid sequences of three components of OMHP recognized by the sera from patients with IgAN revealed homology with those reported for outer membrane protein (OMP) P6 precursor, OMP P5, and P2 porin protein of H. influenzae. Results suggest that patients with IgAN have glomerular deposits of OMP P6 precursor, OMP P5, or P2 porin protein of HP, and a specific increase in the production of IgA antibodies against OMHP via polyclonal activation against these, with switching of production from one isotype to another, e.g. from IgM to IgA.
Beneficial effect of Salmonella typhimurium infection and of immunoglobulins from S. typhimurium-infected mice on the autoimmune disease of (NZB×NZW)F1 mice
Authors: Matsiota-Bernard P , Hentati B , Pié S , Legakis N , Nauciel C , Avrameas S
Published: November 2003

Various infections can precede or aggravate autoimmune diseases. Yet a beneficial effect of infection has also been described and various mechanisms have been postulated to explain this effect. The aim of this study was to examine the hypothesis that infection can have an immunoregulatory effect on the autoimmune process via the increased production of natural polyreactive antibodies. The effect of Salmonella typhimurium infection on the lupus-like disease of (NZB×NZW)F1 (B/W) mice was therefore studied. The effect of IgM and IgG preparations isolated from the serum of S. typhimurium-infected C57Bl/6 and CBA mice on the autoimmune disease of B/W mice was also tested. C57Bl/6 and CBA mice were chosen because they are respectively genetically susceptible and resistant to S. typhimurium infection and they differ in their antibody response during the early phase of infection. CBA mice can mount a specific anti-bacterium antibody response, whereas C57Bl/6 mice present increased production of polyreactive antibodies. The infection effect was evaluated on several disease parameters, i.e. survival, incidence of high grade proteinuria and serum IgM and IgG antibody activity directed against a panel of autoantigens. Our main findings were: (i) infection of B/W mice with an attenuated strain of S. typhimurium delayed the course of the autoimmune disease when performed before the appearance of autoimmune symptoms; and (ii) IgM and IgG preparations from S. typhimurium-infected C57Bl/6 mice had a similar effect, whereas the IgM and IgG preparations from infected CBA mice, as well as from normal C57Bl/6 and CBA mice, were ineffective. These results suggest that S. typhimurium infection can beneficially influence the development of the autoimmune disease of B/W mice. The immunoregulatory effect of the infection seems to be related, at least partially, to the increase of a particular population of antibodies, the polyreactive antibodies.
Structural differences between the human and mouse 52-kD Ro autoantigens associated with poorly conserved autoantibody activity across species
Authors: Keech C L , Gordon T P , McCluskey J
Published: November 2003

Anti-nuclear autoantibodies found in human autoimmune diseases frequently cross-react with homologous autoantigens in distant species, supporting the notion that autoantibodies target conserved functional domains. However, the 52-kD Ro(SS-A) protein is an exception, in that human autoantibodies are not known to recognize any equivalent antigen in the cells of rodents and other non-primate species. To understand this lack of cross-reactivity we have isolated cDNAs encoding the mouse 52-kD Ro molecule. The cDNA encoding mouse 52-kD Ro revealed an open reading frame of 470 amino acids, with 70% sequence identity to the human 52-kD Ro antigen. The putative leucine-zipper and zinc-finger motifs present in human Ro52 were conserved in the mouse protein. Recombinant mouse 52-kD Ro protein reacted with human autoantibodies by ELISA and immunoblot, but with approximately 10-fold lower reactivity than recombinant human 52-kD Ro protein under the same conditions. Detection of both human and mouse 52-kD Ro by immunoblot was dependent on antigen concentration which was limiting in the cell equivalents generally used in immunoblot assays. Differential chaotropic disruption of antibody binding suggested a lower avidity of human autoantibody binding to the mouse 52-kD Ro protein compared with the human antigen. Thus the poor reactivity of native mouse 52-kD Ro with human autoantibodies is associated with species divergence diffusely distributed throughout the primary structure of the 52-kD Ro molecule.
Affinity-purified cardiolipin-binding antibodies show heterogeneity in their binding to oxidized low-density lipoprotein
Authors: Vaarala O , Puurunen M , Lukka M , Alfthan G , Leirisalo-Repo M , Aho K , Palosuo T
Published: November 2003

Antiphospholipid antibodies in autoimmune sera have been shown to react with a complex of phospholipids (cardiolipin) and a plasma phospholipid-binding protein, β2-glycoprotein I (apolipoprotein H). The binding of these antibodies was inhibited by oxidized low-density lipoprotein (LDL) in sera from patients with systemic lupus erythematosus (SLE), suggesting cross-reactivity between antiphospholipid antibodies and antibodies binding to oxidized LDL. We purified antiphospholipid antibodies by cardiolipin-polyacrylamide column from seven SLE sera and studied the reactivity of eluted fractions with cardiolipin–β2-glycoprotein I complex and oxidized LDL (malondialdehyde-conjugated LDL) in solid-phase enzyme immunoassay. In four sera the binding of IgG antibodies to cardiolipin–β2-glycoprotein I complex and to oxidized LDL appeared in the same fractions, whereas in three sera reactivities against cardiolipin and oxidized LDL were observed, at least in part, in separate fractions. The binding to solid-phase cardiolipin was dependent on the presence of exogenous β2-glycoprotein I in all fractions. Our findings show that antiphospholipid antibodies are heterogeneous in their binding to oxidized LDL, indicating that these two antibodies may have different subspecificities. Some eluted fractions reacted only with oxidized LDL, and did not show binding to cardiolipin–β2-glycoprotein I complex, suggesting that the lipid part in the antigenic complex might be responsible for the cross-reactivity of these antibodies. Accordingly, the biological functions of antibodies against phospholipid–β2-glycoprotein I complex and antibodies against oxidized LDL may also be different.