bluuebunny/crossref_metadata_2025 huggingface dataset:
Over‐expression of cyclin D1 in chronic B‐cell malignancies with abnormality of chromosome 11q13
Authors: Delmer Alain , Ajchenbaum‐Cymbalista Florence , Tang Ruoping , Ramond Sylvie , Faussat Anne‐Marie , Marie Jean‐Pierre , Zittoun Robert
Published: July 2010

Accurate identification of B‐cell chronic malignancies is sometimes uncertain, despite careful cytologic and immunophenotypic evaluation. Cytogenetics and molecular biology studies may therefore prove useful, because some of these disorders are associated with nonrandom abnormalities, such as the t(11;14)(q13;q32) translocation and bcl‐1 rearrangement mainly observed in mantle‐cell lymphoma (MCL). We studied the expression of cyclin D1 in malignant lymphoid cells from the peripheral blood of 32 patients with various B‐cell chronic lympho‐proliferative disorders, using Northern blot (NB) and RNA in situ hybridization (ISH). Cytogenetic analysis was informative in 18 cases, and most of the missing karyotype data were from typical B‐CLL cases where a t(11;14) is unlikely to be found. Over‐expression of cyclin D1 mRNA was observed by both NB and ISH in four samples (MCL: two cases; lymphoplasmacytic lymphoma: one case, unclassified B‐cell chronic disorder: one case). In each of these cases there was an abnormality of chromosome 11q13, either a t(11;14)(q13;q32) translocation (three cases) or a del(11)(q13) without evidence of chromosome 14 involvement (one case). Cytogenetic and gene rearrangement studies are not available in all institutions and have some technical pitfalls. Because of its close association with bcl‐1 rearrangement and/or t(11;14), the demonstration of cyclin D1 mRNA over‐expression either by NB, or, more conveniently, by ISH, may represent additional information which could be of help for the identification of B‐cell malignancies.
Desferrioxamine therapy accelerates clearance of iron deposits after bone marrow transplantation for thalassaemia
Authors: Giardini C. , Galimbbrti M. , Lucarelli G. , Polchi P. , Angelucci E. , Baronciani D. , Gaziev D. , Erer B. , Nasa G. La , Barbanti I. , Muretto P.
Published: July 2010

We treated 18 heavily iron‐loaded patients who had become ex‐thalassaemics after bone marrow transplantation with subcutaneous desferrioxamine therapy for 5‐20 months. As determined using serum ferritin concentration, transferrin saturation and stainable liver iron obtained in follow‐up biopsies, marked decreases in body iron stores were observed with this regimen. Moreover, the liver function tests demonstrate a trend to normalization in all cases. Local skin reactions to desferrioxamine were the only toxicities observed. We conclude that pharmacological iron chelation is a safe and effective therapy in the reduction of iron deposits in this clinical situation; it therefore represents a valid alternative to phlebotomy in selected patients.
Evaluation of transfusional iron overload before and during iron chelation by magnetic resonance imaging of the liver and determination of serum ferritin in adult non‐thalassaemic patients
Authors: Jensen P. D. , Jensen F. T. , Christensen T. , Ellegaard J.
Published: July 2010

The ability to quantitate transfusional iron overload is crucial for determining the need for and the efficacy of chelation therapy in patients with long‐standing transfusion‐dependent anaemias. We evaluated the usefulness of some indirect measures of iron overload in estimating the iron concentration in the liver ‐ the most important iron storage organ ‐ in 26 non‐chelated adult non‐thalassaemic patients. Liver Iron concentration was determined noninvasively by magnetic resonance imaging (MRI). The standard error of the estimated liver iron concentration was 80 μmol Fe/g dried liver tissue when using the number of transfused blood units, and 93 μmolFe/g when using a serum ferritin assay. Follow‐up in 11 patients (1248 months) revealed that serum ferritin is a poor measure of the liver iron concentration during iron chelation. However, this discrepancy was individually different and seemed to be dependent on the erythropoietic marrow activity. By monitoring the liver iron concentration by MRI. we compared the efficacy of chelation with desferrioxamine given either by subcutaneous continuous infusions or by bolus injections. Depletion of liver iron stores could be achieved efficiently by both regimens.
The incidence of factor VIII inhibitors in the United Kingdom, 1990‐93
Authors: Colvin B. T. , Hay C. R. M. , Hill F. G. H. , Preston F. E. ,   for the Inhibitor Working Party on behalf of the United Kingdom Haemophilia Centre Directors Organization
Published: July 2010

Reports of all the factor VIII inhibitors arising in the United Kingdom in patients with haemophilia A during the years 1990‐93 have been collated by the United Kingdom Haemophilia Centre Directors Organization. 32 new inhibitors were reported during this period, giving an average incidence of new inhibitors of 1.5 per 1000 patients registered per year. Although most occurred in patients with severe haemophilia under the age of 10. 29% occurred in patients whose VIIIC was >3 iu/dl, and 38% in patients over the age of 10 years. The incidence of inhibitors rose during the study period, but this change was not statistically significant.
Second early allogeneic stem cell transplantations for graft failure in acute leukaemia, chronic myeloid leukaemia and aplastic anaemia
Authors: Guardiola Philippe , Kuentz Mathieu , Garban Frédéric , Blaise Didier , Reiffers Josy , Attal Michel , Buzyn Agnès , Lioure Bruno , Bordigoni Pierre , Fegueux Nathalie , Tanguy Marie‐Laure , Vernant Jean‐Paul , Gluckman Eliane , Socié Gérard ,   French Society of Bone Marrow Transplantation
Published: July 2010

In this retrospective multicentre study, we analysed the results of 82 consecutive second early allogeneic transplants for primary (n = 28) or secondary (n = 54) graft failures performed between 1985 and 1997 in patients with acute leukaemia (n = 33), aplastic anaemia (n = 29) or chronic myeloid leukaemia (n = 20). HLA‐matched siblings were used in 64 cases. The same donors were used for both transplants in 56 cases and the first transplant was T‐cell depleted in 30 cases. The median age at transplant was 25 years and the median intertransplant time interval was 2 months. Estimates of the 3‐year overall survival and day 100 transplant‐related mortality were 30% and 53% respectively. A recipient age < 34 years at transplant, an intertransplant time interval ≥ 80 d and a positive recipient cytomegalovirus serology were predictors of a better outcome. The use of cyclosporin A (CsA) after second transplant had a dramatic impact on outcome, the best results being observed with CsA alone. The day 40 probability of neutrophil recovery was 73%. The use of peripheral blood progenitor cells (PBPCs) was associated with a higher and faster neutrophil recovery. Other factors associated with neutrophil recovery were an intertransplant time interval ≥ 80 d and a positive recipient cytomegalovirus serology. Therefore, second early allogeneic transplantation for graft failure is an effective treatment, especially if patients can receive CsA for graft‐versus‐host disease prevention and are retransplanted more than 80 d from first transplant.
TEL–AML1 fusion gene frequency in paediatric acute lymphoblastic leukaemia in Brazil
Authors: Magalhães I. Q. , Pombo‐de‐Oliveira M. S. , Bennett C. A. , Cordoba J. C. , Dobbin J. , Ford A. M. , Greaves M. F.
Published: July 2010

We analysed 67 samples from Brazilian children of diverse ethnic origins with acute lymphoblastic leukaemia (ALL) for the presence of the TEL–AML1 fusion gene transcripts using reverse transcription polymerase chain reaction (RT‐PCR). All 12 positive cases (20% of the 60 B‐cell precursor ALL) had common (CD10+) ALL with a mean age of 4 years (range 1–10 years). We conclude that the frequency, age, distribution and clinical features of the TEL–AML1 fusion gene‐positive ALL is similar in the diverse ethnic backgrounds of the Brazilian children to that in other countries with predominantly white Caucasian or oriental ethnicity. Apparent exceptions to this generality are discussed.
Follow‐up by cytogenetic and fluorescence in situ hybridization analysis of allogeneic bone marrow transplantation in two children with Fanconi's anaemia in transformation
Authors: Ortega M. , Caballín M. R. , Ortega J. J. , Olive T. , Coll M. D.
Published: July 2010

Results of bone marrow transplantation (BMT) in patients with Fanconi's anaemia (FA) in transformation are very poor and only a few cases with favourable outcome have been reported. We present the follow‐up of two FA‐myelodysplastic syndrome (MDS) patients with monosomy 7 and complex karyotype implicating chromosome 1. Both relapsed with acute myeloid leukaemia (AML) following an allogeneic BMT from an HLA‐identical brother. The patients showed clonal cytogenetic evolution coinciding with the leukaemic transformation. In one patient, fluorescence in situ hybridization using X and Y chromosome probes detected an increase of host cells before clinical relapse. Both patients received a successful second allogeneic BMT from the same donor using a more intensive treatment regimen and remain in clinical and cytogenetic remission more than 3 years later.
Nonsense mutation in exon V of the factor XI gene does not abolish platelet factor XI expression
Authors: Shirk Rebecca A. , Konkle Barbara A. , Walsh Peter N.
Published: July 2010

Factor XI (FXI) is a plasma glycoprotein produced by the liver that plays an essential role in blood coagulation. Individuals deficient in this protein are prone to bleeding following trauma or surgery. Well‐washed platelets possess FXI‐like activity and FXI antigen that differs in molecular weight from plasma FXI and is associated with FXI mRNA from platelets in which exon V is absent. Some individuals deficient in plasma FXI produce the platelet form of the protein. The molecular basis for the presence of platelet FXI in plasma FXI‐deficient patients is unknown. In the current study, to help determine the mechanism for this differential expression, the FXI genotype was determined for three plasma FXI‐deficient individuals who express platelet FXI. All three individuals had a nonsense mutation encoding a stop codon in exon V of the FXI gene that would result in a severely truncated polypeptide. An exon V nonsense mutation in the FXI gene combined with the absence of exon V in platelet FXI mRNA provides a plausible mechanism for the differential expression of platelet FXI in plasma FXI‐deficient patients.
Respiratory burst of neutrophils is not required for stem cell mobilization in mice
Authors: Van Os Ronald , Robinson Simon N. , Drukteinis Dainius , Sheridan Tara M. , Mauch Peter M.
Published: July 2010

It has been suggested that mature neutrophils may play an essential role in the cascade of events leading to egress of stem cells from the bone marrow to the peripheral blood. To investigate further the role of mature neutrophils and of reactive oxygen intermediates (ROIs), known to be involved in the signal transduction of neutrophils, we used mice deficient in respiratory burst, and thus the production of ROIs, to study the involvement of this activation pathway in stem cell mobilization. B6 mice with chronic granulomatous disease (CGD) received either cyclophosphamide (200 mg/kg) on day 1 and granulocyte colony‐stimulating factor (G‐CSF) (250 µg/kg/d) on days 3–6 or a single dose of interleukin 8 (IL‐8; 30 µg/mouse) as a mobilization regimen. On day 7, the number of stem and progenitor cells in blood and bone marrow was compared with control B6 animals (with intact respiratory burst). White blood cell counts, bone marrow cellularity and the frequency of granulocyte–macrophage colony‐forming cells (GM‐CFC), and cobblestone area‐forming cells (CAFC) on days 7 (CAFC‐7) and 28 (CAFC‐28) were determined. After cyclophosphamide and G‐CSF (CY + G), both mouse strains showed considerable mobilization of CAFC‐7 and CFU‐GM to the blood. Normal mice showed up to a 1905‐fold increase in progenitors per ml blood, whereas CGD mice showed up to a 264‐fold increase in blood progenitors. IL‐8 also induced mobilization in both mouse strains. In addition to progenitors, primitive stem cells measured as CAFC‐28 and as CAFC at day 35 were also mobilized by both mobilization protocols in normal as well as in CGD mice. In conclusion, respiratory burst and the subsequent signal transduction pathway do not appear to be required for mobilization of stem cells. Accordingly, neutrophils either are not involved in stem cell mobilization or other signalling pathways within neutrophils must exist that lead to the release of factors which activate stem cell egress from the bone marrow.
Prospective screening by a panfungal polymerase chain reaction assay in patients at risk for fungal infections: implications for the management of febrile neutropenia
Authors: Hebart Holger , Löffler Jürgen , Reitze Heike , Engel Andreas , Schumacher Ulrike , Klingebiel Thomas , Bader Peter , Böhme Angelika , Martin Hans , Bunjes Donald , Kern Winfried V. , Kanz Lothar , Einsele Hermann
Published: July 2010

Invasive fungal infections are a major cause of mortality in neutropenic cancer patients. To determine whether a polymerase chain reaction (PCR)‐based assay enabled the identification of patients at risk for invasive fungal infections, a prospective monitoring once per week was performed during 92 neutropenic episodes in patients receiving chemotherapy for acute leukaemia or high‐dose therapy followed by allogeneic or autologous stem cell transplantation, with the investigators blinded to clinical and microbiological data. PCR positivity was documented in 34 out of 92 risk episodes. All patients developing proven invasive fungal infection were found PCR positive, and PCR was found to be the earliest indicator of invasive fungal infection preceding clinical evidence by a mean of 5·75 d (range 0–14 d). In febrile neutropenic patients without a prior history of invasive fungal infection, a sensitivity of 100% and a specificity of 73% of the PCR assay for the development of proven or probable invasive fungal infection was documented. In conclusion, panfungal PCR performed prospectively once a week enabled the identification of patients at high risk for invasive fungal infections.
Arginine therapy: a novel strategy to induce nitric oxide production in sickle cell disease
Authors: Morris Claudia R. , Kuypers Frans A. , Larkin Sandra , Sweeters Nancy , Simon Julie , Vichinsky Elliott P. , Styles Lori A.
Published: July 2010

To determine the effects of l‐arginine (l‐Arg) supplementation on nitric oxide metabolite (NOx) production, oral l‐Arg was given to normal controls, sickle cell disease (SCD) patients at steady state and SCD patients hospitalized with a vaso‐occlusive crisis (VOC). l‐Arg (0·1 g/kg) increased NOx formation by 18·8 ± 68% in normal controls, whereas steady‐state SCD patients demonstrated a paradoxical decrease in NOx of −16·7 ± 4% (P = 0·004). In contrast, patients with VOC demonstrated a dramatic increase in NOx production by +77·7 ± 103%, a response that was dose dependent. l‐Arg appears to be the rate‐limiting step in NOx production during VOC. Oral arginine may therefore benefit SCD patients by inducing an increase in NO production during VOC.
The importance of locally derived reference ranges and standardized calculation of dilute Russell's viper venom time results in screening for lupus anticoagulant
Authors: Gardiner C. , Mackie I. J. , Malia R. G. , Jones D. W. , Winter M. , Leeming D. , Taberner D. A. , Machin S. J. , Greaves M.
Published: July 2010

Three commercial dilute Russell's viper venom time (DRVVT) kits were evaluated at four UK centres experienced at performing lupus anticoagulant (LA) tests. Each centre established a normal reference range for the DRVVT ratio calculated against local pooled normal plasma from 20 healthy normal subjects. Plasma from LA‐positive patients and LA‐negative thrombophilia patients was also tested. DRVVT ratios and the degree of correction were assessed in a variety of ways to reflect not only the UK national Guidelines, but also the manufacturers' recommendations. The reference range data showed a normal distribution in each case, but considerable variation in the mean and SD between the centres and reagents, with the mean +2SD value ranging from 1·06 to 1·19. The use of an arbitrary DRVVT ratio of < 1·1 as the cut‐off value for normality, which is applied in many laboratories, is therefore inappropriate. Although no single kit had a clear overall advantage in terms of sensitivity and specificity, the way in which the screen and confirmation data were analysed had a major impact on the interpretation of the results. A data analysis method employing a mean plus two standard deviations (SDs) cut‐off for normality, and judgement regarding confirmation of LA based on a percentage correction of DRVVT ratio, was the simplest and most consistent, with overall sensitivity and specificity values of 81% and 94%, respectively, for uncomplicated LA‐positive and LA‐negative thrombophilia samples. We conclude that the 1991 BSCH Guidelines are in need of revision, each laboratory should establish its own normal reference range for the DRVVT ratio and a common method should be used for calculating the degree of correction with confirmation reagents, so that LA results can be correctly interpreted between laboratories. Standardizing DRVVT interpretation in this way should improve the consistency of LA detection.
Abnormalities of adherent layers grown from bone marrow of patients with myelodysplasia
Authors: Tennant G. B. , Walsh V. , Truran L. N. , Edwards P. , Mills K. I. , Burnett A. K.
Published: July 2010

Myelodysplastic syndromes (MDS) are characterized by a clonal disorder of haemopoiesis with defective growth in vitro. The long‐term culture system was used to examine aspects of stromal function in MDS patients. Primary long‐term cultures of MDS bone marrow showed poor myelopoiesis with progenitors being detected for a median 3·5 weeks (n = 12) compared with 18 weeks in cultures of normal marrow (n = 10; P < 0·0001). The haemopoietic function of adherent layers was assessed in secondary co‐cultures seeded with 5 × 106 cord blood mononuclear cells on irradiated normal (n = 27; aged 38–82 years) or MDS (n = 32; aged 41–86 years) adherent layers (> 60% confluent). The median myeloid progenitor number/cord blood co‐culture was 135 in 5‐week‐old cultures with normal adherent layers and 22 in those with MDS layers (P < 0·0001). Myeloid colonies were detectable for a median 11 weeks with normal adherent layers and 6 weeks with MDS adherent layers (P < 0·0001); erythroid colonies were detectable for 7 weeks (normal) compared with 5 weeks (MDS) (P < 0·01). The differences in granulocyte‐macrophage colony forming unit (CFU‐GM) generation were not related to patient age. Cells from adherent layers of at least half of the primary normal (n = 48) and MDS (n = 26) long‐term cultures expressed cytokines [interleukin (IL)‐3, IL‐1β, thrombopoietin (Tpo) and erythropoietin (Epo)] and receptors for retinoic acid (RARα) [IL‐2, IL‐3, macrophage colony stimulating factor (M‐CSF) (Fms) and Tpo (Mpl)]. Only IL‐1β expression was reduced in week‐5 MDS cultures compared with those from normal marrows (P < 0·05). There was also a highly significant decline in IL‐1β expression in normal (but not MDS) adherent layers between week 5 and week 10. Thus, the adherent layers in cultures grown from MDS patients were haemopoietically defective and showed abnormal IL‐1β expression.
Purging of acute myeloid leukaemia cells from stem cell grafts by hyperthermia: enhancement of the therapeutic index by the tetrapeptide AcSDKP and the alkyl‐lysophospholipid ET‐18‐OCH3
Authors: Wierenga Pieter K. , Setroikromo Rita , Vellenga Edo , Kampinga Harm H.
Published: July 2010

Hyperthermia has been shown to be a potential purging modality in autologous stem cell transplantation settings owing to its selective toxicity towards leukaemic cells. We describe two approaches to further increase the therapeutic index of the hyperthermic purging modality by using normal murine bone marrow cells and a murine model for acute myeloid leukaemia. First, the tetrapeptide AcSDKP was used to protect the normal haematopoietic progenitor cells against hyperthermic damage. Pretreatment for 8 h at 37°C with 1 × 10−9 mol/l AcSDKP resulted in a decrease in hyperthermic sensitivity of only normal haematopoietic progenitor cells. This combined treatment protocol revealed a therapeutic index (ratio of surviving fractions of normal vs. leukaemic cells) of > 500, which was considered to be sufficient for purging. This was confirmed in vivo by the survival of lethally irradiated recipients transplanted with purged simulated remission bone marrow (1 × 106 normal bone marrow cells and 5 × 104 leukaemic cells). A further increase of the therapeutic index cells was achieved by the alkyl‐lysophospholipid ET‐18‐OCH3. An incubation for 4 h at 37°C with 25 μg/ml in the presence of 5% fetal calf serum preferentially enhanced the cytotoxic effect towards the leukaemic stem cell. The combination of AcSDKP and ET‐18‐OCH3 with hyperthermia resulted in a therapeutic index of > 5000. This enabled a reduction of the hyperthermic treatment and will further minimize the toxicity to normal haematopoietic stem cell subsets, while a therapeutic index far above the required value is achieved. This tripartite purging treatment therefore offers a safe and fast purging protocol for the elimination of residual leukaemic cells in autografts.
Myeloperoxidase gene expression in non‐infant pro‐B acute lymphoblastic leukaemia with or without ALL1/AF4 transcript
Authors: Serrano J. , Lo Coco F. , Sprovieri T. , Elia L. , Vitale A. , Gregorj C. , Tafuri A. , Sánchez J. , Román J. , Torres A. , Cimino G.
Published: July 2010

In this study, we examined myeloperoxidase (MPO) gene expression in a series of 31 non‐infant pro‐B acute lymphoblastic leukaemia (ALL) patients that included 16 cases with the t(4;11) translocation and/or the resultant ALL1/AF4 chimaeric gene. Sixteen out of 31 cases (51%) were MPO mRNA positive/enzyme negative. MPO mRNA was detected in nine out of 16 (56%) and seven out of 15 (47%) patients with and without the ALL1/AF4 fusion transcript respectively. The comparative study between MPO mRNA positive and negative cases showed statistically significant differences with regard to age and white blood cell (WBC) count, and was 39·5 years vs. 26·3 years (P = 0·016) and 71·4 × 109/l vs. 157·8 × 109/l (P = 0·046) in the MPO mRNA positive and negative groups respectively. The correlation analysis between MPO mRNA expression, age, WBC count and leukaemic relapse according to the presence/absence of the ALL1/AF4 fusion showed that the statistically significant differences observed in the whole group were related mostly to the ALL1/AF4‐positive ALL patients. In fact, in this latter group, the mean WBC count and patients' age were 85 ± 79 × 109/l vs. 289·8 ± 102 × 109/l (P = 0·0005) and 44·8 ± 15·3 years vs. 26·7 ± 13·7 years (P = 0·01) in patients with and without MPO mRNA expression respectively. It appears, therefore, that the assessment of MPO mRNA expression enables a further dissection of leukaemia heterogeneity in apparently homogeneous genetic/immunophenotypic ALL subsets.
Chromosome 13 abnormalities in multiple myeloma are mostly monosomy 13
Authors: Avet‐Loiseau Hervé , Daviet Axelle , Saunier Stéphanie , Bataille on behalf of the Intergroupe Francophone du Myélome Régis
Published: July 2010

Chromosome 13 abnormalities are frequently observed in multiple myeloma (MM). Several reports recently demonstrated the strong prognostic value of these abnormalities, associated with a short survival. Cytogenetic studies have shown that most of these abnormalities are complete monosomies. In order to define the common minimal deletion, we analysed a series of 234 patients with MM using fluorescence in situ hybridization (FISH) with a panel of five probes mapping along the whole chromosome 13. A chromosome 13 abnormality was observed in 98 patients (42%), 90 of whom (92%) displayed a complete monosomy. In seven of the eight remaining patients presenting partial deletions, the three probes specific for the 13q14 region were deleted. Only one patient (1%) displayed a small deletion of the D13S319 locus. In conclusion, FISH should be used for the analysis of chromosome 13 abnormalities, using probes mapping in the 13q14 region.
Bone marrow fibrosis: pathophysiology and clinical significance of increased bone marrow stromal fibres
Authors: Kuter David J. , Bain Barbara , Mufti Ghulam , Bagg Adam , Hasserjian Robert P.
Published: July 2010

In bone marrow biopsies, stromal structural fibres are detected by reticulin and trichrome stains, routine stains performed on bone marrow biopsy specimens in diagnostic laboratories. Increased reticulin staining (reticulin fibrosis) is associated with many benign and malignant conditions while increased trichrome staining (collagen fibrosis) is particularly prominent in late stages of severe myeloproliferative diseases or following tumour metastasis to the bone marrow. Recent evidence has shown that the amount of bone marrow reticulin staining often exhibits no correlation to disease severity, while the presence of type 1 collagen, as detected by trichrome staining, is often associated with more severe disease and a poorer prognosis. It was originally thought that increases in bone marrow stromal fibres themselves contributed to the haematopoietic abnormalities seen in certain diseases, but recent studies suggest that these increases are a result of underlying cellular abnormalities rather than a cause. A growing body of evidence suggests that increased deposition of bone marrow stromal fibres is mediated by transforming growth factor‐β and other factors elaborated by megakaryocytes, but it is likely that other cells, cytokines and growth factors are also involved. This suggests new avenues for investigation into the pathogenesis of various disorders associated with increased bone marrow stromal fibres.
Stimulation of new bone formation by the proteasome inhibitor, bortezomib: implications for myeloma bone disease
Authors: Oyajobi Babatunde O. , Garrett I. Ross , Gupta Anjana , Flores Alda , Esparza Javier , Muñoz Steve , Zhao Ming , Mundy Gregory R.
Published: July 2010

Impaired bone formation contributes to the lack of bone healing in multiple myeloma and there is a need for agents with bone anabolic properties to reverse the bone deficit in patients. Bortezomib, a proteasome inhibitor with antitumour efficacy in myeloma patients, enhanced new bone formation in mouse calvarial cultures; this effect was blocked by dickkopf 1(Dkk1), an antagonist of Wnt signalling implicated in myeloma bone disease. Bortezomib inhibited Dkk1 expression in calvariae and bone marrow‐derived stromal cells, suggesting a novel mechanism by which bortezomib exerts its effects in bone. Clinical trials in patients with myeloma bone disease are needed to validate these results.
Polymorphisms in von Willebrand factor gene promoter influence the glucocorticoid‐induced increase in von Willebrand factor: the lesson learned from Cushing syndrome
Authors: Casonato Alessandra , Daidone Viviana , Sartorello Francesca , Albiger Nora , Romualdi Chiara , Mantero Franco , Pagnan Antonio , Scaroni Carla
Published: July 2010

Cushing syndrome (CS) features high‐glucocorticoid secretion and an associated hypercoagulable state often involving an increase in von Willebrand factor (VWF). To identify any influence of VWF promoter on glucocorticoid haemostatic effects, four polymorphic positions (‐3267, ‐2708, ‐2659 and ‐2525) segregating as haplotypes 1 (GCAG) or 2 (CTGA) were analysed in 50 CS patients with high VWF (group I) and normal VWF (group II) levels, divided by ABO group. Genotype distribution differed significantly between the two groups: in group I, 25·8% had genotype 1/1, 22·6% had 2/2 and 38·7% had 1/2; in group II, 0% had genotype 1/1, 57·9% had 2/2 and 31·6% had 1/2 (P = 0·03). Patients’ genotypes also differed from those of controls (P = 0·003 for group I, P = 0·03 for group II). Haplotype 1 was prevalent in group I, haplotype 2 in group II (P = 0·002), both with frequencies differing from controls (P < 0·001 and P = 0·009). By odds ratio analysis, genotype 1/1 carried a 12 times greater risk of high‐VWF levels than genotype 2/2, and haplotype 1 carried a five times greater risk than haplotype 2. Our findings suggest that VWF promoter haplotypes influence the corticosteroid‐mediated increase in VWF.
Influences on parental evaluation of the content of early intervention following early identification of deafness: a study about parents' preferences and satisfaction
Authors: Gascon‐Ramos M. , Campbell M. , Bamford J. , Young A.
Published: July 2010

Background  Following early identification of deafness attention has now turned to early intervention of deaf children and their families. Improved outcomes associated with early identification and quality early intervention have been long predicated. However, the effectiveness of early intervention is mediated by parents' values, beliefs and preferences. How these mediate the nature of intervention is key to understand how early intervention impacts on outcomes.

Methods  Eighty‐two parents of early identified deaf children participated in a longitudinal study (UK). Data from parents were collected through postal questionnaires. Parents provided repeated data on My Views on Services questionnaire (at 6‐month interval). They also filled in once the Trait Emotional Intelligence questionnaire and a socio‐demographic questionnaire. Statistical analyses were carried out to investigate parents' preferences and satisfaction with content of early intervention.

Results  Families had contact with a median of five professionals and 45 h of early intervention (on a 6‐month basis). Parents view all content of early intervention as important. Content that supported parents to support their child was a priority for them. Length of involvement in early intervention and mothers' personal disposition did not have an effect on ratings of importance. However, mothers' educational background did influence ratings on the Supporting Parents subscale (F= 3.41; d.f. = 3 and 66; P= 0.023). Parents' satisfaction scores with content of early intervention were neutral (mean: 0.2; SD 0.8). Parents were more satisfied with contents that supported them to support their deaf child, than with those aimed to support themselves. Parents' satisfaction with content that supported themselves did increase with length of involvement in early intervention (t=−3.435; P= 0.004). Significant correlations (r= 0.221; r= 0.230; r= 0.252) between mothers' well‐being and satisfaction scores were identified.

Conclusion  These findings underscore the importance of understanding how parents' beliefs, values and perceived need impact on their experience of early intervention.
HLA-DR, DQ and T cell antigen receptor constant beta genes in Japanese patients with ulcerative colitis
Authors: KOBAYASHI K , ATOH M , KONOEDA Y , YAGITA A , INOKO H , SEKIGUCHI S
Published: July 2010

We studied the T cell antigen receptor (TcR) constant beta chain genes on HLA typed Japanese patients with ulcerative colitis (UC). A TcR constant beta EcoRI 6.0-kb fragment was present in all Japanese UC patients (n= 17) but completely absent in the controls (n= 35) (x2= 47.6, P<0.001). The frequency of HLA-DR2 antigen was significantly higher in UC patients (85%versus 28% in controls, P<0.001). Furthermore, HLA-DQwl antigen was also increased in UC patients (96%versus 60% in controls, P<0.001). However, HLA-DR4 antigen was significantly decreased in UC patients (12%versus 37%, P=0.02). HLA-DR1 antigen was not found in UC patients and was present in only 15% of the controls. These results suggest that TcR beta chain and HLA-DQwl antigen may be important in the pathogenesis of Japanese UC.
Identification and characterization of IgA andVicia villosa-binding T cell subsets in rheumatoid arthritis
Authors: FORTUNE F , KINGSTON J , BARNES C S , LEHNER T
Published: July 2010

Autoimmunity may be due to augmentation of immune responses by human CD8 cells which bind the lectin Vicia villosa (VV). We have investigated T cells in rheumatoid arthritis (RA) by double immunofluorescence flow cytometry, in order to assess VV-binding CD8 and CD4 cells from the peripheral blood and synovial fluid. A significant increase in CD8+ VV adherent (P<0.0001) and CD4+ VV adherent cells (P <0.001) was found in synovial fluid, as compared with peripheral blood from patients with RA. A significant increase in VV-binding CD8+ or CD4+ cells was, however, not found in the blood of patients with RA, as compared with controls. We suggest that the lack of VV-binding T cells separated from blood, in contrast to those from synovial fluid, may be due to an inhibiting agent expressing N-acetyl d-galactosamine. Indeed, IgA1 is rich in N-acetyl d-galactosamine, it inhibits VV binding to T cells and is significantly bound to CD8 cells (P<0.001). The IgA1 was then characterized and in about half the patients J chains and secretory component was found, suggesting that the IgA1 is of the polymeric and secretory variety. IgA bound to the T cells engaged the Fcα receptors and a significant decrease in the Fcα receptors was found in CD8 cells (P<0.000l) and CD4 cells (P<0.0l). Desorption studies were then carried out on CD8 and CD4 cells which showed that a loss of cell-bound IgA1 was associated with an increase in VV binding. Conversely, adsorption of IgA to T cells was associated with a loss in VV binding. The results suggest that the failure of VV binding to CD8+ andCD4+ cells from peripheral blood of patients with RA can be ascribed to cell-bound IgA 1. Cytophilic IgA1 may inhibit the function of CD8+ VV binding cells, thereby preventing augmentation of the systemic immune response, consistent with the lack of extra-articular disease in these patients with RA.
Prostaglandin E1 suppresses macrophage infiltration and ameliorates injury in an experimental model of macrophage-dependent glomerulonephritis
Authors: CATTELL V , SMITH J , COOK H T
Published: July 2010

Prostaglandin E1 (PGE1) suppresses macrophage infiltration and ameliorates injury in an experimental model of macrophage dependent glomerulonephritis. Macrophages are mediators of glomerular injury in models of proliferative glomerulonephritis. We have recently shown that macrophages in glomerulonephritis have low prostaglandin E2 (PGE) generation, and other evidence implicates eicosanoids as regulators of macrophage activation. Here we have studied in rats the effect of 15(S)-15-methyl PGE1 (M-PGE1) on accelerated nephrotoxic nephritis, a model of acute macrophage-dependent glomerular injury. M-PGE1 ameliorated proteinuria (day 4; 61 ± 13 mg/24 h, n=9; vehicle treated, 164.17 mg/24 h, n=11; P<0.002) and glomerular hypercellularity; quantification of infiltrating macrophages by isolating glomerular cells showed reduction in the numbers of macrophages (44.9/glomerulus; vehicle treated, 119.15/glomerulus; P <0.02) with inhibition of Ia antigen expression on infiltrating macrophages (8.5%; vehicle treated 25.4%P < 0.05). Glomerular binding of nephrotoxic globulin and levels of autologous antibodies were not affected by M-PGE1. Thus the mechanism of suppression involves inhibition of macrophage accumulation and activation. M-PGE1 administered to normal rats did not affect numbers of resident leucocytes (12.6±1.5/glomerulus; vehicle treated, 13.2±1.3/glomerulus) or alter Ia antigen expression (4.1±0.2 Ia + cells/glomerulus; vehicle treated, 3.9±0.6/glomerulus). This study suggests a therapeutic role for PGE1 in this type of glomerulonephritis, and has implications for the pathophysiology of macrophage-mediated inflammation.
An increase in polymorphonuclear leucocyte chemotaxis accompanied by a change in the membrane fluidity with age during childhood
Authors: YASUI K , MASUDA M , TSUNO T , MATSUOKA T , KOMIYAMA A , AKABANE T , MURATA K
Published: July 2010

Leucocyte membrane fluidity affects cell deformability as well as the accessibility of receptors and the degree of their exposure on the membrane. These effects modulate the subsequent cellular responsiveness. We have assayed membrane fluidity of polymorphonuclear leucocytes of children, using an excimer-forming lipid technique in flow cytometry. and evaluated its relation to their chemotaxis to a formyl peptide. We report that membrane fluidity and chemotaxis of polymorpho nuclear leucocytes are increased with age. These findings may have important implications for the physiological processes in polymorphonuclear leucocyte motility during childhood.
Natural thymocytotoxic autoantibodies in non-obese diabetic (NOD) mice: characterization and fine specificity
Authors: LEHUEN A , ALTMAN J , BACH J-F , CARNAUD C
Published: July 2010

The NOD mouse is a model of human juvenile type I diabetes mellitus. As in humans and in the BB rat model, the development of diabetes in NOD mice is accompanied by evident manifestations of cell-mediated and humoral autoimmunity. Beside autoantibodies directed at putative islet cell antigens, NOD sera contain antibodies with specificity for lymphocyte cell-surface determinants. Here we demonstrate that these anti-lymphocyte antibodies have the same characteristics of target cell specificity, of isotype, and of temperature reactivity, as do natural thymocytotoxic autoantibodies (NTA) from lupic NZB mice, or from mice undergoing polyclonal B cell activation. We also demonstrate that the thymocytotoxic activity of NOD sera is not due to cross-reactive anti-insulin antibodies. Biochemical characterization of the determinants recognized by these anti-lymphocyte antibodies reveals two membrane-associated proteins of 28 and 33 kD, partially similar to the two peptides recognized by NTA from NZB mice (30 and 33 kD). Altogether, these results suggest that NOD mice develop manifestations of polyclonal B cell activation similar to those observed in lupus-prone mice. The relationship of these anomalies with the organ-specific pancreatic disease remains to be properly evaluated.
In vitroproduction of B cell growth factor and B cell differentiation factor by peripheral blood mononuclear cells and bronchoalveolar lavage T lymphocytes from patients with idiopathic pulmonary fibrosis
Authors: EMURA M , NAGAI S , TAKEUCHI M , KITAICHI M , IZUMI T
Published: July 2010

The activation of B lymphocytes and formation of immune complexes have been suggested to play an important role in the pathogenesis of idiopathic pulmonary fibrosis (IPF). To investigate the mechanisms of activation of B lymphocytes, we studied the production of B cell growth factor (BCGF) and B cell differentiation factor (BCDF) in patients with IPF and those with interstitial pneumonia associated with collagen vascular diseases (IP-CVD), in comparison with healthy controls. Culture supernatants of peripheral blood mononuclear cells from patients with IPF induced more IgM and IgA production by B lymphocytes than those from healthy controls, indicating a higher production of BCDF in the patients. Culture supernatants of T lymphocytes obtained from bronchoalveolar lavage fluids (BALF) of patients with IPF induced higher proliferation of B lymphocytes than those from healthy controls, indicating a higher production of BCGF. An increase in production of BCGF and BCDF was not observed in patients with IP-CVD. In the light of these results, it was suggested that there may be an imbalance in T lymphocyte subsets that release lymphokines like BCGF and BCDF in patients with IPF, and that the subsets may differ between blood and BALF. It remains to be elucidated whether the activation of B lymphocytes depending on T lymphocytes determines the development of disease in IPF.
Prolonged circulation of immune complexes due to various altered immune functions contributes to nephritis in MKL/lprmice
Authors: GRANHOLM N A , CAVALLO T
Published: July 2010

To gain some insight into the pathogenesis of protiferativc lupus nephritis in MRL/Ipr mice we investigated the kinetics of removal of immune complexes from the circulation, the carrier stale of blood cells, the uptake of complexes by the mononuclear phagocyte system, and the localization of complexes in kidneys. In nephritic MRL/lpr mice challenged with a subsaturating dose of radiolabelled complexes (2.5 mg bovine serum albumin-anti-bovine serum albumin) liver uptake was profoundly decreased, removal of circulating complexes was delayed, and 12-h kidney localization of complexes was enhanced 7.3-fold, in comparison to control mice. The findings were not encumbered by differences in complement concentration and most likely are attributable to various altered immune functions: spontaneous polyclonal activation of Bcells, enhanced production of endogenous immune complexes, delayed removal of complexes from the circulation, and decreased uptake of complexes by the mononuclear phagocyte system. In concert, such altered functions contribute to prolonged circulation of complexes to result in their enhanced deposition in the microcirculation.
Influence of FK506 and cyclosporin A on alloantibody production and lymphocyte activation following blood transfusion
Authors: WOO J , PROPPER D J , MACLEOD A M , THOMSON A W
Published: July 2010

The effect of administration of cyclosporin A (CyA) or the novel macrolide FK506 was investigated in AO rats given DA blood transfusions. CyA (10 mg/kg, orally) or FK506 (1 mg/kg, intramuscularly) administered for 14 days from the time of transfusion effectively inhibited primary anti-MHC class I alloantibody production. This profound inhibitory effect persisted throughout the 2-month investigation period, with little increase in ‘secondary’ alloantibody production following a challenge injection 28 days after drug withdrawal. Flow cytometrie analysis revealed no significant differences in the absolute numbers of W3/25+ (CD4+), OX-8+ (CD8+) or OX-12+ (B lymphocytes), in either the spleen or peripheral blood of transfused compared with normal, untreated animals. However, a small but significant increase in the numbers of splenocytes expressing the activation marker OX-40 (activated CD4+ cells) was observed in transfused animals. Either CyA or FK506 significantly reduced the number of cells expressing OX-39 (interleukin-2 receptors) and OX-40. Treatment of transfused animals with CyA, but not FK506 for 14 days resulted in minor, transient reduction in peripheral blood OX-19+ and W3/25+ cells, while ‘sparing’ the OX-8+ cells; these changes were not observed in spleens. In contrast, the absolute spleen cell numbers of OX-19+, W3/25+ and OX-8+ cells were significantly reduced in transfused animals given 14 days of FK506 treatment, while the corresponding blood cells were unaffected. Induction of splenic lymphoproliferative responses by the T cell mitogen concanavalin A remained normal in animals receiving transfusion alone or with CyA. In contrast, profound inhibition of mitogenic responses was observed in FK506-treated animals and this inhibitory effect declined gradually following drug withdrawal. No non-specific suppressor cell activity was detected in the spleens of rats given transfusion alone or in CyA or FK506-treated transfused animals.
Experimental murine thyroiditis induced by porcine thyroid peroxidase and its transfer by the antigen-specific T cell line
Authors: KOTANI T , UMEKI K , HIRAI K , OHTAKI S
Published: July 2010

Thyroid peroxidase purified from porcine thyroid (pTPO) was found to induce an experimental murine thyroiditis with genetic restriction which was very different from that induced by mouse thyroglobulin (mTg). C57BL/6 and C57BL/10 (both H-2b) were good responders for thyroiditis, whereas A/J (H.2a), BALB/c (H-2d), DBA/2 (H-2d), CBA (H-2k), C3H/He (H-2k), and SJL/J (H-2s) were poor responders. Genetic analyses using congenic or recombinant strains revealed the following results: The H-2-linked gene (probably the I-A subregion) had a weak association with the induction of thyroiditis, and at least one non-H-2-linked gene controlled the development of thyroid lesions; antibody production to pTPO, porcine thyroglobulin (pTg) and mTg did not correlate with the incidence of thyroiditis in any strain. None of the murine thyroid microsome-specific antibodies tested by the indirect immunofluorescent technique was detected. The T cell line specific for pTPO was successfully transferred to produce thyroid lesions in C57BL/6 mice. Thyroiditis appeared 3 days after the transfer of T cell blasts, and a low concentration of anti-pTPO antibodies was detected concurrently. Thyroid lesions remained up to 48 days with almost the same extent of thyroiditis, but anti-pTPO antibodies gradually increased. In the vaccination experiments using either 0–645 C/kg (2500 rad)-irradiated or 0.3% glutaraldehyde-fixed T cell blasts, the induction of thyroid lesions by transfer was strongly suppressed. Glutaraldehyde fixation was more effective than X-irradiation in preventing thyroiditis after the transfer of T cell blasts. Vaccination also suppressed significantly the development of thyroid lesions after pTPO administration.
Inhibition of interferon-gamma-mediated activation in mouse macrophages treated with lipoarabinomannan
Authors: SIBLEY L D , ADAMS L B , KRAHENBUHL J L
Published: July 2010

Lipoarabinomannan (LAM), purified from the cell walls of Mycobacterium leprae and M. tuberculosis, is a potent inhibitor of interferon-gamma (IFN-γ) mediated activation of macrophages. The capability of LAM to inhibit IFN-γ activation of macrophages in vitro was dose dependent and required a 24-h pre-exposure. Defective activation was evident as a block in IFN-γ-induced cytocidal activity for tumour cell targets and microbicidal capacity for intracellular Toxoplasma gondii. Additionally, LAM treatment blocked the induction of surface la antigens on peritoneal macrophages by IFN-γ. The requirement for pretreatment with LAM was further substantiated by the finding that peritoneal macrophages that were activated in vivo were not affected by LAM treatments and retained full microbicidal function. However, once inhibited by LAM treatment in vitro, macrophages remained fully refractory to IFN-gamma; activation for up to 5 days in culture. Inhibition of IFN-γ activation in macrophages treated with LAM was not overcome by 100-fold increases in the dose of IFN-γ used or by a constant dose of IFN-γ in combination with 100-fold increases in the level of endotoxin used to trigger cytotoxic activity. The defect in IFN-γ unresponsiveness was not due to altered receptor function, as control and LAM-treated macrophages showed similar capacity to bind, internalize, and digest radiolabelled IFN-γ. Based on the in vitro findings reported here, the inhibition of IFN-γ-mediated macrophage activation by exposure to LAM may contribute to defective macrophage function observed in lepromatous granulomas and thus constitutes an important aspect of pathogenesis in mycobacterioses.
Interleukin-6 and ACTH act synergistically to stimulate the release of corticosterone from adrenal gland cells
Authors: SALAS M A , EVANS S W , LEVELL M J , WHICHER J T
Published: July 2010

We investigated whether interleukin-6 (IL-6) could cause the release of corticosterone by a direct interaction with the adrenal gland. Primary cultures of rat adrenal glands were obtained by dispersion with collagenase and incubated for 24 h with different doses of IL-6. Levels of corticosterone were measured by competitive protein binding assay. A significant (P< 0.025) dose-dependent increase in corticosterone levels was seen at all doses used. Time course experiments demonstrated that IL-6 stimulated corticosterone release over a period of 24 h but not after 12 or 3 h. The stimulation of adrenal cells with different doses of ACTH1–24 and 40 U/ml of IL-6 showed a synergistic effect when IL-6 was combined with low concentrations of ACTH1–24 (2 and 20 pmol/l). This effect was not evident at higher doses. Our results suggest that IL-6 may act at different levels of the hypothalmic pituitary adrenal axis. Moreover the finding of a synergistic effect with ACTH1–24 indicates that IL-6 could play a role in the long term response to stress.