bluuebunny/crossref_metadata_2025 huggingface dataset:
Age‐related changes in factor VII proteolysis in vivo
Authors: Ofosu F. A. , Craven S. , Dewar L. , Anvari N. , Andrew M. , Blajchman M. A.
Published: November 2003

Previous studies have reported that pre‐operative plasmas of patients over the age of 40 years who developed post‐operative deep vein thrombosis (DVT) had approximately twice the amount of proteolysed factor VII found in plasmas of patients in whom prophylaxis with heparin or low Mr heparin was successful. These and other studies also reported higher concentrations of thrombin–antithrombin III in pre‐ and post‐operative plasmas of patients who developed post‐operative thrombosis than in plasmas of patients in whom prophylaxis was successful. Whether the extent of factor VII proteolysis seen in the patients who developed post‐operative DVT is related to the severity of their disease or age is not known. This report investigated age‐related changes in the concentrations of total factor VII protein, factor VII zymogen, factor VIIa, tissue factor pathway inhibitor, thrombin–antithrombin III, and prothrombin fragment 1+2 in normal plasmas and the relationships between these parameters. With the exception of thrombin–antithrombin III, statistically significant increases in the concentrations of these parameters with age were found. Additionally, the differences between the concentrations of total factor VII protein and factor VII zymogen, an index factor VII proteolysis in vivo, were statistically significant only for individuals over age 40. Using linear regression analysis, a significant correlation was found to exist between the concentrations of plasma factor VIIa and prothrombin fragment 1+2. Since factor VIIa–tissue factor probably initiates coagulation in vivo, we hypothesize that the elevated plasma factor VIIa (reflecting a less tightly regulated tissue factor activity and therefore increased thrombin production in vivo) accounts for the high risk for post‐operative thrombosis seen in individuals over the age of 40.
Second transplantation using allogeneic peripheral blood stem cells in a β‐thalassaemia major patient featuring stable mixed chimaerism
Authors: OR R. , KAPELUSHNIK J. , NAPARSTEK E. , NAGLER A. , FILON D. , OPPENHEIM A. , AMAR A. , AKER M. , SAMUEL S. , SLAVIN S.
Published: November 2003

Allogeneic bone marrow transplantation (BMT) for β‐thalassaemia major carries the risks of disease recurrence due to residual thalassaemic stem cells or true immune‐mediated rejection. We report a thalassaemic patient who displayed stable mixed chimaerism with only 5% donor‐derived cells for about 5 years after BMT. Displacement of host cells was accomplished by ambulatory non‐myeloablative conditioning and allogeneic G‐CSF mobilized peripheral blood stem cell transplantation from the same donor, resulting in full reconstitution. Patients featuring stable mixed chimaerism after BMT may benefit from allogeneic cell therapy with immunocompetent lymphocytes and stem cells, whilst avoiding supralethal conditioning.
The ultrastructure of mantle cell lymphoma and other B‐cell disorders with translocation t(11;14)(q13;q32)
Authors: Resnitzky Peretz , Matutes Estela , Hedges Michael , Morilla Ricardo , Brito‐Babapulle Vasantha , Khokhar Tariq , Catovsky Daniel
Published: November 2003

We conducted an ultrastructural study in 22 cases of B‐lymphoproliferative disorders in leukaemic phase bearing the t(11;14) translocation. The features of peripheral blood leukaemic cells in nine cases of mantle cell lymphoma (MCL) were compared to those diagnosed as B‐prolymphocytic leukaemia (B‐PLL) (five cases), splenic lymphoma with villous lymphocytes (SLVL) (four cases), lymphoplasmocytic lymphoma (LPL) (one case), chronic lymphocytic leukaemia with >10% prolymphocytes (CLL/PL) (one case) and unclassified B‐non Hodgkin’s lymphoma (B‐NHL) (two cases). The ultrastructural characteristics were also compared to those present in B‐NHL without t(11;14), including cases of follicular centre lymphoma (FCL). This study shows that MCL has distinct ultrastructural features including a cleaved or indented nucleus with an even heterochromatin distribution, an absent or inconspicuous nucleolus, low N/C ratio, abundant mitochondria, a well developed Golgi zone, profiles of endoplasmic reticulum and centrioles. This pattern clearly differs from that found in FCL cells. The nuclear pattern of MCL cells also differed from the cells in the other disorders with t(11;14), but shared an organelle‐rich cytoplasm, and features which were not apparent in cases without t(11;14). The cytoplasmic changes observed in cells bearing t(11;14) suggest increased cellular activity which may relate to the chromosome translocation and the resulting over‐expression of bcl‐1.
Activity of the ligand for c‐mpl, thrombopoietin, in early haemopoiesis
Authors: Itoh Ryugo , Katayama Naoyuki , Kato Takashi , Mahmud Nadim , Masuya Masahiro , Ohishi Kohshi , Minami Nobuyuki , Miyazaki Horoshi , Shiku Horoshi
Published: November 2003

We examined the role of the ligand for c‐mpl, thrombopoietin (TPO), in murine early haemopoiesis, using a serum‐free culture system. TPO in combination with the ligand for c‐kit (SF) or interleukin‐3 (IL‐3) supported colony formation by marrow cells of 5‐fluorouracil (5‐FU)‐treated mice, whereas TPO alone yielded no colony. When blast cell colonies grown in the presence of TPO plus SF or TPO plus IL‐3 were individually replated in suspension cultures containing serum and several growth factors, various combinations of myeloid lineages were seen, indicating that the progenitors supported by TPO plus SF or TPO plus IL‐3 are multipotential. Delayed addition experiments demonstrated that TPO has the potential to effectively support the survival of haemopoietic progenitors. We then studied the effects of TPO on proliferative kinetics of cycling progenitors. TPO hastened IL‐3‐dependent growth of progenitors by shortening the time required for cell cycling. These results suggest that TPO, as a single factor, can support the survival of haemopoietic progenitors and TPO synergizes with SF or IL‐3 to act on early multipotential haemopoietic progenitors.
Clinical and molecular remission after allogeneic blood cell transplantation in a patient with mantle‐cell lymphoma
Authors: CorRadini Paolo , Ladetto Marco , Astolfi Monica , Voena Claudia , Tarella Corrado , Bacigalupo Andrea , Pileri AlessandRo
Published: November 2003

Mantle‐cell lymphoma (MCL) is a B‐cell tumour with a usually poor prognosis, characterized by the proliferation of small cleaved lymphocytes with a diffuse growth pattern. We report a polymerase chain reaction‐based analysis of minimal residual disease in a patient who achieved complete remission after allogeneic blood cell transplantation (BCT). Rearrangement of the immunoglobulin heavy‐chain genes was used to generate a lymphoma‐specific molecular marker. Lymphoma cells were not detectable in a bone marrow sample collected 12 months after BCT. Our findings suggest that allogeneic BCT may offer a curative approach to MCL.
Angioid streaks are part of a familial syndrome of dyserythropoietic anaemia (CDA III)
Authors: Sandström Herbert , Wahlin Anders , Eriksson Mikael , Holmgren Gösta , Lind Lisbeth , Sandgren Ola
Published: November 2003

Congenital dyserythropoietic anaemia type III (CDA III) is a rare disease inherited in an autosomal dominant way and characterized by mild to moderate haemolytic anaemia. Most patients are adapted to their disease and have no or few complaints. Bone marrow examination shows a characteristic picture with erythroid hyperplasia and multinucleate erythroblasts. 20% of patients in a Swedish family affected with the CDA III condition have monoclonal gammopathy or multiple myeloma. By linkage and recombination analysis in the same family, the gene linked to the CDA III condition (CDAN3) has been located to chromosome 15q22.

In this paper we report the observation of visual disturbances with macular degeneration and angioid streaks in six patients with CDA III and discuss the apparent association between CDA III, angioid streaks and monoclonal gammopathy. We suggest that this triad forms a previously unreported syndrome.
Factor VIII gene analysis in Japanese CRM‐positive and CRM‐reduced haemophilia A patients by single‐strand conformation polymorphism
Authors: Morichika Shogo , Shima Midori , Kamisue Seiki , Tanaka Ichiro , Imanaka Yasufumi , Suzuki Hiroshi , Shibata Hironobu , Pemberton Susan , Gale Karen , McVey John , Tuddenham Edward G. D. , Yoshioka Akira
Published: November 2003

Haemophilia A is the most common X‐linked blood coagulation disorder; it is caused by deficiency of factor VIII activity (FVIII:C). Half of the affected patients do not have detectable levels of FVIII protein in their plasma, whereas about 5% have normal levels of the FVIII antigen (FVIII:Ag) (> 50 u/dl), and are called cross‐reacting material (CRM) positive (CRM+ or A+). About 45% of patients have reduced levels of the FVIII:Ag (1–50 u/dl), classified as CRM reduced (CRMR or AR). We screened the FVIII gene of 13 Japanese patients (five CRM+ and eight CRMR) by single‐strand conformation polymorphism, and identified 11 different mutations in 13 patients by analysing all 26 exons (Trp255Cys, Tyr473Cys, Gly479Arg, Arg531His, Thr667Arg, Arg1689Cys, Arg1941Gln, Arg2150His, Arg2159Cys, Thr2245Ala and Gly2285Val). Seven mutations were identified in the A domains (four in the A2 domain). All the mutations are point mutations resulting in missense codons. Four mutations (Trp255Cys, Thr667Arg, Thr2245Ala and Gly2285Val) have not been described previously.
Allogeneic marrow transplantation for primary myelofibrosis and myelofibrosis secondary to polycythaemia vera or essential thrombocytosis
Authors: Anderson Jeanne E. , Sale George , Appelbaum Frederick R. , Chauncey Thomas R. , Storb Rainer
Published: November 2003

Primary myelofibrosis is a clonal haemopoietic disorder, incurable with conventional therapy, and associated with a median survival of 4–5 years. Patients with polycythaemia vera and essential thrombocytosis who progress into a myelofibrotic picture also have a poor prognosis. Between 1980 and 1996, 13 patients with myelofibrosis due to one of these three myeloproliferative disorders (primary myelofibrosis [n=8], essential thrombocytosis [n=3], polycythaemia vera [n=2]) underwent allogeneic marrow transplantation in Seattle. The median age was 40 years (range 18–49). The median time from myeloproliferative diagnosis to transplantation was 39 months (range 5–192). Three patients received preparative regimens containing total body irradiation and 10 received busulphan–cyclophosphamide regimens. Nine patients received marrow from HLA‐matched related donors, one from a one antigen mismatched related donor, and three from HLA‐matched unrelated donors. The median time to both granulocyte and platelet engraftment was 21 d. Nine patients survive between 1.2 and 7.1 years post‐transplant. Two patients relapsed at 1 year post‐transplant, both of whom survive in a chronic myeloproliferative state. Four patients died of transplant‐related complications between 43 d and 2.2 years post‐transplant. At 1 year post‐transplant the majority of the disease‐free survivors have normal peripheral blood counts and none‐to‐minimal marrow fibrosis. These preliminary results are encouraging, and suggest that stem cell transplantation can be curative therapy for selected patients with myelofibrosis.
Risk of venous thromboembolism associated with a G to A transition at position 20210 in the 3′‐untranslated region of the prothrombin gene
Authors: Brown Karen , Luddington Roger , Williamson David , Baker Peter , Baglin Trevor
Published: November 2003

The odds ratio for the FII 20210G/A mutation in 504 patients with venous thromboembolism compared to controls was 2.0 (95% CI 1.0–4.0) and, for factor V Leiden, 5.8 (95% CI 3.3–10.3). 3/504 patients were heterozygous for both mutations. None of the patients had combined natural anticoagulant deficiency and the FII 20210G/A mutation. We conclude that the FII 20210G/A mutation is present in 2.6% of the population and the relative risk of venous thromboembolism in carriers is 2.0.
Analysis of p53 gene deletions in patients with non‐Hodgkin's lymphoma by dual‐colour fluorescence in‐situ hybridization
Authors: Clodi Katharina , Younes Anas , Goodacre Angela , Roberts Mark , Palmer Judy , Younes Mamoun , Cabanillas Fernando , Andreeff Michael
Published: November 2003

The most common tumour suppressor gene altered in human cancers is p53, which is located on the short arm of chromosome 17. Structural abnormalities of the short arm and loss of chromosome 17 have been reported to confer resistance to chemotherapy in patients with non‐Hodgkin's lymphoma (NHL). Therefore we studied the incidence and prognostic value of p53 deletions in patients with NHL by fluorescence in‐situ hybridization using a 40 kb cosmid probe. Specimens obtained from 79 patients with NHL were studied. 46 patients were untreated, and 33 were previously treated. 40 tumours had indolent and 39 had aggressive histologies. p53 deletions were observed in 14 specimens (18%) in 32–90% of the cells. No statistically significant difference in the incidence of p53 deletion was observed between indolent and aggressive NHLs or between untreated and previously treated patients. However, p53 deletions were observed in three of four patients with transformed lymphoma. In the untreated patients, p53 deletion had no effect on response to therapy, time to treatment failure, or survival. We conclude that p53 deletions are uncommon in NHL, and may be frequent in patients with transformed lymphoma. In this study, p53 deletions did not influence treatment outcome or prognosis of NHL. Because monosomy 17 and 17p abnormalities have been reported to confer poor prognosis in NHL, other tumour suppressor genes on 17p should therefore be studied.
Murine endothelial cells support fetal liver erythropoiesis and myelopoiesis via distinct interactions
Authors: Ohneda Osamu , Bautch Victoria L.
Published: November 2003

Endothelial cells are an important component of the haemopoietic microenvironment. To investigate how endothelial cells are involved in haemopoiesis, two established murine endothelial cell lines were assayed in stromal cultures with fetal liver haemopoietic cells. Both endothelial cell lines allowed for the proliferation and differentiation of erythroid and monocyte‐macrophage precursors, suggesting that support for haemopoiesis is a general property of endothelial cells.

Erythropoiesis was dependent on the addition of erythropoietin (Epo), whereas myelopoiesis was independent of added Epo. Haemopoietic colonies developed in close contact with the endothelial cells. Erythroid colonies did not develop when transwell filters were used between the stroma and haemopoietic cells, or when conditioned medium was used in place of stromal cells. In contrast, monocyte‐macrophage colonies formed in the presence of transwell filters or conditioned medium. Thus close cell contact is necessary for erythropoiesis but not myelopoiesis under these conditions. These results suggest that different molecular mechanisms are used by endothelial cells to support erythroid development and myeloid development in the mouse fetal liver.
Sustained proliferation, multi‐lineage differentiation and maintenance of primitive human haemopoietic cells in NOD/SCID mice transplanted with human cord blood
Authors: Cashman Johanne , Bockhold Kathryn , Hogge Donna E. , Eaves Allen C. , Eaves Connie J.
Published: November 2003

Time course studies of sublethally irradiated non‐obese mice with severe combined immunodeficiency (NOD/SCID mice) transplanted intravenously with 107 human cord blood cells showed a rapid and parallel regeneration of human erythroid, granulopoietic, megakaryopoietic and B‐lymphoid progenitors, as well as more primitive subpopulations of CD34+ cells (defined by their multi‐lineage in vitro colony‐forming ability, coexpression of Thy‐1, or functional activity in long‐term culture‐initiating cell [LTC‐IC] assays), in the marrow, spleen and blood. Maximum numbers of human cells were reached within 6 weeks and were then sustained for another 18–20 weeks. 3H‐thymidine suicide studies showed all types of in vitro clonogenic human progenitors tested and the human LTC‐IC to be proliferating in vitro throughout this period. A 2‐week course of injections of human Steel factor, interleukin‐3, granulocyte‐macrophage colony‐stimulating factor and erythropoietin given just prior to assessment of the mice had no effect on any of these human engraftment parameters. 4–6 weeks post‐transplant, the marrow of primary NOD/SCID recipients contained human cells that were able to regenerate lymphopoiesis and/or myelopoiesis in secondary irradiated NOD/SCID mice. These findings establish a baseline for the kinetics of engraftment, multi‐lineage differentiation and self‐renewal of human cord blood stem cells in this xenogeneic transplant model and thus set the stage for future studies of their regulation in vivo.
Sensitive non‐radioactive PCR detection of clonal T‐cell receptor‐gamma gene rearrangements in childhood acute lymphoblastic leukaemia
Authors: Haber Michelle , Kwan Edward , Zhu Ling , Marshall Glenn M. , Norris Murray D.
Published: November 2003

Rearrangement of the T‐cell receptor gamma (TCRγ) gene potentially provides a valuable target for monitoring minimal residual leukaemia by the Polymerase Chain Reaction (PCR). However, existing strategies directed at this locus frequently lack sensitivity or specificity. We describe a novel PCR strategy for improved detection of clonal TCRγ gene rearrangements based on the design of two overlapping clone‐specific reverse PCR primers spanning the TCRγ junctional region, which are used sequentially in conjunction with forward V‐region specific primers. Unlike other strategies, specificity is generated from the initial and subsequent round of PCR amplification. This non‐radioactive technique is highly specific and sensitive, and should prove effective for monitoring minimal residual leukaemia.
Atypical lymphocyte morphology: an adverse prognostic factor for disease progression in stage A CLL independent of trisomy 12
Authors: Oscier D. G. , Matutes E. , Copplestone A. , Pickering R. M. , Chapman R. , Gillingham R. , Catovsky D. , Hamblin T. J.
Published: November 2003

We studied 270 patients with Binet stage A chronic lymphocytic leukaemia looking for adverse prognostic factors. In a multivariate analysis the following features were found to be risk factors for disease progression: atypical lymphocyte morphology (defined as either > 10% prolymphocytes or > 15% lymphocytes with cleaved nuclei or lymphoplasmacytoid cells); more than two karyotypic abnormalities; lymphocyte count > 30 × 109/l; lymphocyte doubling time < 1 year; enlargement of one or more lymph node groups.

In a univariate analysis the presence of trisomy 12 also correlated with progressive disease, but this was largely a consequence of the association between trisomy 12 and atypical lymphocyte morphology. Atypical lymphocyte morphology is an important prognostic factor in stage A CLL, and one which incurs no additional investigational cost.
Monitoring of oral anticoagulant therapy in lupus anticoagulant positive patients with the anti‐phospholipid syndrome
Authors: Lawrie A. S. , Purdy G. , Mackie I. J. , Machin S. J.
Published: November 2003

Introduction of the International Normalized Ratio (INR) has improved the standardization of laboratory control of oral anticoagulant therapy (OAT). However, it has been reported that misleading INR results can be obtained from OAT patients with lupus anticoagulant (LA). To investigate this claim, we studied 35 OAT patients, 14 of whom had anti‐phospholipid syndrome (APS) with a documented LA. Attainment of anticoagulation was confirmed by chromogenic assay of factor VII and factor X. Prothrombin times were performed using eight thromboplastins (five derived from rabbit brain, two recombinant human tissue factor and one made from human placenta) with an International Sensitivity Index (ISI) of <1.40. When using the thromboplastin manufacturers' ISI there was a significant difference (ANOVA, P<0.0001) between INR results obtained with the eight reagents for both APS (average CV = 12.4%) and non‐APS (average CV = 12.5%) patient groups. Variation using the eight thromboplastins was assessed by calculating the CV for each sample; these values were then pooled for each patient group to give the average CV for all samples with all reagents for the two patient groups. Results for both patient groups exhibited markedly reduced variation (APS group average CV = 6.5%, non‐APS group average CV = 5.8%) when locally assigned ISI values were employed in the calculation of INRs. Our data does not support the suggestion that the INR may not reflect the true level of anticoagulation in the long‐term warfarin‐treated patient, in whom lupus anticoagulant was detected. However, there was strong evidence that thromboplastin use should be restricted to those clot detection systems for which the reagent's manufacturer has assigned an ISI, or local ISI assignment must be undertaken. The inappropriate use of a generic (i.e. optical or mechanical clot detection system without regard to specific analyser type) ISI value can lead to ambiguous results.
Distribution of the cytogenetic abnormality +i(3)(q10) in persistent polyclonal B‐cell lymphocytosis: a FICTION study in three cases
Authors: Callet‐Bauchu Evelyne , Renard Nathalie , Gazzo Sophie , Poncet Chantal , Morel Dominique , Pagès Jacqueline , Salles Gilles , Coeur Pierre , Felman Pascale
Published: November 2003

Persistent polyclonal B‐cell lymphocytosis (PPBL) is a rare entity characterized by a moderate but sustained lymphocytosis where some binucleated or bilobulated circulating forms constitute, even if they are not entirely specific, the cytological hallmark of the disease. An additional chromosome long arm i(3)(q10) has recently been reported as a recurrent cytogenetic aberration, contrasting with a usual polyclonal immunoglobulin expression. To determine more precisely the distribution of the chromosomal abnormality within the peripheral lymphocyte population and study the relationship between the +i(3)(q10) and the bilobulated character, we investigated three new cases of PPBL displaying the cytogenetic abnormality on the karyotype, using a technique of simultaneous fluorescence immunophenotyping and interphase cytogenetics (FICTION). We demonstrated that the +i(3)(q10) was restricted to the B lymphocytes, independently of the κ or λ light chain isotype and was present in both bilobulated and non‐bilobulated cells. Therefore it is likely that the cytogenetic abnormality occurs at an early stage of lymphocyte differentiation in a precursor cell already committed to the B‐cell lineage, before any rearrangement of immunoglobulin genes has taken place.
Analysis of p18INK4C in adult T‐cell leukaemia and non‐Hodgkin's lymphoma
Authors: Hatta Yoshihiro , Spirin Konstantin , Tasaka Taizo , Morosetti Roberta , Said Jonathan W. , Yamada Yasuaki , Tomonaga Masao , Koeffler H. Phillip
Published: November 2003

p18INK4C, a cyclin‐dependent kinase inhibitor, is a homologue of p15INK4B and p16INK4A which are frequently altered in a variety of malignancies. We searched for structural alterations of the p18INK4C gene in 44 adult T‐cell leukaemias (ATLs), 101 non‐Hodgkin's lymphomas (NHLs), two polyclonal B‐cell proliferations, seven ATL cell lines and seven leukaemia/lymphoma cell lines, by Southern blot and polymerase chain reaction–single‐strand conformation polymorphism (PCR‐SSCP) analyses. No genomic alterations of the p18INK4C gene were found in any of the samples. By RT‐PCR, p18INK4C was not expressed in three of five ATL cell lines, whereas it was expressed in all the non‐ATL leukaemia/lymphoma cell lines. Tax did not inhibit the expression of p18INK4C in tax‐expressing Jurkat cells.
Antithymocyte globulin for patients with myelodysplastic syndrome
Authors: Molldrem Jeffrey J. , Caples Mary , Mavroudis Dimitrios , Plante Michelle , Young Neal S. , Barrett A. John
Published: November 2003

Twenty‐five transfusion‐dependent myelodysplastic syndrome (MDS) patients (with < 20% blasts) were treated in a phase II study with antithymocyte globulin (ATG) at 40 mg/kg/d for four doses and then followed with blood counts every 2 weeks and clinic visits every 3 months, for a median of 14 months (range 1–38 months). 11 (44%) patients responded and became transfusion‐independent after ATG, including three complete responses, six partial responses, and two minimal responses. Responses were observed in 9/14 patients (64%) with refractory anaemia (RA) and 2/6 patients (33%) with refractory anaemia with excess blasts (RAEB). Median response duration was 10 months (range 3–38 months). The Kaplan‐Meier estimate of overall survival was 84% at 38 months, with one early death due to pneumonia and two deaths from disease progression to leukaemia. Side‐effects consisted mainly of mild serum sickness in all patients. A single course of ATG restored haemopoiesis in some patients with MDS and was well tolerated.
Platelet activity of high‐dose factor VIIa is independent of tissue factor
Authors: Monroe Dougald M. , Hoffman Maureane , Oliver Julie A. , Roberts Harold R.
Published: November 2003

High‐dose recombinant factor VIIa has been successfully used as therapy for haemophiliacs with inhibitors. The mechanism by which high‐dose factor VIIa supports haemostasis is the subject of some controversy. Postulating a mechanism in which activity is dependent on tissue factor at the site of injury explains the localization of activity but not the requirement for high doses. Postulating a mechanism in which factor VIIa acts on available lipid independently of tissue factor explains the requirement for high doses but not the lack of systemic procoagulant activity. We report that factor VIIa bound weakly to activated platelets (Kd ∼ 90 nm). This factor VIIa was functionally active and could initiate thrombin generation in the presence of plasma concentrations of prothrombin, factor X, factor V, antithrombin III and tissue factor pathway inhibitor. The activity was not dependent on tissue factor. The concentration of factor VIIa required for detectable thrombin generation agreed well with the lowest concentration of factor VIIa required for efficacy in patients. High‐dose factor VIIa may function on the activated platelets that form the initial haemostatic plug in haemophilic patients. These observations are in agreement with clinical trials which have shown that high‐dose factor VIIa was haemostatically effective without causing systemic activation of coagulation.
Bone marrow fibrosis and disease activity in multiple myeloma monitored by the aminoterminal propeptide of procollagen III in serum
Authors: Abildgaard N. , Bendix‐Hansen K. , Kristensen J. E. , Vejlgaard T. , Ristelli L. , Nielsen J. L. , Heickendorff L.
Published: November 2003

Simple bone marrow fibrosis is seen in 10–30% of multiple myeloma (MM) patients. We investigated the incidence and characteristics of the bone marrow stromal alterations, in order to characterize the collagens involved by immunohistochemistry, and to evaluate the use of serum aminoterminal propeptide of type III procollagen (PIIINP) as a marker of marrow fibrogenesis and disease activity in MM. 34 consecutive patients with newly diagnosed MM were included prospectively, and followed for 12–30 months. Compared with the findings in 15 normal individuals we found increased interstitial deposits of collagen III in 48% of MM patients, whereas deposits of collagen I were not increased. Interstitial fibrosis appeared to be restricted to areas of severe plasma cell infiltration, but it could also have a more dispersed presentation in the severely infiltrated marrow. There was a high co‐distribution of collagen III fibrils and reticulin fibres. Serum PIIINP levels were elevated in most patients, and in the follow‐up study serum PIIINP showed a good correlation with the response to treatment. Patients with resistant or progressive disease had continually elevated levels of PIIINP. In most patients with responsive disease serum PIIINP normalized, and we observed no relapses in patients who had normal serum PIIINP levels. Other patients who responded to treatment by reduced M‐component level, but had persistently elevated serum levels of PIIINP, had either early relapses or developed progression of osteolytic lesions in spite of unchanged M‐component levels. Therefore an elevated serum PIIINP during treatment might indicate an active malignant clone. Serum PIIINP does not simply follow the M‐component, but gives further information of potential therapeutic value.
The predominant E2F complex in human primary haemopoietic cells and in AML blasts contains E2F‐4, DP‐1 and p130
Authors: WILLIAMS C. D. , LINCH D. C. , SØRENSEN T. S. , LA THANGUE N. B. , THOMAS N. S. B.
Published: November 2003

The E2F family of transcription factors are thought to play an important role in the control of cell cycle progression. There is now also increasing evidence that some family members may act as oncogenes or tumour suppressor genes. The characterization of these proteins in human primary haemopoietic cells and acute myeloid leukaemia (AML) blasts may thus give an insight to the molecular mechanisms governing proliferation and leukaemogenesis in these cells. Therefore we analysed the expression of E2F‐DNA binding activity and the constituent proteins found in the complexes in human primary haemopoietic cells of various lineages. We also studied blasts from 18 patients with acute myeloid leukaemia (AML). On electromobility shift assays (EMSA) a single E2F‐DNA binding complex was detected in T cells, B cells and monocytes which was shown to contain E2F‐4, DP‐1 and p130, indicating that all quiescent haemopoietic cells have the same complex. Examination of 18 AML samples by EMSA revealed the presence of E2F binding and no gross abnormalities were detected. An E2F‐4/p130 complex was detected in representative samples of all FAB types analysed. Thus abnormalities of E2F function are unlikely to play a primary pathogenic role in AML.
Acquired von Willebrand's disease due to aberrant expression of platelet glycoprotein Ib by marginal zone lymphoma cells
Authors: TEFFERI AYALEW , HANSON CURTIS A. , KURTIN PAUL J. , KATZMANN JERRY A. , DALTON ROBERT J. , NICHOLS WILLIAM L.
Published: November 2003

A 69‐year‐old woman presented with splenic marginal zone lymphoma associated with acquired von Willebrand's disease (AVWD). Laboratory abnormalities included markedly decreased plasma levels of factor VIII coagulant (C) activity (VIII:C 28%), von Willebrand’s factor (VWF) antigen (Ag) (vWF:Ag < 6%), and VWF ristocetin cofactor (RCo) activity (VWF:RCo, < 12%). VWF multimer analysis revealed a severe type II defect. Treatment with cryoprecipitate, high‐dose gamma globulin or desmopressin given intravenously was unsuccessful. Clinical bleeding and coagulation abnormalities showed transient improvement after replacement therapy with Humate‐P concentrate. The coagulation abnormalities improved partially after splenectomy and completely after subsequent chemotherapy. The neoplastic lymphocytes in the blood and spleen strongly expressed platelet glycoprotein Ib (CD42) and VWF but not other platelet‐associated antigens.
Dose reduction of asparaginase under pharmacokinetic and pharmacodynamic control during induction therapy in children with acute lymphoblastic leukaemia
Authors: AHLKE ELVIRA , NOWAK‐GO¨TTL ULRIKE , SCHULZE‐WESTHOFF PETRA , WERBER GISELA , BO¨RSTE HEDWIG , WU¨RTHWEIN GUDRUN , JU¨RGENS HERBERT , BOOS JOACHIM
Published: November 2003

The enzyme asparaginase is an important element in the therapy of acute lymphoblastic leukaemia (ALL). The usual asparaginase dose as prescribed in the ALL‐BFM‐86/90 treatment protocol for the therapy of ALL is 10 000 IU/m2 at 3 d intervals and had been developed on the basis of the E. coli asparaginase preparation CrasnitinTM from the Bayer company. Using the described schedule the E. coli asparaginase preparation from the Medac company shows significantly higher biological activity than the Bayer preparation. These findings prompted an attempt to reduce the dose of the Asparaginase medacTM under careful pharmacokinetic and pharmacodynamic monitoring. At the first step of dose reduction in ALL treatment protocol I, 11 children received 5000 IU/m2 of Asparaginase medacTM. Another 15 children were given 2500 IU/m2 of the enzyme at the second step of dose reduction. Prior to each asparaginase dose, blood samples were taken to determine amino acids and trough enzyme activity. Concurrent with the asparaginase monitoring, the coagulation parameters were measured. 96% of samples from the first step of dose reduction (5000 IU/m2 every third day) showed complete L‐asparagine depletion (<0.1 μM), the median trough enzyme acitivity was 265 IU/l. At the second step of dose reduction (2500 IU/m2) complete L‐asparagine depletion was seen in 97% of samples, and the median trough enzyme acitivity was 102 IU/l. Cerebrospinal fluid (CSF) depletion was complete in all samples tested (11/11).  We concluded that an Asparaginase medacTM dose reduced from the usual 10000 IU/m2 down to 5000 IU/m2 or 2500 IU/m2, applied at 3 d intervals, was sufficient to achieve complete L‐asparagine depletion in serum. Changes of the fibrinogen levels was significantly less pronounced in the group on 2500 IU.
Atypical HbH disease in a Surinamese patient resulting from a combination of the −SEA and −α3.7 deletions with HbC heterozygosity
Authors: GIORDANO P. C. , HARTEVELD C. L. , MICHIELS J. J. , TERPSTRA W. , BATELAAN D. , VAN DELFT P. , PLUG R. J. , VAN DER WIELEN M. J. R. , LOSEKOOT M. , BERNINI L. F.
Published: November 2003

The first case of haemoglobin H (HbH) disease in combination with haemoglobin C (HbC) is reported in a man of Surinamese origin. Only haemoglobin A (HbA) and HbC were detected by electrophoresis. The amount of HbC was much less than expected in HbC heterozygotes. The synthesis ratio (βA +βC/α) indicated an α‐thalassaemia defect with two non‐functional α genes, which did not correlate with the degree of haemolysis and anaemia displayed by the patient. The DNA analysis of the α‐genes clusters revealed a defect combination −SEA/−α3.7. The haematological data and the physiopathology of this atypical case are compared with the typical HbH disease found in a first cousin of the propositus. Data on the globin chains expression and on the formation of βA and βC homotetramers in HbH/HbC disease are presented.
Synthetic peptides based on Chlamydia trachomatis antigens identify cytotoxic T lymphocyte responses in subjects from a trachoma-endemic population
Authors: Holland M J , Conway D J , Blanchard T J , Mahdi O M S , Bailey R L , Whittle H C , Mabey D C W
Published: November 2003

CD8+ cytotoxic T lymphocytes (CTL) recognize peptide antigens in the context of class I MHC antigen molecules. To identify peptides capable of eliciting anti-Chlamydia trachomatis CTL responses, 13 synthetic peptides conforming to human leucocyte antigen (HLA)-B8- or -B35-predicted binding motifs were synthesized using sequences based on C. trachomatis major outer membrane protein (MOMP) and heat shock protein 60 (hsp60). Two of 11 HLA-B35-predicted binding peptides were able to stabilize HLA-B35 in an in vitro binding assay. All peptides were tested in CTL assays using peripheral blood mononuclear cells (PBMC) isolated from 26 HLA-B8 or -B35 individuals resident in a trachoma-endemic community. Responses to MOMP and hsp60 peptides were identified in a minority of both HLA-B8 and -B35 individuals. Two of 12 HLA-B8 subjects responded to MOMP and 1/13 to hsp60 peptides. Responses in HLA-B35 subjects were similar, 1/13 subjects responding to MOMP and 2/13 to hsp60 peptides. CTL responses were observed only in children resolving current infection and in adults without scarring of the conjunctiva. These results suggest that anti-chlamydial CTL occur at low levels in peripheral blood, but may be important in the resolution of naturally acquired human ocular chlamydial infection.
Cross-linking of antigen receptor via Ig-β (B29, CD79b) can induce both positive and negative signals in CD40-activated human B cells
Authors: VAN KOOTEN C , GALIBERT L , SEON B K , GARRONE P , LIU Y-J , BANCHEREAU J
Published: November 2003

Antigen-dependent activation of B lymphocytes is mediated through surface immunoglobulins and their associated molecules Ig-α (CD79a, Mb1) and Ig-β (CD79b, B29). Here we show that an antibody directed against the extracellular part of human Ig-β can, when cross-linked by CD32-transfected L cells, induce an IL-2-dependent proliferation of tonsil B cells. With the use of L cells stably transfected with both CD32 and CD40L, anti-Ig-β activation of B cells was combined with CD40 triggering, an important component of the T cell-dependent B cell activation. This dual cellular activation resulted in two different phases, with initially synergistic proliferative effects, both without and with IL-2 or IL-10. Then, after 5–6 days of culture, cells stimulated with both anti-Ig-β and CD40L underwent massive cell death, in contrast to B cells activated with CD40L alone. Cell death was not prevented by the addition of IL-2 or IL-10, but was prevented by the addition of IL-4. These results are discussed in the context of positive and negative selection of mature B cells.
Modulation of Fas-mediated apoptosis of human thyroid epithelial cells by IgG from patients with Graves' disease (GD) and idiopathic myxoedema
Authors: KAWAKAMI A , EGUCHI K , MATSUOKA N , TSUBOI M , URAYAMA S , KAWABE Y , TAHARA K , ISHIKAWA N , ITO K , NAGATAKI S
Published: November 2003

The expression of two autoimmune thyroid diseases, GD and idiopathic myxoedema, is associated with antibodies to the thyroid-stimulating hormone (TSH) receptor. Thyroid stimulating antibodies (TSAb) in GD are TSH agonists and cause hyperthyroidism as well as goitre, whereas thyroid stimulation blocking antibodies (TSBAb) in idiopathic myxoedema are TSH antagonists and cause hypothyroidism and thyroid atrophy. We investigated the effect of antibodies to TSH receptor on Fas-mediated apoptosis of thyroid epithelial cells (thyrocytes). Human IgG was isolated from healthy donors, patients with GD and idiopathic myxoedema. Human thyrocytes were obtained from surgical specimens. Thyrocytes were cultured in the presence or absence of human IgG with or without interferon-gamma (IFN-γ) or IL-1β for a specified time. After incubation, we examined the level of cAMP in cultured supernatants and both Fas and Bcl-2 expression on thyrocytes. In addition, we examined anti-Fas-mediated apoptosis of thyrocytes. Fas expression on thyrocytes was significantly down-regulated by Graves' IgG and TSH, although idiopathic myxoedema IgG did not affect Fas expression on thyrocytes. Idiopathic myxoedema IgG abrogated the effect of TSH on both cAMP production and inhibition of Fas expression on thyrocytes. Treatment of thyrocytes with IL-1β or IFN-γ caused a marked augmentation of Fas expression on thyrocytes. The increase of Fas expression of thyrocytes induced by IL-1β or IFN-γ was significantly suppressed in the presence of TSH or Graves' IgG. Anti-Fas-induced apoptosis of thyrocytes was observed in thyrocytes treated with IL-1β or IFN-γ, but was markedly inhibited in the presence of TSH or Graves' IgG. Furthermore, idiopathic myxoedema IgG abrogated most of the inhibitory effect of TSH on Fas-mediated apoptosis of thyrocytes treated with IL-1β or IFN-γ. Bcl-2 expression of thyrocytes did not change after stimulation with TSH, Graves' IgG, idiopathic myxoedema IgG, IL-1β or IFN-γ. These results suggest that TSAb found in Graves' patients may be potentially involved in the development of goitre by inhibition of Fas-mediated apoptosis of thyrocytes. In addition, TSBAb inhibit the action of TSH and increase the sensitivity toward Fas-mediated apoptosis of thyrocytes, inducing thyroid atrophy seen in patients with idiopathic myxoedema.
Antibodies protect mice against challenge with tick-borne encephalitis virus (TBEV)-infected macrophages
Authors: KREIL T R , BURGER I , BACHMANN M , FRAISS S , EIBL M M
Published: November 2003

TBEV is a flavivirus highly pathogenic for humans. By transfer of antibodies directed to the TBEV surface glycoprotein E into mice, immune protection against subsequent inoculation with free TBEV particles could be achieved. After natural TBEV infection via the skin, however, cells of the monocyte/macrophage lineage were recently demonstrated to represent an important source of local virus replication before viraemia occurs. Whether antibodies can protect against virus challenge when contracted in the form of infected cells, however, is still unclear. In the current study, TBEV antibodies protected mice against challenge with either free virus or TBEV-infected macrophages equally well. This observation may be of more general significance.
N-3 polyunsaturated fatty acids modulate the expression of functionally associated molecules on human monocytes and inhibit antigen presentation in vitro
Authors: HUGHES D A , PINDER A C
Published: November 2003

N-3 polyunsaturated fatty acid (PUFA)-rich diets are associated with suppression of cell-mediated immune responses, but the mechanisms are unclear. Specific immune responses are initiated by antigen-presenting cells (APC). We have previously shown in vitro that the n-3 PUFA, eicosapentaenoic acid (EPA), inhibits the expression of HLA-DR, an MHC class II molecule required for normal APC function on human blood monocytes. In contrast, docosahexaenoic acid (DHA) enhanced the expression of this molecule on unstimulated monocytes, but both n-3 PUFA suppressed its expression on interferon-gamma (IFN-γ)-activated monocytes. In the present study we show that when EPA and DHA were combined at the same ratio as is commonly found in fish oil supplement capsules (3:2) there was no significant effect in vitro on the expression of HLA-DR on unstimulated monocytes, but the expression on IFN-γ-activated monocytes remained significantly inhibited. In the same in vitro system a significant reduction in the ability of IFN-γ-activated monocytes to present tetanus toxoid antigen to autologous lymphocytes was observed following culture with the combined n-3 PUFA. These findings support previous animal studies which suggest that n-3 PUFA can inhibit the antigen-presenting function of mononuclear phagocytes.
Circadian rhythm of leucocytes and lymphocyte subsets and its possible correlation with the function of the autonomic nervous system
Authors: SUZUKI S , TOYABE S , MORODA T , TADA T , TSUKAHARA A , IIAI T , MINAGAWA M , MARUYAMA S , HATAKEYAMA K , ENDOH K , ABO T
Published: November 2003

There are physiological variations in the levels of leucocytes. Among these, the circadian rhythm is very important in terms of the magnitude. Since newly identified lymphocyte subsets (i.e. extrathymic T cells) have recently been detected, a comprehensive study of the circadian rhythm was conducted. All leucocytes were found to vary in number or proportion with a circadian rhythm and were classified into two groups. One group—granulocytes, macrophages, natural killer (NK) cells, extrathymic T cells, γδ T cells, and CD8+ subset—showed an increase in the daytime (i.e. daytime rhythm). The other group—T cells, B cells, αβ T cells, and CD4+ subset—showed an increase at night. Humans are active and show sympathetic nerve dominance in the daytime. Interestingly, granulocytes and lymphocyte subsets with the daytime rhythm were found to carry a high density of adrenergic receptors. On the other hand, lymphocyte subsets with the night rhythm carried a high proportion of cholinergic receptors. Reflecting this situation, exercise prominently increased the number of cells with the daytime rhythm. These results suggest that the levels of leucocytes may be under the regulation of the autonomic nervous system.
Trypanosoma cruzi induces strong IL-12 and IL-18 gene expression in vivo: correlation with interferon-gamma (IFN-γ) production
Authors: MEYER ZUM BÜSCHENFELDE C , CRAMER S , TRUMPFHELLER C , FLEISCHER B , FROSCH S
Published: November 2003

IFN-γ, produced after infection with Trypanosoma cruzi, has been shown to be crucial in the determination of resistance or susceptibility. We have performed a detailed study on the expression of IFN-γ and of the IFN-γ-inducing cytokines IL-12 and IFN-γ-inducing factor (IGIF)/IL-18 with regard to time course and tissue localization. IFN-γ was present in high amounts in the serum and in the supernatants of unseparated spleen cells and isolated CD4+ and CD8+ T cells from the spleens of infected mice which were stimulated ex vivo with T. cruzi. Using the in situ hybridization technique we demonstrate that IL-12 p40 messages were expressed in the spleen and increased during infection, correlating with the expression of IFN-γ transcripts. Furthermore, we show for the first time that the mRNA for the cytokine IL-18 was induced by a parasitic infection and that this expression increased during infection with T. cruzi. Interestingly, the message for IL-18 was produced earlier during infection and already had declined until day 38, when IFN-γ and IL-12 p40 transcripts were optimally expressed. Surprisingly, the changes in IL-12 and IL-18 mRNA production were clearly seen only by in situ hybridization, but less clearly by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). This is possibly due to the extensive activation and proliferation of spleen cells observed during infection leading to a dilution of these specific mRNAs.
Autonomic nervous dysfunction in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA): possible pathogenic role of autoantibodies to autonomic nervous structures
Authors: MAULE S , QUADRI R , MIRANTE D , PELLERITO R A , MARUCCO E , MARINONE C , VERGANI D , CHIANDUSSI L , ZANONE M M
Published: November 2003

Autonomic nervous dysfunction has been previously reported in SLE, RA and systemic sclerosis, but the pathogenesis of such a complication is poorly understood. In the present study, four standard cardiovascular autonomic function tests were performed in 34 female patients with connective tissue diseases and in 25 healthy control subjects, and results expressed as cardiovascular (CV) test scores. Moreover, in each subject the presence of circulating complement-fixing autoantibodies directed against sympathetic and parasympathetic nervous structures, represented by superior cervical ganglia and vagus nerve, respectively, was simultaneously assessed by an indirect immunofluorescent complement-fixation technique, using rabbit tissue as substrate. None of the patients reported autonomic symptoms. However, an abnormal CV test score (≥ 5) was detected in 15% of the patients and in none of the healthy control subjects, approaching statistical significance (P = 0·07). No correlation was found between CV test results and disease duration, type of therapy or presence of conventional autoantibodies. One or two autoantibodies to autonomic nervous structures were detected in six patients (18%) and not in the control subjects (P < 0·05). Values of deep breathing test were significantly lower in autoantibody-positive patients compared with those amongst the control subjects (P < 0·05), and an abnormal CV test score was significantly associated with the presence of autoantibodies to autonomic nervous structures (P < 0·05). In conclusion, we confirm that autonomic nervous function can be impaired in patients with connective tissue diseases, and suggest that autoantibodies directed against autonomic nervous system structures may play a role in the pathogenesis of the autonomic dysfunction.