bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhancement of human platelet activation by the combination of low concentrations of collagen and rabbit anticardiolipin antibodies
Authors: Wang Lai , Su Chen‐Yi , Chou Kuang‐Yu , Wang Cheng‐Teh
Published: March 2003

Summary. Low concentrations of collagen and anticardiolipin antibodies (ACLA), which were raised in rabbits by immunization with cardiolipin (CL), co‐operatively activated human gel‐filtrated platelets (GFP). GFP activated by adding ACLA 5 min prior to collagen (ACLA + Col) showed strong responses in cytosolic Ca2+ mobilization and cell aggregation; the responses decreased after 1 min, however, when collagen was added prior to ACLA (Col + ACLA). Col + ACLA was 30% less effective than the ACLA + Col in: (1) the phosphorylation of pleckstrin and myosin light chain; and (2) the secretion of α‐ and dense granules. Indomethacin inhibited Ca2+ mobilization, pleckstrin phosphorylation and cell aggregation in platelets stimulated by ACLA + Col. The thromboxane B2 level in platelets induced by ACLA + Col was similar to that stimulated by low concentrations of collagen alone. ACLA + Col increased the activities of phospholipase C (PLC) as determined by formation of phosphatidic acid (PA), whereas indomethacin and adenosine 2′,5′‐diphosphate, an antagonist of the ADP P2Y1 receptor, inhibited PA formation. These results suggest that ACLA, thromboxane A2 derived from the collagen pathway and secreted ADP co‐operatively augment PLC activity and lead to platelet aggregation.
Superior effect of 9‐cis retinoic acid (RA) compared with all‐trans RA and 13‐cis RA on the inhibition of clonogenic cellgrowth and the induction of apoptosis in OCI/AML‐2 subclones: is the p53 pathway involved?
Authors: Koistinen Pirjo , Zheng Aiping , Säily Marjaana , Siitonen Timo , Mäntymaa Pentti , Savolainen Eeva‐Riitta
Published: March 2003

Summary. In the present study, the effects of 9‐cis retinoic acid (RA) and 13‐cis RA on acute myeloblastic leukaemia (AML) cell growth and the induction of apoptosis as well as its relationship with bcl‐2 and p53 were compared with those of all‐trans RA (ATRA). The study was performed with the subclones of the retinoid‐sensitive OCI/AML‐2 cell line. The most prominent inhibitory effect on clonogenic cell growth and morphological apoptosis was shown by 9‐cis RA. In addition, Western blotting revealed the most obvious translocation of p53 from cytosol to nucleus in the case of 9‐cis RA, which was the only retinoid able to change the conformation of p53 from mutational to wild type, as demonstrated by flow cytometry. There was no difference between the retinoids in the downregulation of bcl‐2 as analysed by Western blotting and flow cytometry. The RA receptor (RAR)‐α antagonist had no effect on apoptosis in any of the three retinoids studied using the annexin V method. In conclusion, this study shows that 9‐cis RA was a more potent agent than ATRA or 13‐cis RA in inducing growth arrest and apoptosis in the OCI/AML‐2 subclones. The effect was associated with the downregulation of bcl‐2 and was hardly mediated through the RAR‐α receptor, but might be related to the activation of p53.
Many αIIbβ3 autoepitopes in chronic immune thrombocytopenic purpura are localized to αIIb between amino acids L1 and Q459
Authors: Mcmillan Robert , Wang Lei , Lopez‐Dee Jennifer , Jiu Sandra , Loftus Joseph C.
Published: March 2003

Summary. In chronic immune thrombocytopenic purpura (ITP), autoantibodies bind to platelet surface proteins, particularly αIIb, resulting in platelet destruction by the reticulo‐endothelial system. In order to better localize the autoepitopes on αIIb, we studied the binding of antibodies to Chinese hamster ovary (CHO) cells expressing either αIIbβ3 or αIIb‐αvβ3 chimaeras in which a segment of αIIb (either amino acids L1‐Q459, L1‐F223 or F223‐Q459) was substituted for that portion of αv. We evaluated platelet‐associated autoantibodies from 14 ITP patients with αIIb‐dependent antibodies. Ten of 14 bound to αIIb (L1‐Q459)‐αvβ3, showing that autoepitopes were often localized to this region of αIIb. In addition, each of the autoantibodies binding to αIIb (L1‐Q459)‐αvβ3, also bound to CHO cells expressing either αIIb(L1‐F223)‐αvβ3 or αIIb(F223‐Q459)‐αvβ3). In two of the three eluates tested, > 95% of the autoantibody binding to αIIb could be adsorbed using CHO cells expressing any of the three chimaeras, showing that the epitope(s) have contact points on either side of amino acid F223; in the third eluate, only a portion (∼40%) could be adsorbed by the chimaeric cell lines showing that, in this patient, an additional antibody was also present, directed to a site distal to amino acid Q459. The remaining four eluates bound to CHO cells expressing αIIbβ3 but to none of the chimaeras, suggesting that these epitopes are also distal to amino acid Q459. We conclude that the binding of many anti‐αIIbβ3 autoantibodies is dependent on the presence of αIIb amino acids L1‐Q459.
Acquired trisomy 21 and distinct clonal evolution in acute megakaryoblastic leukaemia in young monozygotic twins
Authors: Stark Batia , Jeison Marta , Preudhomme Claude , Fenaux Pierre , Ash Shifra , Korek Yifat , Stein Jerry , Zaizov Rina , Yaniv Issac
Published: March 2003

Summary. An intrauterine origin of childhood acute lymphoblastic leukaemia (ALL) was proven by the identical clonotypic gene rearrangement in the concordant leukaemias of monozygotic twins, arising from a single clonogenic progeny. The monozygotic twins, presented at the age of 22 months with acute megakaryoblastic leukaemia (AML‐M7) in one and myelodysplasia transformed to AML‐M7 in the other. Leukaemic cells in both twins carried trisomy 21 and additional different clonal evolution changes of del(20q) in the first twin and trisomy 8 in the second. AML‐M7 of late infancy with trisomy 21 may be included in the leukaemias of intrauterine origin, possibly a result of genotoxic insult.
Cytotoxic chemotherapy following tandem autotransplants in multiple myeloma patients
Authors: Fassas Athanasios B.‐T. , Spencer Trey , Desikan Raman , Zangari Maurizio , Anaissie Elias , Barlogie Bart , Tricot Guido
Published: March 2003

Summary. High‐dose treatment (HDT) with autologous stem cell transplant (ASCT) is superior to conventional chemotherapy in multiple myeloma. However, relapses eventually occur, especially in the presence of unfavourable cytogenetic abnormalities, high β‐2 microglobulin levels prior to transplant and extensive prior treatment. Cytotoxic consolidation chemotherapy, following tandem transplants (TT), was given to 75 myeloma patients with at least one poor prognostic factor. When their outcome was compared with that of 75 matched controls who received dexamethasone ± interferon post TT, no event‐free or overall survival advantage was observed. Other approaches may be required to improve survival in multiple myeloma.
Influence of the different CD34+ and CD34– cell subsets infused on clinical outcome after non‐myeloablative allogeneic peripheral blood transplantation from human leucocyte antigen‐identical sibling donors
Authors: Menéndez Pablo , Pérez‐Simón Jose A. , Mateos Maria V. , Caballero Maria D. , González Marcos , San‐Miguel Jesus F. , Orfao Alberto
Published: March 2003

Summary.  Currently, no information is available regarding the influence of the different CD34+ cell subsets infused on the haematopoietic recovery, following non‐myeloablative allogeneic peripheral blood stem cell transplantation (allo‐PBSCT). We have explored, in a group of 13 patients receiving non‐myeloablative allo‐PBSCT from human leucocyte antigen‐identical sibling donors, the influence of the total dose of CD34+ haematopoietic progenitor cells (HPC) infused, compared with that of the different CD34+ HPC and CD34– leucocyte subsets in the leukapheresis samples, on both engraftment and clinical outcome. The overall numbers of total CD34+ HPC (P = 0·002) and myelomonocytic‐committed CD34+ HPC infused (P = 0·0002) were strongly associated with neutrophil recovery (> 1 × 109 neutrophils/l), the latter being the only independent parameter influencing neutrophil recovery. Regarding long‐term engraftment, only the number of immature CD34+ HPC infused/kg correlated with the duration of hospitalization in the first 2 years after discharge (r = −0·75, P = 0·005). Both the overall amount of CD34+ HPC and the number of myelomonocytic CD34+ HPC infused showed a significant influence on the risk of graft‐versus‐host disease (GVHD). Thus, the overall probability of GVHD was 100%vs 25% for patients receiving ≥ 5 × 106 CD34+ HPC or ≥ 3·5 × 106 of myelomonocytic‐committed CD34+ HPC vs lower doses (P = 0·013). None of the other CD34+ and CD34– cell subsets analysed correlated with development of GVHD. In summary, our results suggest that in non‐myeloablative allo‐PBSCT, high numbers of CD34+ HPC, especially the myelomonocytic‐committed CD34+ progenitors, lead to rapid neutrophil engraftment. However, they also strongly impair clinical outcome by increasing the incidence of GVHD.
H2‐mismatched transplantation with repetitive cell infusions and CD40 ligand antibody infusions without myeloablation
Authors: Lambert Jean‐François , Colvin Gerald A. , Zhong Suju , Wang Han , D'Hondt Lionel , Abedi Mehrdad , Frimberger Angela E. , Stewart F. Marc , Quesenberry Peter J.
Published: March 2003

Summary. Graft rejection and graft‐versus‐host disease are major problems in mismatched marrow transplants along with toxicity from standard myeloablative host treatments. We have established a tolerization model, using 1 Gy irradiation, which reduces stem cell capacity to < 10% of control while causing minimal myelosuppression, donor antigen pre‐exposure (spleen cells), CD40‐ligand antibody blockade and high levels of marrow (40 × 106 cells), which allows for stable long‐term multilineage engraftment in H2‐mismatched murine marrow transplants. We now show that the establishment of ‘microchimaerism’ (0·5–3·8%) sets the stage for macrochimaerism, with subsequent marrow infusions in H2‐mismatched mice with CD40‐ligand blockade only. Neither irradiation nor spleen cell exposure were necessary. When 40 x 106 bone marrow cells were infused on weeks 0, 12, 14 and 16, blood engraftment was about seven times the single 40 × 106 control. When marrow cells were given on weeks 0, 3, 4, 5 and 6, engraftment at 24 weeks post transplant was 17·9 ± 1·2%, compared with 2·7 ± 0·8% for the single 40 × 106 control (P = 0·009). We have shown stable, long‐term multilineage chimaerism and established that the schedule of marrow administration, not the total cell dose, is critical for tolerization. This approach indicates that microchimaerism can tolerize for subsequent marrow infusions and produce macrochimaerism. This strategy could be applied in clinical human transplants.
Molecular and flow cytometric characterization of the CD4 and CD8 T‐cell repertoire in patients with myelodysplastic syndrome
Authors: Melenhorst J. Joseph , Eniafe Rhoda , Follmann Dean , Nakamura Ryo , Kirby Martha , Barrett A. John
Published: March 2003

Summary. We studied 18 patients with myelodysplastic syndrome (MDS), measuring clonality and T‐cell receptor Vbeta (TCRBV) expression of CD4 and CD8 T cells by polymerase chain reaction and by flow cytometric analysis of TCRBV families. The CD4 and CD8 T‐cell repertoire in most MDS patients is characterized by an abnormal TCRBV‐restricted expansion of T cells in CD4 and CD8 cells, and increased expression of the CD8 effector marker CD57 of multiple TCRBV in CD8 cells. Clonality analysis of CD4 and CD8 cells showed that seven of 10 patients analysed had a major clone in the CD8 cells but not in CD4 cells. Furthermore, in one patient we found that both the CD57– and CD57+ fraction contained the clone (which was absent from the TCRBV‐negative fraction). These data suggest that, in MDS, multiple T‐cell expansions can be found in both helper and cytotoxic T cells, and that, in the CD8 cells, T cells functionally differentiate in vivo from memory to effector T cells. Together, these data support the hypothesis of the involvement of T cells in the pathogenesis of MDS.
Developmental changes in the expression of transcription factors GATA‐1, ‐2 and ‐3 during the onset of human medullary haematopoiesis
Authors: Dame Christof , Sola Martha C. , Fandrey Joachim , Rimsza Lisa M. , Freitag Patricia , Knöpfle Gisela , Christensen Robert D. , Bungert Jörg
Published: March 2003

Summary. Regulation of gene expression during the ontogeny of haematopoiesis in the human fetal bone marrow is poorly understood. Studies in mice demonstrated that GATA‐1, ‐2 and ‐3 play pivotal roles in haematopoiesis. In this study, we identified GATA‐1‐, GATA‐2‐ and GATA‐3‐expressing cells in bone marrow sections and analysed the expression of GATA‐transcription factors during the development of human fetal bone marrow haematopoiesis using semiquantitative reverse transcription‐polymerase chain reaction (RT‐PCR). We showed that GATA‐1, ‐2 and ‐3 were expressed only in haematopoietic cells in the bone marrow. RT‐PCR analysis demonstrated that (1) GATA‐1 expression significantly increased during gestation; (2) GATA‐2 expression peaked at the onset of medullary haematopoiesis, declined thereafter, and remained at a constant level after 30 weeks post conception; and (3) GATA‐3 expression revealed no changes during development. The results indicated that the onset of medullary haematopoiesis in humans is accompanied by high expression of GATA‐2, reflecting high proliferation rates of early haematopoietic progenitor cells, whereas expression of GATA‐1 mirrors haematopoietic activity.
A novel DKC1 mutation, severe combined immunodeficiency (T+B–NK– SCID) and bone marrow transplantation in an infant with Hoyeraal–Hreidarsson syndrome
Authors: Cossu Fausto , Vulliamy Tom J. , Marrone Anna , Badiali Manuela , Cao Antonio , Dokal Inderjeet
Published: March 2003

Summary.  X‐linked Hoyeraal–Hreidarsson syndrome (XL‐HHS) is the severe infantile variant of X‐linked dyskeratosis congenita (XL‐DC) and both are due to mutations in the DKC1 gene within Xq28. We report a novel missense mutation in DKC1 exon 3 (T113→C, Ile38Thr) in a Sardinian infant with XL‐HHS in whom the disease was characterized by ‘T+B–NK–’ severe combined immunodeficiency and bone marrow failure. He underwent sibling bone marrow transplantation using a conditioning regimen (fludarabine, rabbit antithymocyte globulin, low‐dose melphalan) selected according to the HHS/DC phenotype. This was associated with low toxicity, prompt engraftment with adequate immune reconstitution and full donor haemopoiesis.
Chronic leg ulceration in homozygous sickle cell disease: the role of venous incompetence
Authors: Clare Andrea , FitzHenley Michael , Harris June , Hambleton Ian , Serjeant Graham R
Published: March 2003

Summary. Chronic leg ulceration is a common cause of morbidity in Jamaican patients with homozygous sickle cell (SS) disease. Ulcers heal more rapidly on bed rest and deteriorate on prolonged standing, suggesting a role of venous hypertension in their persistence. This hypothesis has been tested by Doppler detection of venous competence in SS patients and in matched controls with a normal haemoglobin (AA) genotype in the Jamaican Cohort Study. Venous incompetence was significantly more frequent in SS disease [137/183 (75%)] than in non‐pregnant AA controls [53/137 (39%)]. Past or present ulceration occurred in 78 (43%) SS patients, with a highly significant association between leg ulceration and venous incompetence in the same leg (P < 0·001). Prominence and/or varicosities of the veins and spontaneous leg ulcers were more common among patients with multiple sites of incompetence. The association of venous incompetence with chronic leg ulceration identifies a further pathological mechanism contributing to the morbidity of SS disease. The cause of venous incompetence is unknown but the sluggish circulation associated with dependency, turbidity and impaired linear flow at venous valves, hypoxia‐induced sickling, the rheological effects of high white cell counts, and activation of components of the coagulation system may all contribute. Venous hypertension in SS patients with leg ulceration suggests that firm elastic supportive dressings might promote healing of chronic leg ulcers.
Antibodies to factor VIIa in patients with haemophilia and high‐responding inhibitors
Authors: Astermark Jan , Ekman Maj , Berntorp Erik
Published: March 2003

Summary. The haemostatic effect of by‐passing agents such as activated prothrombin complex concentrates (aPCC) and recombinant factor VIIa is inadequate in some patients with severe haemophilia and high‐responding inhibitors. Theoretically, this could be due to antibody reactivity to procoagulant proteins other than the deficient factor. To evaluate this hypothesis, immunoglobulin (Ig) fractions from six multi‐transfused patients (three haemophilia A and three haemophilia B) were purified on protein A sepharose and then subjected to immunoaffinity chromatography on factor IX sepharose and factor VIIa sepharose. All three Ig fractions from the haemophilia B patients, but not commercially available Ig, contained antibodies that bound to both gels. None of the haemophilia A patients had antibodies to factor IX but all three had antibodies towards factor VIIa. The immunoaffinity purified antifactor IX and VIIa antibodies from the haemophilia B patients inhibited thrombin formation in vitro using Feiba® as active enzyme, but had no significant effect in the presence of NovoSeven®. In contrast, no inhibitory effect was observed with the antifactor VIIa antibodies from the haemophilia A patients. Cross‐reactivity to factor IX was seen for the antifactor VIIa antibodies from the patients with haemophilia B. Our findings show that antibody reactivity to other procoagulant factors such as factor VIIa exists in patients with high‐responding inhibitors and that these antibodies may have an inhibitory potential that correlates to the amount of active enzyme present. The characterization of the antibody profile may facilitate an optimal treatment with by‐passing agents in severe bleeding events.
Comparison of a specialist haematological malignancy database against a regional cancer registry: case ascertainment and diagnostic accuracy
Authors: Phekoo Karen , Møller Henrik , Richards Mike , Schey Steve
Published: March 2003

Summary. No large‐scale study has been performed to assess the problem of registering all subtypes of haematological malignancies. We compared registration of haematological malignancies between 1994 and 1996 by haematologists in 14 National Health Service Trusts in the Eastern part of the South Thames Region with data for the same area recorded by the Thames Cancer Registry (TCR). Case ascertainment and diagnostic accuracy were the two main outcome measures. A combined total of 4714 haematological malignancies were recorded over the 3‐year period. Of these, 1329 (28%) were common to both databases, 1975 (42%) were recorded only by the TCR and 1410 (30%) were recorded only by the haematologists. Nearly one‐third (31%) of all cases recorded by the TCR were death certificate‐only registrations. The TCR records were obtained from 30 clinical specialities. Haematology only accounted for 35% of these cases. Discordant diagnoses were recorded in 20% of the cases that were recorded in both databases. Our data suggests that both registers have deficiencies in collecting and validating data on the incidence of haematological malignancies. To address this, a partnership was established in 1998 between haematologists in the South Thames Region and the Thames Cancer Registry. It is anticipated that engaging clinicians in the collection and validation of data will enhance the completeness of case ascertainment and improve the quality of data on haematological malignancies.
Analysis of factor VIII inhibitors in a haemophilia A patient with an Arg593→Cys mutation using phage display
Authors: Bril Wendy S. , Turenhout Ellen A. M. , Kaijen Paul H. P. , Van Den Brink Edward N. , Koopman Maria M. W. , Peters Marjolein , Voorberg Jan
Published: March 2003

Summary. We characterized anti‐factor VIII antibodies in a mild haemophilia A patient with an Arg593→Cys mutation in the A2 domain, using V gene phage‐display technology. All isolated single‐chain variable‐domain antibody fragments were directed against residues Arg484‐Ile508, a binding site for factor VIII inhibitors in the A2 domain. After a further period of replacement therapy, a transient rise in inhibitor titre was observed. These antibodies were directed against the A2 domain. Activation of a pre‐existing pool of B cells, which express antibodies against residues Arg484‐Ile508, could explain the rapid anamnestic response.
Long‐term ex vivo expansion of human fetal liver primitive haematopoietic progenitor cells in stroma‐free cultures
Authors: Peters Rowayda , Leyvraz Serge , Faes‐van't Hull Eveline , Jaunin Philippe , Gerber Stefan , Rollini Pierre
Published: March 2003

Summary. Successful expansion of haematopoietic cells in ex vivo cultures will have important applications in transplantation, gene therapy, immunotherapy and potentially also in the production of non‐haematopoietic cell types. Haematopoietic stem cells (HSC), with their capacity to both self‐renew and differentiate into all blood lineages, represent the ideal target for expansion protocols. However, human HSC are rare, poorly characterized phenotypically and genotypically, and difficult to test functionally. Defining optimal culture parameters for ex vivo expansion has been a major challenge. We devised a simple and reproducible stroma‐free liquid culture system enabling long‐term expansion of putative haematopoietic progenitors contained within frozen human fetal liver (FL) crude cell suspensions. Starting from a small number of total nucleated cells, a massive haematopoietic cell expansion, reaching > 1013‐fold the input cell number after ∼300 d of culture, was consistently achieved. Cells with a primitive phenotype were present throughout the culture and also underwent a continuous expansion. Moreover, the capacity for multilineage lymphomyeloid differentiation, as well as the recloning capacity of primitive myeloid progenitors, was maintained in culture. With its better proliferative potential as compared with adult sources, FL represents a promising alternative source of HSC and the culture system described here should be useful for clinical applications.
Plasma matrix metalloproteinase and tissue inhibitor of metalloproteinase in patients with agnogenic myeloid metaplasia or idiopathic primary myelofibrosis
Authors: Wang Jen C. , Novetsky Akiva , Chen Chi , Novetsky Allan D.
Published: March 2003

Summary. Agnogenic myeloid metaplasia (AMM) is characterized by bone marrow fibrosis with abnormal accumulation of extracellular matrix components (ECM), which is dependent on the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Twenty‐five patients with AMM, 30 with essential thrombocythemia (ET), 12 with polycythemia vera (PV) and 20 normal control subjects were studied. AMM patients had decreased plasma levels of MMP‐3 and marked elevated levels of TIMP‐1, but MMP‐1, MMP‐2 and MMP‐9 levels were not significantly different from control subjects. Elevated levels of plasma TIMP‐1, but not MMPs, were found in ET and PV. Reduced MMP activity together with increased TIMP‐1 activity may be essential in fibrosis formation.
Intraclonal diversity in a Sezary syndrome with a differential response to 2‐deoxycoformycin of the two lymphoma cell populations
Authors: Granjo Elisa , Lima Margarida , Lopes José Manuel , Cunha Nória , Teixeira Maria Dos Anjos , Santos Filipe , Candeias Jorge , Resende Carlos , Santos Ana Helena , Balanzategui Ana , Orfão Alberto , Matutes Estella
Published: March 2003

Summary. We report a case of Sezary syndrome with two abnormal CD4+ T‐cell populations detected in the peripheral blood by flow cytometry immunophenotyping and DNA cell content, suggesting a biclonal T‐cell lymphoproliferative disorder. Despite these findings, molecular analysis of the T‐cell receptor genes was consistent with a monoclonal T‐cell proliferation, supporting the existence of intraclonal diversity rather than a true biclonal disease. The patient achieved a transient response with 2‐deoxycoformycin, with a selective decrease of the larger/hyperploid T‐cell population; later on, an increased representation of this T‐cell population was observed concomitantly with clinical relapse.
Engraftment of early erythroid progenitors is not delayed after non‐myeloablative major ABO‐incompatible haematopoietic stem cell transplantation
Authors: Maciej Zaucha J. , Mielcarek Marco , Takatu Alessandra , Little Marie‐Terese , Gooley Theodore , Baker Jennifer , Maloney David G. , Sandmaier Brenda M. , Maris Michael , Chauncey Thomas , Storb Rainer , Torok‐Storb Beverly
Published: March 2003

Summary. We hypothesized that patients undergoing major ABO‐incompatible non‐myeloablative haematopoietic stem cell transplantation (nm‐HSCT) might experience prolonged haemolysis after transplant due to the delayed disappearance of host plasma cells producing anti‐donor isohaemagglutinins (HAs). To address this question, we analysed data from 107 consecutive patients transplanted with allogeneic peripheral blood stem cells from human leucocyte antigen‐matched (related, n = 84; unrelated, n = 23) donors after non‐myeloablative conditioning (200 cGy total body irradiation ± fludarabine). In total, 23 out of the 107 patients received major or major/minor ABO‐incompatible transplants. Red blood cell (RBC) transfusion requirements during the first 120 d post transplant were higher in major ABO‐mismatched than in ABO‐matched recipients (0·12 vs 0·03 median units RBC concentrate/d, P = 0·04). Two patients developed transient pure red cell aplasia, which had resolved spontaneously by 9 months after transplant. Major ABO incompatibility did not influence rates of engraftment. Patients with sustained engraftment experienced gradual declines of anti‐donor HAs, and the estimated median time to reaching IgM and IgG titres of < 1:1 was at least 133 d in evaluable patients, approximately twice longer than reported after myeloablative conditioning. There was a strong correlation between degrees of donor chimaerism in erythroid burst‐forming units, granulocyte macrophage colony‐forming units and granulocytes, indicating that donor erythroid engraftment, defined by early erythroid progenitors, was as prompt as myeloid engraftment. In conclusion, our data suggest that major ABO‐incompatibility is not a barrier to successful non‐myeloablative HSCT.
A patient with paroxysmal nocturnal haemoglobinuria in whom granulocyte colony‐stimulating factor administration resulted in improvement of recurrent enterocolitis and its associated haemolytic attacks
Authors: Fujimi Akihito , Matsunaga Takuya , Kogawa Katsuhisa , Ohnuma Takayuki , Takahira Naoki , Abe Tomoyuki , Kitaoka Keisuke , Kogawa Takahiro , Tanaka Ikuta , Morii Kazuhiro , Terui Takeshi , Sakamaki Sumio , Kato Junji , Kura Toshiro , Maeda Takeshi , Niitsu Yoshiro
Published: March 2003

Summary. We report an elderly patient with paroxysmal nocturnal haemoglobinuria (PNH), having recurrent enterocolitis and haemolytic attacks associated with cellular immunodeficiency. On admission, the patient had normal neutrophil count and function but a decreased T‐cell count, decreased mitogenic reactions, and a negative tuberculin test. Granulocyte colony‐stimulating factor (G‐CSF) was administered, resulting in an increased T‐cell count, normalization of T‐cell function, increased blood levels of helper T cell (Th)1 and Th2 cytokines and improvement in the enterocolitis and haemolytic attacks. This suggests that G‐CSF may be useful in the treatment of elderly PNH patients with cellular immunodeficiency.
Expression of stanniocalcin‐1 in megakaryocytes and platelets
Authors: Serlachius Martina , Alitalo Riitta , Olsen Henrik S. , Andersson Leif C.
Published: March 2003

Summary. Stanniocalcin‐1 (STC) is a 56‐kDa homodimeric glycoprotein hormone originally found in fish, in which it regulates calcium/phosphate homeostasis and protects against toxic hypercalcaemia. The recently characterized human STC is 80% similar to fish STC. We have earlier reported a high expression of STC in terminally differentiated human and rodent brain neurones, and found that STC contributes to the maintenance of their integrity. Here, we report that mature megakaryocytes and platelets display high STC content. K562 cells, induced to megakaryocytoid differentiation in vitro, acquired expression of STC, which was not seen in untreated K562 cells or cells induced to erythroid differentiation.
A study on 289 consecutive Korean patients with acute leukaemias revealed fluorescence in situ hybridization detects the MLL translocation without cytogenetic evidence both initially and during follow‐up
Authors: Kim Hee Jin , Cho Han Ik , Kim Eui Chong , Ko Eun Kyong , See Cha Ja , Park Seon Yang , Lee Dong Soon
Published: March 2003

Summary. Translocations involving the MLL gene on the chromosome 11 (11q23) are frequently observed in acute leukaemia. The detection of this genetic change has a unique significance as a result of its implication of poor prognosis. To reveal the utility of fluorescence in situ hybridization (FISH) in detecting the MLL translocation, we analysed 289 consecutive Korean patients (children and adults) with acute leukaemias using both conventional cytogenetic analysis (CC) and FISH, placing an emphasis on the result discrepancies. Twenty‐two of 289 patients (7·6%) had the 11q23/MLL translocation. In nine of 22 patients (41%), only FISH detected the translocation. In eight of these 22 patients, a total of 19 follow‐up examinations were performed, of which FISH detected a significant level of leukaemic cells harbouring the MLL translocation in five patients (26%) without cytogenetic evidence. In addition to the MLL translocation, FISH detected submicroscopic amplification, partial deletion of the MLL gene and trisomy 11 in 12 patients without cytogenetic evidence. In summary, up to 41% of the MLL translocations at initial work‐up and 26% during follow‐up were detected by FISH without cytogenetic evidence. Thus, we recommend that MLL FISH should be performed in the diagnosis and monitoring of acute leukaemias in combination with CC.
Activation of integrin αIIbβ3 expressed in Chinese hamster ovary cells is required for interaction with solid‐phase von Willebrand factor
Authors: Mekrache Médina , Legendre Paulette , Kieffer Nelly , Baruch Dominique
Published: March 2003

Summary. Dithiothreitol (DTT) is known to induce an active conformation of αIIbβ3 integrin and to promote the aggregation of Chinese hamster ovary (CHO)‐αIIbβ3 cells in the presence of soluble fibrinogen (Fg). The aim of this study was to compare adhesion and spreading with Fg or von Willebrand factor (VWF) of CHO‐αIIbβ3 cells in the presence or absence of DTT. Our results indicate that DTT treatment was required to induce cell spreading on VWF. In contrast, CHO‐αIIbβ3 cell spreading on Fg was already optimal in the absence of DTT. We used a small perfusion chamber coupled to videomicroscopy to demonstrate that CHO‐αIIbβ3 cells that were adherent and spread on VWF required DTT activation to resist to detachment under increasing shear rates (50–1600/s). In contrast, untreated or DTT‐treated cells spread on Fg were able to resist to extremely high flow rates. These data provide novel evidence that activated αIIbβ3 is absolutely required for spread cells to resist detachment and strengthens the importance of the αIIbβ3 activation step for adhesion and spreading to VWF.
Human bone marrow angiogenesis: in vitro modulation by substance P and neurokinin A
Authors: Pelletier Laurent , Angonin Regis , Regnard Jacques , Fellmann Dominique , Charbord Pierre
Published: March 2003

Summary. We have previously described a culture system for human bone marrow endothelial cells that organize into capillary tubes associated to pericytes. In the present work, we used this model to assess the angiogenic properties of tachykinins, which have been demonstrated to be involved in neuro–immuno–haematopoietic interactions. The substance P (SP) and neurokinin A (NKA) were similarly potent at increasing in vitro angiogenesis, via NK1 and NK2 receptors respectively. These mediators were not produced by cells in culture, suggesting that in vivo they may be released by nerve fibres in the bone marrow. Therefore, we looked for in situ innervation of the human bone marrow, unknown to date, using immunohistochemistry techniques. As in rodents, arterioles were largely innervated, associated with between one and 10 nerve fibres. Capillary innervation was more restrictive as a unique thin nerve fibre was found in the vicinity of only 6% of these vessels. Finally, no nerve fibres were observed in the vicinity of sinus walls. In conclusion, both in vitro results and the anatomical display of nerve fibres suggest a role in human bone marrow for the vasoactive neuropeptides SP and NKA, which were secreted into a perivascular location. These neural mediators might modulate blood flow in the bone marrow both in the short term by adjusting vascular tone and in the long term by inducing angiogenesis.
A 3′UTR mutation affects β‐globin expression without altering the stability of its fully processed mRNA
Authors: Bilenoglu Onur , Basak A. Nazli , Russell J. Eric
Published: March 2003

Summary. Determinants of mRNA stability are frequently positioned in the 3′UTR where they are not subject to disruption by actively translating ribosomes. Two related individuals with β thalassaemia who carry a β‐globin gene containing a 13 nt deletion in its 3′UTR have recently been described. Its position within the 3′UTR, as well as its relative distance from other known functionally important elements, suggested that the deletion might overlay previously unrecognized determinants of β‐globin mRNA stability. We studied the impact of the Δ13 mutation on β‐globin gene expression in vitro and in vivo. The adverse effect of the Δ13 mutation on β‐globin expression was confirmed in studies utilizing reticulocytes from a βΔ13 heterozygote, which indicated a sixfold reduction in the relative level of the mutant mRNA. Additional in vitro analysis indicated that the deletion did not affect the capacity of the βΔ13 mRNA to assemble an mRNA‐stabilizing mRNP ‘β‐complex’. Unexpectedly, functional tests in both primary erythroid cells and in a transgenic mouse model demonstrated that the βΔ13 mRNA was fully stable, suggesting that the Δ13 mutation affects accumulation of the fully processed mRNA at an earlier step. Consistent with this, there was a relative excess of unprocessed βΔ13 mRNA in erythroid progenitors from a βΔ13 heterozygote. Taken together, these results define a new thalassaemic determinant, which acts to decrease β‐globin mRNA levels by inhibiting the efficiency of nuclear processing events, and suggest a previously unanticipated complexity to the role of the 3′UTR elements in the regulation of β‐globin gene expression.
Thrombus precursor protein, endogenous thrombin potential, von‐Willebrand factor and activated factor VII in suspected deep vein thrombosis: is there a place for new parameters?
Authors: Bucek Robert A. , Reiter Markus , Quehenberger Peter , Weltermann Ansgar , Kyrle Paul A. , Minar Erich
Published: March 2003

Summary. Because of its high negative predictive value, D‐Dimer is an important parameter in the exclusion of deep vein thrombosis (DVT) but it produces a high number of false‐positive results. We therefore evaluated different blood parameters in 74 consecutive patients with suspected DVT, whose final diagnosis was based on the results of Duplex ultrasound or venography. DVT was diagnosed in 52·7%. While D‐Dimer, thrombin–antithrombin complexes, prothrombin fragment 1+2, von‐Willebrand factor and thrombus precursor protein were significantly increased in patients with DVT, there was no influence concerning endogenous thrombin potential and activated factor VII. There was no significant correlation between the thrombus extension or the duration of symptoms with any of these parameters. D‐Dimer showed the best sensitivity (94·9%) to specificity (45·7%) ratio and neither the sole nor the additional evaluation of any other investigated parameter increased its diagnostic performance. We, therefore, conclude that the determination of D‐Dimer remains the ‘gold standard’ in the laboratory testing of patients with suspected DVT.
Common expression of an unusual CD45 isoform on T cells from patients with large granular lymphocyte leukaemiaand autoimmune lymphoproliferative syndrome
Authors: Bleesing Jack J. H. , Janik John E. , Fleisher Thomas A.
Published: March 2003

Summary. Patients with T‐cell large granular lymphocyte (T‐LGL) leukaemia and autoimmune lymphoproliferative syndrome (ALPS) share many features, including autoimmunity and an expansion of (cytotoxic) T cells, which in ALPS patients express an unusual (B220) isoform of CD45, corresponding to an altered O‐glycosylation profile. Here we showed that T‐LGL leukaemia cells also expressed this B220 isoform. We hypothesize that B220+ T cells constitute proliferating T cells that have become competent to undergo apoptosis, but that constitutive (ALPS) or functional (T‐LGL) defects prevent this process. Altered O‐glycosylation of the extracellular domains of CD45 may have consequences for this tyrosine phosphatase as a regulator of cell proliferation and survival.
Follicular lymphoma lacking the t(14;18)(q32;q21): identification of two disease subtypes
Authors: Horsman Douglas E. , Okamoto Ichiro , Ludkovski Olga , Le Nhu , Harder Lana , Gesk Stefan , Siebert Reiner , Chhanabhai Mukesh , Sehn Laurie , Connors Joseph M. , Gascoyne Randy D.
Published: March 2003

Summary. The clinical and pathological features, including karyotype data and BCL2 protein expression pattern, of follicular lymphoma without a t(14;18)(q32;q21) have not been well defined. We have identified and conducted a detailed analysis of 50 cases with follicular lymphoma who lack the t(14;18). Fluorescent in situ hybridization (FISH) analysis was used to exclude cases with a cryptic IGH/BCL2 rearrangement. BCL2 protein expression level was assessed by immunohistochemistry. The karyotypes were assessed for recurrent sites of structural rearrangement, duplications and deletions on a band‐by‐band basis, and compared with a large cohort of cases with t(14;18). A distinct pattern of chromosomal alterations was identified in the cases without t(14;18). BCL2 protein overexpression was detected in 33% of 49 tested cases. In this minority, the karyotypes frequently showed increased copies of chromosome 18. The majority of cases (67%) did not show BCL2 overexpression and were characterized prominently by the presence of t(3;14)(q27;q32), implying a role for BCL6. Follicular lymphomas that lack a t(14;18) were segregated into two subgroups with distinct cytogenetic, phenotypic and possibly clinical features: one with BCL2 protein overexpression not related to an IGH/BCL2 rearrangement and a second without BCL2 overexpression. Objective identification of BCL2 expression level as well as BCL2 and BCL6 status by cytogenetic or FISH analysis has potential clinical utility and may yield insights into alternative genetic mechanisms associated with B‐cell lymphomas with a follicular growth pattern.
Expansion of CD8+ T cells that express low levels of the B cell‐specific molecule CD20 in patients with multiple myeloma
Authors: Katopodis Ourania , Liossis Stamatis‐Nick , Viglis Vassilios , Pouli Anastasia , Dimopoulos Meletios‐Athanasios , Sfikakis Petros P.
Published: March 2003

Summary. An expanded cytotoxic/memory T‐cell subpopulation expressing low levels of the B‐cell‐specific CD20 molecule was found in peripheral blood and bone marrow of patients with multiple myeloma at the time of diagnosis, but returned to normal levels following treatment. CD3+CD20dim cells were also increased in monoclonal gammopathy of unknown significance albeit at lower levels. Lower CD3+CD20dim cell numbers at baseline may be associated with lack of response to treatment and a poor outcome. Because expansion of these T cells may be related to disease status, further studies should investigate their potentially unique function in plasma cell dyscrasias.
DDAVP enhances the ability of blood monocytes to form rosettes with activated platelets by increasing the expression of P‐selectin sialylated ligands on the monocyte surface
Authors: Pereira Arturo , Del Valle Onorato María , Sanz Cristina
Published: March 2003

Summary. The mechanism through which DDAVP (1‐deamino‐8‐d‐arginine vasopressin) promotes blood coagulation is not completely understood. As blood monocytes have been identified as a target for DDAVP, we investigated whether this drug increased monocyte adhesion to activated platelets, which would result in the close intercellular contact that is necessary for a juxtacrine effect on platelets and/or endothelium at sites of vascular injury. Monolayers of non‐confluent monocytes adhered to glass slides were incubated with thrombin‐activated, formaldehyde‐fixed platelets before and after the adherent monocytes were stimulated with DDAVP or n‐formyl‐methyl‐leucyl‐phenylalanine (fMLP). The number of platelets involved in rosettes with monocytes was quantified, and the effect of DDAVP or fMLP on the monocyte surface expression of P‐selectin ligands and CD11b/CD18 was assessed. DDAVP or fMLP increased the number of activated platelets involved in rosettes with monocytes by 2·8‐ and 4·9‐fold respectively. EDTA and inhibitors of the P‐selectin/counter‐receptor interaction decreased the platelet numbers in rosettes by 80–90%, whereas inhibitors of the integrin‐mediated adhesion reduced rosettes by 40–50%. Blocking the P‐selectin glycoprotein ligand‐1 (PSGL‐1) with the monoclonal antibody, Pl‐1, decreased the platelet numbers in rosettes by only 50%. In contrast, surface expression of the sialylated ligands of P‐selectin and, to a lesser extent, of CD11b/CD18 increased upon monocyte activation with DDAVP or fMLP, whereas it decreased slightly with PSGL‐1. These results indicate that DDAVP enhanced the ability of blood monocytes to bind activated platelets, mainly by increasing the expression of P‐selectin sialylated ligands on the monocyte surface. A similar effect was achieved with fMLP.
Novel approach to the treatment of intestinal fistula in the inaccessible abdomen: transbursal end-to-side duodenogastrostomy
Authors: Bosscha K , van Vroonhoven Th J M V
Published: March 2003

Background

Treatment of enterocutaneous fistula in patients with intra-abdominal sepsis and a surgically inaccessible abdomen is frequently unsuccessful.

Methods

A new approach has been devised: total disconnection of the proximal digestive tract, which can be performed through the bursa omentalis without entering the scarred abdomen.

Results

The procedure was carried out in four patients with high-output small bowel fistula and an inaccessible abdomen. Output of fistulas stopped promptly, recovery from intra-abdominal sepsis was achieved, the abdomens became accessible again and continuity of the digestive tract could be restored in all patients after intervals of 2–5·5 months.

Conclusion

Transbursal end-to-side duodenogastrostomy is a useful procedure when traditional surgical interventions have failed or cannot be applied.
Laparoscopic stoma formation for faecal diversion
Authors: Hollyoak M A , Lumley J , Stitz R W
Published: March 2003

Background

Laparoscopic creation of an intestinal stoma may be preferable to open operation when intervention is required solely for faecal diversion.

Methods

Experience with laparoscopic intestinal stoma formation for faecal diversion from a single institution is presented.

Results

A total of 55 stomas were studied, 40 laparoscopic and 15 open. The conversion rate from laparoscopic to open operation was 5 per cent. Mean(s.e.m.) operating time was significantly reduced for laparoscopic stomas (54(4·7) versus 72(8·7) min). Time to return of bowel function was significantly reduced (1·6(0·3) versus 2·2(0·2) days). Mean(s.e.m.) hospital stay was significantly reduced in the laparoscopic group (7·4(0·5) versus 12·6(2·5) days).

Conclusion

Morbidity and mortality appeared to be reduced in patients undergoing laparoscopic stoma formation. The technique was found to be safe, suitable for the majority of patients and to give results superior to those of open surgery.
Flap advancement and core fistulectomy for complex rectal fistula
Authors: Miller G V , Finan P J
Published: March 2003

Background

The treatment of low fistula in ano is well accepted but controversy surrounds the management of high trans-sphincteric fistulas and more complex fistulas. This study assesses the clinical results of advancement flap techniques in association with core fistulectomy for complex fistula in ano.

Methods

A retrospective analysis of the use of advancement flap techniques together with core fistulectomy in 25 patients (26 fistulas) was performed. Clinical outcome was assessed in terms of fistula healing, continence, failure and technical problems.

Results

Successful healing of 20 of the 26 complex fistulas was achieved using this technique with no disturbance of continence and minimal (technical) complications.

Conclusion

Flap advancement and core fistulectomy is a safe, effective procedure for complex rectal fistulas with good functional results and minimal or no disturbance of continence, and should be considered for the treatment of complex perianal fistulas.
Expression of c-myc oncoprotein represents a new prognostic marker in cutaneous melanoma
Authors: Ross D A , Wilson G D
Published: March 2003

Background

Overexpression of the c-myc oncoprotein has been observed to be of prognostic significance in several human cancers. This study was undertaken to establish whether c-myc has any prognostic significance in melanoma.

Methods

Expression of c-myc oncoprotein was studied in a prospective series of primary and secondary malignant melanomas. Ethanol-fixed tissue was digested into nuclei and stained for flow cytometric analysis using a pan-myc antibody.

Results

Overexpression of the c-myc oncoprotein was prevalent in both disease stages. In primary disease, overexpression was associated with tumour thickness and with the presence of ulceration, age and reduced disease-free interval. Of most importance, high expression of c-myc predicted poor outcome in both primary and metastatic disease. In addition, c-myc was able to discriminate prognosis in melanomas uniformly stratified as thick (greater than 3 mm) lesions.

Conclusion

These results indicate an important role for c-myc in melanoma which may lead to better prognostic information and identification of new therapeutic strategies.
Experience with transcranial Doppler monitoring reduces the incidence of particulate embolization during carotid endarterectomy
Authors: Smith J L , Evans D H , Gaunt M E , London N J M , Bell P R F , Naylor A R
Published: March 2003

Background

The aim of this study was to investigate whether the introduction of routine transcranial Doppler (TCD) ultrasonography during carotid endarterectomy reduces the incidence of microembolization by altering operative technique.

Methods

The number and nature of microemboli detected during the first 75 consecutive carotid endarterectomies performed with TCD monitoring during 1992–1993 (group 1) were compared with those in a similar series of 75 consecutive patients undergoing carotid endarterectomy in 1995 (group 2), after substantial experience (210 patients) with TCD monitoring. Emboli were classified as either particulate or gaseous.

Results

In patients with evidence of particulate emboli during the dissection phase of the operation, the total number of particulate emboli fell significantly in patients in group 2 (P = 0·019). Similarly, in patients in whom microembolization was detected on immediate opening of the shunt, the total number of microemboli also fell significantly in group 2 (P = 0·003). Overall, the median (95 per cent confidence interval) number of particulate emboli detected during the entire procedure fell significantly from 21 (16–29) in group 1 to 9 (7–14) in group 2 (P = 0·0008).

Conclusion

TCD monitoring plays an important role in the training and quality control of carotid endarterectomy and helps significantly to reduce the amount of microembolization.