bluuebunny/crossref_metadata_2025 huggingface dataset:
Multilineage glycosylphosphatidylinositol anchor‐deficient haematopoiesis in untreated aplastic anaemia
Authors: Mukhina Galina L. , Buckley J. Thomas , Barber James P. , Jones Richard J. , Brodsky Robert A.
Published: March 2003

Aplastic anaemia and paroxysmal nocturnal haemoglobinuria (PNH) are closely related disorders. In PNH, haematopoietic stem cells that harbour PIGA mutations give rise to blood elements that are unable to synthesize glycosylphosphatidylinositol (GPI) anchors. Because the GPI anchor is the receptor for the channel‐forming protein aerolysin, PNH cells do not bind the toxin and are unaffected by concentrations that lyse normal cells. Exploiting these biological differences, we have developed two novel aerolysin‐based assays to detect small populations of PNH cells. CD59 populations as small as 0·004% of total red cells could be detected when cells were pretreated with aerolysin to enrich the PNH population. All PNH patients displayed CD59‐deficient erythrocytes, but no myelodysplastic syndrome (MDS) patient or control had detectable PNH cells before or after enrichment in aerolysin. Only one aplastic anaemia patient had detectable PNH red cells before exposure to aerolysin. However, 14 (61%) had detectable PNH cells after enrichment in aerolysin. The inactive fluorescent proaerolysin variant (FLAER) that binds the GPI anchors of a number of proteins on normal cells was used to detect a global GPI anchor deficit on granulocytes. Flow cytometry with FLAER showed that 12 out of 18 (67%) aplastic anaemia patients had FLAER‐negative granulocytes, but none of the MDS patients or normal control subjects had GPI anchor‐deficient cells. These studies demonstrate that aerolysin‐based assays can reveal previously undetectable multilineage PNH cells in patients with untreated aplastic anaemia. Thus, clonality appears to be an early feature of aplastic anaemia.
Molecular studies in patients with chronic myeloid leukaemia in remission 5 years after allogeneic stem cell transplant define the risk of subsequent relapse
Authors: Mughal Tariq I. , Yong Agnes , Szydlo Richard M. , Dazzi Francesco , Olavarria Eduardo , Van Rhee Frits , Kaeda Jaspal , Cross Nick C. P. , Craddock Charles , Kanfer Ed , Apperley Jane , Goldman John M.
Published: March 2003

We identified 103 consecutive patients who, 5 years after allogeneic transplantation for chronic myeloid leukaemia (CML), were in molecular remission (MR). The 103 patients were divided into three groups on the basis of reverse transcription–polymerase chain reaction (RT–PCR) studies for BCR‐ABL transcripts in the first 5 years post transplant: Group A comprised 63 patients who had been continuously PCR negative; Group B comprised 20 patients with one or more positive PCR result but only at a low level; and Group C comprised 20 patients who had fulfilled the criteria for molecular relapse, been treated with donor lymphocyte infusions (DLI) and had thereafter regained complete MR within the 5‐year post‐transplant period. The median follow‐up for all 103 patients was 8·4 years from transplant (range 5–17·6 years). In group A only one patient relapsed at 9·2 years. In group B eight patients (40%) relapsed: six at molecular, one at cytogenetic and one haematological levels. The actuarial probabilities of survival at 10 years for patients in Groups A, B and C were 97·4%, 92·9% and 100% respectively; the probabilities of relapse were 3%, 54% and 0% respectively. We conclude that molecular studies during the first 5 years post transplant can help to predict long‐term leukaemia‐free survival and, possibly, cure of CML.
Comparison of megakaryopoiesis in vitro of paired peripheral blood progenitor cells and bone marrow harvested during remission in patients with acute myeloid leukaemia
Authors: Briggs Mark , Adams Julie A. , Brereton Michelle L. , Burgess Robert , Hyde Keith , Lenehan Helen , Yin John A. Liu
Published: March 2003

We have studied paired peripheral blood progenitor cells (PBPC) and bone marrow (BM) samples from 12 acute myeloid leukaemia (AML) patients following intensive chemotherapy, and assessed direct granulocyte–macrophage colony‐forming units (CFU‐GM), erythroid burst‐forming units (BFU‐E), megakaryocyte CFU (CFU‐Mk) numbers and the production of CD61+ (platelet glycoprotein IIIa) cells in suspension culture in response to various haemopoietic growth factor combinations. We found that CFU‐GM and BFU‐E numbers per 105 mononuclear cells were similar in both AML PBPC and BM harvests; CFU‐Mk numbers, however, were significantly higher in PBPC than BM. In addition, the higher total white cell count of the PBPC harvests meant that PBPC have much higher numbers of total progenitors per collection. CD61+ cell numbers in suspension cultures of AML PBPC and BM were lower than those of harvested normal marrow. However, response to pegylated recombinant human megakaryocyte growth and development factor (PEGrHuMGDF) both alone and in combination with other growth factors was qualitatively similar to that of normal BM. As with normal BM, response to PEGrHuMGDF alone did not increase further with addition of granulocyte colony‐stimulating factor (G‐CSF), granulocyte–macrophage CSF (GM‐CSF), interleukin 6 (IL‐6) or erythropoietin (EPO) in the AML PBPC and BM. Further responses over PEGrHuMGDF alone were seen when added with stem cell factor (SCF) or with a combination of SCF + IL‐3 + EPO in both AML PBPC and BM cultures; however, the magnitude of the response was greater in the PBPC cultures. Response to PEGrHuMGDF + IL‐3 was seen in the PBPC cultures but not in the AML BM. These data suggest that, in AML patients, there are proportionally more megakaryocyte progenitor cells in the mobilized PBPC than in the BM harvests, which would explain the more rapid platelet recovery following PBPC autografts.
Plasma of patients with Upshaw–Schulman syndrome, a congenital deficiency of von Willebrand factor‐cleaving protease activity, enhances the aggregation of normal platelets under high shear stress
Authors: Yagi H. , Konno M. , Kinoshita S. , Matsumoto M. , Ishizashi H. , Matsui T. , Titani K. , Fujimura Y.
Published: March 2003

Upshaw–Schulman syndrome (USS) is an autosomal recessive disorder characterized by repeated episodes of chronic thrombocytopenia and microangiopathic haemolytic anaemia (MAHA) that responds dramatically to infusions of fresh frozen plasma (FFP). Recent studies have provided consistent evidence that USS is a congenital deficiency of plasma von Willebrand factor‐cleaving protease (VWF‐CPase) activity and, therefore, unusually large VWF multimers (UL‐VWFMs) are present in the plasma. However, the molecular mechanism of the clinical symptoms of USS is not well understood. We studied the relationship between UL‐VWFMs and thrombocytopenia in two USS patients by analysing platelet aggregation using a mixture of the patient's plasma and normal washed platelets under high shear stress. Our results clearly showed a remarkably enhanced high shear stress‐induced platelet aggregation (H‐SIPA) by the patient's plasma. At 24 h after FFP infusion (≈10 ml/kg body weight), the enhanced H‐SIPA became almost completely normalized but, 2 d later, it began to return to the preinfusion level. These results were in accordance with the change in VWFM patterns. The specific effects of enhanced H‐SIPA on VWF, platelet glycoprotein Ib and endogenous ADP released from platelets upon stimulation were confirmed using reagents that specifically inhibit their respective functions. Our present results clearly indicate that thrombocytopenia in USS patients is caused by a combination of the presence of UL‐VWFMs, platelets and high shear stress generated in the microcirculation.
Effects of combinations of therapeutic agents on the proliferation of progenitor cells in chronic myeloid leukaemia
Authors: Marley Stephen B. , Davidson R. John , Goldman John M. , Gordon Myrtle Y.
Published: March 2003

Summary. Combination of STI571, a tyrosine kinase inhibitor, with other drugs may be beneficial in the treatment of chronic myeloid leukaemia (CML). We measured the effects of STI571, AG490, farnesyltransferase inhibitor (FTI), interferon alpha (IFN‐α), cytosine arabinoside (Ara‐C) and all‐trans retinoic acid (ATRA), singly and in combination, on clonogenic leukaemic cell proliferation. STI571, IFN‐α and ATRA each reduced proliferation by 50–60%; AG490, FTI and Ara‐C had less effect. Comparing the observed and expected (i.e. additive) effects of drug combinations showed STI571 + FTI, STI571 + AG490 and IFN‐α+ ATRA were additive; STI571 + IFN‐α, IFN‐α+ Ara‐C and STI571 + AG490 + FTI were less than additive. Thus, STI571 + FTI, STI571 + AG490 and IFN‐α+ ATRA may be better combination therapies for CML than STI571 +  IFN‐α, IFN‐α+ Ara‐C or STI571 + AG490 + FTI.
T‐cell analysis in identical twins reveals an impaired anti‐follicular lymphoma immune response in the patient but not in the healthy twin
Authors: Gitelson Elena , Spaner David , Buckstein Rena , Franssen Edmee , Hewitt Karen , Lim Megan S. , Pennell Nancy , Schultze Joachim L. , Berinstein Neil L.
Published: March 2003

Summary. In lymphomas an innate defect in the T‐cell repertoire could account for the impaired tumour‐specific immune response; alternatively, the tumour itself could exert an inhibitory effect on the immune system. To address this issue we analysed the T‐cell responses against follicular lymphoma (FL) in identical twins as it can be postulated that their overall T‐cell repertoire is identical. While differences between the T‐cell response of the patient and the healthy twin would point to a tumour‐induced T‐cell unresponsiveness, impaired responses in both would point to a defective T‐cell repertoire. We demonstrated an impaired tumour‐specific proliferation (P = 0·035 and P = 0·013) and cytokine release (P = 0·004 and P = 0·0008) of both peripheral blood and tumour‐derived T‐cells, respectively, in the FL patient compared with the T‐cell response of the healthy twin. Moreover, only syngeneic primed T cells were able to directly lyse unmodified FL cells of the patient. These data support previous findings in murine lymphomas and suggest that inhibitory mechanisms during tumour growth, rather than a defective T‐cell repertoire, are responsible for the insufficient T‐cell response in lymphoma.
Establishment and characterization of a Kaposi's sarcoma‐associated herpesvirus‐ and Epstein–Barr virus‐negative malignant lymphoma cell line (OHK) with primary effusion lymphoma immunophenotype
Authors: Kuwabara Hiroko , Nagai Masami , Kawakami Kimihiro , Ohmori Minoru , Tasaka Taizo , Funamoto Yasunobu , Yamaoka Genji , Nishio Hajime , Suzuki Koichi , Mori Hiroshi
Published: March 2003

Summary. A novel cell line, designated OHK, was established from ascites of a 59‐year‐old Japanese woman with diffuse large B‐cell lymphoma showing a peculiar serosal tropism, as seen in primary effusion lymphomas (PEL). OHK exhibited a large pleomorphic morphology with irregular nuclei and distinct nucleoli, and included immunoblastic and Reed–Sternberg‐like giant cells. On ultrastructural examination, rich intermediate filaments, and well‐developed Golgi apparati and rough endoplasmic reticulum, were seen. Immunophenotypically, OHK lacked T and B cell‐associated antigens, and had CD10, CD30, CD33 and CD138 antigens. Although OHK cells did not express immunoglobulin (Ig) protein, Southern blot analysis demonstrated clonal rearrangements of Ig heavy and light chain genes. These observations suggest that OHK cells are derived from preterminally differentiated B cells, and that they have features of PEL. Kaposi's sarcoma‐associated herpesvirus and Epstein–Barr virus were not detected. OHK displayed hyperploid karyotypes with multiple structural abnormalities, and produced some cytokines such as macrophage‐colony‐stimulating factor (M‐CSF), granulocyte‐CSF, interleukin 6 and transforming growth factor β1. In particular, vascular endothelial growth factor (VEGF), whose stimulation of vascular permeability is thought to be critical to the pathogenesis of PEL, was also produced in large quantities. These results indicate that OHK may be a useful tool for the investigation of PEL.
CD34+/CD90+ cells infused best predict late haematopoietic reconstitution following autologous peripheral blood stem cell transplantation
Authors: Sumikuma Toshiya , Shimazaki Chihiro , Inaba Tohru , Ochiai Naoya , Okano Akira , Hatsuse Mayumi , Ashihara Eishi , Nakagawa Masao
Published: March 2003

Summary.  This study aimed to identify which graft product subset of cells might be the most predictive of late haematopoietic recovery (three to 12 months) following autologous peripheral blood stem cell transplantation (PBSCT). The relationships between the numbers of reinfused CD34+ cells and their immature subsets such as CD34+/CD90+, CD34+/AC133+, CD34+/CD38– and CD34+/HLA‐DR– cells, and haemoglobin, white blood cell (WBC) and platelet counts at 3, 6, 9 and 12 months after PBSCT, were studied in 25 patients with haematological and solid malignancies. The total CD34+ cell number, as well as CD34+/CD90+ and CD34+/AC133+ cell numbers, correlated with platelet counts at 3, 6, 9 and 12 months after PBSCT, but the CD34+/CD90+ cells infused best predicted platelet recovery during the first 12 months after PBSCT (P < 0·0238 at any time‐point). The CD34+/AC133+ cell dose also correlated with WBC counts at 3 months post PBSCT. In addition, all patients receiving more than 80 × 104 CD34+/CD90+ cells/kg showed platelet counts greater than 100 × 109/l at all points after PBSCT, suggesting that this value of the CD34+/CD90+ cells infused was a threshold dose for durable haematopoietic engraftment after PBSCT.
Semiquantitative measurement of IgG subclasses and IgM of platelet‐specific antibodies in a glycoprotein‐specific platelet–antigen capture assay
Authors: Kelsch Reinhard , Hutt Kerstin , Cassens Uwe , Sibrowski Walter
Published: March 2003

Summary. The detection of platelet‐specific antibodies is of high clinical interest in diseases with immune thrombocytopenia. The glycoprotein‐specific platelet–antigen capture (PAC)‐assay developed in this study is especially suited to the differentiation of platelet‐specific immunoglobulin (Ig) G subclasses and the determination of platelet‐specific IgM in serum or on platelets. The problems with unspecific signals or low sensitivity usually seen with the detector antibodies available are effectively overcome, as unbound detector antibodies are removed at an early stage in the assay. We investigated 14 maternal alloantisera from cases of neonatal alloimmune thrombocytopenia (NAITP) and six sera from patients with autoimmune thrombocytopenic purpura (AITP). In NAITP sera, we found IgG1 alone in 57%, IgG1 + IgG3 in 21% and IgG1 + IgG2 in 14% of cases. One serum contained IgG1 + IgG2 + IgG3. In AITP, three out of the six sera contained IgG1 alone. One serum contained IgG1 + IgG2. One patient, with highly refractory AITP, had platelet‐specific IgG1 + IgG2 + IgG3 in his serum. A patient with AITP in remission and normal platelet counts only showed platelet‐bound IgG2. The detection of platelet‐specific ‘whole IgG’ is possible too. However, at this time the commonly used monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) method should not be replaced for this purpose, as it is well standardized and used with similar results in many laboratories. The PAC assay sensitively detects the subclasses of platelet‐specific IgG and human leucocyte antigen (HLA)‐antibodies independently. It is easy to perform and takes less time than other platelet glycoprotein‐specific methods.
Assessment of thrombocytopenic disorders using the Platelet Function Analyzer (PFA‐100®)*
Authors: Carcao Manuel D. , Blanchette Victor S. , Stephens Derek , He Lijing , Wakefield Cindy D. , Butchart Sheila , Christie Douglas J. , Rand Margaret L.
Published: March 2003

Summary.  The Platelet Function Analyzer (PFA‐100®) was used to measure platelet function in paediatric patients with destructive versus underproduction thrombocytopenia. Closure time (CT) and total volume (TV) measurements with standard 150 μm apertures discriminated between patients with similar platelet counts from 30 to 150 × 109/l. However, at platelet counts < 30 × 109/l, a 100‐μm aperture (experimental) gave the best assessment of platelet function. TV results could be analysed even when CTs were indeterminate. Further investigations are warranted to more fully understand the relationships among platelet function as measured by the PFA‐100® in standard/experimental modes, bleeding and transfusion outcome in thrombocytopenia.
A wider molecular spectrum of β‐thalassaemia in Myanmar
Authors: Ne‐Win  , Harano Teruo , Harano Keiko , Thein‐Thein‐Myint  , Rai‐Mra  , Okada Shigeru , Shimono Kunio , Aye‐Aye‐Myint
Published: March 2003

Summary.  Two hundred and nine beta‐thalassaemia (β‐Thal) alleles of 158 unrelated Myanmar patients (107 HbE‐β‐Thal; 51 β‐Thal major) were analysed for β‐globin gene mutations. Amplification refractory mutation system (ARMS) characterized six β‐thal mutations known to Myanmar [βIVSI‐1(G→T), codon 41/42(–TCTT), βIVSI‐5(G→C), codon 17(A→T), βIVS II‐654(C→T), and −28 Cap (A→G)] in 166/209 (79·4%) alleles. DNA sequencing of 24 alleles from 43 ARMS‐negative samples (20·6%) identified an additional 12 new mutations, to produce a total of 18 different mutations. Nineteen alleles (9·1%) remained for further characterization. The molecular spectrum of Myanmar β‐Thal is wider and more heterogeneous than previously reported.
Protein C/S ratio, an accurate and simple tool to identify carriers of a protein C gene mutation
Authors: Libourel Eduard J. , Meinardi Johan R. , De Kam Pieter‐Jan , Veenstra Rense , Van Der Schaaf Wim , Ruiters Marcel H. J. , Van Der Meer Jan
Published: March 2003

Summary.  Hereditary protein C deficiency is demonstrated by lowered protein C plasma levels in a patient and at least one first‐degree relative. This approach is insufficient in some cases owing to overlapping protein C levels in carriers and non‐carriers of a protein C gene mutation. The protein C/S ratio is a simple and more accurate tool to detect carriers. In four families, 62% of 29 carriers, compared with none of 39 non‐carriers, were protein C deficient. Sensitivity and specificity of the protein C/S ratio were 90% and 92% respectively. The protein C/S ratio seems to be more accurate than the measurement of protein C levels alone to identify carriers of a protein C gene mutation.
Spontaneous remission of low‐grade B‐cell non‐Hodgkin's lymphoma following withdrawal of methotrexate in a patient with rheumatoid arthritis: case report and review of the literature
Authors: Baird Richard D. , Nellis van Zyl‐Smit Richard , Dilke Timothy , Scott Sara E. , Rassam Saad M. B.
Published: March 2003

Summary. A 69‐year‐old woman, who had suffered from deforming rheumatoid arthritis since the age of 40 years, had been treated with methotrexate for 3 years. She presented with a 7 week history of neck lymphadenopathy. Biopsy revealed low‐grade marginal‐zone B‐cell non‐Hodgkin's lymphoma. Computerized tomography and bone marrow biopsy confirmed stage IIIA disease. Spontaneous complete remission of the lymphoma was achieved 14 months after withdrawing immune suppression with methotrexate.
SAB – a promising new treatment to improve remission rates in AML in the elderly?
Authors: Wheatley Keith
Published: March 2003

Summary. This short report presents the results of a comparison of complete remission rates and reasons for failure, between two series of patients aged 60 years or over with acute myeloid leukaemia (AML), and discusses their interpretation.
Protein kinase C‐ζ overexpression induces erythroid phenotype in the monocytic leukaemia cell line U937
Authors: Mansat‐De Mas Véronique , De Thonel Aurélie , Gaulin Vanessa , Demur Cécile , Laurent Guy , Quillet‐Mary Anne
Published: March 2003

Summary. Previous studies have established that protein kinase C‐ζ (PKC‐ζ) is critical for neuronal cell differentiation. However, the role of PKC‐ζ in haematopoietic cell differentiation is less clear. In this study, we have investigated the influence of PKC‐ζ overexpression on the phenotype of the human monocytic U937 leukaemic cells. In two PKC‐ζ‐overexpressing clones (U937 ζJ and U937 ζB), PKC‐ζ expression levels and activity were three to fourfold higher, and the enzyme accumulated both in the cytoplasm and in the nucleus compared with U937 control cells. PKC‐ζ‐overexpressing U937 cells exhibited an erythroid phenotype characterized by high levels of glycophorin A, cell haemoglobinization, increased GATA‐1 transcripts and protein expression, compared with controls. Immunoprecipitation studies revealed that GATA‐1 protein was constitutively phosphorylated in PKC‐ζ‐overexpressing cells. Moreover, GATA‐1 did not interact with PKC‐ζ but interacted with ERK1, which was constitutively activated and accumulated in the nucleus of U937 ζJ. However, ERK1 phosphorylation inhibition by PD098059 did not influence either GATA‐1 phosphorylation or GATA−1/ERK1 interaction. Collectively, these results suggest a model in which PKC‐ζ induces MEK‐dependent ERK1 activation, ERK1 translocation to the nucleus, GATA−1/ERK1 interaction and ERK1‐independent GATA‐1 phosphorylation resulting in GATA‐1 accumulation. To conclude, this study provides evidence for the role of PKC‐ζ in erythroid gene regulation.
Differentiation between non‐Hodgkin's lymphoma recurrence and second primary lymphoma by VDJ rearrangement analysis
Authors: Libra Massimo , De Re Valli , Gasparotto Daniela , Gloghini Annunziata , Marzotto Alessandra , Milan Isabella , Tirelli Umberto , Stivala Franca , Carbone Antonino , Boiocchi Mauro
Published: March 2003

Summary. Relapses in non‐Hodgkin's lymphomas (NHL) could be due to the reappearance of the initial neoplasm or new primary tumours. Discrimination between the two events would allow a more targeted therapeutic approach. VDJ rearrangement was used as marker of clonality in metachronous biopsy specimens from 10 patients with relapsed B‐NHL. Complimentarity determining region 3 was amplified and sequenced. D‐JH was identical in eight matched primary/secondary tumours, confirming the diagnosis of recurrence. In contrast, primary and secondary tumours in two patients were of different clonal origin. Our data indicate that VDJ analysis is a fundamental tool for identificaton of relapses in NHL.
Use of a non‐depleting anti‐CD4 antibody to modulate the immune response to coagulation factors VIII and IX
Authors: Salooja Nina , Kemball‐Cook Geoffrey , Tuddenham Edward G. D. , Dyson Julian
Published: March 2003

Summary. The generation of antibodies to therapeutic factors VIII or IX is a major problem in the management of haemophilia and places potential limitations on the application of gene therapy. We have investigated the administration of a non‐depleting anti‐CD4 antibody for modulation of the immune response to human recombinant coagulation factors VIII and IX. In mice given these clotting factors, co‐administration of anti‐CD4 antibody significantly reduced the appearance of factor‐specific antibodies. These data provide evidence that the neutralizing antibody response to exogenous coagulation factors may be controllable if non‐depleting anti‐CD4 antibody is co‐administered at the time of initial replacement therapy.
Etilefrine for the prevention of priapism in adult sickle cell disease
Authors: Okpala Iheanyi , Westerdale Neill , Jegede Tina , Cheung Betty
Published: March 2003

Summary. Priapism is a common complication of sickle cell disease (SCD) that could lead to erectile dysfunction and psychosocial problems. Treatment of established fulminant priapism is usually not satisfactory. It is therefore important to prevent this complication of SCD. The alpha‐adrenergic agonist etilefrine (50–100 mg/d) produced a good clinical response in 13 of 18 (72%) adults who have recurrent priapism; 17 had SCD and one sickle cell trait. After a follow‐up of 1–48 months, none of the 18 people on etilefrine developed hypertension or sexual dysfunction. Similar efficacy and safety profiles of the drug have been reported previously.
Allogeneic stem cell transplantation in the myelodysplastic syndromes: interim results of outcome following reduced‐intensity conditioning compared with standard preparative regimens
Authors: Parker Jane E. , Shafi Tariq , Pagliuca Antonio , Mijovic Aleksandar , Devereux Stephen , Potter Mike , Prentice H. Grant , Garg Mamta , Yin John A. , Byrne Jenny , Russell Nigel H. , Mufti Ghulam J.
Published: March 2003

Summary. Conventional allogeneic stem cell transplantation (SCT) for myelodysplastic syndrome (MDS) is associated with excessive procedure‐related mortality. The outcome following volunteer‐unrelated donor (VUD) or sibling allogeneic SCT was therefore evaluated in 23 MDS patients conditioned with reduced‐intensity regimens (fludarabine/busulphan/Campath‐1H) because of advanced age (48 vs 37 years, P = 0·002) and/or co‐morbidity (19 vs 3, P < 0·0001) which precluded conventional transplantation, and compared with 29 treated with standard protocols [busulphan/cyclophosphamide (Bu/Cy); Bu/Cy/total‐body irradiation/Campath‐1G]. Graft‐versus‐host disease (GVHD) prophylaxis comprised of cyclosporine/methotrexate. One hundred per cent donor engraftment (variable number tandem repeat analysis/cytogenetics/fluorescence in situ hybridization) was achieved in 18/19 (95%) evaluable patients receiving reduced‐intensity regimens, although six (32%) have subsequently shown mixed chimaerism. Reduced‐intensity conditioning was associated with significantly reduced duration of aplasia, less mucositis, fever, antibiotic, analgesia, parenteral nutrition use, less acute and chronic GVHD, and lower early procedure‐related mortality [two (9%) vs nine (31%), P < 0·05]. Six patients relapsed (two standard, four reduced‐intensity) and two (reduced‐intensity) experienced late graft failure. The 2 year actuarial overall/disease‐free survival (OS/DFS) was 48/39% in the reduced‐intensity arm and 44/44% in the standard group. The 2 year non‐relapse mortality was 31% and 50% respectively. In VUD recipients, OS was superior in the reduced‐intensity arm (49%vs 34%). Predictors of DFS included good/intermediate‐risk karyotype, low/intermediate‐1 International Prognostic Scoring system score, human leucocyte antigen compatibility and attainment of complete remission. Our data demonstrates that VUD or sibling allogeneic SCT following reduced‐intensity conditioning is feasible in high‐risk MDS patients considered unsuitable for standard transplantation and is associated with comparable 3·5 year DFS to those receiving conventional regimens.
Frequent co‐expression of HoxA9 and Meis1 genes in infant acute lymphoblastic leukaemia with MLL rearrangement
Authors: Imamura Toshihiko , Morimoto Akira , Takanashi Mami , Hibi Shigeyoshi , Sugimoto Tohru , Ishii Eiichi , Imashuku Shinsaku
Published: March 2003

Summary. We studied the expression of HoxA9 and Meis1 by reverse transcriptase‐polymerase chain reaction analysis in leukaemic cells from cases of infant acute lymphoblastic leukaemia (ALL, n = 27) and childhood ALL (n = 29). These two genes were co‐expressed significantly more frequently in infant ALL than in childhood ALL (19/27 vs0/29 cases, P < 0·001) and were highly associated with MLL gene rearrangement in infant ALL cases (P < 0·001). These findings indicate that the HoxA9 and Meis1 genes are closely associated with MLL gene rearrangement in the development of infant ALL, which represents a distinct entity of childhood ALL.
Response to thalidomide in chemotherapy‐resistant mantle cell lymphoma: a case report
Authors: Wilson Edward A. , Jobanputra Shialesh , Jackson Robert , Parker Anne N. , McQuaker I. Grant
Published: March 2003

Summary. Mantle cell lymphoma is an aggressive B‐cell lymphoma with a poor median survival despite conventional therapy. Here, we present the case of a patient with multiply relapsed mantle cell lymphoma, having failed treatment with chemotherapy, steroids and rituximab. He was treated with single‐agent thalidomide at a dose of 800 mg daily and entered a good partial remission which was maintained for the next 6 months. There is clearly a need for further studies of thalidomide in mantle cell lymphoma to confirm this promising initial result.
Factor VIII deficiency not induced by FVIII gene mutation in a female first cousin of two brothers with haemophilia A
Authors: Mazurier Claudine , Parquet‐Gernez Armelle , Gaucher Christine , Lavergne Jean‐Maurice , Goudemand Jenny
Published: March 2003

Summary. In this study, we reinvestigated a 20‐year‐old woman, the first cousin of two brothers with severe haemophilia A. This patient was previously assumed to be a carrier of haemophilia A due to her FVIII deficiency. We identified a novel FVIII gene mutation in the family and demonstrated that the FVIII deficiency in this female patient did not result from this gene mutation, but was linked to molecular defects in the von Willebrand factor gene.
Pamidronate causes apoptosis of plasma cells in vivo in patients with multiple myeloma
Authors: Gordon Sharon , Helfrich Miep H. , Sati Hamdi I. A. , Greaves Michael , Ralston Stuart H. , Culligan Dominic J. , Soutar Richard L. , Rogers Michael J.
Published: March 2003

Summary. Anti‐resorptive bisphosphonates, such as pamidronate, are an effective treatment for osteolytic disease and hypercalcaemia in patients with multiple myeloma, but have also been shown to cause apoptosis of myeloma cell lines in vitro. In this study, we found that a single infusion of pamidronate, in 16 newly diagnosed patients with multiple myeloma, caused a marked increase in apoptosis of plasma cells in vivo in 10 patients and a minimal increase in four patients (P < 0·05). The nitrogen‐containing bisphosphonates pamidronate and zoledronic acid also induced apoptosis of authentic, human bone marrow‐derived plasma cells in vitro. Apoptosis of plasma cells in vitro was probably caused by inhibition of the mevalonate pathway and loss of prenylated small GTPases, as even low concentrations (≥ 1 µmol/l) of zoledronic acid caused accumulation of unprenylated Rap1A in cultures of bone marrow mononuclear cells in vitro. GGTI‐298, a specific inhibitor of geranylgeranyl transferase I, also induced apoptosis in human plasma cells in vitro, suggesting that geranylgeranylated proteins play a role in signalling pathways that prevent plasma cell death. Our results suggest that pamidronate may have direct and/or indirect anti‐tumour effects in patients with multiple myeloma, which has important implications for the further development of the more potent nitrogen‐containing bisphosphonates, such as zoledronic acid, in the treatment of myeloma.
Helicobacter pylori infection and monoclonal gammopathy of undetermined significance
Authors: Rajkumar S. Vincent , Kyle Robert A. , Plevak Matthew F. , Murray Joseph A. , Therneau Terry M.
Published: March 2003

Summary. A recent report found resolution of monoclonal gammopathy of undetermined significance (MGUS) in nearly 30% of patients upon eradication of concomitant Helicobacter pylori (H. pylori) infection. We performed serologicalal testing for H. pylori on 93 MGUS patients and 98 control subjects. Seroprevalence of H. pylori was not significantly different between the two groups, 30% and 32% respectively. A retrospective review of Mayo Clinic records revealed identical diagnosis rates of H. pylori infection (33%) by serology, breath or stool testing between patients with MGUS and those with negative monoclonal protein studies. There was no evidence of resolution of MGUS with H. pylori therapy.
Two double heterozygous mutations in the F7 gene show different manifestations
Authors: Nagaizumi Keiko , Inaba Hiroshi , Suzuki Takashi , Hatta Yoshihiro , Hagiwara Takeshi , Amano Kagehiro , Arai Morio , Fukutake Katsuyuki
Published: March 2003

Summary. We sequenced the factor VII gene (F7) in two unrelated Japanese patients with factor VII (FVII) deficiency. In the first (an asymptomatic 46‐year‐old man with FVII activity and antigen levels of 1·2% and 21% of normal respectively), novel E25K and H348Q mutations were identified in the doubly heterozygous state. In transiently transfected HEK293 cells, the level of FVII‐E25K mutant activity in the culture media was significantly lower than that of FVII wild type, whereas the antigen levels of both proteins were similar. This suggests that the E25K mutation is associated with a dysfunctional FVII molecule. In the second patient (a 47‐year‐old woman with FVII activity and antigen levels of less than 1% and 6% respectively), an IVS4+1 mutation and a novel −96C to T transition were detected in the double heterozygous state. In electrophoretic mobility shift assays, the −96T mutation was shown to disrupt binding of Sp1.
Acquired immunodeficiency syndrome‐associated lymphomas are efficiently lysed through complement‐dependent cytotoxicity and antibody‐dependent cellular cytotoxicity by rituximab
Authors: Golay Joseè , Gramigna Rosanna , Facchinetti Valeria , Capello Daniela , Gaidano Gianluca , Introna Martino
Published: March 2003

Summary. Rituximab (Mabthera) and alemtuzumab (Campath®, Mabcampath®) are non‐conjugated IgG1 therapeutic monoclonal antibodies directed against the CD20 and CD52 surface antigens respectively. They are presently used in the therapy of indolent B‐cell non‐Hodgkin's lymphoma (B‐NHL) and of B‐cell chronic lymphocytic leukaemia, and are thought to act mainly through complement‐dependent cytotoxicity (CDC) and antibody‐dependent cellular cytotoxicity (ADCC). Here we have analysed the capacity of these two monoclonal antibodies to lyse cell lines of acquired immunodeficiency syndrome (AIDS)‐related B‐NHL through either complement activation or antibody‐dependent cytotoxicity. Rituximab strongly activated both CDC and ADCC against CD20‐positive AIDS‐NHL cells lines, inducing up to 60–98% and 20% specific lysis respectively. In contrast, alemtuzumab was a poor activator of CDC, even in the AIDS‐NHL cell lines expressing high amounts of CD52, leading to a lysis of only 1–30%, whereas it was at least as strong as rituximab in inducing ADCC of the same lines (up to 30% specific lysis). Altogether, these data offer a first in vitro rationale supporting the therapeutic use of rituximab for CD20‐positive AIDS‐NHL.
Does isochromosome 7q mandate bone marrow transplant in children with Shwachman–Diamond syndrome?
Authors: Cunningham Joan , Sales Mark , Pearce Andrew , Howard Julie , Stallings Ray , Telford Nicholas , Wilkie Rosalie , Huntly Brian , Thomas Angela , O'Marcaigh Aengus , Will Andrew , Pratt Norman
Published: March 2003

Summary. We report on nine children with Shwachman–Diamond syndrome (SDS), eight of whom had clonal abnormalities of chromosome 7. Seven children had an isochromosome 7 [i(7)(q10)] and one a derivative chromosome 7, all with an apparently identical (centromeric) breakpoint. Children with SDS are predisposed to myelodysplasia (MDS) and acute myeloid leukaemia (AML) often with chromosome 7 abnormalities. Allogeneic transplants have been used to treat these children, however, they are a high‐risk transplant group and require careful evaluation. Three of the children were transplanted but only one survived, who to our knowledge remains the longest surviving SDS transplant patient (4·5 years +). The six non‐transplanted children are well. In classic MDS, chromosome 7 abnormalities are associated with rapid progression to acute leukaemia; however, we present evidence to suggest that isochromosome 7q may represent a separate disease entity in SDS children. This is a particularly interesting finding given that the SDS gene has recently been mapped to the centromeric region of chromosome 7. Our studies indicate that i(7)(q10) is a relatively benign rearrangement and that it is not advisable to offer allogeneic transplants to SDS children with i(7)(q10) alone in the absence of other clinical signs of disease progression.
Long‐term follow‐up of granulocyte colony‐stimulating factor receptor mutations in patients with severe congenital neutropenia: implications for leukaemogenesis and therapy
Authors: Ancliff Phil J. , Gale Rosemary E. , Liesner Ri , Hann Ian , Linch David C.
Published: March 2003

Summary. Severe congenital neutropenia (SCN) is characterized by profound neutropenia, recurrent severe bacterial infections and maturation arrest in the myeloid lineage. Granulocyte colony‐stimulating factor (G‐CSF) treatment results in clinical improvement in over 90% of cases. Point mutations of the G‐CSF receptor (G‐CSFR) have been implicated in the progression of SCN to acute myeloid leukaemia (AML). Data are presented here on the 9‐year follow‐up of seven patients and the further screening of 18 other cases. One of the two original cases with a G‐CSFR mutation has improved clinically; nevertheless, mutant DNA could still be detected at a very low level > 8 years after identification. The second child with a mutation progressed to myelodysplasia/AML 5 years after her mutation was detected. No mutations were found in the 18 new cases. One of three transformed cases had a G‐CSFR mutation. This work is in agreement with the suggestion that G‐CSFR mutations may provide a survival advantage to haemopoietic stem cells, but argues against the inevitability of leukaemic progression in their presence. Furthermore, the low frequency of G‐CSFR mutations in SCN and the importance of regular screening and close clinical and laboratory follow‐up if a mutation is found were demonstrated.
Upregulation of CXCR1 by proliferating cells in patients with lymphoproliferative disease of granular lymphocytes
Authors: Zambello Renato , Trentin Livio , Facco Monica , Carollo Davide , Cabrelle Anna , Tosoni Alicia , Cannas Giovanna , Nicolardi Linda , Agostini Carlo , Semenzato Gianpietro
Published: March 2003

Summary. The expression and the functional activities of different chemokine receptors (CC motif: CCR1, CCR2, CCR3, CCR5, CCR6; CXC motif: CXCR1, CXCR2, CXCR3, CXCR4, CXCR5) were investigated in 12 patients with lymphoproliferative disease of granular lymphocytes (LDGL). Six patients were characterized by the proliferation of CD3+ve GL and six patients by the expansion of CD3‐ve GL. The interleukin 8 (IL‐8/CXCL8) receptor CXCR1 was expressed in 12/12 patients, the CXCR4 in 6/12 patients (four CD3+ve and two CD3‐ve) and the CXCR3 in 3/12 patients (one CD3+ve and two CD3‐ve). CXCR1 was expressed only by proliferating GL. Other CC and CXC receptors were not expressed on proliferating GL (< 2%). In functional assays, purified GL from the patients displayed significant migration in response to specific chemokines, indicating that CXCR1, CXCR3 and CXCR4 were functionally active in these patients. In addition, a significant reduction of IL‐8/CXCL8‐mediated cell migration was reported in the presence of anti‐CXCR1 monoclonal antibody. Our results indicate that expanding cells from patients with LDGL express specific CXCR. These data may help to define functional properties of proliferating GL in patients with LDGL and contribute toward the understanding of the complex clinical features of this disease. In particular, as CXCR1 was expressed in all of the patients studied, we speculate that abnormal expression of this receptor on proliferating GL might play a role in the pathogenesis of neutropenia, which represents a common feature in LDGL patients.
Radical and nerve-preserving surgery for rectal cancer in the Netherlands: a prospective study on morbidity and functional outcome
Authors: Maas C P , Moriya Y , Steup W H , Kiebert G M , Klein Kranenbarg W M , van de Velde C J H
Published: March 2003

Background

Operative procedures for primary rectal cancer from Japan combine pelvic nerve-preserving techniques with radical tumour resection to ensure optimal local tumour control with minimal bladder and sexual dysfunction. A prospective study was undertaken to evaluate morbidity and functional outcome of such a technique in Dutch patients.

Methods

Forty-seven patients were operated on by a Japanese surgeon. Postoperative course was monitored. Voiding and sexual function were analysed using questionnaires completed by patients.

Results

After operation, only prolonged paralytic ileus (five of 47 patients) and perineal wound dehiscence (five of 18) occurred more frequently than reported in literature. There were no deaths. No patient developed urinary incontinence. Three of 11 women and 19 of 30 men were sexually active. Two men were impotent after operation. Impotence was related to sacrifice of the inferior hypogastric plexus (P = 0·037). Preservation of the superior hypogastric plexus was crucial for ejaculation (P = 0·003).

Conclusion

A relationship between sacrifice of specific nerve structures and accompanying dysfunction was established. The nerve-preserving technique yields good results in terms of morbidity and functional outcome, and should be considered for adoption as a standard surgical procedure for primary rectal cancer.
Laparoscopic management of acute small bowel obstruction
Authors: Bailey I S , Rhodes M , O'Rourke N , Nathanson L , Fielding G
Published: March 2003

Background

Laparoscopic management of acute small bowel obstruction is hypothetically attractive but little is known of its clinical potential.

Methods

A retrospective study was undertaken of patients with acute small bowel obstruction requiring surgery, managed by a laparoscopic unit (LU; n = 69) and a general unit (GU; n = 70).

Results

Laparoscopy was performed in 55 patients (80 per cent) in the LU compared with ten (14 per cent) in the GU. Laparoscopic surgery completed treatment in 31 patients (45 per cent) in the LU and assisted in a further 15 (22 per cent). Patients treated laparoscopically were discharged earlier than those treated by laparotomy (median 3 (range 1–15) versus median 8 (range 1–46) days). Patients treated laparoscopically had a higher chance of early unplanned reoperation than those treated by laparotomy (five of 35 versusfour of 88) (P < 0·05).

Conclusion

Laparoscopy can be performed in a high percentage of patients requiring surgery for acute small bowel obstruction. Hospital stay was reduced but the risk of early unplanned reoperation was increased in patients managed laparoscopically.
Evaluation of ultrasonography and clinical diagnostic scoring in suspected appendicitis
Authors: Galindo Gallego M , Fadrique B , Nieto M A , Calleja S , Fernández-Aceñero M J , Ais G , González J , Manzanares J J
Published: March 2003

Background

Several diagnostic aids have been developed to improve diagnosis in suspected appendicitis including ultrasonography and clinical diagnostic scoring. The aim of this study was to elaborate a new scoring system and to measure its accuracy in the preoperative diagnosis of appendicitis, comparing it with the available scoring systems.

Methods

The clinical, radiological and ultrasonographic data of 192 patients with suspected appendicitis were collected prospectively.

Results

Only six of the 12 variables analysed were shown to have prognostic significance. Using Bayesian methodology, a weight was given to each criterion and two overall scores were calculated (ultrasonographic and classical scores). A cut-off point was identified to separate patients who needed surgery and those for observation. The ultrasonographic score showed an 81 per cent sensitivity and a 96 per cent specificity, compared with 60 and 73 per cent respectively for the classical score.

Conclusion

Ultrasonography increases the diagnostic accuracy in patients with suspected acute appendicitis.
Non-invasive measurement of colonic blood flow distribution using laser Doppler imaging
Authors: Hajivassiliou C A , Greer K , Fisher A , Finlay I G
Published: March 2003

Background

This study tested a prototype laser Doppler scanner for the measurement of human colonic blood flow.

Methods

Blood flow distribution was assessed in human colon during operation in six controls and in six patients with inflammatory bowel disease undergoing colectomy. Image processing software analysed several hundred reading points, expressing average flow in perfusion units.

Results

Blood flow in the colon was not significantly lower in the control group than in patients with inflammatory bowel disease (mean(s.e.m.) 297·8(24·5) versus347.2(59·0) perfusion units, P=0·12). Intestinal colonic mural blood supply was demonstrated up to a distance of 6 cm and ischaemia demarcation lines were identified before the onset of visible changes.

Conclusion

This prototype laser Doppler flowscanner overcomes the previous limitations of laser Doppler flowmeters and may have many clinical and research applications.
Functional results after transanal rectal advancement flap repair of trans-sphincteric fistula
Authors: Kreis M E , Jehle E C , Ohlemann M , Becker H D , Starlinger M J
Published: March 2003

Background

Transanal rectal advancement flap repair is an operation to treat trans-sphincteric fistula which leaves the external sphincter muscle essentially untouched. Anal sphincter function was evaluated prospectively before and after this procedure.

Methods

Anorectal manometry was performed in 24 patients before operation and 3 months after surgery. A detailed standardized questionnaire on faecal continence was answered before surgery, then at 3 and 48 months after surgery.

Results

No significant differences were seen between mean(s.e.m.) preoperative and postoperative values for maximum squeeze pressure (100·0(9·7) versus118·0(12·7) mmHg), maximum resting pressure (56·6(4·3) versus 52·8(4·1) mmHg), rectal compliance (4·4(0·6) versus 3·5(0·5) ml/mmHg) or any other parameter of anorectal manometry. The questionnaire revealed the occurrence of minor incontinence in two patients following surgery, which remained unchanged for 4 years. Three other patients had continence disturbances 4 years after surgery which were probably unrelated to the procedure.

Conclusion

In addition to high success rates, transanal rectal advancement flap repair also yields excellent functional results. This procedure should be performed for trans-sphincteric fistula in place of alternative treatments whenever feasible.
Management of malignant oesophageal obstruction with self-expanding metallic stents
Authors: Cowling M G , Hale H , Grundy A
Published: March 2003

Background

The use of self-expanding metal stents for palliation of malignant dysphagia is increasing. Experience in 70 patients was reviewed with respect to the value of stenting and management of the complications encountered.

Methods

Oesophageal stents were inserted in 70 patients (42 men) of mean age 73 years with malignant oesophageal obstruction. Data regarding stent insertion and degree of dysphagia were gathered prospectively.

Results

Seventy-six stents were placed in 70 patients. By the end of the study 57 patients had died and 13 were still alive. Three patients died within 3 days of stent insertion and dysphagia was relieved in 64 of the 67 patients remaining. Stent migration, tumour ingrowth and overgrowth, and food impaction were encountered during follow-up in eight patients.

Conclusion

Insertion of self-expanding metal stents for the palliation of malignant oesophageal obstruction is a successful therapy which can be carried out with relative ease. Palliation of dysphagia with an appropriate stent can be expected in up to 95 per cent of patients.