bluuebunny/crossref_metadata_2025 huggingface dataset:
Lansoprazole 30 mg daily versus ranitidine 150 mg b.d. in the treatment of acid‐related dyspepsia in general practice
Authors: Jones R. H. , Baxter G.
Published: November 2003

Aim:

To compare lansoprazole 30 mg daily with ranitidine 150 mg b.d. in the treatment of acid‐related dyspepsia in general practice.

Methods:

In a double‐blind, parallel group, randomized, multicentre study conducted in 32 general practices in the UK, 213 patients were randomized to receive lansoprazole 30 mg daily, and 219 to receive ranitidine 150 mg b.d., for 4 weeks. All patients had experienced symptoms of reflux‐like or ulcer‐like dyspepsia on at least 4 of the 7 days prior to the study; 75% had experienced dyspepsia in the past, and 74 of the lansoprazole patients and 77 of the ranitidine patients had documented histories of acid‐related disorders, investigated by either radiology or endoscopy.

Results:

After 2 weeks 55% of the lansoprazole patients and 33% of the ranitidine group were symptom‐free (P = 0.001, χ2 = 7.12) with corresponding 4‐week figures of 69% and 44%, respectively (P = 0.001, χ2 = 18.03). Similar figures were found at both 2 and 4 weeks for daytime and night‐time heartburn and epigastric pain scores; in the lansoprazole group, at 4 weeks, 80% of patients were free of daytime heartburn and 81% of night‐time epigastric pain, compared with 55% (P = 0.001, χ2 = 15.44) and 65% (P = 0.01, χ2 = 6.10) in the ranitidine group.

Conclusion:

Superior symptom relief for patients presenting with ulcer‐like and reflux‐like symptoms in general practice is provided by lansoprazole 30 mg daily compared with ranitidine 150 mg twice daily.
Colonic range of 5‐ASA enemas in healthy individuals, with a comparison of their physical and chemical characteristics. An open, single‐dose, randomized, cross‐over study
Authors: OTTEN M. H. , HAAS G. DE , ENDE R. VAN DEN
Published: November 2003

Background: 5‐Aminosalicylic acid (5‐ASA) is widely used as topical treatment in patients with distal inflammatory bowel disease. The enema spread and retention time are considered to be important factors in the efficacy of this therapeutic agent. Whereas colonic spread is widely investigated in selected patient populations and volunteers, much less attention has been given to the in vitro differences of physical and chemical properties, although they may influence the in vivo characteristics.

Methods: Two different brand enemas, Salofalk and Asacol, both containing 2 g mesalazine, were compared with respect to their in vivo and in vitro characteristics. The retrograde spread, maximum distribution and wall adhesion, as well as the retention time of the enemas, was examined by the addition of a technetium tracer dose in 12 healthy volunteers on two separate occasions. In addition, several chemical properties such as pH, viscosity, particle size, dispersion rate, specific surface area and residual volume after application were analysed and compared.

Results: With its larger volume and higher viscosity the Asacol preparation reached a substantially larger proportion of the colon and produced a significantly higher retention time in the proximal parts of the large intestine. In addition, more than double the amount of 5‐aminosalicylic acid was not expelled from the semi‐rigid Salofalk enema container after application. With respect to chemical properties it was demonstrated that the Asacol preparation showed a significantly smaller size of micronized 5‐aminosalicylic acid particles, better homogeneity and much less aggregation of the drug. This resulted in an almost threefold higher specific surface area per g active compound.

Conclusions: The Asacol enema appears to be superior in several aspects of the galenical formulation. The better dispersion and larger specific surface area, in conjunction with a larger distribution, better bowel wall adherence and retention time in vivo, constitute a clear theoretical and possible clinical advantage.
A serologic survey of Helicobacter pylori infection in 3281 Italian patients endoscoped for upper gastrointestinal symptoms
Authors: PALLI D. , VAIRA D. , MENEGATTI M. , SAIEVA C.
Published: November 2003

Aim:To assess the seroprevalence of Helicobacter pylori infection in a large series of patients with upper gastrointestinal symptoms referred for their first upper gastrointestinal endoscopy, and to evaluate any association with disease.

Methods: A large survey of 3281 dyspeptic patients undergoing their first endoscopic examination was undertaken, involving 93 centres in Italy. A blood sample was taken from each participant, for measuring IgG antibodies against H. pylori using a commercially available kit, and two biopsies of the antral gastric mucosa were obtained for evaluating a Giemsa‐stained specimen.

Results: Endoscopic macroscopic diagnoses included normal mucosa (25.3%), gastroduodenitis (51.6%), gastric and duodenal ulcers (3.7 and 14.9%, respectively) and other conditions, including gastric cancer (0.8%). Overall, the seroprevalence result was 71.3% with a strong positive association with increasing age and male sex and a negative one with educational level. According to endoscopic diagnoses, the association with H. pylori seropositivity was highest for duodenal and gastric ulcer (multivariate odds ratio: 6.1 and 2.2) and lowest for carcinoma. The comparison between the results of serology and the single Giemsa‐stained specimen showed good reliability of H. pylori IgG, particularly in a subgroup (n = 2056) for which the interpretation was performed by a single dedicated pathologist: sensitivity and specificity were 92 and 78%, respectively.

Conclusion: Commercial IgG serology is a reliable tool for the assessment of H. pylori infection in large‐scale multicentre surveys. A very high seroprevalence among dyspeptic patients was confirmed, particularly in the presence of peptic disease. Factors associated with the infection were very similar to those usually reported in the general population, but male patients showed a significantly higher prevalence.
Low and high doses of rabeprazole vs. omeprazole for cure of Helicobacter pylori infection
Authors: Kositchaiwat C. , Ovartlarnporn B. , Kachintorn U. , Atisook K.
Published: November 2003

Background : Low‐dose rabeprazole‐based triple therapy was effective for Helicobacter pylori eradication in a few Japanese studies.

Aim : To compare the effectiveness of 1‐week low‐dose and high‐dose rabeprazole‐based triple therapy with those of omeprazole.

Methods : One hundred and sixty‐two H. pylori‐infected dyspeptic patients were randomized to receive twice daily for 1 week either rabeprazole 10 mg (R10), rabeprazole 20 mg (R20) or omeprazole 20 mg (O) in combination with amoxicillin 1000 mg (A) and clarithromycin 500 mg (C). H. pylori status assessment was by the CLO test and histology at entry and by the 13C‐urea breath test at 4–6 weeks after cessation of therapy.

Results : H. pylori eradication rates in intention‐to‐treat groups were 85%, 96% and 83% for R10AC, R20AC and OAC, respectively. Eradication rates in per protocol groups were 86%, 96% and 90% for R10AC, R20AC and OAC, respectively. On an intention‐to‐treat analysis, the R20AC group had a significantly higher eradication rate than did R10AC or OAC (P < 0.05). However, the higher eradication rate with R20AC did not reach statistical significance in the per protocol analysis. Drug intolerance was found in three OAC patients.

Conclusion : High‐dose rabeprazole‐based triple therapy is more effective than its low‐dose equivalent or omeprazole in eradicating H. pylori infection.
Proto‐oncogene c‐mpl is involved in spontaneous megakaryocytopoiesis in myeloproliferative disorders
Authors: LI Yan , Hetet Gilles , Kiladjian Jean‐Jacques , Gardin Claude , Grandchamp Bernard , Briere Jean
Published: November 2003

Spontaneous megakaryocytic colonies (CFU‐MK) formation without the addition of Meg‐CSA in myeloproliferative disorders (MPD) has been reported by many laboratories. The mechanism by which this occurs is still unknown. In our previous work we have found that the spontaneous colonies persisted in serum‐free agar culture although the colony cells were smaller and the colony numbers fewer than in plasma clot culture and that monoclonal antibodies against IL3, IL6 and GM‐CSF had no inhibitory effect on spontaneous CFU‐MK in both semi‐solid cultures. Recently, proto‐oncogene c‐mpl and c‐mpl ligand, thrombopoietin (TPO), have been shown to specifically participate in the regulation of normal human megakaryocytopoiesis. In order to test the hypothesis that c‐mplc‐mpl ligand pathway is involved in the spontaneous growth of megakaryocyte progenitors, we investigated mRNA expressions of c‐mpl and TPO in cells grown in serum‐free liquid culture using RT‐PCR. The c‐mpl expression was detected in the cultured cells from all nine patients (six with ET, two with PV, one with PMF) who had spontaneous CFU‐MK in clonal assays. However, none of the patients expressed TPO mRNA in these cells. Pre‐incubation of nonadherent mononuclear cells with thioester‐modified antisense oligodeoxynucleotide to c‐mpl at a concentration of 6μM significantly decreased the cloning efficiency of spontaneous megakaryocyte growth by 42.5% (P<0.05) in plasma clot assay (seven with ET, one with PV) and 69.6% (P<0.05) in serum‐free agar culture (six with ET, one with PV). In control experiments the introduction of a scrambled oligomer to antisense oligodeoxynucleotide had no such effect on spontaneous colony formation. These results indicate that c‐mpl exerts an important effect in the growth of spontaneous megakaryocytopoiesis in MPD.
Mature micromegakaryocytes: an unusual developmental pattern in term infants
Authors: Levine R. F. , Olson T. A. , Shoff P. K. , Miller M. K. , Weisman L. E.
Published: November 2003

We sought to gain perspective on platelet production in the fetus and the newborn by counting and characterizing megakaryocytes from available cord blood. Elutriation was used to isolate circulating megakaryocytes from umbilical arteries and veins obtained at scheduled caesarean sections of nine normal term fetuses. Megakaryocytes were identified by established criteria, their diameters measured, and maturation stages recorded. Large numbers of megakaryocytes, mostly mature, were found in both the umbilical arteries and veins, many times more than previously observed circulating in adult blood. In term infants more than a third of the mature megakaryocytes had unusually decreased nuclear lobation and were dwarf cells with diameters as small as 13 μm, which we considered to be micromegakaryocytes. The atypicality of these small but mature cells is seen as merely a leftward skewing in the development of megakaryocyte ploidies. We believe that in normal fetuses the extent of megakaryocyte ploidization and development is distinctive and probably regulated differently to the adult pattern.
Effects of myeloid cell growth factors on alkaline phosphatase, myeloperoxidase, defensin and granulocyte colony‐stimulating factor receptor mRNA expression in haemopoietic cells of normal individuals and myeloid disorders
Authors: Tsuruta Toshihisa , Tani Kenzaburo , Shimane Miyuki , Ozawa Keiya , Takahashi Satoshi , Tsuchimoto Daisuke , Takahashi Keisuke , Nagata Shigekazu , Sato Noriharu , Asano Shigetaka
Published: November 2003

The mRNA expressions of alkaline phosphatase (ALP), myeloperoxidase (MPO), defensin and G‐CSF receptor (G‐CSFR) in bone marrow cells of normal individuals and myeloid disorders, with or without in vitro stimulation by myeloid cell growth factors, i.e. G‐CSF, GM‐CSF and IL‐3, were examined as markers for myeloid cell differentiation in both mononuclear cell (MNC) and polymorphonuclear cell (PMN) fractions. Without any stimulation, ALP mRNA was expressed only in PMNs, G‐CSFR mRNA in PMNs were expressed stronger than in MNCs; both MPO and defensin mRNA were expressed to the same degree in both fractions. With stimulation, the ALP mRNA expression in both fractions was strongly enhanced by G‐CSF, but the expression was inhibited by GM‐CSF and/or IL‐3. MPO mRNA expression was stimulated by G‐CSF and/or GM‐CSF in MNCs. G‐CSFR mRNA expression was enhanced by G‐CSF in both fractions. Defensin mRNA expression was inhibited by G‐CSF. In cases of myelodysplastic syndrome and chronic myelogenous leukaemia which display a suppressed maturation of myeloid cells, our results demonstrated an almost normal response to these growth factors. Our results suggest that studies on these myeloid marker mRNA expressions would provide more knowledge about the differentiation state and cytokine reactivity of myeloid cells in normal individuals as well as various disorders.
Characterization of peripheral blood progenitor cells (PBPC) mobilized by filgrastim (rHuG‐CSF) in normal volunteers: dose–effect relationship for filgrastim with the character of mobilized PBPC
Authors: Tanaka Ryuhei , Matsudaira Tadahiro , Aizawa Jun , Ebihara Yasuhiro , Muraoka Kenji , Tsuji Kohichiro , Ikebuchi Kenji , Kodama Kazuo , Takaku Fumimaro , Nakahata Tatsutoshi
Published: November 2003

Filgrastim (rHuG‐CSF)‐mobilized peripheral blood progenitor cells (PBPC) in healthy Japanese volunteers were characterized in detail using two clonal cell culture systems and double‐colour flow cytometry to detect multilineage colony‐forming cells and subsets of CD34+ cells. The kinetics of PBPC during the administration of filgrastim was studied, and possible differences in the character of progenitor cells relative to given doses of filgrastim were investigated. Filgrastim was administered subcutaneously to normal volunteers for 7 d at doses of 100, 200 or 400 μg/m2 (10 per cohort). Treatment with 100 or 200 μg/m2 filgrastim was well tolerated; however, the 400 μg/m2 dose level was not completed because of bone pain and myalgia. The treatment strikingly mobilized various types of progenitor cells, including highly proliferative megakaryocytic colony‐forming cells. The number of progenitor cells peaked on days 5 and 6. The fold increase of circulating progenitor cells from the baseline value in the volunteers treated with 200 μg/m2 filgrastim was more pronounced than in those treated with 100 μg/m2. Treatment with 200 μg/m2 also released the less mature progenitor cells (i.e. mixed colony‐forming cells, CD34+/33− cells, and CD34+/HLA‐DR− cells) into circulation better than the 100 μg/m2 dose. These results suggest that daily subcutaneous injection with 200 μg/m2 filgrastim for 5 d will effectively mobilize, both qualitatively and quantitatively, PBPC in healthy donors.
A flow cytometric platelet immunofluorescence crossmatch for predicting successful HLA matched platelet transfusions
Authors: SINTNICOLAAS K. , LÖWENBERG B.
Published: November 2003

Platelet crossmatching may provide a useful way of selecting donors for effective platelet transfusions in patients refractory to random donor platelet concentrates due to alloimmunization. We assessed the predictive value of a flow cytometric platelet immunofluorescence crossmatch test for the outcome of HLA matched platelet transfusions in a group of alloimmunized patients. Platelet immunofluorescence (PIFT) crossmatches were performed for 104 HLA‐matched platelet transfusions administered to 30 patients. A negative PIFT crossmatch correctly predicted a successful platelet transfusion (1 h post‐transfusion platelet recovery >20%) in 56/75 (75%) cases. We also considered non‐immunological factors that, in combination with alloimmunization, might have contributed to an unsuccessful transfusion result, i.e. fever, septicaemia, splenomegaly, disseminated intravascular coagulation and bleeding. The predictive value of a negative PIFT crossmatch was better when these non‐immunological factors were absent [48/59 (81%) correct predictions] than when these factors were present [8/16 (50%) correct predictions] (P=0.01; chi‐square test).  The effect of ABO incompatibility between donor and recipient on the predictive value of the PIFT crossmatch was also analysed. Positive PIFT crossmatches occurred more frequently in ABO incompatible donor–recipient combinations [in 18/28 (64%) cases] than in ABO‐compatible donor–recipient combinations [in 11/76 cases (14%)] (P<0.001, chi‐square test). Successful platelet transfusions were observed on 53/76 (70%) occasions in ABO compatible transfusions as compared to 16/28 (57%) in ABO incompatible transfusions. This difference was not statistically significant (P=0.23; chi‐square test). Consequently, a negative PIFT crossmatch appeared to be non‐predictive for the transfusion outcome in cases of ABO incompatibility between donor and recipient.  We conclude that the PIFT crossmatch for platelet donor selection in addition to matching for HLA antigens, is predictive for the outcome of ABO compatible transfusions in alloimmunized recipients and prediction levels are increased when non‐immunological causes for platelet refractoriness are absent.
Serum erythropoietin levels in paroxysmal nocturnal haemoglobinuria: implications for therapy
Authors: McMullin M. F. , Hillmen P. , Elder G. E. , Lappin T. R. J. , Luzzatto L.
Published: November 2003

In order to assess the rationale and possible indications for the use of recombinant erythropoietin in paroxysmal nocturnal haemoglobinuria (PNH), we have measured endogenous erythropoietin (Epo) levels in 18 patients with PNH and in 44 patients with iron deficiency anaemia (IDA). In both groups of patients we found a significant inverse correlation between Epo and haemoglobin (Hb). However, the mean Epo level was significantly higher in the PNH group (385 mU/ml) than in the IDA group (136 mU/ml). The range of Epo levels at any given Hb was greater in the PNH group than in the IDA group. There was a significant positive correlation between Epo and absolute reticulocyte count. Since Epo administration is unlikely to benefit patients with high levels of endogenous Epo, we conclude that in the majority of patients with PNH there is no indication for treatment with Epo.
A case of refractory anaemia with p53 point mutation at codon 249 (AGG to ATG)
Authors: Kikukawa Masayuki , Aoki Naoto , Mori Mayumi
Published: November 2003

Although mutations of the p53 tumour‐suppressor gene have been observed occasionally in the advanced stages of myelodysplastic syndromes (MDS), they have not been detected in early refractory anaemia (RA) or RA with ring sideroblasts phases.

Using single‐strand conformation polymorphism analysis and direct sequencing, we searched for p53 mutations in a patient who progressed from RA to overt leukaemia. A p53 mutation at codon 249 (AGG to ATG) was observed in RA, RA with excess of blasts in transformation and overt leukaemia.

We describe a case of MDS with p53 mutation at codon 249 detected in the RA phase.
The Role of Intensive Therapy and Autologous Blood and Marrow Transplantation for Chemotherapy‐Sensitive Relapsed and Primary Refractory Non‐Hodgkin’s Lymphoma: Identification of Major Prognostic Groups
Authors: Prince H. Miles , Imrie Kevin , Crump Michael , Stewart A. Keith , Girouard Caroline , Colwill Richard , Brandwein Joseph , Tsang Richard W. , Scott J. Gerald , Sutton David M. C. , Pantalony Dominic , Carstairs Kelvin , Sutcliffe Simon B. , Keating Armand
Published: November 2003

Patients with intermediate grade non‐Hodgkin's lymphoma (NHL) who relapse or fail to achieve a complete remission after anthracycline‐containing induction regimens have a poor outcome with conventional‐dose salvage treatment. This outcome may be improved with intensive therapy and autologous transplantation (ABMT) but even in patients with proven chemotherapy‐sensitive disease, relapse rates of up to 60% are observed. Reliable and powerful prognostic indicators are needed to identify appropriate patients for this expensive procedure and those subjects to whom alternative or additional treatment should be offered. We were interested in testing the hypothesis that tumour burden, and hence remission status immediately prior to transplant, is an important prognostic indicator of survival. We aggressively treated patients with conventional‐dose salvage chemotherapy to maximum tumour response, and tested, by multivariate regression analysis, predictors of outcome post‐transplant. We studied 81 consecutive patients with intermediate grade and immunoblastic NHL who achieved either a partial (PR) or complete remission (CR) following repetitive cycles of conventional‐dose salvage therapy. Intensive therapy consisted of etoposide (60 mg/kg) and intravenous melphalan (160–180 mg/m2) with or without total body irradiation (TBI) followed by infusion of autologous unpurged bone marrow and/or blood cells. The predicted 4‐year survival and progression‐free survival (PFS) with a median follow‐up of 37 months was 58% and 48% (95% confidence interval (CI) 37–55%), respectively. The only factor predictive of outcome was remission status at transplant (P = 0.0001). The PFS at 4 years for the CR group was 61% (95% CI 53–75%). In contrast, only 25% (95% CI 11–40%) of patients undergoing autotransplant in PR were progression free at 4 years. We conclude that remission status at transplant after maximum tumour reduction is a powerful prognostic indicator.
Transient normal platelet counts and decreased requirement for interferon during pregnancy in essential thrombocythaemia
Authors: Shpilberg Ofer , Shimon Ilan , Sofer Orit , Dolitski Mordechai , Ben‐Bassat Isaac
Published: November 2003

We report on the remarkable decrease in the platelet counts during pregnancy in two women with essential thrombocythaemia following treatment with recombinant interferon‐α (r‐IFN‐α). Prior to pregnancy, the first patient was treated for 10 months with r‐IFN‐α 3×106 units/d six times per week, and the platelet count ranged between 750 and 800×109/l. Starting from the sixth week of gestation, the platelet count decreased to normal levels and remained so, resulting in a lower r‐IFN‐α requirement. Following successful delivery of a healthy newborn an abrupt rise of the platelet count to pre‐gestation values was observed, necessitating increased r‐IFN‐α dosage as before pregnancy. The second patient when she became pregnant had been treated with r‐IFN‐α 3×106 units/d six times per week for 10 weeks. Starting from the 24th week of gestation the platelet count decreased, and despite reduction in the dose of r‐IFN‐α reached normal values at the time of delivery. The exact mechanism for the platelet count normalization during pregnancy is unclear, and several possibilities are discussed.
Eosinophilia associated with proliferation of CD3+4−8−αβ+ T cells with chromosome 16 anomalies
Authors: Kitano Kiyoshi , Ichikawa Naoaki , Mahbub Bilkis , Ueno Mayumi , Ito Toshiro , Shimodaira Sigetaka , Kodaira Hiroshi , Ishida Fumihiro , Kobayashi Hikaru , Saito Hiroshi , Okubo Yoshio , Enokihara Hideo , Kiyosawa Kendo
Published: November 2003

We describe a patient with eosinophilia and an abnormal CD3+4−8−αβ+ T‐cell population. Chromosomal analysis of sorted CD3+4−8− cells revealed abnormal karyotypes on chromosome 16. In the presence of IL‐2 the production of IL‐5 from CD3+4−8− cells was higher than that from CD3+4+/8+ cells. Eosinophil survival‐enhancing activity in the patient serum was inhibited by a combination of anti‐IL‐5 and anti‐GM‐CSF monoclonal antibodies. These data suggest that increased production of IL‐5 and GM‐CSF from the abnormal CD3+4−8− cells might cause eosinophilia.
Bone marrow histology and CD34 immunostaining in the prognostic evaluation of primary myelodysplastic syndromes
Authors: Oriani A. , Annaloro C. , Soligo D. , Pozzoli E. , Cortelezzi A. , Lambertenghi Deliliers G.
Published: November 2003

The prognostic impact of bone marrow biopsy (BMB) histology and CD34 immunoreactivity was compared with that of the more conventional parameters (the FAB diagnosis, peripheral blood values, percentage of BM blasts and some common prognostic scores) in 100 MDS patients. Statistical correlations among the cytological, haematological, histological and immunohistochemical parameters and their relationship with clinical outcome were searched for. At univariate analysis, FAB classification (P < 0.001), pattern of blastic infiltration at BMB (P < 0.005), presence of CD34+ aggregates (P < 0.0005), percentage of blasts in BM aspirate (P < 0.0001) and percentage of CD34 positivity (P < 0.0001) proved to be linked to leukaemic transformation and, except for FAB classification, retained a high degree of prognostic significance in terms of survival. Leukaemic transformation occurred in 16/18 patients simultaneously presenting ‘large’ blastic infiltrates at BMB and CD34+ aggregates (P < 0.00001); 9/17 evaluable patients died within 12 months of diagnosis (P < 0.001). Discriminant functions for leukaemic transformation and survival did not offer any advantage over univariate analysis in the prognostic work‐up. The results indicate that the size of blastic aggregates and CD34 positivity allowed patients with a worse prognosis to be identified irrespective of their FAB subtype, but the prognostic impact is considerably greater when both parameters are simultaneously taken into account, as testified by the restricted and homogenous subgroup of patients with both ‘large’ and CD34‐positive aggregates.
Haematological abnormalities in Shwachman‐Diamond syndrome
Authors: Smith Owen P. , Hann Ian M. , Chessells Judith M. , Reeves Brian R. , Milla Peter
Published: November 2003

We have analysed the haematological parameters in 21 patients with Shwachman‐Diamond syndrome (SDS) seen over a 25‐year period at our institution. Neutropenia, although present in all patients, was intermittent in two‐thirds, constant in the rest and was associated with impaired chemotaxis in all of those patients tested. Fetal haemoglobin (HbF) was elevated in 80% of the patients at some stage, and anaemia and thrombocytopenia was documented in 66% and 24% respectively. Bone marrow samples were taken in over half of the patients. Myelodysplastic syndrome (MDS) developed in seven (33%) patients, five of whom had acquired clonal structural chromosome abnormalities in their bone marrows. In five of the patients with MDS (24%) transformation to acute myeloid leukaemia occurred. Like other constitutional bone marrow failure syndromes, SDS has a predilection to leukaemic transformation hitherto assumed to be in the region of 5–10%. The data presented here suggest that this figure probably represents an underestimate. Shwachman‐Diamond syndrome is an interesting model of leukaemia development and greater understanding of the clinical spectrum of this rare disorder should produce further insights into its pathobiology.
Elevated platelet count features the variant type of BCR/ABL junction in chronic myelogenous leukaemia
Authors: YAMAGATA TETSUYA , MITANI KINUKO , Kanda Yoshinobu , Yazaki Yoshio , Hirai Hisamaru
Published: November 2003

A variant form of BCR/ABL junction was identified in a patient with chronic myelogenous leukaemia (CML). The BCR/ABL fusion mRNA of this patient showed in‐frame junction between BCR exon c3 and ABL exon 2. Although the diagnosis of CML was made, the patient showed clinical features of essential thrombocythaemia (ET) rather than that of typical CML. Treatment with interferon‐α showed no cytogenetic response. The c3‐a2 type of BCR/ABL junction seems to be associated with elevated platelet count and thus could form a novel clinical entity different from typical CML.
THE RETINOBLASTOMA GENE (RB1) IN ACUTE MYELOID LEUKAEMIA: ANALYSIS OF GENE REARRANGEMENTS, PROTEIN EXPRESSION AND COMPARISON OF DISEASE OUTCOME
Authors: Jamal Rahman , Gale Rosemary E. , Thomas N. Shaun B. , Wheatley Keith , Linch David C.
Published: November 2003

The occurrence of retinoblastoma gene abnormalities in a large subset of various malignancies suggests an important role for this tumour suppressor gene in carcinogenesis, but this varies considerably from one tumour type to another and results in patients with acute myeloid leukaemia (AML) have been controversial. We analysed 106 AML patients and 18 normal controls for RB1 gene rearrangements and 86 AML patients for RB protein (pRB) expression. Southern blot analysis detected no gross gene rearrangements, but several restriction enzyme polymorphisms were observed. By Western blot analysis, 20 patients (23%) had no detectable pRB protein and seven (8%) had truncated pRB bands. Discordance between the DNA and protein data suggests that there may be minor deletions and point mutations in the RB1 gene or abnormalities in the proteins regulating the expression of pRB. No significant differences in the frequency of attainment of complete remission or length of survival were observed between patients with normal and abnormal pRB.
Expression and function of receptors for extracellular matrix molecules in the differentiation of human megakaryocytes in vitro
Authors: Mossuz Pascal , Schweitzer Annie , Molla Annie , Berthier Rolande
Published: November 2003

The role of adhesive interactions with the extracellular matrix components of the bone marrow (BM) stroma has been widely studied in the differentiation of erythroid and myelomonocytic cells, but not in the megakaryocytic lineage. The development of efficient culture techniques for the production of megakaryocytes (Mks) from CD34+ purified BM cells, enables the study of the expression and function of adhesion receptors for collagen (VLA‐2), fibronectin (VLA‐4 and VLA‐5) and laminin (VLA‐6) during the maturation of Mks. We have shown that a significant percentage of CFU‐MK (roughly 20%) adhere to fibronectin but not to collagen and laminin. The expression and adhesion of Mks developing in liquid culture from BM‐CD34+ cells were tested at days 4, 7 and 10 of incubation. The expression of VLA‐2, VLA‐5 and VLA‐6 on day 10 cultured Mks enabled purification of intermediate and large polyploid Mks by FACS sorting whereas VLA‐4 appeared to label only immature Mks and myeloid cells. We observed that only a small proportion of mature Mks was able to adhere to collagen without spreading at day 10 of culture, whereas 30% of Mks adhered to fibronectin as early as day 4 of incubation, 40% of which also attached to laminin. Our data suggest that VLA‐4 may be involved in the adhesion of CFU‐MK and immature Mks on fibronectin, then replaced by VLA‐5 in the final stages of maturation. The expression of VLA‐6 and the number of adherent Mks on laminin increased sharply between day 7 and 10 of incubation. A number of mature polyploid Mks found in day 10 of culture exhibited characteristic features of intense spreading on laminin and fibronectin which were not observed on collagen.
Leukaemia presenting as marrow hypoplasia: molecular detection of the leukaemic clone at the time of initial presentation
Authors: Morley A. A. , Brisco M. J. , Rice M. , Snell L. , Peng L‐M. , Hughes E. , Neoh S‐H. , Sykes P. J.
Published: November 2003

Occasional cases of transient marrow hypoplasia in childhood evolve into acute leukaemia. We studied two children who presented with marrow hypoplasia following infection and who developed acute lymphoblastic leukaemia 2–3 months later. A simple polymerase‐chain‐reaction (PCR) test for monoclonality showed that immunoglobulin heavy‐chain gene rearrangements of the same size were present at the times of both hypoplasia and leukaemia, and DNA sequencing confirmed identity of these rearrangements. PCR‐based quantification, using patient‐specific primers, showed in both patients that the leukaemic clone made up 20–25% of the marrow cells during hypoplasia. In contrast, four patients with typical aplastic anaemia showed only polyclonal B‐cell populations in the marrow. We conclude that the leukaemic clone was already present at the time of hypoplasia in the two index patients and that in future a simple PCR test for monoclonality could be used to screen patients with marrow aplasia or hypoplasia for the presence of a monoclonal B‐cell population. Patients with monoclonal populations could then be monitored carefully for subsequent development of leukaemia.
Mycosis fungoides and Sezary syndrome are not associated with HTLV‐I infection: an international study
Authors: Bazarbachi A. , Soriano V. , Pawson R. , Vallejo A. , Moudgil T. , Matutes E. , Peries J. , Molina A. , De The H. , Schulz T. F. , Catovsky D. , Gill P. S.
Published: November 2003

Association between mycosis fungoides (MF), its leukaemic variant Sezary syndrome (SS) and the human T‐cell lymphotropic virus type‐I (HTLV‐I) has been controversial, with the reported incidence of infection varying between 0% and nearly 100%. We studied 127 patients (85 MF, 28 SS, five Sezary cell leukaemia, four lymphomatoid papulosis, and five unspecified cutaneous T‐cell lymphomas (CTCL)) originating from Europe (France, Spain, U.K., Portugal) or from U.S.A. (California) for the presence of HTLV‐I infection markers. HTLV‐I and ‐II serology were performed on 78 patients using standard immunological methods. Reverse transcriptase (RT) assay was also performed in 26 cases using an RT‐PCR‐based method of high sensitivity. Molecular analyses were performed on 215 DNA samples (121 from fresh PBMCs, 26 from PBMCs after short‐term culture and 68 from skin lesions) by PCR amplification using HTLV‐I and ‐II gag, pol, env, pX and LTR specific primers.

Immunological tests were negative except for two sera which were indeterminate. PCR with all HTLV‐I and ‐II primer pairs showed negative results in all 215 samples investigated. No RT activity was detected in short‐term PBMC cultures of any of the 26 cases studied. The results of this large study from five different countries clearly indicate that MF and SS are not associated with HTLV‐I infection.
Magnetic resonance imaging patterns in patients with multiple myeloma
Authors: Kusumoto Shuya , Jinnai Itsuro , Itoh Katsuro , Kawai Nobutaka , Sakata Tohru , Matsuda Akira , Tominaga Kazunori , Murohashi IKUO , Bessho Masami , Harashima Kunitake , Heshiki Atsuko
Published: November 2003

Sixty‐one consecutive patients with multiple myeloma were studied with magnetic resonance (MR) imaging of the spine. Sagittal T1‐weighted and short inversion time (TI) inversion recovery (STIR) images were obtained. The MR patterns of the bone marrow were classified as diffuse (D) (n = 26), nodular (N) (n = 11), D + N (n = 13) or normal (n) (n = 11). Abnormal patterns were seen in 50 (82%) of the 61 patients. Correlations were found between the MR imaging patterns and some laboratory findings (WBC, haematocrit, platelet count, serum albumin, and percentage of marrow plasmacytosis). The survival of the patients with abnormal MRI patterns was significantly poorer than that of the patients with normal patterns. However, the survival of patients with a nodular pattern did not differ from those with a normal pattern. The MR imaging pattern of the bone marrow in patients with multiple myeloma is a useful factor in the assessment of prognosis.
Total absence of protein 4.2 and partial deficiency of band 3 in hereditary spherocytosis
Authors: Kanzaki Akio , Hayette Sandrine , Morlé Laurette , Inoue Fumihide , Matsuyama Reiko , Inoue Takafumi , Yawata Ayumi , Wada Hideho , Vallier Agnes , Alloisio Nicole , Yawata Yoshihito , Delaunay Jean
Published: November 2003

Unlike previously reported cases with total protein 4.2 deficiency due to mutations in the EPB42 gene, we describe a total deficiency in protein 4.2 with normal EPB42 alleles. Hereditary spherocytosis (HS) was observed in a Japanese woman (unsplenectomized) and her daughter (splenectomized). The mother showed a partial deficiency in band 3 and a proportional reduction in protein 4.2. She was heterozygous for a novel allele of the EPB3 gene, allele Okinawa, which contains the two mutations that define the Memphis II polymorphism (K56E, AAG → GAG, and P854L, CCG → CTG) and, additionally, the mutation: G714R, GGG → AGG, located in a highly conserved position of transmembrane segment 9. The latter change was responsible for HS. In trans to allele Okinawa, the daughter displayed allele Fukuoka: G130R, GGA → AGA, an allele known to alter the binding of protein 4.2 to band 3. The daughter presented with a more pronounced decrease of band 3, and lacked protein 4.2, resulting in aggravated haemolytic features. Although the father was not available for study, heterozygosity for allele Fukuoka has been documented in another individual who showed no clinical or haematological signs, and a normal content of band 3. We suggest that band 3 Okinawa binds virtually all the protein 4.2 in red cell precursors, band 3 Fukuoka being unable to do so, and that the impossibility of band 3 Okinawa incorporation into the membrane leads to degradation of the band 3 Okinawa protein 4.2 complex. In contrast, band 3 Fukuoka, free of bound protein 4.2, could then incorporate normally into the bilayer. Thus, protein 4.2 would not appear in the daughter's red cell membrane.
CD34+ peripheral blood progenitor cell and monocyte derived dendritic cells: a comparative analysis
Authors: Herbst Birgit , Köhler Gabriele , Mackensen Andreas , Veelken Hendrik , Mertelsmann Roland , Lindemann Albrecht
Published: November 2003

Dendritic cells (DC) have been generated in vitro from either CD34+ haemopoietic progenitor cells (HPC) or peripheral blood monocytes (Mo) in the presence of specific cytokine combinations, including granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Since differences between DC from either source may be important for the clinical use of these antigen‐presenting cells (APC), a comparative analysis was performed. HPC were expanded in the presence of interleukin (IL)‐3, IL‐6 and stem cell factor (SCF) (days 1–7) and subsequently induced by IL‐4 + GM‐CSF (days 8–26) to differentiate to Langerhans‐type cells (pLC). The latter cytokines were similarly used to generate Mo‐derived LC (mLC). Maturation of both cell types, pLC and mLC, to interdigitating DC‐type cells (iDC) was induced by tumour necrosis factor‐α (TNF‐α) or lipopolysaccharide (LPS). Analysis of mLC/pLC and miDC/piDC with respect to morphology, phenotype, antigen uptake and presentation revealed a high similarity of DC from either source. The majority of mLC, however, exhibited a more mature differentiation stage, compared to pLC, evidenced from lower numbers of multilaminar MHC class II compartments and less efficient APC function for extracellular protein antigens. Although macropinocytosis was performed by LC, neither LC nor iDC from either source were able to take up 0.5 μm latex beads. However, phagocytosis of 0.5 μm and 1 μm beads was performed by Mo that could subsequently be induced to become iDC, thus providing the unique opportunity to present phagocytosed material in DC‐type fashion. Mo may be the preferential source for clinical use of iDC‐type cells since preparation and culture are easier to perform and are less costly while APC function is similar to HPC‐derived iDC.
The effect of thrombin on the dynamic exchange between intraplatelet and extraplatelet fibrinogen
Authors: Vila Virtudes , Martínez‐Sales Vicenta , Réganon Edelmiro , Aznar Justo
Published: November 2003

We examined the distribution of platelet fibrinogen and the exchange between intra‐ and extra‐platelet fibrinogen in unstimulated and thrombin‐stimulated platelets. In unstimulated platelets 60% of platelet fibrinogen was found in the soluble platelet fraction and 40% in the insoluble one. In platelets activated with thrombin, changes took place in the distribution of intraplatelet fibrinogen but not in the total fibrinogen content. At 0.5 U/ml of thrombin the fibrin(ogen) content of the insoluble and soluble fractions was approximately 80% and 20%, respectively. When we evaluated how extraplatelet fibrinogen affects the content and distribution of intraplatelet fibrinogen, we found that when unlabelled fibrinogen was added to unstimulated and thrombin‐stimulated platelets the content and distribution of intraplatelet fibrinogen remained unaltered. However, when 125I‐fibrinogen was added, it was incorporated into unstimulated and thrombin‐stimulated platelets. In unstimulated platelets, 70% of the incorporated 125I‐fibrinogen was in the soluble fraction and 30% in the insoluble. In thrombin‐stimulated platelets the distribution of the incorporated 125I‐fibrinogen was 62% and 38% in soluble and insoluble fractions respectively. MoAb to GPIIb–IIIa produced 80% and 60% inhibition of 125I‐fibrinogen incorporation by unstimulated and thrombin‐stimulated platelets. Our data showed dynamic exchange between intraplatelet and extraplatelet fibrinogen both in unstimulated and thrombin‐stimulated platelets mediated mainly by GPIIb–IIIa.
Activity of TNF‐related apoptosis‐inducing ligand (TRAIL) in haematological malignancies
Authors: Snell Virginia , Clodi Katharina , Zhao Shourong , Goodwin Raymond , Thomas Elaine K. , Morris Stephan W. , Kadin Marshall E. , Cabanillas Fernando , Andreeff Michael , Younes Anas
Published: November 2003

T‐cell cytotoxicity is primarily mediated by two cell surface proteins, Fas ligand (FasL) and tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL), and intracellular perforin and granzyme granules. FasL‐deficient and perforin‐deficient T lymphocytes maintain cytotoxicity but fail to induce graft‐versus‐host disease (GVHD) when transplanted into mice, suggesting that GVHD and graft‐versus‐tumour (GVT) effects can be dissociated, and that TRAIL is not involved in the pathogenesis of GVHD. Because TRAIL could mediate a favourable GVT effect it became important to study the spectrum of its activity and to investigate factors that can dissociate its expression from FasL. TRAIL induced apoptosis in 11/41 (27%) tumour specimens of haematological origin compared to 16/41 (39%) induced by FasL. Although eight specimens were sensitive to both FasL and TRAIL, no synergism was observed between these two ligands. TRAIL induced apoptosis in a dose and time dependent manner with an ED50 of 0.5 μg/ml and EDmax of 1 μg/ml. TRAIL activity was not reduced by the over‐expression of the multidrug resistant (MDR) protein, and was not enhanced by 9‐cis retinoic acid (RA), which can down‐regulate bcl‐2 protein. Both ligands were simultaneously up‐regulated in normal peripheral blood lymphocytes in response to IL‐2, IL‐15 and anti‐CD3 antibody, whereas IL‐10 had no effect. Together, our data show that (1) TRAIL can mediate cell death in a variety of human haematological malignancies, (2) resistance to TRAIL is not mediated by MDR protein, (3) the lack of synergy between TRAIL and FasL suggests that either one is sufficient to mediate T‐cell cytotoxicity, and (4) within the panel of cytokines tested, the expression of TRAIL and FasL could not be dissociated.
Relationship of T leukaemias with cerebriform nuclei to T‐prolymphocytic leukaemia: a cytogenetic analysis with in situ hybridization
Authors: BRITO‐BABAPULLE VASANTHA , HODA MALJAIE S. , MATUTES ESTELLA , HEDGES MICHAEL , YUILLE MARTIN , CATOVSKY DANIEL
Published: November 2003

Sezary cell leukaemia (SCL) is a mature T‐cell leukaemia with characteristic cerebriform nuclei, whereas Sezary syndrome (SS) involves a mature T‐cell lymphoma with a similar nuclear morphology. We have examined these diseases by cytogenetics, chromosome painting and fluorescence in situ hybridization (FISH). Both diseases had complex cytogenetic abnormalities. All three cases of SCL investigated had inv(14)(q11;q32) and two had iso(8q). No case of SS had these abnormalities but, instead, iso(17q) or 17p+ was present in the three cases of SS investigated and FISH indicated loss of heterozygosity due to deletion of a region at 17p13 that included the tumour suppressor gene P53, implicating it in this malignancy. One case of SCL had iso(17q). The abnormalities of chromosomes 8 and 14 in SCL are commonly observed in T‐prolymphocytic leukaemia (T‐PLL) and suggest that SCL may be a variant of T‐PLL rather than of SS.
Unrelated donor bone marrow transplantation for children and adolescents with aplastic anaemia or myelodysplasia
Authors: DAVIES STELLA M. , WAGNER JOHN E. , DEFOR TODD , BLAZAR BRUCE R. , KATSANIS EMMANUEL , KERSEY JOHN H. , ORCHARD PAUL J. , MCGLAVE PHILLIP B. , WEISDORF DANIEL J. , RAMSAY NORMA K. C.
Published: November 2003

Allogeneic transplantation from an HLA‐matched family member has been shown to be effective in reconstituting normal haemopoiesis in young people with severe cytopenias, classified as myelodysplastic syndrome (MDS) or severe aplastic anaemia (SAA). Unrelated donor transplant is a therapeutic choice for patients without a suitable family member donor. We report the outcome of seven patients < 20 years old with SAA and 10 with MDS treated with BMT from an HLA A,B DRB1 matched (n =8) or A or B locus mismatched (n =9) unrelated donor at the University of Minnesota between March 1988 and August 1995. Primary graft failure occurred in two patients and secondary graft failure in one, who was subsequently successfully engrafted with a second donor marrow infusion. Grades II–IV GVHD occurred in 10/16 (63%), and grades III–IV in 6/16 (37%) evaluable patients. Nine of the 17 patients (six with MDS and three with SAA) survive with full donor chimaerism, a median of 1.2 years post‐BMT (range 3 months to 7 years). We recommend early referral for consideration of unrelated donor BMT for young patients with MDS, and patients with SAA without response to immunosuppression.
The relative importance of the X‐linked FCP locus and β‐globin haplotypes in determining haemoglobin F levels: a study of SS patients homozygous for βS haplotypes
Authors: CHANG YEN‐PEI C. , MAIER‐REDELSPERGER MICHELINE , SMITH KIRBY D. , CONTU LICINIO , DUCROCQ ROLANDE , DE MONTALEMBERT MARIANE , BELLOY MARIE , ELION JACQUES , J. DOVER GEORGE , GIROT ROBERT
Published: November 2003

Five factors have been hypothesized to influence the 20‐fold variation in fetal haemoglobin (Hb F) levels in sickle cell anaemia (SS): age sex, α‐globin gene number, β‐globin haplotype, and the X‐linked F‐cell production locus (FCP) that regulates the production of Hb F containing erythrocytes (F cells). We analysed the association of these factors with Hb F levels in 112 SS patients living in France who are homozygous for the three common African β‐globin haplotypes (Benin, Bantu or Central African Republic and Senegal). We found that: (1) FCP accounts for about 40% of the overall variation in Hb F levels, (2) when the FCP influence is removed, β‐globin haplotype is associated with 14% of the remaining Hb F variation, and (3) the other factors have little influence. Comparison with our previous study of SS individuals in Jamaica leads to the following conclusions: (1) the X‐linked FCP locus is a major determinant of Hb F levels in SS disease, (2) factors linked to the β‐globin haplotype have only a small effect on the variation in Hb F levels, in either the homozygous or heterozygous state, and (3) approximately half of the variation in Hb F levels still remains to be explained.
Comparison of immunofiltration assay of plasma D‐dimer with diagnostic imaging in deep vein thrombosis
Authors: KILLICK S. B. , PENTEK P. G. , MERCIECA J. E. , CLARKE M. F. , BEVAN D. H.
Published: November 2003

This prospective study was designed to assess the diagnostic sensitivity, specificity and negative predictive value of the NycoCard D‐dimer plasma immunofiltration assay in patients with suspected deep vein thrombosis (DVT) confirmed by ultrasonography/venography. 84 medical patients were recruited: 43 patients (51%) had proven venous thrombosis, 33 by venography and 10 by ultrasonography. The sensitivity of NycoCard D‐dimer in patients with DVT was 95.3%, the specificity was 22.0% and the negative predictive value was 81.8%. An algorithm including the NycoCard D‐dimer test for the acute management of DVT is proposed. This would enable low‐risk patients to be discharged early from hospital, without imaging or anticoagulant therapy.