bluuebunny/crossref_metadata_2025 huggingface dataset:
Seven‐day treatment for Helicobacter pylori infection: ranitidine bismuth citrate plus clarithromycin and tetracycline hydrochloride
Authors: WILLIAMS M. P. , HAMILTON M. R. , SERCOMBE J. C. , POUNDER R. E.
Published: November 2003

Background: Dual therapy with ranitidine bismuth citrate plus clarithromycin twice daily for 14 days is an effective regimen for eradicating Helicobacter pylori infection.

Aim: To determine whether this regimen can be improved by the addition of a second antibiotic, tetracycline hydrochloride, whilst reducing the duration of treatment to 7 days.

Methods: Sixty‐one out‐patients were enrolled to this open treatment study. All had H. pylori infection, as determined by 13C‐urea breath test and, for those undergoing endoscopy, by rapid urease test. Patients were treated with ranitidine bismuth citrate 400 mg, clarithromycin 500 mg and tetracycline hydrochloride 500 mg all twice daily for 7 days. Eradication of H.␣pylori was assessed by two separate 13C‐urea breath tests, the first 28–68 days after the completion of treatment, the second 28–162 days later. H. pylori infection was considered cured if both tests were negative.

Results: All 61 patients were included in the intention‐to‐treat efficacy analysis. Successful eradication of H.␣pylori was achieved in 55/61 patients (90 %; 95% CI: 82–98%). Fifty‐nine out of sixty‐one patients reported 100% compliance; one patient missed a single dose of medication and the other withdrew at 48 h due to nausea and vomiting. Minor adverse events were reported by 30/61 patients.

Conclusion: One‐week triple therapy with ranitidine bismuth citrate, clarithromycin and tetracycline, all twice daily, is a safe and well‐tolerated regimen which eradicates H. pylori in 90% of infected patients.
Influence of tegaserod on proximal gastric tone and on the perception of gastric distension
Authors: Tack J. , Vos R. , Janssens J. , Salter J. , Jauffret S. , Vandeplassche G.
Published: November 2003

Background: Tegaserod, a 5‐hydroxytryptamine‐4 receptor agonist, enhances gastric emptying, but its effects on proximal stomach function have not been studied.

Aim: To study the effect of tegaserod on gastric compliance, accommodation and perception of distension in humans.

Methods: Nineteen healthy volunteers (10 females; mean age, 23.9 years) were studied on three separate occasions after 7 days of treatment with placebo, tegaserod 2 mg b.d. or tegaserod 6 mg b.d. in a double‐blind, randomized, three‐way cross‐over design. After the introduction of a barostat bag, stepwise distensions were performed to determine gastric compliance and sensitivity, and a mixed liquid meal was administered in isobaric mode to assess accommodation.

Results: Tegaserod had no effect on the pressures or volumes inducing first perception or discomfort. Tegaserod 6 mg b.d. enhanced fasting gastric compliance compared with placebo. Pre‐prandial and post‐prandial intra‐balloon volumes were significantly higher after 6 mg b.d. than after placebo. Both tegaserod 2 and 6 mg b.d. shortened the time to maximum post‐prandial intra‐balloon volume. The amplitude of meal‐induced gastric relaxation (post‐prandial minus pre‐prandial volumes) did not differ between the treatment arms.

Conclusion: In humans, tegaserod allows for larger intra‐balloon volumes both before and after a meal. These findings warrant the investigation of the therapeutic potential of tegaserod in dyspeptic patients with impaired accommodation.
Ursodeoxycholic acid — adverse effects and drug interactions
Authors: Hempfling W. , Dilger K. , Beuers U.
Published: November 2003

Background : Ursodeoxycholic acid is increasingly being used for the treatment of chronic cholestatic liver diseases. It appears to be generally well tolerated, but a systematic review on drug safety is lacking.

Aim : As experimental data suggest a role of bile acids in the regulation of hepatic drug metabolism at both the transcriptional and post‐transcriptional level, the literature was screened for adverse drug reactions and drug interactions related to ursodeoxycholic acid.

Methods : A systematic review of the literature was performed using a refined search strategy to evaluate the adverse effects of ursodeoxycholic acid and its interactions with other drugs.

Results : Ursodeoxycholic acid caused diarrhoea in a small proportion of patients. Rare skin reactions were due to drug adjuvants rather than the active substance. Decompensation of liver cirrhosis was reported after the administration of ursodeoxycholic acid in single cases of end‐stage primary biliary cirrhosis. Recurrent right upper quadrant abdominal pain was incidentally observed. The absorption of ursodeoxycholic acid was impaired by colestyramine, colestimide, colestipol, aluminium hydroxide and smectite. Metabolic drug interactions were reported for the cytochrome P4503A substrates, ciclosporin, nitrendipine and dapsone.

Conclusions : Ursodeoxycholic acid is generally well tolerated. Drug absorption interactions with anion exchange resins deserve consideration. Metabolic interactions with compounds metabolized by cytochrome P4503A are to be expected.
Is there any association between myocardial infarction, gastro‐oesophageal reflux disease and acid‐suppressing drugs?
Authors: Johansson S. , Wallander M.‐A. , Ruigómez A. , García Rodríguez L. A.
Published: November 2003

Background : A link between gastro‐oesophageal reflux disease and coronary heart disease has been suggested.

Aim : To estimate the incidence of myocardial infarction in patients with newly diagnosed gastro‐oesophageal reflux disease in comparison with that in the general population.

Methods : A population‐based cohort study was performed in the UK. Patients aged 18–79 years with a first diagnosis of gastro‐oesophageal reflux disease (n = 7084) were identified and a group of 10 000 patients free of gastro‐oesophageal reflux disease were sampled. A nested case–control analysis was performed to assess the risk factors for myocardial infarction.

Results : The incidence of myocardial infarction in the general population was 4.0 per 1000 person‐years [95% confidence interval (CI), 3.2–4.9] and 5.1 per 1000 person‐years (95% CI, 4.1–6.4) in patients with gastro‐oesophageal reflux disease. The relative risk of myocardial infarction in patients with gastro‐oesophageal reflux disease was 1.4 (95% CI, 1.0–1.9). The increased risk of myocardial infarction was limited to the immediate days after the diagnosis of gastro‐oesophageal reflux disease. Previous chest pain was an important predictor of myocardial infarction in patients free of gastro‐oesophageal reflux disease. No association was found between the use of acid‐suppressing drugs and the risk of myocardial infarction.

Conclusion : Our results suggest that gastro‐oesophageal reflux disease is not an independent predictor of myocardial infarction. Rather, the increased risk of myocardial infarction in patients with gastro‐oesophageal reflux disease in the immediate days after diagnosis indicates that prodromal ischaemic symptoms were misinterpreted as reflux symptoms.
Therapeutic drug monitoring: antiarrhythmic drugs
Authors: Campbell T. J. , Williams K. M.
Published: November 2003

Antiarrhythmic agents are traditionally classified according to Vaughan Williams into four classes of action. Class I antiarrhythmic agents include most of the drugs traditionally thought of as antiarrhythmics, and have as a common action, blockade of the fast‐inward sodium channel on myocardium. These agents have a very significant toxicity, and while they are being used less, therapeutic drug monitoring (TDM) does significantly increase the safety with which they can be administered. Class II agents are antisympathetic drugs, particularly the β‐adrenoceptor blockers. These are generally safe agents which do not normally require TDM. Class III antiarrhythmic agents include sotalol and amiodarone. TDM can be useful in the case of amiodarone to monitor compliance and toxicity but is generally of little value for sotalol. Class IV antiarrhythmic drugs are the calcium channel blockers verapamil and diltiazem. These are normally monitored by haemodynamic effects, rather than using TDM. Other agents which do not fall neatly into the Vaughan Williams classification include digoxin and perhexiline. TDM is very useful for monitoring the administration (and particularly the safety) of both of these agents.
Single dose and steady state pharmacokinetics of temocapril and temocaprilat in young and elderly hypertensive patients
Authors: Püchler K. , Sierakowski B. , Roots I.
Published: November 2003

Aims
 The aim of this study was to determine the potential impact of age on the pharmacokinetics of temocapril and its pharmacologically active diacid metabolite, temocaprilat, in hypertensive patients.

Methods
 Male and female patients with mild to moderate essential hypertension (DBP 95–114 mmHg inclusive) were allocated to two age groups: young, ≤40 years; elderly, ≥69 years, (n=18 per group). In Part I of the study, subjects took a single oral tablet dose of 20 mg temocapril hydrochloride following an overnight fast. In Part II they took seven once daily oral tablet doses of 20 mg temocapril hydrochloride. Pharmacokinetic profiles were determined after the single and the last dose. Trough plasma samples were taken before each dose in Part II. Urine was collected for 24 h following the single and the last dose.

Results
 Steady state was reached within 1 week in both groups. Statistically significant differences were detected in AUC and AUCss for temocaprilat as well as in CLR for temocapril and temocaprilat, respectively, after a single dose and at steady state. All other pharmacokinetic parameters for temocapril and temocaprilat did not show any significant difference.

Conclusions
 The pharmacokinetic differences detected in the elderly do not require a dose adjustment per se. Nonetheless, a lower starting dose may be appropriate as elderly hypertensive patients are usually considered to be at an increased risk of first dose hypotension at the onset of treatment with an ACE inhibitor.
Second‐trimester pregnancy loss is associated with activated protein C resistance
Authors: Rai Raj , Regan Lesley , Hadley Elizabeth , Dave Manisha , Cohen Hannah
Published: November 2003

We have investigated whether activated protein C resistance (APCR) is associated with second‐trimester miscarriage. The prevalence of APCR was significantly higher amongst women with a history of second‐trimester miscarriage (10/50; 20%) compared with either women with a history of first‐trimester miscarriages only (4/70; 5.7%) or a control group of parous women with no previous history of pregnancy losses (3/70; 4.3%) (P < 0.02). These results suggest that APCR may be an important mechanism of second‐trimester pregnancy loss, possibly related to the increase in intravascular coagulation that occurs during pregnancy.
Serum neural cell adhesion molecule in multiple myeloma and other plasma cell disorders
Authors: Smith S. R. , Auerbach B. , Morgan L.
Published: November 2003

Serum embryonic neural cell adhesion molecule (eNCAM) levels were measured at diagnosis in 92 patients with plasma cell disorders. Significantly elevated levels of serum eNCAM were detected in patients with multiple myeloma when compared to both normal controls and patients with monoclonal gammopathy of uncertain significance (MGUS). Very high levels of serum eNCAM were seen in patients with high tumour burdens. There was a significant correlation between serum eNCAM and β2‐microglobulin (β2m) (r = 0.33; P = 0.002), but not between serum eNCAM and C‐reactive protein or serum albumen. There was a trend towards longer survival for patients with low serum NCAM. The median survival of the low serum eNCAM group (eNCAM < 20 U/ml) was 36 months compared to 16 months for the high serum eNCAM group (log rank test χ2 = 2.42, P = 0.1). Serum eNCAM is a new marker of tumour mass in multiple myeloma and correlates with clinical stage and β2m. Patients with low serum eNCAM levels may have a survival advantage. Serum eNCAM warrants further evaluation as a tumour marker and prognostic factor in multiple myeloma.
Erythropoietin production and erythropoiesis in compensated and anaemic states of hereditary spherocytosis
Authors: Guarnone Roberta , Centenara Esther , Zappa Manuela , Zanella Alberto , Barosi Giovanni
Published: November 2003

A compensated haemolytic state is defined by decreased red cell life‐span without anaemia, i.e. by increased erythropoiesis in the absence of the physiological stimulus for erythropoietin (Epo) production. We evaluated s‐Epo levels and the expansion of erythropoiesis (as measured by circulating transferrin receptor, s‐TfR) in 32 patients with hereditary spherocytosis (HS) with the aim of verifying whether the enhanced erythropoiesis of compensated haemolysis was Epo‐dependent. 20 of the patients (62.5%) had normal Hb values (> 12 g/dl in females and > 13 g/dl in males). Their compensated haemolytic state was the result of up to 8.2 times normal s‐Epo and up to 3.9 times normal s‐TfR levels, which were maintained by physiological regulation of erythropoiesis, as documented by the inverse dependence of Hb on s‐Epo levels. Considering that patients with iron‐deficiency anaemia represented the predicted physiological Epo response to anaemia, the observed/predicted ln s‐Epo ratio (O/P ratio) was calculated in HS patients with anaemia and was used as an index of the adequateness of Epo production. All the anaemic HS patients had an O/P ratio > 1, documenting inappropriately high s‐Epo levels. This work demonstrates that the compensated haemolytic state of HS patients is produced by an inappropriately high s‐Epo level, and that the pattern of Epo overproduction is a biological characteristic of the disease.
Further evidence for the clonal nature of the idiopathic hypereosinophilic syndrome: complete haematological and cytogenetic remission induced by interferon‐alpha in a case with a unique chromosomal abnormality
Authors: Malbrain Manu , Van den Bergh Herman , Zachée Pierre
Published: November 2003

A 49‐year‐old man with the idiopathic hypereosinophilic syndrome (HES) and a unique chromosomal abnormality 46,XY,t(5;9)(q32;q33) is reported. Complete cytogenetic remission was induced by interferon alpha‐2b (IFN‐α). The beneficial action of IFN‐α in different stem‐cell disorders such as CML, HES, multiple myeloma and solid tumours such as hypernephroma or malignant melanoma suggests a common regulatory effect possibly by immunomodulation or other (immune‐mediated) mechanisms, but the exact pathophysiological mechanisms remain hypothetic and unresolved. Since it has been known for some years that the genes encoding for GM‐CSF, IL‐3 and IL‐5 reside on the long arm of chromosome 5, it could be possible that the chromosomal translocation in our patient resulted in excess production of these cytokines, hence causing the hypereosinophilia. This case report and the results obtained from the literature review support the growing body of evidence that IFN‐α has a major place in the long‐term treatment of HES, especially in those cases resistant to conventional treatment, with cytogenetic abnormalities, or presenting as a myeloproliferative variant of HES. In those cases IFN‐α results in lower morbidity, lower mortality and long‐term survival.
A novel mutation (Leu817Pro) causing type 2A von Willebrand disease
Authors: Gemmati D. , Serino M. L. , Moratelli S. , Ballerini G. , Furbetta M. , Lunghi B. , Marchetti G. , Bernardi F.
Published: November 2003

We studied a patient affected by von Willebrand disease type 2A who experienced several mild bleeding episodes and was characterized by markedly reduced haemostatic parameters. In the exon 28 of von Willebrand factor (vWF) gene a T to C transition at nucleotide 8680, resulting in the missense mutation Leu817Pro, was found in the heterozygous form in the patient and in two affected relatives.

As suggested by the presence in platelets of a complete spectrum of vWF multimers as well as by the increased vWF antigen levels and improved haemostasis after DDAVP treatment, the mutation is compatible with normal multimerization, and could be responsible for a reduced stability or an impaired physiological secretion of vWF.
Human bone marrow stromal cells prevent apoptosis and support the survival of chronic lymphocytic leukaemia cells in vitro
Authors: Panayiotidis P. , Jones D. , Ganeshaguru K. , Foroni L. , Hoffbrand A. V.
Published: November 2003

Leukaemic cells from most cases of B‐chronic lymphocytic leukaemia die rapidly by apoptosis in vitro unless they are cultured in the presence of interleukin‐4 or interferon α or γ. We now report prolonged survival of purified CLL cells cultured on bone marrow (BM) derived stromal cells in the absence of exogenous growth factors. In 10 cases of CLL examined 0–61% (mean 14.7%) of the cells were viable after 10 d culture in medium alone, whereas in the presence of BM stromal cells 10–102% (mean 47.0%) of cells were recovered alive (P < 0.005) in 7/10 cases of CLL, cells remained viable after 30 d of culture in BM stromal cells with cell recovery of 12–65%. These long‐term cultured CLL cells were Epstein Barr virus negative, shown by the failure to detect the ENBA‐2 and BZLF1 genes of EBV by PCR analysis. Identity between day 0 and day 30 CLL cells was demonstrated by sequence analysis of their clonal IgH CDR3 region. Adherence of CLL cells to BM stromal cell layers was critical for their protection from apoptosis. Separation of CLL cells from stroma by 0.45 μm culture filters resulted in loss of the protective effect of the stromal cells. Stromal cells were also able to protect CLL cells from hydrocortisone‐induced apoptotic cell death. Our findings provide an in vitro system that can be used to analyse the growth requirements of CLL cells and their chemosensitivity in an in vitro environment that mimics the in vivo milieu.
Sodium butyrate delays neutrophil apoptosis: role of protein biosynthesis in neutrophil survival
Authors: Stringer Rebecca , Hart C. , Edwards Steven
Published: November 2003

Sodium butyrate, which directly affects chromatin structure and function in many cells, is as effective as granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) in delaying apoptosis in human neutrophils. Both butyrate and GM‐CSF preserved the ability of neutrophils cultured for 22 h in vitro to generate reactive oxidants and express receptors such as CD16. They also delayed apoptotic morphology and DNA fragmentation, and stimulated de novo biosynthesis: newly‐labelled polypeptides detected by 2D‐polyacrylamide gel electrophoresis after treatment with GM‐CSF and butyrate were very similar. Cycloheximide abrogated the effects of both GM‐CSF and butyrate. Exposure to butyrate for 1 h did not prime oxidant production and did not up‐regulate expression of CD11b. Hence, unlike GM‐CSF, butyrate does not stimulate translocation of granules to the plasma membrane. These data suggest that active gene expression is involved in the regulation of neutrophil apoptosis and changes in chromatin structure and function may control apoptosis in these cells.
The human CD40 gene lies within chromosome 20q deletions associated with myeloid malignancies
Authors: Asimakopoulos Fotios A. , White Nicholas J. , Nacheva Elisabeth P. , Green Anthony R.
Published: November 2003

Deletions of chromosome 20q are associated with myeloid malignancies and have been previously shown to arise in a multipotent progenitor of both myeloid and B cells. However, B‐cell differentiation from the abnormal progenitor was impaired. The CD40 antigen is a surface glycoprotein which is expressed in B cells and haemopoietic stem cells and is important for B‐cell growth and development. Following the recent mapping of CD40 to chromosome 20q we sought to determine its position relative to 20q deletions. Analysis of lymphoblastoid cell lines carrying 20q deletions placed CD40 within a 19–21 cM interval which is almost coincidental with the common deleted region defined by previous analysis of patient samples. Our results raise the possibility that genetic alteration of this locus may contribute to the pathogenesis of myeloid disorders associated with 20q deletions.
Molecular evidence for a common leukaemic progenitor in acute mixed lymphoid and myeloid leukaemia
Authors: Lim Seah H. , Culligan Dominic , Couzens Steve , Fisher Janet , John Susan , Pollard Peter , Burnett Alan , Whittaker Jack
Published: November 2003

De novo acute leukaemia presenting with a mixed lymphoid and myeloid leukaemic population has rarely been described. We have used the consensus Ig heavy chain primers and DNA isolated from these two distinct populations of cells in polymerase chain reactions. We demonstrated that both populations of cells exhibited Ig heavy chain gene rearrangements. Cloning and subsequent DNA sequencing of the amplified products showed a common V‐D‐J junctional nucleotide sequence. This work therefore provides the first evidence that the leukaemic cells in de novo acute leukaemia with a mixed lymphoid and myeloid population are derived from a common progenitor clone and may offer an explanation for the poor prognosis in these patients.
Carboxy‐terminal peptides (C1–24 and C13–24 but not C1–13) of platelet factor 4 inhibit murine megakaryocytopoiesis, an activity which is neutralized by heparin
Authors: Lebeurier Isabelle , Basara Nada , Aïdoudi Sallouha , Amiral Jean , Caen Jacques , Chao Han Zhong
Published: November 2003

Negative regulation of megakaryocytopoiesis is a complex process involving various cytokines. One of these cytokines is platelet factor 4 (PF4), a megakaryocyte/platelet specific protein. PF4 and a carboxy‐terminal peptide related to PF4 have been reported to inhibit human and murine megakaryocytopoiesis. The growth of several megakaryoblastic cell lines: human erythroleukaemia cell line (HEL), Meg‐01 and Dami, was also inhibited by PF4 and a 13–24 carboxy‐terminal peptide related to PF4. We report that peptides corresponding to the 1–24 and 13–24 but not 1–13 carboxy‐terminal region of PF4 inhibit murine megakaryocytopoiesis both in vivo (5 μg/inj) and in vitro (2.5 and 5 μg/ml). Moreover, such an inhibitory activity of PF4‐related peptides is abrogated by heparin (5 IU/dish). These overall data indicate that carboxy‐terminal PF4‐related peptides retain the inhibitory effect of PF4 on both murine single MK and CFU‐MK in vivo and in vitro by acting on an early stage of megakaryocytopoiesis and strongly suggest that the inhibitory activity of the multi‐functional PF4 might be localized in a short carboxy‐terminal region which might include, in part, the PF4 heparin binding domain.
All‐trans‐retinoic acid (ATRA) responsive skin relapses of acute promyelocytic leukaemia followed by ATRA‐induced pseudotumour cerebri
Authors: Selleri Carmine , Pane Fabrizio , Notaro Rosario , Catalano Lucio , Santoro Lidia E. F. , Luciano Luigiana , Frigeri Ferdinando , Salvatore Francesco , Rotoli Bruno
Published: November 2003

A 30‐year‐old woman with acute promyelocytic leukaemia (APL) went into complete remission following idarubicin and cytarabine chemotherapy; 18 months later she developed repeated skin relapse, with no bone marrow involvement. DNA and RNA analysis of skin lesions revealed the presence of the PML/RARα hybrid gene, which was not detected at the same time in bone marrow. The skin relapses were successfully treated by all‐trans‐retinoic acid (ATRA) as single agent over 2 years. However, prolonged administration of ATRA caused pseudotumour cerebri, which disappeared upon drug withdrawal. The absence of the hybrid gene in the bone marrow by RT‐PCR analysis led to the patient being autografted.
Plateau phase in multiple myeloma: an analysis of long‐term follow‐up of 432 patients
Authors: Oivanen Tiina M.
Published: November 2003

Data from an 8‐year follow‐up of 432 myeloma patients were analysed for incidence, duration and prognostic value of plateau phase and factors favouring its achievement. The first‐line chemotherapy was melphalan and prednisolone in 121 cases, combination chemotherapy in 311 cases. The survival times were similar despite different response rates. Any response resulted in survival significantly (P<0.001) better than in patients with progressive disease, but the level of response had no influence in this respect.  A plateau of at least 3 months was achieved in 81%, at least 6 months in 74%, at least 12 months in 59% and at least 24 months in 33%. Groups with significantly different (P<0.001) survivals were identified: a plateau of <3 months, 3–11 months, 12–23 months, and 24 months or longer, with median survivals of 10, 27, 46 and 81 months, respectively.  In the multivariate analysis of pretreatment variables, only haemoglobin (Hb) (P<0.001) and creatinine at 2 months (P<0.01) were significant for a plateau >12 months. After inclusion of chemotherapy data, Hb and time taken to reach the best response were still significant (P=0.002). The predictive power of high Hb and slow response for achieving a plateau of 6 or 12 months was 79%.  Accordingly, the criteria for response of treatment for multiple myeloma should include a stable period of at least 3 months; the criteria for a plateau a stable period of at least 12 months in order to have real prognostic significance.
Molecular detection of t(8;21)/AML1‐ETO in AML M1/M2: correlation with cytogenetics, morphology and immunophenotype
Authors: ANDRIEU VALÉRIE , RADFORD‐WEISS ISABELLE , TROUSSARD XAVIER , CHANE CATHERINE , VALENSI FRANÇOISE , GUESNU MARTINE , HADDAD ELIE , VIGUIER FRANK , DREYFUS FRANÇOIS , VARET BRUNO , FLANDRIN GEORGES , MACINTYRE ELIZABETH
Published: November 2003

The t(8;21) identifies a subgroup of acute myeloid leukaemia (AML) with a relatively good prognosis which may merit different treatment. It is associated predominantly, but not exclusively, with AML M2, and corresponds to rearrangements involving the AML1 and ETO genes. AML1‐ETO positive, t(8;21) negative cases are well recognized but their incidence is unknown. In order to determine optimal prospective AML1‐ETO RT‐PCR screening strategies, we analysed 64 unselected AML M1 and M2 cases and correlated the results with other biological parameters. Molecular screening increased the overall detection rate from 8% to 14%. AML1‐ETO was found in 3% (1/32) of AML M1 and 25% (8/32) of M2, including three patients without a classic t(8;21) but with chromosome 8 abnormalities. It was more common in younger patients. Correlation with morphology enabled development of a scoring system which detected all nine AML1‐ETO‐positive cases with a false positive rate of 7% (4/55). Although certain AML1‐ETO‐positive cases demonstrated characteristic immunological features (CD19 and CD34 expression, CD33 negativity), each of these markers was insufficiently specific to permit prediction in an individual case. We conclude that initial routine prospective molecular screening for AML1‐ETO in all AMLs, combined with standardized morphological and immunological analysis, is desirable in order to produce improved prognostic stratification and to determine whether screening can ultimately be restricted to appropriate subgroups.
Human herpesvirus‐6‐associated exanthema in a patient with acute lymphocytic leukaemia
Authors: FUJITA HIROYUKI , MARUTA ATSUO , TOMITA NAOTO , TAGUCHI JUN , SAKAI RIKA , SHIMIZU AKIO , HARADA MASAOKI , OGAWA KOUJI , KODAMA FUMIO , OKUBO TAKAO
Published: November 2003

We report the first case of human herpesvirus‐6 (HHV‐6) associated exanthema in a patient with acute lymphocytic leukaemia (ALL). We analysed DNA extracted from an exanthematous lesion using the polymerase chain reaction (PCR). DNA was positive for HHV‐6 but negative for herpes simplex virus, varicella zoster virus, and cytomegalovirus. Immunohistochemical staining of the skin with monoclonal antibody against HHV‐6 confirmed the infection. The possibility of HHV‐6 infection should be considered when an atypical skin rash is seen in patients with ALL.
Predominant expression of the long isoform of Bcl‐x (Bcl‐xL) in human lymphomas
Authors: Xerri Luc , Parc Patricia , Brousset Pierre , Schlaifer Daniel , Hassoun Jacques , Reed John C. , Krajewski Stanislas , Birnbaum Daniel
Published: November 2003

Bcl‐x is a member of the bcl‐2 family of proteins which are characterized by their ability to modulate apoptosis. Alternative splicing results in two distinct bcl‐x mRNAs encoding a long isoform, bcl‐xL, which acts as a bcl‐2 agonist; and a short isoform, bcl‐xS, which inhibits bcl‐2 effects. The aim of the study was to determine whether bcl‐x is expressed in lymphoma tissues and to characterize the respective production of bcl‐xS and bcl‐xL. We investigated the expression of bcl‐x mRNA in a series of 50 non‐Hodgkin's lymphomas (NHL) and Hodgkin's disease (HD) cases using a RT‐PCR method in order to amplify both transcripts simultaneously, and to estimate their relative abundance. The rearrangements of the bcl‐2 gene were analysed by RT‐PCR expression of the hybrid bcl‐2–IgH mRNA. In addition, 20 PCR‐positive NHL cases and three HD cases were analysed by immunohistochemistry using bcl‐x polyclonal antisera. RT‐PCR showed bcl‐x expression in 43/45 NHLs and 5/5 HD cases. The bcl‐xL transcript was predominant in all positive cases and was associated with variable amounts of bcl‐xS. There was no significant correlation between the profile of bcl‐xL/bcl‐xS expression and the histological and immunological subtyping. Bcl‐x immunodetection was positive in the neoplastic cell component in all analysed cases, but the degree of staining was highly variable between cases. Expression of the hybrid bcl‐2–IgH gene was detected by RT‐PCR in five cases of follicular NHL and in one case of HD, but this group of tumours did not display a particular profile of bcl‐xL/bcl‐xS expression. We conclude that bcl‐x is commonly expressed by malignant cells in various types of malignant lymphomas, with a predominance of the bcl‐xL transcript. Since the corresponding bcl‐xL isoform can block the cell death machinery and potentialize bcl‐2 effects, it may be involved in some pathways of lymphomagenesis.
Induction of nitric oxide synthase is involved in the mechanism of Fas‐mediated apoptosis in haemopoietic cells
Authors: Selleri Carmine , Sato Tadatsugu , Raiola Anna Maria , Rotoli Bruno , Young Neal S. , Maciejewski Jaroslaw P.
Published: November 2003

Induction of nitric oxide synthase (iNOS) and production of the toxic metabolite nitric oxide (NO) is one of the interferon‐γ (IFN‐γ) and tumour necrosis factor‐α (TNF‐α) regulated effector mechanisms that can lead to apoptosis of haemopoietic progenitor cells. Fas‐receptor (Fas‐R) expression can be stimulated by IFN‐γ and TNF‐α. Transactivation of iNOS, and possibly Fas‐R promoters, by interferon regulatory factor‐1 expressed in response to IFN‐γ may be a part of the iNOS transduction pathway. We investigated whether the effects of Fas‐R triggering in haemopoietic cells were mediated by NO. On Western blotting, we observed that Fas‐receptor agonist, monoclonal antibody CH11, enhanced expression of iNOS. As shown by the reverse transcription polymerase chain reaction, CH11 also induced iNOS mRNA expression in purified CD34+ cells. To determine whether NO was involved in Fas‐mediated apoptosis we inhibited iNOS‐catalysed production of NO using anti‐sense (AS) oligodeoxynucleotides (ODN) directed against iNOS mRNA. After culture of haemopoietic cells in the presence of AS‐ODN, iNOS expression decreased and was no longer enhanced by Fas. This effect was associated with the prevention of Fas‐mediated apoptosis, as determined by a DNA fragmentation and terminal deoxynucleotidyl transferase staining. In colony assays, specific AS‐oligonucleotides prevented FAS‐mediated inhibition of colony formation by total bone marrow and CD34+ progenitor cells. Our data suggest that the inhibitory effects of Fas, including induction of apoptosis, are mediated by effector mechanisms that may be similar to those described for IFN‐γ and TNF‐α.
Splenectomy as an adjuvant measure in the treatment of severe aplastic anaemia
Authors: Speck B. , Tichelli A. , Widmer E. , Harder F. , Kissling M. , Würsch A. , Stebler Gysi Ch , Signer E. , Bargetzi M. , Orth B. , Gratwohl A. , Nissen C.
Published: November 2003

The role of splenectomy in aplastic anaemia (AA) is controversial. The hazards of operating on a severely pancytopenic patient, the fear of compromising the patient’s immune function, and the improvement of non‐surgical treatment have made splenectomy unpopular in this disease.

We have evaluated positive and adverse effects of splenectomy in 80 patients with severe aplastic anaemia (SAA) treated with antilymphocyte globulin (ALG) (group A), using 52 nonsplenectomized ALG patients as controls (group B). All patients survived the operation. Nonfatal complications of surgery occurred in 10 (12.5%). Splenectomy induced a significant increase of peripheral blood neutrophils, reticulocytes and platelets within 2 weeks, followed by a continuous increase of all values over the following weeks. 28/132 patients (21%) developed a late clonal disorder of haemopoiesis, paroxysmal nocturnal haemoglobinuria (PNH) or myelodysplastic syndrome (MDS), or both. Their incidence was identical in groups A and B. 13/28 (59%) died, 10/17 (59%) in group A and 3/11 (27%) in group B (not significant (n.s.)). Overall probability of survival at 18 years after ALG was 51 ± 6% for group A and 61 ± 7% for group B (n.s.).

We conclude that splenectomy in AA is safe. It induces an immediate increase of peripheral blood counts and, thereafter, a continuous improvement of haemopoiesis. It does not increase the incidence of late clonal complications but has a borderline effect on mortality from these disorders. Splenectomy should be reconsidered in selective nontransplanted patients who have prolonged transfusion requirements despite otherwise optimal treatment.
Regulated plasma levels of colony‐stimulating factors, interleukin‐6 and interleukin‐10 in patients with acute leukaemia and non‐Hodgkin's lymphoma undergoing cytoreductive chemotherapy
Authors: Reisbach Gilbert , Kamp Torsten , Welzl Gerhard , Giez Christiane , Abedinpour Fariborz , Lodri Antal , Kaboth Werner , Dörmer Peter , Nerl Christoph
Published: November 2003

Endogenous plasma levels of granulocyte colony stimulating factor (G‐CSF), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), IL‐6 and IL‐10 were measured in a total of 70 patients undergoing cytoreductive chemotherapy for treatment of acute leukaemia or non‐Hodgkin’s lymphomas. The diagnoses were acute myeloid leukaemia (AML; n = 30), acute lymphoblastic leukaemia (ALL; n = 6), non‐Hodgkin’s lymphomas (NHL; n = 11) and other malignant haematological disorders including myelodysplastic syndromes (n = 23). After chemotherapy, plasma G‐CSF was elevated (mean 5.6 ng/ml; range 1.2–10 ng/ml), and was inversely correlated with white blood cell counts (WBC) (r = −0.7, P < 0.001). Occurrence of fever (T>38.0°C) during severe myelosuppression (WBC<1 × 109/l) was associated with an additional increase of G‐CSF levels (P < 0.001). Plasma IL‐6 correlated significantly with fever (range <1 to 1100 pg/ml, mean 130 pg/ml; r = 0.5, P < 0.001) but revealed only a weak association with WBC or platelet counts. In patients treated with recombinant G‐CSF (n = 9), an association between IL‐6 and fever was still observed after chemotherapy. During the nonfebrile status (total n = 242; AML n = 124), IL‐6 levels remained <9 pg/ml in 90% of cases, whereas G‐CSF increased with leucopenia (r = −0.72; P < 0.001). In contrast, endogenous GM‐CSF remained normal and IL‐10 showed only a slight increase (21% of samples; maximum 22 pg/ml) in severe leucopenia. In particular, IL‐10 levels did not correlate with G‐CSF or IL‐6 levels.

We conclude that systemic release of G‐CSF and IL‐6 is obviously not abrogated by cytoreductive chemotherapy in acute leukaemia and NHL and may add to the therapeutic efficacy of recombinant cytokines. Also, plasma levels of G‐, GM‐CSF or IL‐6 appear to be regulated by separate mechanisms.
Sustained complete cytologic and molecular remission induced by donor leucocyte infusions alone in an acute myeloblastic leukaemia in relapse after bone marrow transplantation
Authors: Buzyn‐Veil Agnès , Belanger Coralie , Audat Françoise , Hermine Olivier , Bodemer Christine , Ribrag Vincent , Radford Isabelle , Poirel Hélène , Chane Catherine , Valensi Françoise , Macintyre Elisabeth , Varet Bruno
Published: November 2003

Therapeutic options for treatment of recurrence of leukaemia after allogeneic bone marrow transplantation (BMT) are limited. A beneficial effect of donor lymphocyte infusions (DLI) has not previously been described in acute myeloid leukaemia (AML) relapse. We report a case of AML with t(8;21), relapsing 3 months after BMT, who received DLI without adjuvant chemotherapy or growth factors. The patient developed acute GVHD and achieved a rapid complete remission of his AML by both cytologic and molecular criteria of at least 14 months duration, thereby showing that DLI for AML in relapse after BMT is an alternative therapeutic option.
Trisomy 12 defines a group of CLL with atypical morphology: correlation between cytogenetic, clinical and laboratory features in 544 patients
Authors: Matutes E. , Oscier D. , Garcia‐Marco J. , Ellis J. , Copplestone A. , Gillingham R. , Hamblin T. , Lens D. , Swansbury G. J. , Catovsky D.
Published: November 2003

We have analysed the clinical and laboratory features in 544 patients with chronic lymphocytic leukaemia (CLL) with available cytogenetics and fluorescence in‐situ hybridization (FISH) analysis for trisomy 12 in half of them, to examine the correlation between chromosome abnormalities and clinical or laboratory parameters. Five chromosome groups were defined: (1) trisomy 12 (18%), detected as the sole abnormality or associated with other changes; (2) del(13)(q12–14) (7%); (3) other abnormal karyotypes (20%); (4) normal karyotype (41%); and (5) no divisions (14%). There were no differences in the age distribution between the five groups. Clinical stages (Binet) were: A (74%), B (12%) and C (14%). Stage A was common in cases with del(13q)(82%), normal (84%) and other abnormal karyotypes (74%), whereas it was less common in trisomy 12 cases (64%) and those with no divisions (48%). Typical CLL morphology was found in 83% of cases; 10% had more than 10% prolymphocytes (CLL/PL) and 7% had other atypical features. CLL with trisomy 12 was the only group with a high frequency of either CLL/PL (31%) or atypical morphology (24%). Atypical morphology and CLL/PL were even more frequent when trisomy 12 was associated with other chromosomal abnormalities (70% v 46%). The incidence of cases with CLL/PL and other atypical morphology was significantly lower in the other chromosome groups (P<0.001. There were no differences in immunophenotype among the various groups except for a higher frequency of stronger Smlg and FMC7 expression in cases with trisomy 12, particularly those with CLL/PL and other atypical morphology. Our findings confirm that trisomy 12 defines a subgroup of CLL with more frequent atypical morphology, including CLL/PL, stronger SmIg and FMC7 expression, more advanced stages (B and C in 18%) and possibly worse prognosis.
Feasibility of peripheral blood progenitor cell harvest and transplantation in patients with poor‐risk myelodysplastic syndromes
Authors: Demuynck H. , Delforge M. , Verhoef G. E. G. , Zachee P. , Vandenberghe P. , Van Den Berghe H. , Boogaerts M. A.
Published: November 2003

Myelodysplastic syndromes (MDS) are a group of clonal haematological disorders with a highly unfavourable prognosis. Allogeneic bone marrow transplantation offers the sole possibility for cure and prolonged survival, but is only available for a minority of patients. Therefore, we investigated the feasibility of PBPC collection and transplantation in 11 patients with high‐risk myelodysplasia who were not eligible for allogeneic bone marrow transplantation. In six patients, PBPC were harvested after mobilization with G‐CSF alone. Five patients were harvested during the recovery phase of intensive chemotherapy combined with G‐CSF. This resulted in seven patients in an adequate CD34 progenitor yield >1 × 106/kg. Six patients obtained a CFU‐GM content of the PBPC harvest >10 ×104/kg. Five patients were subsequently transplanted following a standard BuCy4 regimen. The median to ANC (absolute neutrophil count) ≥0.5 and 1.0 × 109/l was respectively 14 d (range 10–18) and 16 d (range 11–25). Platelets were self‐supporting ≥20 × 109/l after a median of 41 d (range 8–144). One patient had a persistent lack of platelet engraftment unresponsive to infusion of back‐up bone marrow.

These data demonstrate that in selected patients with high‐risk MDS, adequate PBPC collection appears feasible, enabling the harvest of sufficient cell numbers required for rapid and stable engraftment after reinfusion. Improvement in mobilization efficiency may enable the collection of higher CD34+ progenitor cell numbers required for more rapid platelet engraftment. PBPC transplantation may be an alternative treatment option for patients who lack an allogeneic marrow donor. Follow‐up is, however, still too limited to draw any conclusion regarding the long‐term cure rate.
Serum interleukin‐6 (IL‐6) and interleukin‐4 (IL‐4) in patients with multiple myeloma (MM)
Authors: Kyrstsonis Maria‐Christina , Dedou ssis Georges , Bax evanis Constantin , Stamate lou Marina , Maniatis Alice
Published: November 2003

In this study we determined, in patients with multiple myeloma (MM), serum levels of IL‐4 and IL‐6 at diagnosis and during the course of the disease, seeking a correlation with disease activity and prognosis. We studied 54 MM patients, 41 of whom responded to chemotherapy whilst 11 were resistant. At diagnosis, IL‐6 was increased in 66% of patients (median 35.5 pg/ml) whereas IL‐4 was low (median 4 pg/ml) in 75% of patients. In responding patients, IL‐4 increased in remission (median 25 pg/ml), whereas IL‐6 decreased (median 4 pg/ml). In chemotherapy‐resistant patients, IL‐6 and IL‐4 values remained stable during the course of the disease.
Trisomy 12 and structural abnormalities of 13q14 occurring in the same clone in chronic lymphocytic leukaemia
Authors: Mould S , Gardiner A. , Corcoran M. , Oscier D. G.
Published: November 2003

Trisomy 12 and deletions or translocations of 13q14 are the commonest cytogenetic abnormalities in chronic lymphocytic leukaemia but rarely co‐exist in the same patient. We describe eight patients from a series of > 400 patients with CLL in whom trisomy 12 and t or del 13 occur in the same clone. Using FISH we have identified clones with trisomy 12 alone, t or del 13q14 alone and both abnormalities, in each of the patients studied. This implies that neither trisomy 12 nor t or del 13q14 is the initiating event in leukaemogenesis, but does not exclude the possibility of a submicroscopic abnormality of 13q14 occurring as an early event.
Diamond Blackfan anaemia: differential pattern of in vitro progenitor response to macrophage inflammatory protein 1‐alpha
Authors: McGuckin Colin P. , Liu Wai M. , Ball Sarah E. , Gordon‐Smith Edward C. , Uhr Mario R.
Published: November 2003

The congenital disorder of erythropoiesis Diamond Blackfan anaemia (DBA) exhibits a defect in the stem/progenitor cell compartment, located at the erythroid progenitor level (CFU‐GEMM, BFU‐E, CFU‐E). Treatment of DBA with interleukin‐3 (IL‐3) has had limited effect, despite in vitro studies suggesting that progenitor cells were capable of responding to IL‐3. Whether IL‐3 is not reaching the appropriate defective target cell, the cells cannot respond, or the marrow humoral inhibitory system is overriding it, is not clear. To investigate humoral inhibitory activities we examined the response of 15 DBA bone marrows in vitro to the inhibitory chemokine macrophage inflammatory protein 1‐α (MIP1‐α) in the presence of the stimulatory cytokines erythropoietin, granulocyte‐macrophage colony‐stimulating factor, IL‐3, and stem cell factor. In vitro data agreed with our previous work showing that our patients formed three statistically different groups in response to stimulatory cytokines (type I DBA erythroid colony numbers≈ normal > type II DBA > type III DBA). Addition of MIP1‐α to cultures caused average erythroid and myeloid suppression, which sequentially increased with DBA type (type I inhibition < type II < type III). The differential level of inhibition shown by MIP1‐α in these DBA patients lends further evidence for the presence of distinct subgroups in this disorder.
The combination of erythropoietin and granulocyte colony‐stimulating factor increases the rate of haemopoietic recovery with clinical benefit after peripheral blood progenitor cell transplantation
Authors: Pierelli Luca , Scambia Giovanni , Menichella Giacomo , D'Onofrio Giuseppe , Salerno Giovanna , Panici Pierluigi Benedetti , Foddai Maria Laura , Vittori Mariangela , Lai Marco , Ciarli Marina , Puglia Giovanni , Mancuso Salvatore , Bizzi Bruno
Published: November 2003

In order to investigate the effects of erythropoietin (EPO) plus granulocyte colony‐stimulating factor (G‐CSF) administration after peripheral blood progenitor cell transplantation (PBPCT) we performed a phase I/II study in patients with high‐risk cancer. 15 consecutive patients were treated with recombinant human G‐CSF (rhG‐CSF) at the dose of 5 μg/kg subcutaneously (s.c.) every 24 h until day +12 and with recombinant human EPO (rhEPO) at the dose of 150 IU/kg s.c. every 48 h until day +11 following PBPCT. Their haemopoietic recovery was compared to that obtained in eight historic control patients who did not receive any cytokines after PBPCT. No side‐effects were observed during EPO plus G‐CSF treatment and the treatment was not discontinued in any of the patients before completion of the treatment plan. The administration of EPO plus G‐CSF after PBPCT produced a significant increase in the rate of white blood cell (WBC) (P = 0.0005), polymorphonuclear leucocyte (PMN) (P = 0.0005) and platelet (PLT) (P = 0.0105) recovery compared to the control group. The acceleration in haemopoietic recovery observed in the EPO plus G‐CSF‐treated patients produced a significant reduction of the days with WBC < 1×109/l (P = 0.0009), PMN  < 0.2×109/l (P = 0.0030) and PMN < 0.5×109/l (P = 0.0006). EPO plus G‐CSF‐treated patients required a significantly lower number of single donor PLT transfusions (P = 0.0142) and did not experience neutropenic fever, but historic control patients experienced fever > 38°C for a median period of 4 d (0–12) with a median period of parenteral antibiotic administration of 7.5 d (0–17). The length of the hospital stay was significantly shorter in the study group than in the historic control group (P = 0.0264). In conclusion, we can confirm that EPO plus G‐CSF treatment is feasible and potentiates the haemopoietic recovery after PBPCT, thus simplifying the clinical management of cancer patients who undergo high‐dose chemotherapy.