bluuebunny/crossref_metadata_2025 huggingface dataset:
The Relationship Between the Internal Audit Function and Corporate Governance in the EU – a Survey
Authors: Paape Leen , Scheffe Johan , Snoep Pim
Published: November 2003

Based on a survey carried out at the behest of the European Confederation of Institutes of Internal Auditors (ECIIA), this article rather ambitiously attempts to sketch out the relationship between the Internal Audit Function (IAF) and Corporate Governance (CG) issues among the top listed companies in the European Union (EU). ‘Attempts’ is used advisedly, because the authors are far from convinced that the results they obtained – in terms of numbers and percentages of responses – have provided them with a solid basis for the textured and nuanced overview they were aiming at. Having said that, it is, nevertheless, clear that there are surprising, and even startling, differences in the way Internal Auditors operate in the EU, even in the largest companies, and their awareness of, and opinions about, their role in CG concerns. Put succinctly, there is plenty of room for evolution in both mind set and daily practice.
The effect of decaffeination of coffee on gastro‐oesophageal reflux in patients with reflux disease
Authors: Pehl C. , Pfeiffer A. , Wendl B. , Kaess H.
Published: November 2003

Background:

Patients with reflux disease often complain of heartburn after ingestion of coffee. Induction of gastro‐oesophageal reflux has been demonstrated by pH‐metry following the intake of coffee in healthy volunteers. The reflux was reduced when the coffee had undergone a decaffeination process. The aim of this study was to investigate the effect of decaffeination of coffee on reflux in patients with reflux disease.

Methods:

Seventeen reflux patients underwent two oesophageal 3‐h pH measurements. The patients received, in a double‐blind study design in a randomized order, 300 mL of either regular or decaffeinated coffee together with a standardized breakfast. The fraction time oesophageal pH < 4 was calculated during the three postprandial hours.

Results:

For regular coffee the fraction time was calculated to a median of 17.9% with a range of 0.7–56.6%. The fraction time was significantly reduced to 3.1% (0–49.9%) after ingestion of decaffeinated coffee.

Conclusion:

The amount of gastro‐oesophageal reflux induced by the intake of regular coffee in patients with reflux disease can be reduced by the decaffeination of coffee.
The absorption of low‐dose methotrexate in patients with inflammatory bowel disease
Authors: Moshkowitz M. , Oren R. , Tishler M. , Konikoff F. M. , Graff E. , Brill S. , Yaron M. , Gilat T.
Published: November 2003

Background:

Recent clinical trials have demonstrated that methotrexate may have an important therapeutic role in the treatment of patients with inflammatory bowel disease, who are either refractory or intolerant to traditional medical therapy. The aim of this study was to evaluate the pharmacokinetics of low‐dose oral methotrexate in patients with inflammatory bowel disease.

Methods:

Methotrexate (12.5 mg) was given orally to nine patients with inflammatory bowel disease: five with Crohn's disease, and four with ulcerative colitis, and to six patients with rheumatoid arthritis who served as a control group. Blood samples were drawn at specific intervals to evaluate methotrexate plasma levels.

Results:

Methotrexate was rapidly absorbed in all patients. Peak concentrations (Cmax) varied considerably, ranging from 0.25–0.87 μM . The mean Cmax values were similar in all patient groups (0.59 ± 0.12, 0.69 ± 0.16 and 0.54 ± 0.18 μM, P not significant) for Crohn's disease, ulcerative colitis and rheumatoid arthritis, respectively. The mean area under curve in 120 min (AUC0–120) was also similar in all patient groups (32.9 + 11.3, 43.6 + 9.9 and 41.8 + 14.9 ng.min/mL, P not significant) for Crohn's disease, ulcerative colitis and rheumatoid arthritis, respectively. The mean time to reach Cmax, (tmax), varied between patient groups (84, 112 and 95 min, respectively, with a significant difference, P<0.02, between the Crohn's disease and ulcerative colitis groups. A negative correlation was found between methotrexate dosage/kg and Cmax (r=−0.74) only in Crohn's disease patients but not in the other patient groups.

Conclusions:

Orally administered methotrexate is well absorbed in patients with inflammatory bowel disease including those with severe small bowel disease or resection. If methotrexate is proven to be effective in inflammatory bowel disease, it should be administered orally.
Treating the symptoms of gastro‐oesophageal reflux disease: a double‐blind comparison of omeprazole and cisapride
Authors: Galmiche J.‐P. , Barthelemy P. , Hamelin B.
Published: November 2003

Background: Few studies have specifically addressed the management of the symptoms of gastro‐oesophageal reflux disease, and there are no comparative data in this respect for acid pump inhibitors and prokinetic agents.

Methods: Following endoscopy 424 patients presenting with heartburn as the predominant symptom of gastro‐oesophageal reflux disease were randomized to treatment with omeprazole 20 or 10 mg once daily, or cisapride 10 mg four times daily, in a double‐blind, double‐dummy, parallel group, multicentre study. Symptoms and quality of life were assessed at 4 weeks. Patients still experiencing heartburn continued therapy for a further 4 weeks and the assessments were repeated.

Results: At 4 weeks, heartburn was resolved in 65% (95% CI: 57–73%), 56% (48–64%) and 41% (32%–49%) of patients treated, respectively, with omeprazole 20 mg and 10 mg once daily, and cisapride. Both omeprazole doses were significantly more effective than cisapride (P < 0.01). The same order of efficacy was observed regardless of the presence of erosive oesophagitis. Regurgitation and epigastric pain also improved to a greater degree with omeprazole than with cisapride. Quality of life was improved in all treatment groups, and the improvement in the reflux dimension of the Gastrointestinal Symptom Rating Scale (GSRS) score was significantly different between groups (P = 0.002).

Conclusions: Omeprazole 20 or 10 mg once daily is significantly more effective than cisapride in the resolution of heartburn, regardless of the presence of erosive oesophagitis, and this is accompanied by an improvement in patient quality of life.
The management of severe ulcerative colitis
Authors: Hyde G. M. , Jewell D. P.
Published: November 2003

Severe ulcerative colitis is a potentially life‐threatening condition but the mortality has fallen dramatically over the past 30–40 years. It is now less than 2%, including surgical mortality, and should only be seen in patients with significant co‐existing disease. Early recognition of the severity of the colitis, intensive medical therapy, close liaison between physician and surgeon, and prompt surgery when necessary have all contributed to this improved outcome. Despite the use of high‐dose intravenous corticosteroids, 20–30% of patients will make a poor response and will require urgent surgery. The use of intravenous cyclosporin has proved effective at reducing the immediate surgical rate in this group of unresponsive patients and appears safe. Whether cyclosporin reduces the need for surgery in the longer term is much less certain. Clinical, radiological, endoscopic and laboratory parameters can now be used to predict the course of a severe attack. These help in the timing of urgent surgery and are potentially helpful in determining when to begin other therapies such as cyclosporin.
Nicotine tartrate liquid enemas for mildly to moderately active left‐sided ulcerative colitis unresponsive to first‐line therapy: a pilot study
Authors: SANDBORN W. J. , TREMAINE W. J. , LEIGHTON J. A. , LAWSON G. M. , ZINS B. J. , COMPTON R. F. , MAYS D. C. , LIPSKY J. J. , BATTS K. P. , OFFORD K. P. , HURT R. D. , GREEN J.
Published: November 2003

Background: Ulcerative colitis is predominantly a disease of non‐smokers, and transdermal nicotine is therapeutic but often results in side‐effects. Administration of nicotine as a liquid rectal enema results in less systemic nicotine absorption.

Aim: To determine the safety and clinical response of nicotine tartrate liquid enemas for active left‐side ulcerative colitis in a pilot study.

Methods: Ten non‐smoking patients with mildly to moderately active left‐sided ulcerative colitis unresponsive to first‐line therapy were treated in an open protocol with nightly nicotine tartrate liquid enemas at a dose of 3 mg nicotine base for 1 week then 6 mg for 3 weeks. Clinical assessments were determined at baseline and 4 weeks by endoscopy, physician assessment and a patient diary of daily symptoms. Peak and trough serum nicotine and trough plasma cotinine were determined by gas chromatography/mass spectrometry and high performance liquid chromatography, respectively.

Results: After 4 weeks of treatment, 5/7 patients (71%) showed clinical and sigmoidoscopic improvement (per protocol analysis). The other three patients discontinued therapy within 7 days because of inability to retain the liquid enemas. No patients showed histologic improvement. Six of the patients who completed the 4‐week study had peak and trough serum nicotine concentration determined, only 1 of 6 patients had a detectable peak nicotine concentration (value 2.3 ng/mL), and all six patients had undetectable trough nicotine concentrations. The mean trough plasma cotinine concentration was 13 ± 10 ng/mL. Transient and mild adverse events occurred in 4/10 patients (nausea, lightheadedness, tremor, sleep disturbance). Given the low or undetectable serum nicotine concentrations, these adverse events are not likely to be related to the nicotine enemas.

Conclusions: Nicotine tartrate liquid enemas administrated at a dose of 3 mg nicotine base/day for 1 week and then 6 mg/day for 3 weeks are safe and appear to result in clinical improvement in some patients with mildly to moderately active, left‐sided ulcerative colitis unresponsive to first‐line therapy. Placebo‐controlled trials are warranted to confirm these preliminary findings.
Age‐dependent eradication of Helicobacter pylori with dual therapy
Authors: TREIBER G. , AMMON S. , KLOTZ U.
Published: November 2003

Background: Combined treatment using an acid‐inhibiting drug with antibiotics can cure Helicobacter pylori infection. However, eradication rates are highly variable, especially if a proton pump inhibitor is used with amoxycillin. Therefore it is important to define factors/predictors of the clinical outcome.

Methods: In a single‐blind study, 60 H. pylori‐positive patients prospectively matched for diagnosis (erosive gastritis, duodenal and gastric ulcer), age (above and below 50 years) and smoking habits were randomly treated (each group n = 20) for 2 weeks with amoxycillin (1 mg b.d.) and either omeprazole (20 mg b.d.), lansoprazole (30 mg b.d.) or ranitidine (300 mg b.d.). Intragastric pH and plasma levels of the administered drugs were monitored over a dosing interval of 12 h.

Results: The overall eradication rates were 45% (intention‐to‐treat, ITT, 27/60) or 47% (per protocol 27/58); they did not differ (ITT) between omeprazole (50%), lansoprazole (40%) and ranitidine (45%). Median pH and time at which intragastric pH was above 4 was slightly lower for ranitidine (4.0 ± 1.7; 51 ± 25%) than for omeprazole (5.4 ± 1.1; 77 ± 25%; P < 0.05) or lansoprazole (4.4 ± 1.6; 68 ± 32%). Plasma concentrations of amoxycillin were comparable in all three treatment groups. Post‐treatment H. pylori status was not dependent on those levels, or the drug‐induced extent or duration of increased intragastric pH. However, H. pylori‐eradicated patients were significantly (P < 0.05) older (56 ± 13 years) than patients still H. pylori‐positive (47 ± 14 years). In addition, in patients older than 50 years (n = 33), eradication was higher (P < 0.01) than in patients (n = 25) below 50 years (65 vs. 24%). Eradication rate was highest (75–83%) in subgroups of patients (>50 years and history of peptic ulcer or smokers). Neither activity/grade of peptic ulcer or erosive gastritis nor initial diagnosis were predictors for clinical outcome.

Conclusion: The age of patients must be regarded as a major determinant of H. pylori eradication rate and may represent an important factor contributing to the highly variable clinical results.
Ferrous sulphate interacts with captopril
Authors: Schaefer J. P. , Tam Y. , Hasinoff B. B. , Tawfik S. , Peng Y. , Reimche L. , Campbell N. R. C.
Published: November 2003

Aims
 To determine if iron binds strongly to captopril and reduces captopril absorption.

Methods
 A variety of in vitro experiments was conducted to examine iron binding to captopril and a randomized, double‐blind, placebo controlled, cross‐over study design was used to assess the in vivo interaction. Captopril (25 mg) was coingested with either ferrous sulphate (300 mg) or placebo by seven healthy adult volunteers. Subjects were phlebotomized and had blood pressure measured at 0, 0.25, 0.5, 1, 2, 4, 6, 8, and 12 h post ingestion. A 1 week washout period was used.

Results
 The coingestion of ferrous sulphate and captopril was associated with a 37% (134 ng ml−1 h, 95% CI 41–228 ng ml−1 h, P=0.03) decrease in area under the curve (AUC) for unconjugated plasma captopril. There were no substantial changes in Cmax (mean difference;–32; 95% CI −124–62 ng ml−1 (P=0.57)) or in tmax (mean difference; 0; 95% CI −18–18 min (P=0.65)) for unconjugated captopril when captopril was ingested with iron. There was a statistically insignificant increase in AUC for total plasma captopril of 43% (1312 ng ml−1 h, 95% CI −827–3451 ng ml−1 h P=0.27) when captopril was ingested with iron. The addition of ferric chloride to captopril resulted in the initial rapid formation of a soluble blue complex which rapidly disappeared to be replaced by a white precipitant. The white precipitate was identified as captopril disulphide dimer. There were no significant differences in systolic and diastolic blood pressures between the treatment and placebo groups.

Conclusions
 Co‐administration of ferrous sulphate and iron results in decreased unconjugated captopril levels likely due to a chemical interaction between ferric ion and captopril in the gastrointestinal tract. Care is required when coprescribing captopril and iron salts.
Growth of macroscopic human megakaryocyte colonies from cord blood in culture with recombinant human thrombopoietin (c‐mpl ligand) and the effects of gestational age on frequency of colonies
Authors: Nishihira Hirokazu , Toyoda Yasunori , Miyazaki Hiroshi , Kigasawa Hisato , Ohsaki
Published: November 2003

We investigated the effects of recombinant human thrombopoietin (rhTPO) on the growth of megakaryocytic (MK) colony derived MK progenitors from human cord blood (CB) in vitro and the effects of gestational age on the number of MK colonies. The results demonstrated that rhTPO alone supports the growth of MK colonies and induces not only proliferation but also differentiation of MK progenitors. CB shows a high frequency of MK colonies; most of which are very large and equivalent to high proliferative potential colony‐forming unit‐megakaryocyte. The colonies could be macroscopically observed as white spots in the culture dish. Preterm neonates showed greater numbers of MK progenitors than term neonates and there was an inverse correlation between gestational age and concentration of MK progenitors of CB. The effects of gestational age was an important factor on the proliferative capacity of MK progenitors and on the response to rhTPO.
Characterization of a large chromosomal deletion in the PROS1 gene of a patient with protein S deficiency type I using long PCR
Authors: Holmes Zandra R. , Bertina Rogier M. , Reitsma Pieter H.
Published: November 2003

A long‐PCR‐based technique was developed to investigate a possible deletion in the protein S gene, PROS1, in a family with type I protein S (PS) deficiency (pedigree PS62). Long‐PCR across introns produced an unexpected 2 kb PCR product between exon VII and XII not seen in control individuals, in addition to the expected 2.5 kb exon VII–VIII product. This result suggested that a deletion had occurred between exons VII and XII in the PS‐deficient family members. All were heterozygous for the deletion. Sequencing of the cloned 2 kb fragment gave the precise location of the breakpoints within introns 7 and 11. Significant similarity existed in both introns to repetitive sequences, e.g. Alu and Mer12, but no significant similarity was evident between the two introns themselves. The technique of long‐PCR is simple and more informative than Southern blotting in detecting and characterizing large intragenic deletions.
Modulation of anti‐idiotypic immune response by immunization with the autologous M‐component protein in multiple myeloma patients
Authors: Bergenbrant Susanne , Yi Qing , Ö Sterborg Anders , Björkholm Magnus , Ö Sby Eva , Mellstedt HÅkan , Lefvert Ann Kari , Holm Göran
Published: November 2003

Multiple myeloma is characterized by a proliferation of clonal B lymphocytes and plasma cells. The idiotypic structure of clonal immunoglobulin (Ig) expressed on the tumour B‐cell surface can be regarded as a tumour‐specific antigen and, as such, a potential target for anti‐idiotypic T and B cells in an immune regulation of the tumour‐cell clone. Active immunization using the autologous monoclonal Ig as a ‘vaccine’ was shown to induce tumour‐specific immunity in murine B‐cell tumours and in human B‐cell lymphoma. With the aim to induce or amplify an anti‐idiotypic response in multiple myeloma, five stage I–III patients were repeatedly immunized with the autologous monoclonal IgG. Induction of idiotype‐specific cellular immunity was analysed in vitro by an enzyme‐linked immunospot assay (interferon‐γ and interleukin‐4 secreting cells). B cells secreting anti‐idiotypic IgM antibodies were also analysed. An anti‐idiotypic T‐cell response was amplified 1.9–5‐fold in three of the five patients during immunization. The number of B cells secreting anti‐idiotypic antibodies also increased in these three patients. In two of the patients induction of idiotype‐specific immunity was associated with a gradual decrease of blood CD19+ B cells. The induced T‐cell response was eliminated during repeated immunization. Further studies are warranted to optimize the immunization schedule in order to achieve a long‐lasting T‐cell immunity against idiotypic determinants on the tumour clone. A role for immunity in controlling the tumour clone remains to be established.
Evidence for a single origin of factor V Leiden
Authors: Cox M. J. , Rees D. C. , Martinson J. J. , Clegg J. B.
Published: November 2003

Factor V Leiden is the most common hereditary blood clotting disorder so far identified, with an allele frequency of 4%. The low prevalence of the mutation outside of Europe suggests it occurred as a single event in the European founding population. In this study four polymorphisms have been identified defining different haplotypes in the exon 13 region of the factor V gene. One of these polymorphisms predicts a novel amino acid change, threonine to serine, in the B‐domain of factor V. Statistical evidence for a single origin of factor V Leiden is provided through the association of the mutation with a single exon 13 haplotype.
Plasmin accelerates platelet‐dependent prothrombinase formation without activating the platelets
Authors: Liu Longbin , Freedman John , Hornstein Adriana , Dewar Lori , Blajchman Morris , Ofosu Frederick
Published: November 2003

Patients with acute myocardial infarction who undergo thrombolytic therapy may shortly thereafter present evidence for increased platelet activation and thrombin activity, and recurrent thrombosis. This study investigated whether plasmin activates platelets and prothrombin in recalcified platelet‐rich plasma (RPRP) to cause (at least in part) these side‐effects of thrombolytic therapy. Plasmin (0.1 and 1.0 CU/ml) addition to RPRP with 1 μM r‐tick anticoagulant peptide (the latter a factor Xa inhibitor which abrogates prothrombin activation by prothrombinase at the concentration used) resulted in no change in the concentration of prothrombin fragment 1 + 2, or in the expression of GMP‐140, the resting and activated GP IIb–IIIa conformers, and GPIb on platelets. Thus, plasmin neither activates platelets nor prothrombin in RPRP. However, plasmin accelerated platelet activation and secretion, and prothrombin fragment 1 + 2 production in RPRP. When combined with 1 μM r‐tick anticoagulant peptide and 1 or 10 nMα‐thrombin to RPRP, plasmin also increased the number of GMP‐140 molecules expressed/platelet without enhancing α‐thrombin binding to the platelets. Additionally, plasmin accelerated prothrombin activation when it was added to washed platelets resuspended in factor V depleted plasma simultaneously with 10 mM CaCl2, 10 nMα‐thrombin for 10 s (to activate platelets and platelet factor V), followed by 4 μM hirudin and 1 nM factor Xa. Thus, plasmin potentiates the platelet release reaction in response to α‐thrombin (probably by increasing the availability of factor V on the platelets) to enhance prothrombin activation in RPRP. These actions of plasmin may contribute to the increased platelet activation and thrombotic side‐effects that can occur after thrombolytic therapy.
Heparin‐induced skin lesions
Authors: Warkentin Theodore E.
Published: November 2003

Six patients who developed heparin‐induced skin lesions were tested for the presence of heparin‐dependent, platelet‐activating IgG irrespective of whether they developed thrombocytopenia. All six patients' sera contained potent heparin‐dependent, platelet‐activating IgG. However, only two of the six patients developed thrombocytopenia, and both of these patients developed thrombotic complications. One of the four patients who developed skin lesions without thrombocytopenia also developed a characteristic complication of heparin‐induced thrombocytopenia (adrenal haemorrhagic infarction). These observations indicate that formation of heparin‐dependent, platelet‐activating IgG is a consistent feature of patients who develop heparin‐induced skin lesions irrespective of whether thrombocytopenia occurs.
Rebound after cessation of oral anticoagulant therapy: the biochemical evidence
Authors: Genewein Urs , Haeberli André , Straub P. Werner , Beer Jürg H.
Published: November 2003

The existence of a phenomenon of rebound hypercoagulability after cessation of oral anticoagulant therapy is controversial. The sensitive procoagulant markers for in vivo thrombin and fibrin formation are potential tools for the reassessment of the presence of such a phenomenon. We examined 19 patients anticoagulated for 6 ± 2 months (SD, range 3–12) because of venous thromboembolism or myocardial infarction as follows: twice during stable, oral anticoagulation (INR 3.1–3.7) and then on days 1, 2, 3, 4, 5, 7, 9, 11, 13, 15 and >30 after cessation of oral anticoagulation. Thrombin–antithrombin III complexes (TAT) and fibrinopeptide A (FPA) were measured in addition to the prothrombin times and factors II, V, VII, and X. None of the 19 patients developed clinically manifest thromboembolism within the following 9–18 months. However, the patients’ TAT levels increased transiently: rising from 1.5 ± 0.1 ng/ml (SEM) to 3.0 ± 0.2 ng/ml on day 4 (P < 0.001), and returned to 1.7 ± 0.1 ng/ml after day 30 (normals 1.8 ± 0.33). 17/19 patients showed TAT peak levels above the upper limit of normal between days 3 and 11 (average: day 4), which normalized again after 30 d. 8/19 patients also had transient FPA levels above the upper normal limits (<1.81). We conclude that our patients increased their thrombin and fibrin formation transiently and that a subpopulation reached values consistent with a prethrombotic state.
The effect of iron chelation on haemopoiesis in MDS patients with transfusional iron overload
Authors: Jensen P. D. , Heickendorff L. , Pedersen Bent , Bendix‐Hansen K. , Jensen F. T. , Christensen T. , Boesen A. M. , Ellegaard J.
Published: November 2003

Long‐term follow‐up data are presented on changes in peripheral blood counts and Hb requirements of 11 patients with myelodysplastic syndromes (MDS) during iron chelation treatment with desferrioxamine for up to 60 months. The erythroid marrow activity was indirectly evaluated by repeated determinations of the serum transferrin receptor concentration. The efficacy of iron chelation was evaluated by repeated quantitative determination of the liver iron concentration by magnetic resonance imaging.

Reduction in the Hb requirement (≥50%) was seen in 7/11 (64%) patients. Five patients (46%) became blood transfusion independent. Platelet counts increased in 7/11 (64%) patients and the neutrophil counts in 7/9 (78%) evaluable patients. All patients in whom iron chelation was highly effective showed improvement of erythropoietic output accompanied by an increase in the serum transferrin receptor concentration.

It is concluded that reduction in cytopenia in MDS patients may be accomplished by treatment with desferrioxamine, if the iron chelation is efficient and the patients are treated for a sufficiently long period of time. Exactly how treatment with desferrioxamine works remains a challenge for further investigation.
Treatment of acute myelogenous leukaemia in patients aged 50–65: idarubicin is more effective than zorubicin for remission induction and prolonged disease‐free survival can be obtained using a unique consolidation course
Authors: Pignon B. , Witz F. , Desablens B. , Leprise P. Y. , Francois S. , Linassier C. , Berthou C. , Caillot D. , Lioure B. , Cahn J. Y. , Casassus P. , Sadoun A. , Audhuy B. , Guyotat D. , Briere J. , Vilque J. P. , Baranger L. , Polin V. , Berthaud P. , Hurteloup P. , Herve P. , Harousseau J. L.
Published: November 2003

From December 1987 to June 1992, 251 patients aged 50–65 with de novo acute myelogenous leukaemia (AML) were recruited to a multi‐institutional randomized clinical trial. Induction therapy consisted of Ara‐C (200 mg/m2, continuous infusion, days 1–7) with either zorubicin (ZRB) (200 mg/m2, i.v., days 1–4) or idarubicin (IDR) (8 mg/m2, i.v., days 1–5). Consolidation therapy consisted of a single course of intensive chemotherapy with high‐dose Ara‐C (3 g/m2, 3 h infusion, q 12 h, days 1–4) and m‐Amsa (100 mg/m2/d, i.v., days 5–7).

The complete remission (CR) rate was (73%) with Ara‐C/IDR versus (60%) with Ara‐C/ZRB (P = 0.033). In multivariate analysis, factors found to be significant in predicting CR were normal karyotype and treatment with IDR. With a median follow‐up of 73 months, the median disease‐free survival (DFS) duration of all CR patients and the probability of CR at 6 years were 17 months and 29%. In multivariate analysis the only factor associated with an increased DFS duration was a normal karyotype. The median event‐free survival (EFS) duration for all evaluable patients and the median overall survival duration for all eligible patients were respectively 7 and 12 months without any difference between induction arms.

The study shows that in patients aged 50–65 idarabicin is more effective than zorubicin for remission induction. However, the type of anthracycline did not influence overall survival duration. Using a unique consolidation course, we observed a prolonged DFS which compares favourably with results obtained with more prolonged consolidation therapy or maintenance treatment.
Characterization of P2x1 purinoreceptors on rat platelets: effect of clopidogrel
Authors: Savi Pierre , Bornia Josette , Salel Veronique , Delfaud Marianne , Herbert Jean‐Marc
Published: November 2003

This study aimed to determine the binding characteristics of [3H]α,β‐Me‐ATP, a specific ligand of the P2x1 receptors to rat platelets, and to investigate the effect of clopidogrel, a thienopyridine compound which has been found to selectively inhibit ADP‐induced platelet aggregation and adenylyl cyclase ex vivo. Binding of [3H]α,β‐Me‐ATP to rat platelets was time‐dependent and saturable. Scatchard analysis of the saturation binding data indicated that [3H]α,β‐Me‐ATP bound to one population of specific binding sites with high affinity (KD  =  23.6 ± 1.6 nm; Bmax = 690 ± 24 fmole/108cells) (n=3). Unlabelled α,β‐Me‐ATP as well as 2‐MeS‐ADP and ADP competitively inhibited the specific binding of [3H]α,β‐Me‐ATP with IC50 values of 19.0 ± 6.6, 103 ± 20 and 1120 ± 80 nm respectively (n=3). Other nucleotide analogues such as ATP, ATP‐γS, UTP and GTP also antagonized [3H]α,β‐Me‐ATP binding. When administered orally (10 mg/kg, p.o.), clopidogrel inhibited ADP‐ or 2‐MeS‐ADP‐induced platelet aggregation but did not affect the binding of [3H]α,β‐Me‐ATP to rat platelets ex vivo. In vitro, α,β‐Me‐ATP did not induce the aggregation or shape change of rat platelets and did not interfere with ADP‐induced platelet aggregation.
Rapid genetic screening for haemochromatosis using heteroduplex technology
Authors: Jackson Helen A. , Bowen Derrick J. , Worwood Mark
Published: November 2003

The recently described association between haemochromatosis and mutations in the HFE or HLA‐‘H’ gene has prompted the need for a simple and rapid genetic test capable of detecting multiple mutations simultaneously. Heteroduplex analysis, a new diagnostic technique, fulfills such criteria and we have investigated the potential for the detection of the Cys282Tyr mutation.

100 subjects were genotyped using the heteroduplex approach. The results showed clear distinction between individuals who did not carry the mutation, individuals who were heterozygous for the mutation and homozygous individuals. Heteroduplex results obtained by both silver staining and capillary electrophoresis showed 100% concordance with those obtained by restriction digestion of PCR product.
Clotting factor IX levels in C/EBPα knockout mice
Authors: Davies N. , Austen D. E. G. , Wilde M. D. , Darlington G. J. , Brownlee G. G.
Published: November 2003

Previous studies have indicated that C/EBPα is involved in the regulation of factor IX and mutations of a C/EBP recognition element in the factor IX promoter result in haemophilia B. We now report that mice homozygous for the deletion of the c/ebpα gene are significantly deficient in factor IX transcription.
The rapid diagnosis of acute promyelocytic leukaemia using PML (5E10) monoclonal antibody
Authors: O'Connor Sheila J. M. , Forsyth Peter D. , Dalal Surita , Evans Paul A. , Short Michael A. , Shiach Caroline , Jack Andrew S. , Morgan Gareth J.
Published: November 2003

Acute promyelocytic leukaemia (APL) is characterized cytogenetically by t(15;17)(q22;q21) which results in the production of a PML/RARα fusion protein. Detection of the translocation or the fusion gene product is required for objective diagnosis of APL. This can be accomplished by conventional cytogenetic methods, fluorescence in situ hybridization or RT‐PCR. Such techniques are time consuming and not universally available. The intracellular distribution of the PML protein in promyelocytes is characteristically altered in APL and this can be detected by immunocytochemistry. We have assessed two immunocytochemical methods, immunofluorescence and alkaline phosphatase–anti‐alkaline phosphatase staining (APAAP), with regard to sensitivity, specificity and rapidity of diagnosis. 85 patients with AML including 15 cases of APL were studied. Immunofluorescence PML detection was concordant with RT‐PCR for t(15;17) in 14/15 (93.3%) cases with no false positives. The negative APL case in our series was a patient with a 5′ PML breakpoint who did not express the reciprocal t(17;15) fusion product. APAAP was concordant in only 6/13 (46%) APL cases with one false positive. In conclusion, immunofluorescent localization of PML using 5E10 monoclonal antibody is a rapid, sensitive and specific diagnostic tool for APL.
Activated protein C resistance in homozygous sickle cell disease
Authors: WRIGHT J. G. , COOPER P. , MALIA R. G. , KULOZIK A. E. , VETTER B. , THOMAS P. , PRESTON F. E. , SERJEANT G. R.
Published: November 2003

The activated protein C resistance (APC‐R) ratios in 50 patients with steady state homozygous sickle cell (SS) disease and 59 healthy AA controls was measured. There was a significant reduction in median APC‐R ratio in sickle cell disease compared to controls. This reduction in APC‐R ratio was not explained by (1) the presence of the factor V Leiden, found in only one of 165 patients with SS disease including those tested for APC‐R, or (2) the presence of lupus anticoagulants. However, the raised levels of factor VIIIC in SS patients in this study may be contributing to increased resistance to APC, which in turn may contribute to the vaso‐occlusive complications of SS disease.
Expression of mdr1 and mrp in the normal B‐cell homologue of B‐cell chronic lymphocytic leukaemia
Authors: WALL D. M. P. , EL‐OSTA S. , TZELEPIS D. , BERTONCELLO I. , KANTHARIDIS P. , CHOU S. T. , ZALCBERG J. R. , PARKIN J. D.
Published: November 2003

B‐cell chronic lymphocytic leukaemia (CLL) cells commonly express the multidrug resistance phenotype. The aim of this study was to establish whether the normal homologue in B‐cell ontogeny of B‐CLL also expressed the multidrug resistance (mdr) phenotype. Human tonsillar lymphocytes were sorted to yield two B‐cell subsets based on the expression of CD19, CD5 and CD10. The normal homologue was represented by a population of B cells that was CD19 positive, CD10 negative and weakly expressed CD5. Based upon functional analysis and the detection of mdr1 mRNA by semi‐quantitative PCR, these cells expressed the mdr phenotype. In contrast, functional multidrug resistance could not be demonstrated in CD19‐positive CD10‐positive cells with strong expression of CD5, nor could mdr1 mRNA be found in these cells. MRP was variably expressed in both B‐cell subsets with no discernable differences in the pattern of expression. We conclude that normal B cells with a phenotype resembling that of B‐CLL cells express the multidrug resistance phenotype.
Analysis of blood T‐cell cytokine expression in B‐chronic lymphocytic leukaemia: evidence for increased levels of cytoplasmic IL‐4 in resting and activated CD8 T cells
Authors: MU XI , KAY NEIL E. , GOSLAND MICHAEL P. , JENNINGS C. DARRELL
Published: November 2003

The cytoplasmic cytokines of purified blood T cells (CD4/C]D8) in B‐CLL patients (n = 5) and controls (n =5) were evaluated by flow cytometry. The mean levels of cytoplasmic IL‐4 were significantly elevated in resting and activated B‐CLL CD8 cells compared to control CD8 cells. IL‐4 cytoplasmic levels were comparable for resting B‐CLL and control CD4 cells but greater for B‐CLL activated CD4 cells. The mean fluorescence intensity of B‐CLL CD8 cytoplasmic IL‐4 was 4–5‐fold greater, indicating higher IL‐4 density per CLL CD8 than control CD8 cells. Both CLL CD4 and CD8 cells post‐activation had higher levels of cells double positive for cytoplasmic IL‐4 and interferon. These data indicate that freshly isolated CD8 and CD4 blood T cells from B‐CLL patients have significantly elevated (above control) levels of commitment to expression of IL‐4. Since IL‐4 has an important modulatory impact on CLL and normal B cells, this observation has implications regarding the biology of B‐CLL.
Phenotypic characterization and analysis of allogeneic T cell responses in ZAP-70 dominant negative transgenic mice
Authors: GLADUE R P , ALLEN M , CUNNINGHAM A , GARDNER J , LAQUERRE A M , CONNELLY P A , SHAW A S , MCNEISH J
Published: November 2003

Antigen stimulation of T cells results in a series of biochemical events including the interaction of both SH2 domains of ZAP-70 with phosphorylated ITAMS on the T cell receptor. In order to study the physiological relevance of decreasing native ZAP-70–SH2 interaction in vivo, we generated transgenic mice expressing a T cell-specific, dominant negative form of ZAP-70 consisting of only the tandem SH2 domains (ZAP-NC). Phenotypically, these animals had a comparable distribution of lymphocyte subsets in the thymus and spleen compared with the wild-type (WT) controls. However, examination of peripheral blood revealed a slow but progressive decrease in the number of lymphocytes, particularly CD4+ cells, with age (17% reduction by 3 months, 58% reduction by 6 months). Allogeneic responses were then evaluated in vitro as well as in vivo using a subcutaneous sponge matrix implant. Although spleen cells cultured for 4 days in vitro with alloantigen developed normal functional responses, allogeneic responses generated in vivo within a subcutaneous sponge matrix were impaired. This was characterized by a depression in cytotoxic T lymphocyte (CTL) activity, a 82% reduction in the frequency of helper T cells, and a 78% reduction in the capacity of sponge-infiltrating lymphocytes to produce IL-2 in response to secondary antigen stimulation. These results indicate that although overt lymphocyte development and in vitro function were unremarkable, expression of a truncated ZAP-70 affected the in vivo survival of peripheral lymphocytes and altered the in vivo generation of functional activity to alloantigen.
Inhibition of nitric oxide synthase (NOS) aggravates Staphylococcus aureus septicaemia and septic arthritis
Authors: SAKINIENE E , BREMELL T , TARKOWSKI A
Published: November 2003

The aim of this study was to assess the role of NO and its metabolites in bacterial arthritis. The murine model of haematogenously acquired septic arthritis was used. Swiss mice treated with NOS inhibitors (NG-monomethyl-l-arginine or Nω-nitro-l-arginine methyl ester) were injected intravenously with toxic shock syndrome toxin-1 (TSST-1) producing Staphylococcus aureus LS-1. Arthritis was evaluated clinically and histopathologically. Serum cytokine levels, bacterial isolates and intracellular capacity of macrophages to kill bacteria were also analysed. The frequency of arthritis in mice treated with NOS inhibitors was three to four-fold higher than that in non-treated controls (75% versus 20%). The severity of arthritis, expressed as mean arthritic index, was 1·4 and 0·4, respectively. Cartilage and/or bone destruction occurred in 63% of NOS inhibitor-treated mice, but only in 10% of controls. Also, the cumulative septicaemia-induced mortality was clearly higher in mice treated with NOS inhibitors compared with non-treated controls. Intracellular killing capacity of the peritoneal macrophages, treated in vitro with NOS inhibitors, was decreased. Thus, 24 h after bacterial inoculation peritoneal macrophages pretreated with NOS inhibitors killed more than 10 times less bacteria than the control ones (P < 0.01). We conclude that NOS inhibitors aggravate S. aureus arthritis, possibly by inducing impairment of the intracellular killing capacity of macrophages.
Anti-native and recombinant myeloperoxidase monoclonals and human autoantibodies
Authors: Audrain M A P , Baranger T A R , Moguilevski N , Martin S J , Devys A , Lockwood C M , Muller J Y , Esnault V L M
Published: November 2003

Myeloperoxidase (MPO) is one of the main antigen targets of anti-neutrophil cytoplasmic antibodies (ANCA) in systemic vasculitides. It has been suggested that anti-MPO antibodies may recognize a single epitope on recombinant MPO. If confirmed on native MPO, this might allow specific therapeutic intervention with anti-idiotypic MoAbs to prevent antibody–antigen interaction which is thought to cause activation of neutrophils and vasculitis. We searched for restriction in the epitope recognition profile in 50 patients with anti-MPO autoantibodies, using both native and recombinant MPO. Mouse monoclonals were purified and tested in competition assays. At least four epitopes were identified on native MPO using these monoclonals and only two were conserved on recombinant MPO. We found that human MPO autoantibody response was not restricted to a single epitope on native MPO, as all sera tested did not show the same profile in competitive studies with monoclonals. Furthermure, 30% of human anti-native MPO sera failed to recognize rMPO.
Thiol compounds inhibit mercury-induced immunological and immunopathological alterations in susceptible mice
Authors: Hu H , Moöller G , Abedi-Valugerdi M
Published: November 2003

In vitro mercury induces a high proliferative response in splenic lymphocytes and in vivo it induces a systemic autoimmune disease in susceptible mouse strains. This disease is characterized by increased serum levels of IgE and IgG1 antibodies, by the production of anti-nucleolar antibodies and by the formation of renal immune complex deposits. We have previously found that the presence of 2-mercaptoethanol (2-ME) inhibited mercury-induced cell proliferation in vitro. In this study, we tested the effects of four other thiol compounds, namely dithiothreitol (DTT), L-cysteine, meso-2,3-dimercaptosuccinic acid (meso-DMSA) and 2,3-dimercapto-1-propanesulfonic acid, Na salt (DMPS) on mercury-induced immunological changes both in vitro and in vivo. We found that in vitro, the addition of all thiol compounds abrogated mercury-induced cell aggregation and proliferation. In vivo, injection of meso-DMSA and/or DMPS (s.c. or i.p.) immediately following exposure to mercury markedly decreased IgG1 synthesis in spleen cells and serum IgE levels in mercury-susceptible SJL mice. Treatment with DMPS also prevented mercury-induced IgG1 anti-nucleolar antibody synthesis and the development of mesangial IgG1 immune complex deposits in SJL mice.
Serum concentrations of free and total insulin‐like growth factor‐I, IGF binding proteins ‐1 and‐3 and IGFBP‐3 protease activity in boys with normal or precocious puberty
Authors: Juul Anders , Flyvbjerg Allan , Frystyk Jan , Müller Jørn , Skakkebæk Niels E.
Published: November 2003

OBJECTIVE Circulating IGF‐I and IGF binding protein‐3 (IGFBP‐3) levels both increase in puberty where growth velocity is high. The amount of free IGF‐I is dependent on the IGF‐I level and on the concentrations of the specific IGFBPs. Furthermore, IGFBP‐3 proteolysis regulates the bioavailability of IGF‐I. However, the concentration of free IGF‐I and possible IGFBP‐3 proteolytic activity in puberty has not previously been studied.

SUBJECTS AND MEASUREMENTS We investigated serum levels of easily dissociable IGF‐I concentrations and ultrafiltrated free IGF‐I levels by specific assays in 60 healthy boys and in 5 boys with precocious puberty before and during GnRH agonist treatment. In addition, total serum IGF‐I, IGFBP‐1 and IGFBP‐3 levels as well as IGFBP‐3 protease activity were determined.

RESULTS Free (dissociable and ultrafiltrated) IGF‐I concentrations were significantly higher in pubertal boys than in prepubertal children and correlated significantly with the molar ratio between IGF‐I and IGFBP‐3 (r=0.69, P<0.0001 and r=0.54, P=0.0008, respectively) and inversely with IGFBP‐1 (r=−0.47, P<0.0001 and r=−0.43, P=0.0003, respectively). Multiple regression analysis suggested that IGFBP‐3 level, and not IGFBP‐1, was the major determinant of the free IGF‐I serum level in normal boys. Free IGF‐I levels were elevated in boys with precocious puberty and decreased during GnRH treatment. IGFBP‐3 proteolysis was constant throughout puberty (mean 20%).

CONCLUSIONS We conclude that easily dissociable and ultrafiltrated free IGF‐I serum levels are increased in boys with normal and precocious puberty and suggest that the increased free IGF‐I serum concentration in puberty primarily reflects changes in total concentrations of IGF‐I and IGFBPs secondary to increased GH secretion, but that it is not influenced by changes in IGFBP‐3 proteolysis.
Glandular lesions of the cervix—cytological and histological correlation
Authors: JACKSON S. R. , HOLLINGWORTH T. A. , ANDERSON M. C. , JOHNSON J. , HAMMOND R. H.
Published: November 2003

In this retrospective review we assessed the frequency with which atypical glandular cells in cervical smears predict cervical glandular lesions. Asymptomatic patients (n=34) with one or more smears showing atypical glandular cells and subsequent histopathological assessment were studied. Independent cytological and histological review was undertaken. Cytological review confirmed atypical glandular cells in 29 cases, 17 of which had coexisting squamous dyskaryosis. Histological review of these 29 cases revealed glandular neoplasia in 13 (45%) and microglandular hyperplasia (MEH) in an additional four (14%). Initial reporting had underestimated the prevalence of glandular neoplasia.
CO2 flux estimates tend to overestimate ecosystem C sequestration at elevated CO2
Authors: Niklaus P. A. , Stocker R. , Körner CH. , Leadley P. W.
Published: November 2003

1. Instantaneous leaf photosynthesis and land area‐based net ecosystem CO2 exchange (NEC) are almost universally increased at elevated CO2 concentrations, at least in the short term and under high light conditions. This raises the possibility that terrestrial ecosystems sequester extra C in response to elevated CO2, and it has been hypothesized that part of this extra C is stored in soils.

2. Attempts to quantify ecosystem C sequestration experimentally are based on (i) ecosystem CO2 exchange measurements; (ii) C isotope tracking; and (iii) direct C stock measurements.

3. Because direct C stock measurements are insensitive to increases in ecosystem C storage in the range expected, and C isotope tracking is methodologically difficult because of the need to account for new and old soil C pools, NEC measurements were considered to be a more direct and unbiased method to estimate net ecosystem production and C sequestration at elevated CO2.

4. Here we present a case study in calcareous grassland under long‐term CO2 enrichment in which we demonstrate that calculated C balances are extremely sensitive to systematic experimental biases inherent in any CO2 flux‐based study at elevated CO2. A sensitivity analysis demonstrates that these systematic errors tend to result in severe overestimation of ecosystem C accretion under elevated CO2.

5. Carbon isotope data and soil pool C and N measurements from the same study add to the evidence that the C balance derived from CO2 flux measurements at elevated CO2 is overestimated.

6. Based on this evidence, and the wide attention given to C sequestration at elevated CO2, we suggest a critical reconsideration of the appropriateness of NEC measurement in CO2‐enriched ecosystems as a basis for calculating ecosystem C balances at elevated CO2.