bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of a short postgraduate course in rational pharmacotherapy for general practitioners
Authors: Akici Ahmet , Kalaça Sibel , Ugurlu M. Ümit , Karaalp Atila , Çali Şanda , Oktay Şule
Published: December 2004

Aims  The impact of a short postgraduate course on rational pharmacotherapy planning behaviour of general practitioners (GP) was investigated via a face‐to‐face interview with 25 GPs working at health centres in Istanbul.

Methods  GPs were randomly allocated to control and intervention groups. Intervention group attended a 3‐day‐training program preceded and followed by a written exam to plan treatment for simulated cases with a selected indication. The participants’ therapeutic competence was also tested at the post‐test for an unexposed indication to show the transfer effect of the course. In addition, patients treated by these GP's were interviewed and the prescriptions were analysed regarding rational use of drugs (RUD) principles at the baseline, 2 weeks and 4 months after the course.

Results  At the baseline there was not any significant difference between the control and intervention groups in terms of irrational prescribing habits. The questionnaires revealed that the GPs were not applying RUD rules in making their treatment plans and they were not educating their patients efficiently. Training produced a significant improvement in prescribing habits of the intervention group, which was preserved for 4 months after the course. However, very low scores of the pretest indicate the urgent necessity for solutions.

Conclusions  Training medical doctors on RUD not only at the under‐ but also at the postgraduate level deserves attention and should be considered by all sides of the problem including academia, health authorities and medical associations.
The effect of hypertension and hypercholesterolemia screening with subsequent intervention letter on the use of blood pressure and lipid lowering drugs
Authors: Atthobari Jarir , Monster Taco BM , De Jong Paul E , De Jong‐van den Berg Lolkje TW ,   Prevend Study Group
Published: December 2004

Aim  To evaluate the effect of a letter intervention that was send to both the participants of a population screening and their general practitioners. We also tested what predicting variables influenced the GP to actually prescribe blood pressure lowering drugs (BPLD) or lipid lowering drugs (LLD).

Method  The study design was cross sectional, in the PREVEND outpatient clinic in Groningen University Hospital, the Netherlands. We used the clinical data of the 8592 subjects that participated in the first screening of the PREVEND study. Data on drug use was collected from community pharmacies. Drug use was measured the year before and after the screening with the subsequent intervention letter. As control population without intervention, we used the data from the InterAction DataBase (IADB) standardized for the population characteristics of the intervention group. The letter intervention was sent to participants who had shown after screening to have either an elevated blood pressure or plasma cholesterol, and the letter contained the advice to use a BPLD or LLD. Main outcome measures were proportion of patients prescribed BPLD and/or LLD in the year before and after the intervention, and variables that influence the GP to prescribe BPLD and LLD.

Results  Data from the community pharmacy were available from 7567 (88%) subjects. 397 participants (5.2%) received a letter with advice to start a BPLD, and 326 participants (4.3%) received a letter with advice to start a LLD. The prevalence of patients who were using BPLD and LLD before the intervention was not significantly different between the intervention and control group, 16.6 (CI 95% 15.8 –17.5) vs 16.0 and 4.8 (4.4–5.3) vs 4.6, respectively. After the letter intervention, the prevalence of BPLD use was higher in the intervention group compared with the control group (19.4 [18, 5–20, 3]vs 17.0%), as was the prevalence of LLD use (7.1[6.5–7.7) vs 5.4%). The same held true for the incidence of BPLD (3.4[3.0–3.8]vs 2.5%) and LLD use (2.1 [1, 6–2, 4]vs 1.0%), respectively, in the year after the intervention. Univariate and multivariate analysis showed that a higher blood pressure and cholesterol level, but not the presence of other cardiovascular risk factors, were associated to with a greater percentage use of a BPLD and a LLD.

Conclusion  A population survey followed by a letter of intervention to both the patient and GP are effective to improve the use of blood pressure and lipid lowering drugs as a primary prevention in patients with hypertension and hyperlipidemia. Our therapeutic advice however, was followed only in about one of the three subjects with hypertension and one of the four subjects with hyperlipidemia. The levels of blood pressure and plasma total cholesterol are important variables influencing the GP to prescribe a BPLD and/or LLD.
Have COX‐2 inhibitors influenced the co‐prescription of anti‐ulcer drugs with NSAIDs?
Authors: Teeling Mary , Bennett Kathleen , Feely John
Published: December 2004

Aims  To quantify usage of COX‐2 inhibitors compared with nonselective NSAIDs and to determine their impact (including financial) on the co‐prescription of antipeptic ulcer (anti‐PU) drugs.

Methods  The Irish General Medical Services prescription database (covering 1.2 million people) was examined for NSAID prescriptions during December 1999‐November 2001. NSAID users were excluded during the first 6 months. During the next 12 months (study period) patients on NSAIDs (≥ 3 prescriptions) were identified. The study period and final 6 months provided data on co‐prescription of anti‐PU drugs. Age, gender, number of concomitant prescriptions, co‐prescribing of anti‐PU drugs and monthly cost were evaluated for 8 NSAIDs (n= 4 non‐selective NSAIDs and n= 4 COX‐2 inhibitors) and odds ratios (OR) calculated using logistic regression.

Results  COX‐2 inhibitors were prescribed more frequently in older, female patients and those receiving multiple medications. After adjustment for age, gender and polypharmacy, anti‐PU drugs were prescribed more frequently with COX‐2 inhibitors (OR = 1.31 (1.23,1.40)). COX‐2 inhibitors were up to 10‐fold more expensive, median monthly costs (including anti‐PU drugs) ranging from €34.61 (COX‐2 inhibitors) to €3.26 (nonselective NSAIDs).

Conclusions  Since COX‐2 inhibitors are associated with increased rates of co‐prescription of anti‐PU drugs and are more expensive than non‐selective NSAIDs, these results suggest that the expected cost‐savings with COX‐2 inhibitors may not be occurring in practice.
Pharmacokinetic evaluation of a new oral sustained release dosage form of tramadol
Authors: Malonne H. , Sonet B. , Streel B. , Lebrun S. , De Niet S. , Sereno A. , Vanderbist F.
Published: December 2004

Aims To compare the pharmacokinetic profile of a new modified release formulation of tramadol (Tramadol LP 200 mg, SMB Technology, Marche‐en‐Famenne, Belgium) with that of an immediate release capsule (Topalgic® 50 mg, Grünenthal, Aachen, Germany) after single and multiple dosing and to assess the potential effect of food on its relative bioavailability.

Methods The first study had an open, single‐dose, three‐treatment, three‐period, six‐sequence, randomised, crossover design with at least a five‐day wash‐out. The second study had an open, steady‐state, two‐treatment, two‐period, two‐sequence, randomised crossover design with at least a seven‐day wash‐out. Both studies contained 30 healthy subjects. Both enantiomers of tramadol and O‐demethyl‐tramadol (the only active metabolite of tramadol) were assayed in the plasma using an LC‐MS/MS method. AUC∞, AUCt, Cmax, Tmax, and T1/2 were estimated. Statistical analysis was performed using univariate anova, the Wilcoxon nonparametric method or Friedman's nonparametric anova where appropriate.

Results Tramadol had a significantly lower Cmax and longer Tmax than the conventional formulation. Thus, the mean (± sd) Cmax of tramadol were 646 ± 192 and 300 ± 94 ng ml−1 for Topalgic® 4 × 50mg and Tramadol LP 200 mg, respectively (95% confidence interval on the difference expressed as a percentage 42–51). AUC of tramadol from both formulations was comparable (similar AUC∞ and AUCt). Thus, the mean AUC∞ of (+/–)tramadol obtained after multiple dosing were 4611 ± 1944 and 5105 ± 2101 ngh ml−1 after Topalgic® 4 × 50mg and Tramadol LP 200 mg, respectively (95%CI 102–123%). We also demonstrate that the pharmacokinetics of the drug are not influenced by the intake of food. Thus, the mean AUC∞ of (+/–) tramadol were 5444 ± 1637 and 5169 ± 1580 ngh ml−1 after Tramadol LP 200 mg given in the fasting and fed states, respectively (95%CI = 88–103%).

Conclusions The new sustained release form of tramadol exhibits adequate properties for once a day administration. Furthermore, its pharmacokinetic profile is not affected by the intake of food.
Trends in accessibility to medicines for children in New Zealand: 1998–2002
Authors: Chui Jocelyn , Tordoff June , Kennedy Julia , David Reith
Published: December 2004

Background Reported rates of unlicensed and off‐label use of medicines in children raise concerns regarding overall access of children to medicines

Objective To assess changes in availability of proprietary formulations suitable for infants and young children; licensing of medicines; and subsidization of medicines for children in New Zealand.

Methods Review of the New Ethicals Catalogue, New Ethicals Compendium, product data sheets and the New Zealand Pharmaceutical Schedule covering the years 1998–2002 inclusive.

Results There was a decrease in the total number of medicines licensed in New Zealand from 2014 to 1840; but there was an increase in the number and percentage of suitable formulations that were licensed for paediatric use from 616[31%] to 642[35%]. The number of suitable paediatric formulations that were subsidized decreased from 281[13.9%] to 260[14.1%]. The number of orally available chemical entities with suitable formulations, licensed and subsidized for paediatric use declined from 101[5.0%] to 94[5.1%], but all of these chemical entities that were withdrawn had therapeutic alternatives that were licensed and subsidized. Only 36% of new medicines that had licensing for children were licensed for the 0–23 month age group.

Conclusion There have been modest improvements in licensing of medicines for children in New Zealand from 1998 to 2002.
Optimization of drug prescribing
Authors: Jackson S. H. D. , Mangoni A. A. , Batty G. M.
Published: December 2004

The hazards of prescribing many drugs, including side‐effects, drug interactions, and difficulties of compliance, have long been recognized as particular problems when prescribing for elderly people. The need for appropriate and rational prescribing for elderly patients has been prioritized in the National Service Framework for Older People. This review addresses the research evidence on epidemiology of prescribing in elderly patients, methods of measuring the quality, and the role of the prescriber and the multidisciplinary team in the day‐to‐day optimization of drug therapy.
The presence of emphysema does not affect the systemic bioactivity of inhaled fluticasone in severe chronic obstructive pulmonary disease
Authors: Lee Daniel K. C. , Lipworth Brian J.
Published: December 2004

Aims

To assess the systemic bioactivity of fluticasone proprionate (FP) 2000 µg daily on sensitive adrenal and bone markers in severe chronic obstructive pulmonary disease (COPD) patients with or without significant emphysema.

Methods

Ten patients without emphysema (COPD group: age 55 years, FEV1 51% predicted and DLCO 83% predicted) and 10 patients with emphysema (COPDE group: age 59 years, FEV1 43% predicted and DLCO 49% predicted) received FP 2000 µg daily via a spacer for 2 weeks. There was a 1‐week washout period prior to FP treatment where patients were given salmeterol and oxitropium, after stopping their usual inhaled corticosteroids for the duration of the study. Measurements including overnight 10 h urinary cortisol excretion corrected for creatinine (OUCC) and serum osteocalcin concentrations were performed at baseline following washout and after 2 weeks of FP.

Results

Values for OUCC and serum osteocalcin concentrations pre‐ and post‐FP were not significantly different between the COPD and COPDE groups. There was significant suppression of OUCC (nmol mmol−1) by FP treatment within the COPD group (P = 0.03): 7.86 vs 4.64 (95% CI on the difference 0.47, 5.98), and within the COPDE group (P = 0.006): 7.13 vs 4.27 (95% CI on the difference: 1.03, 4.69). Likewise, there was significant suppression of osteocalcin concentration (nmol l−1) by FP treatment within the COPD group (P = 0.04): 7.24 vs 6.34 (95% CI on the difference: 0.01, 1.78), and within the COPDE group (P = 0.03): 6.92 vs 5.72 (95% CI on the difference: 0.12, 2.29).

Conclusions

Severe COPD patients who are receiving high dose FP are susceptible to the development of systemic adverse effects, irrespective of the presence of emphysema.
The CYP2C8 inhibitor trimethoprim increases the plasma concentrations of repaglinide in healthy subjects
Authors: Niemi Mikko , Kajosaari Lauri I. , Neuvonen Mikko , Backman Janne T. , Neuvonen Pertti J.
Published: December 2004

Aims

Our aim was to investigate the effect of the CYP2C8 inhibitor trimethoprim on the pharmacokinetics and pharmacodynamics of the antidiabetic drug repaglinide, and to examine the influence of the former on the metabolism of the latter in vitro.

Methods

In a randomized, double‐blind, crossover study with two phases, nine healthy volunteers took 160 mg trimethoprim or placebo orally twice daily for 3 days. On day 3, 1 h after the last dose of trimethoprim or placebo, they ingested a single 0.25 mg dose of repaglinide. Plasma repaglinide and blood glucose concentrations were measured for up to 7 h post‐dose. In addition, the effect of trimethoprim on the metabolism of repaglinide by human liver microsomes was investigated.

Results

Trimethoprim raised the AUC(0,∞) and Cmax of repaglinide by 61% (range, 30–117%; P= 0.0008) and 41% (P = 0.005), respectively, and prolonged the t½ of repaglinide from 0.9 to 1.1 h (P = 0.001). Trimethoprim had no significant effect on the pharmacokinetics of its aromatic amine metabolite (M1), but decreased the M1 : repaglinide AUC(0,∞) ratio by 38% (P = 0.0005). No effect of trimethoprim on the blood glucose‐lowering effect of repaglinide was detectable. In vitro, trimethoprim inhibited the metabolism of (220 nm) repaglinide in a concentration‐dependent manner.

Conclusions

Trimethoprim raised the plasma concentrations of repaglinide probably by inhibiting its CYP2C8‐mediated biotransformation. Although the interaction did not significantly enhance the effect of repaglinide on blood glucose concentration at the drug doses used, the possibility of an increased risk of hypoglycaemia should be considered during concomitant use of trimethoprim and repaglinide in patients with diabetes.
A prospective multicentre study of pharmacist initiated changes to drug therapy and patient management in acute care government funded hospitals
Authors: Dooley Michael J. , Allen Karen M. , Doecke Christopher J. , Galbraith Kirsten J. , Taylor George R. , Bright Jennifer , Carey Dianne L.
Published: December 2004

Aims

To determine the cost savings of pharmacist initiated changes to hospitalized patients’ drug therapy or management in eight major acute care government funded teaching hospitals in Australia.

Methods

This was a prospective study performed in eight hospitals examining resource implications of pharmacists’ interventions assessed by an independent clinical panel. Pharmacists providing clinical services to inpatients recorded details of interventions, defined as any action that directly resulted in a change to patient management or therapy. An independent clinical review panel, convened at each participating centre, confirmed or rejected the clinical pharmacist's assessment of the impact on length of stay (LOS), readmission probability, medical procedures and laboratory monitoring and quantified the resultant changes, which were then costed.

Results

A total of 1399 interventions were documented. Eight hundred and thirty‐five interventions impacted on drug costs alone. Five hundred and eleven interventions were evaluated by the independent panels with three quarters of these confirmed as having an impact on one or more of: length of stay, readmission probability, medical procedures or laboratory monitoring. There were 96 interventions deemed by the independent panels to have reduced LOS and 156 reduced the potential for readmission. The calculated savings was $263 221 for the eight hospitals during the period of the study. This included $150 307 for length of stay reduction, $111 848 for readmission reduction.

Conclusions

The annualized cost savings relating to length of stay, readmission, drugs, medical procedures and laboratory monitoring as a result of clinical pharmacist initiated changes to hospitalized patient management or therapy was $4 444 794 for eight major acute care government funded teaching hospitals in Australia.
Effects of a nonpeptide motilin receptor antagonist on proximal gastric motor function
Authors: Kamerling Ingrid M. C. , Van Haarst Aernout D. , Burggraaf Jacobus , Schoemaker Rik C. , De Kam Marieke L. , Heinzerling Hartmut , Cohen Adam F. , Masclee Ad A. M.
Published: December 2004

Aim

To assess the effects of the motilin receptor antagonist RWJ‐68023 on basal and motilin‐stimulated proximal gastric volume.

Methods

Eighteen healthy male volunteers received RWJ‐68023 in two different doses or placebo for 135 min. After 45 min, subjects received a motilin infusion for 90 min. Proximal gastric volume was measured with a barostat at constant pressure and during isobaric distensions. Abdominal symptoms were scored using visual analogue scales. Motilin and RWJ‐68023 concentrations were assessed by radioimmunoassay and liquid chromatography‐mass spectrometry, respectively.

Results

Both dosages of RWJ‐68023 were safe and well tolerated. The most common adverse events were of gastrointestinal origin. RWJ‐68023 did not affect basal proximal gastric volume, but the high‐dose RWJ‐68023 reduced the contractile effect of motilin on the stomach. This antagonizing effect of RWJ‐68023 was only significant (P = 0.014) during the distension procedure.

Conclusions

The RWJ‐68023 doses used in this study were selected to accomplish plasma concentrations that would block the motilin effect entirely. However, the antagonizing effect of RWJ‐68023 was partial and only present when the tonic condition of the stomach was modulated by motilin.
Low body mass index is associated with an increased risk of neuropsychiatric adverse events and concentration impairment in women on mefloquine
Authors: Van Riemsdijk M. M. , Sturkenboom M. C. J. M. , Ditters J. M. , Tulen J. H. M. , Ligthelm R. J. , Overbosch D. , Stricker B. H.
Published: December 2004

Aims

We performed a prospective cohort study to gain more insight into risk factors for neuropsychiatric effects of mefloquine among tourists travelling to tropical areas.

Methods

We enrolled all patients who consulted the Travel Clinic of the Havenziekenhuis & Institute for Tropical Diseases Rotterdam for mefloquine prophylaxis during the period between 1 May 1999 and 7 March 2000. Each patient was followed from baseline (prior to starting mefloquine) up to 3 weeks after starting weekly intake of 250 mg mefloquine. We compared the intraindividual change in scores between baseline and follow‐up visit on the Dutch shortened Profile of Mood States, and on the Continuous Performance Test (CPT) which measures sustained attention.

Results

The final cohort consisted of 151 subjects with a mean age of 38 years. In this population, a significant impairment of mood state was observed in those with a body mass index (BMI) ≤ 20 kg m−2. Stratification for gender showed that the total mood disturbance in females in the lowest BMI category significantly increased by 8.42 points [95% confidence interval (CI) 3.33, 13.50], whereas BMI did not affect the risk in males. Stratification for history of use of mefloquine showed that the risks were highest in first‐time users. Analyses of the CPT showed that reaction time in women with a BMI ≤ 20 kg m−2 increased significantly by 22.5 ms (95% CI 7.80, 37.20), whereas reaction time in men showed a slight and nonsignificant decrease.

Conclusion

Risk factors for mefloquine‐associated neuropsychiatric adverse events and concentration impairment are female gender, low BMI, and first‐time use. The frequency of neuropsychiatric effects is highest in women with a BMI ≤ 20 kg m−2.
Overanticoagulation associated with combined use of lactulose and acenocoumarol or phenprocoumon
Authors: Visser Loes E. , Penning‐van Beest Fernie J. A. , Wilson J. H. P. , Vulto Arnold G. , Kasbergen A. A. Harrie , De Smet Peter A. G. M. , Hofman Albert , Stricker Bruno HCh
Published: December 2004

Some medical textbooks on drug interactions take note of the potential interaction between laxatives and coumarin anticoagulants, but epidemiological evidence that this interaction is of practical importance is lacking. We conducted a follow‐up study in a large population‐based cohort to investigate which laxatives are associated with overanticoagulation during therapy with coumarins. Of the 1124 patients in the cohort, 351 developed an International Normalized Ratio ≥ 6.0. The only laxative with a moderate but significantly increased relative risk of overanticoagulation was lactulose (relative risk 3.4, 95% confidence interval 2.2, 5.3). In view of the widespread use of lactulose, especially among the elderly, awareness of this potential drug interaction is required.
Hypereosinophilic syndrome in a child mosaic for a congenital triplication of the short arm of chromosome 8
Authors: EGESTEN ARNE , HA¨GERSTRAND INGA , KRISTOFFERSSON ULF , GARWICZ STANISLAW
Published: December 2004

An 11‐year‐old boy with mental retardation, malformations, and the mosaic karyotype 47,XY,+i(8p)/46,XY presented with fever, headache and petechiae. Peripheral blood WBC was 190 ×109/l; and contained > 90% mature eosinophils. Cytogenetic analysis of the eosinophils revealed no aberrations except the constitutional karyotype. The patient was diagnosed as having a hypereosinophilic syndrome. Shortly after initiation of therapy he died from extensive mural thrombi of the heart and thrombi of several other organs. This is the first case of congenital triplication of the short arm of chromosome 8 associated with hypereosinophilic syndrome, suggesting involvement of genes on chromosome 8p in the regulation of eosinopoiesis.
Disappearance of prognostic significance of histopathological grading of nodular sclerosing Hodgkin's disease for unselected patients, 1972–92
Authors: VAN SPRONSEN D. J. , VRINTS L. W. , HOFSTRA G. , CROMMELIN M. A. , COEBERGH J. W. W. , BREED W. P. M.
Published: December 2004

The importance of the subclassification of nodular sclerosing Hodgkin's disease (NSHD) according to the British National Lymphoma Investigation Group (BNLI) criteria, which had an independent prognostic value for 90 unselected patients diagnosed in our region in the period 1972–83, is still equivocal. Because survival of patients with Hodgkin's disease improved in our region during the period 1972–92 and because treatment may modify prognostic factors, we re‐evaluated the prognostic value of NSHD subclassification up to 1993. A registry‐based study was performed with data on all 345 Hodgkin patients diagnosed in the period 1972–92. Available histology was reviewed. The prognostic value of nodular sclerosis (NS) grading was evaluated for two periods, 1972–80 and 1981–92, designated as the seventies and the eighties, respectively. NSHD was diagnosed in 57% (n = 195) of all registered cases of Hodgkin's disease, 17 of which could not be evaluated. NS stage I (NSI; 73%) and NS stage II (NSII; 27%) patients exhibited the same distribution of stage during both periods; NSII patients were older than NSI patients in the seventies. NSII patients presented with an elevated ESR and B symptoms more frequently during the eighties and subsequently received combined modality therapy more often. The crude 5‐year survival rate for grade I v grade II NSHD was 85% v 38% (P < 0.05) for the seventies and 84% v 83% for the eighties. Subclassification of NSHD was not an independent prognostic factor after adjustment for age, stage, gender, B symptoms and ESR, though it remained of independent prognostic value when ESR was left out of the Cox model. The most important factors adversely influencing survival were advanced age, advanced stage and male gender. The independent prognostic value of the subclassification of NSHD has disappeared, as reflected by the clearly improved survival of NSII patients, probably due to more intensive treatment in the eighties, whereas survival of NSI patients did not change.
analysis of CD9, CD32 and p67 signalling: use of degranulated platelets indicates direct involvement of CD9 and p67 in integrin activation
Authors: SLUPSKY JOSEPH R. , CAWLEY JOHN C. , KAPLAN CÉCILE , ZUZEL MIRKO
Published: December 2004

The use of agonist monoclonal antibodies (mAbs) to probe the signalling function of platelet membrane proteins is severely limited by the dependence of the mAb effect on Fc–FcγRII interaction. Furthermore, in addition to its anchoring role, the FcγRII receptor itself generates a stimulation signal resulting in granule secretion. Platelet stimulation by the released granule contents can then further obscure the original activation signal. Here we demonstrate that these problems are largely overcome by the use of platelets which had been degranulated with thrombin prior to stimulation with mAbs. We found that, like intact cells, degranulated platelets could also be activated and induced to aggregate by mAbs against a 67 kD membrane protein (known as PTA1) and CD9, and by crosslinked CD32 (FcγRII). However, the signal generated by crosslinked FcγRII was weak compared with that induced by the other monoclonal antibodies. Thus, by diminishing the FcγRII signal contribution, we have succeeded for the first time to clearly dissect the target antigen signal from that generated by FcγRII. In addition to differences in the degree of aggregation, analysis of the signals generated by each mAb showed differences in Ca2+ fluxes and protein phosphorylation. Moreover, the signals generated by CD9 and PTA1 antigens differed significantly in their sensitivity to PKC inhibition or ADP‐ribosylation of the small GTP‐binding protein rhoA. Despite these differences, the signals initiated by all three antigens converged to a common signalling pathway which included activation of tyrosine kinase(s). The pattern of protein phosphorylation strongly resembled that induced by gpIIb/IIIa‐mediated platelet interaction with macromolecular ligands and by mutual cell contact. The multiple intercellular links formed by mAb would have a similar effect since the Fc‐receptor anchorage required for antigen stimulation is already known to be provided by adjacent cells. The present findings suggest that the function of both CD9 and PTA1 antigens is closely associated with gpIIb/IIIa activation.
Peripheral blood stem cell apheresis in normal donors: feasibility and yield of second collections
Authors: Anderlini P. , Lauppe J. , Przepiorka D. , Seong D. , Champlin R. , Ko¨rbling M.
Published: December 2004

We report 13 normal peripheral blood stem cell (PBSC) donors who had a second PBSC collection for allogeneic transplantation performed after the first. The median interval between the first and second collection was 5 months. Mobilization was achieved with filgrastim (12 μg/kg/d). No significant difference was found in the median pre‐apheresis leucocyte count (×109/l) between the two donations (40.2 v 38.5; P = 0.91). The median apheresis yield (×106 CD34+ cells/litre blood processed, first apheresis) was also similar (28 v 27.3; P = 0.91). Filgrastim‐related adverse events were comparable. These data suggest that second PBSC collections are feasible, similarly tolerated and provide comparable apheresis yields.
Agranulocytosis, arthritis and systemic vasculitis in a patient receiving the oral iron chelator L1 (deferiprone)
Authors: CASTRIOTA‐SCANDERBEG ALESSANDRO , SACCO MICHELE
Published: December 2004

A thalassaemic girl presented with agranulocytosis, arthritis of both ankles and clinical and laboratory features consistent with the diagnosis of systemic vasculitis, during oral iron chelator L1 (deferiprone) treatment. Changes in the humoral and cell‐mediated immune function, including antinuclear antibodies (ANA), anti‐DNA and extractable‐nuclear antigens (ENA) antibodies positivity, increased immunoglobulin values, decreased T suppressor and the presence of circulating immune complexes, suggest a cause‐and‐effect relationship with the observed clinical manifestations.

A careful monitoring of the immune function is recommended in patients who are receiving the oral iron chelator L1.
Iron metabolism in transgenic mice with hypoplastic anaemia due to incomplete deficiency of erythropoietin
Authors: RAJA K. B. , MAXWELL P. H. , RATCLIFFE P. J. , SALISBURY J. R. , SIMPSON R. J. , PETERS T. J.
Published: December 2004

Iron overload is a serious complication of many forms of anaemia, arising in part from mechanisms associated with compensatory increases in erythropoiesis. To investigate other mechanisms by which anaemia itself may perturb iron metabolism, without the confounding effects of compensatory erythropoiesis, we studied transgenic mice with a partially disabling insertion in the erythro‐poietin gene, which manifested as incomplete erythropoietin deficiency.

Mice were studied aged 7–8 weeks. Haemoglobin concentrations were 6.6 ± 0.8 g/dl in mice homozygous for the modified erythropoietin gene, 12.9 ± 2.2 g/dl in heterozygous mice and 14.1 ± 1.0 g/dl in controls. Homozygous mice showed significant hepatic iron loading (2‐fold increase in liver non‐haem iron, compared with heterozygous mice and normal controls, with iron staining principally in the periportal hepatocytes). Absorption studies using 59Fe showed increased uptake from the lumen of an in vivo isolated duodenal segment in homozygous mice, although at this point in time overall transfer of radioiron to the circulation and other tissues (mucosal transfer) was not different from controls. These observations demonstrate that anaemia can lead to hepatic iron loading even in the absence of increased erythropoiesis, and are consistent with the possibility that anaemic hypoxia can enhance mucosal iron uptake by the duodenal enterocyte.
Molecular defects in Hb H hydrops fetalis
Authors: CHAN VIVIAN , WAN‐YIN CHAN V. , TANG MARY , LAU K. , TODD D. , CHAN T. K.
Published: December 2004

The molecular defect in two unique cases of Hb H hydrops fetalis has been characterized. Both cases are due to co‐inheritance of a ‘non‐deletion’ defect affecting the α2 gene: at codon 30 (ΔGAG, Glu) and codon 59 (G  →  A, Gly →  Asp) respectively, and a ζ‐α thalassaemia (thal) 1 or α thal 1 genotype. These two non‐deletion defects, unlike previously described cases, resulted in severe anaemia of the fetuses and emphasize the importance of performing prenatal diagnosis for these families.
Expression of intercellular adhesion molecule 1 (ICAM‐1) in childhood acute lymphoblastic leukaemia: correlation with clinical features and outcome
Authors: MIELCAREK MARCO , SPERLING CHRISTIAN , SHRAPPE MARTIN , MEYER ULRIKE , RIEHM HANSJO¨RG , LUDWIG WOLF‐DIETER
Published: December 2004

To investigate whether expression of intercellular adhesion molecule 1 (ICAM‐1, CD54) in childhood acute lymphoblastic leukaemia (ALL) has an impact on the biological behaviour of the disease, we evaluated 751 children of the ALL‐BFM 90 trial with B‐cell precursor‐ (n = 6777) or T‐cell‐ALL (n = 74) for CD54 expression within immunological subgroups, its correlation to certain clinical features, and therapy outcome.

The highest percentage of patients expressing CD54 was found in common‐ and pre‐B‐ALL (76.1% and 61.4%, respectively). There was intermediate expression in pre‐pre‐B‐ALL (47.8%), and the lowest expression was detected in T‐ALL (12.2%).

A significant positive correlation could be demonstrated between low CD54 expression (< 20% stained blasts) and high peripheral leucocyte counts, central nervous system (CNS) involvement, and splenomegaly at the time of diagnosis (P < 0.01). In addition, CD54 expression was a favourable but not independent prognostic factor. Event‐free survival estimate at 4.5 years was 86% for CD54+ patients (n = 463), compared with 78% for CD54− patients (n = 241) (P < 0.01). These findings demonstrate that CD54 expression has an impact on dissemination patterns and outcome of childhood ALL, and emphasizes the potential relevance of adhesion mechanisms in influencing clinical characteristics and prognosis of haematological malignancies.
P‐glycoprotein (PGP) and lung resistance‐related protein (LRP) expression and function in leukaemic blast cells
Authors: MICHIELI MARIAGRAZIA , DAMIANI DANIELA , ERMACORA ANNA , RASPADORI DONATELLA , MICHELUTTI ANGELA , GRIMAZ STEFANIA , FANIN RENATO , RUSSO DOMENICO , LAURIA FRANCESCO , MASOLINI PAOLA , BACCARANI MICHELE
Published: December 2004

P‐glycoprotein (PGP) lung resistance protein (LRP) and multidrug resistance associated protein (MRP) expressions and function were evaluated by flow cytometry in 65 leukaemic patients (38 acute non‐lymphocytic leukaemias, eight acute lymphocytic leukaemias, 19 Ph‐positive chronic myeloid leukaemias in blastic phase). By using the MRK‐16, the LRP‐56 and the MRPm6 MoAbs, 34% of the cases did not over‐express any proteins (−); 24.5% over‐expressed (+) only PGP, 11% only LRP, 1.5% only MRP, 24.5% both PGP and LRP, and 4.5% both PGP and MRP. The mean intracellular daunorubicin accumulation (IDA) and rhodamine 123 (Rh123) retention in the presence or absence of the reversal agent SDZ PSC 833 (PSC) of the PGP−/LRP−/MRP− cases were comparable to the ones observed in normal leucocytes. With respect to the non‐over‐expressing cases, the PGP−/LRP+/MRP− cases showed only an impaired IDA (mean 204 ± 29; P < 0.001). The PGP+/LRP+/MRP− cases had a defect both in IDA (mean 166 ± 47, P < 0.001) and Rh123 retention (mean 0.42 ±0.14; P < 0.001), which were both corrected by PSC. All the PGP+/LRP+/MRP− cases had a defect in IDA (mean daunorubicin (DNR) accumulation 192 ± 44; P < 0.001. However, only in 8/16 of them an evident defect in Rh123 retention was found. In conclusion, both PGP and LRP over‐expression were common in leukaemia. An impaired IDA was found in all cases over‐expressing PGP, LRP or both. The study of Rh123 retention could give incorrect information about the blast cells’ ability to accumulate cytotoxic drugs in patients over‐expressing both PGP and LRP.
Molecular analysis of the Turkish form of deletion‐inversion (δβ)° thalassaemia
Authors: O¨NER C. , O¨NER R. , BALKAN H. , GU¨RGEY A. , YALC¸IN A. , AVCU F. , ALTAY C¸.
Published: December 2004

Two unrelated (δβ)°‐thalassaemia patients from Southern Turkey are presented. DNA studies indicated that both of them are homozygous for the Turkish type of (δβ)°‐thalassaemia characterized by one large deletion of 11.5 kb including the δ and β globin genes at the 5′ end and one small deletion of 1.6 kb at the 3′ end, which are separated by an inverted 7.6 kb long DNA segment that includes L1 repetitive sequence. In the present study a PCR‐based method was performed to produce a unique deletion‐specific product and subjected to sequence analysis for the determination of the breakpoint. DNA polymorphisms in the β‐globin gene cluster of deletion‐inversion type of (δβ)°‐thalassaemia, IVS‐I‐6 and β‐39 globin genes were examined. Analysis of sequence variations in regulatory regions including the 5′ hypersensitive site‐2 of the locus control region (LCR), the δ, Gγ and Aγ 5′ flanking regions and the second intervening sequence (IVS‐II) of Aγ and Gγ genes indicated the presence of close similarities between the chromosome carrying the Turkish form of deletion‐inversion δβ)°‐thalassaemia and the chromosome associated with β‐39 nonsense mutation in haplotype II. These two chromosomes are characterized by the presence of a 4 base pair deletion in the AγT globin gene promoter. A C → T alteration at position −199 5′ to the δ gene was also found to be associated with the Turkish type of (δβ)°‐thalassaemia and β‐39 chromosome.
A low but functionally significant MDR1 expression protects primitive haemopoietic progenitor cells from anthracycline toxicity
Authors: SMEETS MARIE¨LLE , RAYMAKERS REINIER , VIERWINDEN GERTY , PENNINGS ARIE , VAN DE LOCHT LOUIS , WESSELS HANS , BOEZEMAN JAN , DE WITTE THEO
Published: December 2004

Pgp is expressed on normal haemopoietic progenitor cells. The significance of the efflux pump in protecting normal progenitors for anthracycline toxicity is not defined and is the subject of this study. Pgp was measured in CD34+ progenitors with a rhodamine efflux assay. A high efflux, modulated by verapamil, was only found in a distinct subpopulation (20–30%). Pgp measured by the monoclonal antibody antibody (MoAb) MRK‐16 was low in the rhodamine dull, but significantly (P < 0.04) higher than in the rhodamine bright cells. Reverse transcriptase polymerase chain reaction (RT‐PCR) of MDR1 mRNA showed a very weak signal in both populations. In a single‐cell clonogenic assay, rhodamine dull cells appeared less sensitive to anthracyclines (IC50 daunorubicin 0.005 μg/ml; adriamycin 0.03 μg/ml) compared to rhodamine bright cells (IC50 daunorubicin 0.0025 μg/ml; adriamycin 0.01 μg/ml). Furthermore, verapamil significantly (P < 0.05) potentiated anthracycline toxicity only in the rhodamine dull cells, proving its Pgp‐specific modulating effect. Rhodamine dull cells gave larger and more mixed colonies compatible with a more primitive origin. Although detection with MoAbs and RT‐PCR revealed a low Pgp level, functionally this Pgp appeared to be very important in protecting primitive progenitors against anthracycline toxicity. This protection can be jeopardized by administration of Pgp modulators.
Two novel alleles of the ATCT variable number tandem repeat locus VWF.VNTR I in intron 40 of the von Willebrand factor gene
Authors: HADDAD ALBERT P. , SPARROW ROSEMARY L.
Published: December 2004

Two new alleles, allele 9 and allele 15, of the ATCT variable number tandem repeat locus, VWF.VNTR I, of intron 40 of the von Willebrand factor (VWF) gene are described. Both alleles occurred in low frequencies, being detected in only two and one of 285 random Australian Caucasians respectively. In all, 10 alleles were observed representing between six and 15 repeats of the ATCT motif. The allele frequency distribution in this Australian population was similar to other VWF.VNTR I population studies. The additional alleles described here for the VWF.VNTR I further enhance the usefulness of this VNTR locus in human identification work.
Clonal expansion of Vγ3/Vδ3‐expressing γδ T cells in an HIV‐1/2‐negative patient with CD4 T‐cell deficiency
Authors: KABELITZ DIETER , HINZ THOMAS , DOBMEYER THOMAS , MENTZEL ULRICH , MARX SIBYLLE , BO¨HME ANGELIKA , ARDEN BERNHARD , ROSSOL RITA , HOELZER DIETER
Published: December 2004

Immunological investigations were carried out in an HIV‐1/2//HTLV‐1‐negative patient with CD4 T‐cell deficiency (0.357–0.6 × 109/l) and expansion of γδ T cells which accounted for 26–42% of peripheral blood lymphocytes during an observation period of 3 years. Flow cytometry analyses with a panel of available Vγ/Vδ‐specific monoclonal antibodies indicated that the pathologically expanded γδ population expressed Vγ2 or Vγ3 paired with Vδ3 on the surface but lacked the expression of activation antigens such as CD38 or CD71. Cloning and sequencing of RT‐PCR products obtained after amplification of cDNA with Vγ‐Cγ and Vδ‐Cδ specific primers confirmed the presence of a clonally expanded Vγ3/Vδ3 population in the peripheral blood of this patient. Cytotoxicity assays performed with purified γδ T cells as effectors and resting or preactivated autologous CD4 T cells as targets failed to reveal evidence for autoreactive cytotoxicity of Vγ3/Vδ3 cells as a possible mechanism of CD4 T‐cell deficiency in this patient.
Defective response to thrombopoietin and impaired expression of c‐mpl mRNA of bone marrow cells in congenital amegakaryocytic thrombocytopenia
Authors: MURAOKA KENJI , ISHII EIICHI , TSUJI KOHICHIRO , YAMAMOTO SHUICHI , YAMAGUCHI HIDETO , HRA TSHIRO , KOGA HIROYUKI , NAKAHATA TATSUTOSHI , MIYAZAKI SUMIO
Published: December 2004

Congenital amegakaryocytic thrombocytopenia (CAMT) is an uncommon disorder in newborns and infants, characterized by isolated thrombocytopenia and megakaryocytopenia in the first year without physical anomalies. The defect of thrombopoiesis is not well understood. Recently, thrombopoietin (TPO), the ligand for the c‐mpl receptor, was cloned. Accumulating evidence from in vitro and in vivo studies indicate that TPO plays a key role in the regulation of megakaryocytopoiesis. In this study we examined the effect of TPO on megakaryocyte colony formation from a patient with CAMT using a plasma‐containing methylcellulose clonal culture. The in vitro results demonstrated a defective response to TPO in megakaryocyte colony formation from bone marrow mononuclear cells (MNC) of the patient, although interleukin‐3 (IL‐3) but not stem cell factor (SCF) induced only a small number of megakaryocyte colonies. These findings indicated that thrombocytopenia in CAMT could not be corrected by administration of TPO in vitro. Additionally, clonal cultures containing SCF, IL‐3, IL‐6 and erythropoietin showed decreased numbers of erythroid and myelocytic progenitors in the bone marrow of the patient. The serum TPO level measured by enzyme‐linked immunosorbent assay was significantly higher than that in healthy controls. By PCR, marrow MNC from healthy children and from a patient with essential thrombocytosis expressed c‐mpl mRNA, whereas no c‐mpl mRNA was detected in marrow MNC from the patient with CAMT. There was no difference in the CD34 expression and c‐kit mRNA between the CAMT patient and healthy children. The results of this study suggest that the pathophysiology in CAMT may be a defective response to TPO in haemopoietic cells through impaired expression of c‐mpl mRNA.
Inadequate Erythropoietin Response to Anaemia in HIV Patients: Relationship to Serum Levels of Tumour Necrosis Factor‐Alpha, Interleukin‐6 and Their Soluble receptors
Authors: KREUZER KARL‐ANTON , ROCKSTROH JU¨RGEN KURT , JELKMANN WOLFGANG , THEISEN ALBERT , SPENGLER ULRICH , SAUERBRUCH TILMANN
Published: December 2004

Severe anaemia is a frequent complication in advanced HIV infection. In our study we investigated the interaction between cytokine network, HIV infection and erythropoietin (Epo) response with increasing anaemia levels. No correlations could be established between circulating tumour necrosis factor (TNF)‐alpha and any of the examined parameters. However, a negative correlation was found between haemoglobin values and soluble TNF receptor levels (sTNF‐R‐I: r = −0.54; P < 0.001; sTNF‐R II: r = −0.47; P < 0.001) as well as interleukin‐6 levels (r = −0.29; P < 0.001). In contrast, no significant increase in log[Epo], counterbalancing haemoglobin decline and paralleling the rise in sTNF receptors, was found. In patients classified as stage III, according to the Centers for Disease Control (CDC) classification, the erythropoietin response was significantly more impaired than in patients from CDC groups I and II (P < 0.001). The results of this study suggest that similar to its action in vitro, activation of the TNF/TNF‐R system may impair erythropoietin production in HIV‐associated anaemia. Due to the brief half‐life of TNF‐α, this activation is particularly reflected by elevations of soluble TNF receptor levels.
Acute lymphoblastic leukaemia in childhood: cell proliferation without rest
Authors: HIRT ANDREAS , ANTIC VLADAN , WANG EUGENIA , RIDOLFI LU¨THY ANNETTE , LEIBUNDGUT KURT , VON DER WEID NICOLAS , TOBLER ANDREAS , WAGNER HANS‐PETER
Published: December 2004

The percentage of non‐cycling blast cells in children with untreated acute lymphoblastic leukaemia (ALL) was investigated by staining smears for statin, a nuclear protein specifically present in non‐growing resting cells. Results were compared with purified normal CD34‐positive progenitors. A low fraction of ALL and CD34‐positive cells expressed statin (2.0 ± 3.8% and 2.8 ±3.1%, respectively), the growth fraction assessed by staining for the nucleolar antigen p120 was 94% in both ALL and CD34‐positive cell samples. From this analysis it can be concluded that the compartment of non‐replicating cells in ALL as well as in normal CD34‐positive precursor cells collected from peripheral blood is very small and that most cells are cycling.
An antisense Bcr‐Abl phosphodiester‐tailed methylphosphonate oligonucleotide reduces the growth of chronic myeloid leukaemia patient cells by a non‐antisense mechanism
Authors: SMETSERS TOON F. C. M. , LINDERS ELLEN H. P. , VAN DE LOCHT LOUIS T. F. , DE WITTE THEO M. , MENSINK EWALD J. B. M.
Published: December 2004

The specificity of antisense oligonucleotides targeted to the mRNA breakpoint region of the Bcr‐Abl oncogene, found in leukaemic cells from patients with chronic myeloid leukaemia, remains controversial due to non‐specific effects. To prevent protein binding of oligonucleotides we designed and tested a methylphosphonate oligonucleotide with an attached 3′ soluble phosphodiester tail. Growth of chronic myeloid leukaemia (CML) cell lines BV173, KCL‐22 and cells of CML patients tested was inhibited by the b2a2 type antisense Bcr‐Abl oligonucleotide and not with controls. Also the growth of control CD34+ cells of two healthy donors, control cell lines and cells from AML patients was only moderately affected or not affected. Bcr‐Abl protein studies in combination with growth‐determination experiments indicated that the antisense methylphosphonate Bcr‐Abl oligonucleotide tested is a potent inhibitor of the growth of CML cells but works in a non‐antisense manner.
The mechanism of red cell (dis)aggregation investigated by means of direct cell manipulation using multiple optical trapping
Authors: BRONKHORST P. J. H. , GRIMBERGEN J. , BRAKENHOFF G. J. , HEETHAAR R. M. , SIXMA J. J.
Published: December 2004

We used multiple optical trapping to study the mechanism of red cell (dis)aggregation. Two sets of optical ‘tweezers’ were used to bring two red blood cells together to form a two‐cell aggregate and then to pull them apart, to study the interaction between the cells.

We found that cross‐bridging occurred in normal reversible aggregation as we observed binding and the occurrence of small tethers between opposite cell membranes. Furthermore, the cells could only be parted by sliding them side by side with a maximum velocity in the order of μm/s indicating accumulation of the cross‐bridges.