bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcomes of a randomized controlled trial of a clinical pharmacy intervention in 52 nursing homes
Authors: Roberts Michael S. , Stokes Julie A. , King Michelle A. , Lynne Teresa A. , Purdie David M. , Glasziou Paul P. , Wilson D. Andrew J. , McCarthy Sean T. , Brooks Geoffrey E. , De Looze Ferdinandus J. , Del Mar Christopher B.
Published: March 2003

Aims  To evaluate whether a year long clinical pharmacy program involving development of professional relationships, nurse education on medication issues, and individualized medication reviews could change drug use, mortality and morbidity in nursing home residents.

Methods  A cluster randomised controlled trial, where an intervention home was matched to three control homes, was used to examine the effect of the clinical pharmacy intervention on resident outcomes. The study involved 905 residents in 13 intervention nursing homes and 2325 residents in 39 control nursing homes in south‐east Queensland and north‐east New South Wales, Australia. The outcome measures were: continuous drug use data from government prescription subsidy claims, cross‐sectional drug use data on prescribed and administered medications, deaths and morbidity indices (hospitalization rates, adverse events and disability indices).

Results  This intervention resulted in a reduction in drug use with no change in morbidity indices or survival. Differences in nursing home characteristics, as defined by cluster analysis with SUDAAN®, negated intervention‐related apparent significant improvements in survival. The use of benzodiazepines, nonsteroidal anti‐inflammatory drugs, laxatives, histamine H2‐receptor antagonists and antacids was significantly reduced in the intervention group, whereas the use of digoxin and diuretics remained similar to controls. Overall, drug use in the intervention group was reduced by 14.8% relative to the controls, equivalent to an annual prescription saving of $A64 per resident (approximately £25).

Conclusions  This intervention improved nursing home resident outcomes related to changes in drug use and drug‐related expenditure. The continuing divergence in both drug use and survival at the end of the study suggests that the difference would have been more significant in a larger and longer study, and even more so using additional instruments specific for measuring outcomes related to changes in drug use.
Trans‐3′‐hydroxycotinine: Disposition kinetics, effects and plasma levels during cigarette smoking
Authors: Benowitz N. L. , Jacob P.
Published: March 2003

Aims 
(3′R,5′S)‐trans‐3′‐hydroxycotinine (3‐HC) is a major metabolite of nicotine. The aim of this study was to characterize the disposition kinetics of 3‐HC in healthy smokers, including metabolism to (3′R,5′S)‐trans‐3′‐hydroxycotinine glucuronide (3‐HC‐Gluc). We also studied pharmacologic responses to 3‐HC and plasma levels of 3‐HC in a group of smokers.

Methods 
Eight cigarette smokers were studied on a clinical research ward. After 5 days of supervised nonsmoking, each subject received an intravenous infusion of 3‐HC, 4 µg kg−1 min−1 for 60 min. Plasma and urine levels of 3‐HC and 3‐HC‐Gluc and cardiovascular and subjective responses were examined. Plasma levels of 3‐HC, nicotine, and cotinine were measured in 62 smokers on up to three occasions.

Results 
The total plasma clearance of 3‐HC averaged 1.3 ml min−1 kg−1, of which 63% was renal excretion of unchanged drug. An average of 29% of the dose was excreted as 3‐HC‐Gluc. 3‐HC did not have nicotine‐like cardiovascular effects.

Conclusions 
These findings extend our understanding of the quantitative nature of nicotine metabolism. Such data may be of use in quantitating human exposure to nicotine from tobacco and in studying individual variability in nicotine metabolism.
Carbamazepine is not a substrate for P‐glycoprotein
Authors: Owen Andrew , Pirmohamed Munir , Tettey Justice N. , Morgan Paul , Chadwick David , Park B. Kevin
Published: March 2003

Aims  To determine whether the anticonvulsant carbamazepine (CBZ), a known CYP3A4 substrate, is also a substrate for the multidrug efflux transporter P‐glycoprotein (Pgp).

Methods  The role of Pgp in the transport of CBZ was assessed in three systems: (a) in mdr1a/1b(−/−) and wild‐type mice after administration of 2 mg kg−1 and 20 mg kg−1, which served as a model for brain penetration; (b) in Caco‐2 cells, an in vitro model of the intestinal epithelium that is known to express high Pgp levels; and (c) by flow cytometry in lymphocytes using rhodamine 123, a fluorescent substrate for PgP.

Results  Brain penetration of both doses of CBZ at 1 h and 4 h was comparable in wild‐type and mdr1a/1b(−/−) mice. Transport across the Caco‐2 cell monolayer was Pgp‐independent, and was not affected by the Pgp inhibitor PSC‐833. CBZ had no effect on rhodamine 123 efflux from lymphocytes, in contrast to verapamil, which increased fluorescence intensity fivefold.

Conclusion  CBZ is not a substrate for Pgp. Its efficacy is unlikely to be affected by Pgp over‐expression in the brain. Furthermore, the interaction of CBZ with drugs that modulate both CYP3A4 and Pgp function such as verapamil is probably due to inhibition of CYP3A4 and not Pgp.
Simultaneous administration of a cocktail of markers to measure renal drug elimination pathways: absence of a pharmacokinetic interaction between fluconazole and sinistrin, p‐aminohippuric acid and pindolol
Authors: Gross A. S. , McLachlan A. J. , Minns I. , Beal J. B. , Tett S. E.
Published: March 2003

Aims  Previous studies suggest that estimated creatinine clearance, the conventional measure of renal function, does not adequately reflect changes in renal drug handling in some patients, including the immunosuppressed. The aim of this study was to develop and validate a cocktail of markers, to be given in a single administration, capable of detecting alterations in the renal elimination pathways of glomerular filtration, tubular secretion and tubular reabsorption.

Methods  Healthy male subjects (n = 12) received intravenously infused 2500 mg sinistrin (glomerular filtration) and 440 mg p‐aminohippuric acid (PAH; anion secretion), and orally administered 100 mg fluconazole (reabsorption) and 15 mg rac‐pindolol (cation secretion). The potential interaction between these markers was investigated in a pharmacokinetic study where markers (M) or fluconazole (F) were administered alone or together (M + F). Validated analytical methods were used to measure plasma and urine concentrations in order to quantify the renal handling of each marker. Plasma protein binding of fluconazole was measured by ultrafiltration. All subjects had an estimated creatinine clearance within the normal range. The renal clearance of each marker (mean± s.d.) was calculated as the ratio of the amount excreted in urine and the area‐under‐the‐concentration‐time curve. Statistical comparisons were made using a paired t‐test and 95% confidence intervals were reported.

Results  The renal clearances of sinistrin (M: 119 ± 31 ml min−1; M + F: 130 ± 40 ml min−1; P = 0.32), PAH (M: 469 ± 145 ml min−1; M + F: 467 ± 146 ml min−1; P = 0.95), R‐pindolol (M: 204 ± 41 ml min−1; M + F: 190 ± 41 ml min−1; P = 0.39; n = 11), S‐pindolol (M: 225 ± 55 ml min−1; M + F: 209 ± 60 ml min−1; P = 0.27; n = 11) and fluconazole (F: 14.9 ± 3.8 ml min−1; M + F: 13.6 ± 3.4 ml min−1; P = 0.16) were similar when the markers or fluconazole were administered alone (M or F) or as a cocktail (M + F).

Conclusions  This study found no interaction between markers and fluconazole in healthy male subjects, suggesting that a single administration of this cocktail of markers of different renal processes can be used to simultaneously investigate pathways of renal drug elimination.
Pharmacokinetics and pharmacodynamics of midazolam administered as a concentrated intranasal spray. A study in healthy volunteers
Authors: Knoester P. D. , Jonker D. M. , Van Der Hoeven R. T. M. , Vermeij T. A. C. , Edelbroek P. M. , Brekelmans G. J. , De Haan G. J.
Published: March 2003

Aims  To investigate the pharmacokinetic and pharmacodynamic profile of midazolam administered as a concentrated intranasal spray, compared with intravenous midazolam, in healthy adult subjects.

Methods  Subjects were administered single doses of 5 mg midazolam intranasally and intravenously in a cross‐over design with washout period of 1 week. The total plasma concentrations of midazolam and the metabolite 1‐hydroxymidazolam after both intranasal and intravenous administration were described with a single pharmacokinetic model. β‐band EEG activity was recorded and related to midazolam plasma concentrations using an exponential pharmacokinetic/pharmacodynamic model.

Results  Administration of the intranasal spray led to some degree of temporary irritation in all six subjects, who nevertheless found intranasal administration acceptable and not painful. The mean (±s.d.) peak plasma concentration of midazolam of 71 (±25 ng ml−1) was reached after 14 (±5 min). Mean bioavailability following intranasal administration was 0.83±0.19. After intravenous and intranasal administration, the pharmacokinetic estimates of midazolam were: mean volume of distribution at steady state 1.11±0.25 l kg−1, mean systemic clearance 16.1±4.1 ml min−1 kg−1 and harmonic mean initial and terminal half lives 8.4±2.4 and 79±30 min, respectively. Formation of the 1‐hydroxymetabolite after intranasal administration did not exceed that after intravenous administration.

Conclusions  In this study in healthy volunteers a concentrated midazolam nasal spray was easily administered and well tolerated. No serious complications of the mode of administration or the drug itself were reported. Rapid uptake and high bioavailability were demonstrated. The potential of midazolam given via a nasal spray in the acute treatment of status epilepticus and other seizure disruptions should be evaluated.
MDR1 gene polymorphisms and disposition of the P‐glycoprotein substrate fexofenadine
Authors: Drescher Siegfried , Schaeffeler Elke , Hitzl Monika , Hofmann Ute , Schwab Matthias , Brinkmann Ulrich , Eichelbaum Michel , Fromm Martin F.
Published: March 2003

Aims  The C3435T polymorphism in the human MDR1 gene is associated with lower intestinal P‐glycoprotein expression, reduced protein function in peripheral blood cells and higher plasma concentrations of the P‐glycoprotein substrate digoxin. Using fexofenadine, a known P‐glycoprotein substrate, the hypothesis was tested whether this polymorphism also affects the disposition of other drugs in humans.

Methods  Ten Caucasian subjects homozygous for the wild‐type allele at position 3435 (CC) and 10 individuals homozygous for T at position 3435 participated in this study. A single oral dose of 180 mg fexofenadine HCl was administered. Plasma and urine concentrations of fexofenadine were measured up to 72 h using a sensitive LC/MS method. In addition, P‐glycoprotein function was assessed using efflux of the P‐glycoprotein substrate rhodamine 123 from CD56+ cells.

Results 
Fexofenadine plasma concentrations varied considerably among the study population. However, fexofenadine disposition was not significantly different between the CC and TT groups (e.g. AUC(0,∞) CC vs TT: 3567.1±1535.5 vs 3910.1±1894.8 ng ml−1 h, NS; 95% CI on the difference −1364.9, 2050.9). In contrast, P‐glycoprotein function was significantly decreased in CD56+ cells of the TT compared with the CC group (rhodamine fluorescence CC vs TT: 45.6±7.2% vs 61.1±12.3%, P<0.05; 95% CI on the difference 5.6, 25.5).

Conclusions 
In spite of MDR1 genotype‐dependent differences in P‐glycoprotein function in peripheral blood cells, there was no association of the C3435T polymorphism with the disposition of the P‐glycoprotein substrate fexofenadine in this German Caucasian study population. These data indicate that other mechanisms including uptake transporter function are likely to play a role in fexofenadine disposition.
Topographic pharmaco‐EEG mapping of the effects of the South American psychoactive beverage ayahuasca in healthy volunteers
Authors: Riba Jordi , Anderer Peter , Morte Adelaida , Urbano Gloria , Jané Francesc , Saletu Bernd , Barbanoj Manel J.
Published: March 2003

Aims  Ayahuasca is a traditional South American psychoactive beverage used in Amazonian shamanism, and in the religious ceremonies of Brazilian‐based syncretic religious groups with followers in the US and several European countries. This tea contains measurable amounts of the psychotropic indole N,N‐dimethyltryptamine (DMT), and β‐carboline alkaloids with MAO‐inhibiting properties. In a previous report we described a profile of stimulant and psychedelic effects for ayahuasca as measured by subjective report self‐assessment instruments. In the present study the cerebral bioavailability and time‐course of effects of ayahuasca were assessed in humans by means of topographic quantitative‐electroencephalography (q‐EEG), a noninvasive method measuring drug‐induced variations in brain electrical activity.

Methods  Two doses (one low and one high) of encapsulated freeze‐dried ayahuasca, equivalent to 0.6 and 0.85 mg DMT kg−1 body weight, were administered to 18 healthy volunteers with previous experience in psychedelic drug use in a double‐blind crossover placebo‐controlled clinical trial. Nineteen‐lead recordings were undertaken from baseline to 8 h after administration. Subjective effects were measured by means of the Hallucinogen Rating Scale (HRS).

Results  Ayahuasca induced a pattern of psychoactive effects which resulted in significant dose‐dependent increases in all subscales of the HRS, and in significant and dose‐dependent modifications of brain electrical activity. Absolute power decreased in all frequency bands, most prominently in the theta band. Mean absolute power decreases (95% CI) at a representative lead (P3) 90 min after the high dose were −20.20±15.23 µV2 and −2.70±2.21 µV2 for total power and theta power, respectively. Relative power decreased in the delta (−1.20±1.31% after 120 min at P3) and theta (−3.30±2.59% after 120 min at P3) bands, and increased in the beta band, most prominently in the faster beta‐3 (1.00±0.88% after 90 min at P3) and beta‐4 (0.30±0.24% after 90 min at P3) subbands. Finally, an increase was also seen for the centroid of the total activity and its deviation. EEG modifications began as early as 15–30 min, reached a peak between 45 and 120 min and decreased thereafter to return to baseline levels at 4–6 h after administration.

Conclusions  The central effects of ayahuasca could be objectively measured by means of q‐EEG, showing a time pattern which closely paralleled that of previously reported subjective effects. The modifications seen for the individual q‐EEG variables were in line with those previously described for other serotonergic psychedelics and share some features with the profile of effects shown by pro‐serotonergic and pro‐dopaminergic drugs. The q‐EEG profile supports the role of 5‐HT2 and dopamine D2‐receptor agonism in mediating the effects of ayahuasca on the central nervous system.
Association between the N‐acetylation genetic polymorphism and bronchial asthma
Authors: Nacak Muradiye , Aynacioglu A. Sükrü , Filiz Ayten , Cascorbi Ingolf , Erdal M. Emin , Yilmaz Necat , Ekinci Erhan , Roots Ivar
Published: March 2003

Aims  Since polymorphic N‐acetyltransferase 2 (NAT2) has been suggested as a susceptibility factor for atopic diseases, the study was undertaken to investigate whether an association exists between acetylation polymorphism and asthma patients with atopy.

Methods  The frequencies of NAT2 alleles and genotypes were determined by PCR/RFLP in a total of 210 asthma patients (extrinsic (n = 108) and intrinsic (n = 102) asthmatics) and 240 control subjects. Presence of the NAT2*4 (wild‐type) allele defined a NAT2 genotype as rapid and combinations of mutant alleles NAT2*5 A, *5B, *5C, *6 A, and *7B as slow.

Results  Genotypes coding for slow acetylation were detected in 70.4, 58.4 and 58.3% of extrinsic asthmatics, but intrinsic asthmatics and control subjects, respectively. The frequency of slow acetylators was higher among extrinsic asthmatics than intrinsic asthmatics, this difference did not reach statistical significance (odds ratio 1.02, 95% confidence interval 0.64, 1.63, P = 0.085). However, we found a relatively moderate, but significantly higher, increased frequency of slow acetylators among extrinsic asthma patients compared with control subjects (odds ratio 1.70, 95% confidence interval 1.04, 2.76, P = 0.042).

Conclusions  This study shows an association between acetylation polymorphism and susceptibility to extrinsic asthma, but not to intrinsic asthma, suggesting a minor role of the NAT2 polymorphism in the development of atopic asthma.
Spatial distribution of 8‐methoxypsoralen penetration into human skin after systemic or topical administration
Authors: Grundmann‐Kollmann Marcella , Podda Maurizio , Bräutigam Lutz , Hardt‐Weinelt Katja , Ludwig Ralf J. , Geisslinger Gerd , Kaufmann Roland , Tegeder Irmgard
Published: March 2003

Aims  Photochemotherapy employing psoralens combined with UVA irradiation (PUVA) is a standard therapy for a variety of dermatoses. Psoralens can be administered orally or topically in the form of bath or cream preparations. Recommendations for the time of UVA irradiation are mainly based on the time course of minimal phototoxic doses. However, the time course and depth of skin penetration of psoralens is not well characterized.

Methods  We assessed the time course of 8‐MOP concentrations in horizontal epidermal and dermal skin sections in 10 patients undergoing oral (n = 3), cream (n = 4) and bath (n = 3) PUVA therapy. Punch biopsies (4 mm) were taken from ‘healthy’ skin sites. A highly sensitive LC‐MS‐MS method was employed for 8‐MOP analysis.

Results  Epidermal concentrations following cream or bath were highest at the end of the application period (time zero) when irradiation is performed. At this time, 8‐MOP cream provided significantly higher epidermal concentrations (mean ± s.e. mean 128.0 ± 22.6 pg mm−3; 95% CI: 77.6, 178.4) than oral 8‐MOP (27.0 ± 25.3 pg mm−3; 95% CI: 29.3, 83.3 at 1 h; P = 0.025). Conversely, concentrations in the papillary dermis were significantly higher with oral 8‐MOP (20.2 ± 3.1 and 16.2 ± 2.2 pg mm−3 at 1 and 2 h, respectively) than with 8‐MOP cream (7.1 ± 2.8 and 8.4 ± 2.0 pg mm−3 time zero and 0.5 h, respectively; P = 0.020 and 0.045, respectively) or bath (8.8 ± 3.1 and 7.7 ± 2.2 pg mm−3; P = 0.050 and 0.039, respectively). The observed time courses of 8‐MOP concentrations correspond to time courses of photosensitivity found previously with the different treatment modalities.

Conclusions  The higher epidermal 8‐MOP concentrations found after topical 8‐MOP may explain the lower UVA doses needed with the topical route. These results suggest that topical 8‐MOP may be superior in patients where the pathology is localized in the epidermis. In sclerosing diseases, which mainly affect the dermis oral PUVA might be advantageous because dermal concentrations are highest with this route of administration.
Faster absorption of ethanol and higher peak concentration in women after gastric bypass surgery
Authors: Klockhoff H. , Näslund I. , Jones A. W.
Published: March 2003

AimsTo investigate the absorption, distribution and elimination of ethanol in women with abnormal gut as a result of gastric bypass surgery. Patients who undergo gastric bypass for morbid obesity complain of increased sensitivity to the effects of alcohol after the operation.

MethodsTwelve healthy women operated for morbid obesity at least 3 years earlier were recruited. Twelve other women closely matched in terms of age and body mass index (BMI) served as the control group. After an overnight fast each subject drank 95% v/v ethanol (0.30 g kg−1body weight) as a bolus dose. The ethanol was diluted with orange juice to 20% v/v and finished in 5 min. Specimens of venous blood were taken from an indwelling catheter before drinking started and every 10 min for up to 3.5 h post‐dosing. The blood alcohol concentration (BAC) was determined by headspace gas chromatography.

Results    The    maximum    blood‐ethanol    concentration    (Cmax)    was   0.741 ± 0.211 g l−1(± s.d.) in the operated group compared with 0.577 ± 0.112 g l−1in the controls (mean difference 0.164 g l−1, 95% confidence interval (CI) 0.021, 0.307). The median  time  to  peak  (tmax) was 10 min in the bypass patients compared with 30 min in controls (median difference −15 min (95% CI −10, −20 min). At 10 and 20 min post‐dosing the BAC was higher in the bypass patients (P < 0.05) but not at 30 min and all later times (P > 0.05). Other pharmacokinetic parameters of ethanol were not significantly different between the two groups of women (P > 0.05).

Conclusions The higher sensitivity to ethanol after gastric bypass surgery probably reflects the more rapid absorption of ethanol leading to higherCmaxand earliertmax. The marked reduction in body weight after the operation might also be a factor to consider if the same absolute quantity of ethanol is consumed.
Inhibition of basal nitric oxide synthesis increases aortic augmentation index and pulse wave velocity in vivo
Authors: Wilkinson Ian B. , MacCallum Helen , Cockcroft John R. , Webb David J.
Published: March 2003

Aims  To investigate the role of basal nitric oxide (NO) production in regulating large artery stiffness in vivo.

Methods  Incremental doses of the NO synthase inhibitor L‐NG‐monomethyl arginine (LNMMA: 0.1, 0.3, 1.0 and 3.0 mg kg−1 min−1) or placebo were infused in eight healthy men. Arterial stiffness was assessed noninvasively by pulse wave analysis.

Results  Compared with placebo, infusion of LNMMA led to a dose‐dependent increase in mean arterial pressure, peripheral vascular resistance, and aortic and systemic arterial stiffness. There was an accompanying reduction in heart rate and cardiac index. The highest dose of LNMMA resulted in an increase of 25% in AIx (95% confidence limits; 12, 38) and of 16 mmHg in mean arterial pressure (9, 23) compared with infusion of saline.

Conclusions  These data indicate functional regulation of large artery stiffness in vivo by NO, and may provide new therapeutic strategies for cardiovascular risk reduction.
Pharmacokinetics of diclofenac and inhibition of cyclooxygenases 1 and 2: no relationship to the CYP2C9 genetic polymorphism in humans
Authors: Kirchheiner Julia , Meineke Ingolf , Steinbach Nadine , Meisel Christian , Roots Ivar , Brockmöller Jürgen
Published: March 2003

Aims The cytochrome P450 enzyme CYP2C9 catalyses the 4′‐hydroxylation of the nonsteroidal analgesic drug diclofenac in humans. We studied the influences of the known amino acid variants, CYP2C9*2 (Arg144Cys) and CYP2C9*3 (Ile359Leu), on diclofenac pharmacokinetics after a 50‐mg oral dose of diclofenac in healthy volunteers. As a surrogate marker of diclofenac activity, the ex vivo formation of prostaglandin E2 and thromboxane B2, which reflects COX‐2 and COX‐1 activity, was measured.

Methods Genotyping was performed in 516 healthy volunteers to obtain 20 participants with all allelic combinations of the two CYP2C9 variants Arg144Cys (*2) and Ile359Leu (*3). Diclofenac and 4′‐hydroxydiclofenac were quantified in plasma by reversed phase h.p.l.c. after oral intake of 50 mg diclofenac. Concentrations of thromboxane B2 (TxB2) and prostaglandin E2 (PGE2) were measured by immunoassays.

Results There was no evidence of impaired metabolism of oral diclofenac in heterozygous and homozygous carriers of the CYP2C9 alleles *2 and *3 compared with the wild type (mean CL/F (95% CI) 20.5 (11, 30) l h−1 for *1/*1, 29.9 (19, 40) l h−1 for *1/*2, 30.0 (4, 56) l h−1 for *2/*2, 22.6 (12, 33) l h−1 for *1/*3, 23.5 (11, 37) l h−1 for *3/*3 and 37.3 (−15, 89) l h−1 in *2/*3). Furthermore, plasma concentrations of the metabolite 4′‐hydroxydiclofenac were not lower in carriers of the CYP2C9 low‐activity alleles *2 and *3 compared with carriers of the CYP2C9*1/*1 genotype. Marked diclofenac mediated inhibition of COX‐1‐ and COX‐2 activity was detected in all individuals independent of CYP2C9 genotype.

Conclusions Polymorphisms of the CYP2C9 gene had no discernible effect on the pharmacokinetics and pharmacodynamics of diclofenac. The question of whether enzymes other than CYP2C9 play a major role in diclofenac 4′‐hydroxylation in vivo or whether 4′‐hydroxylation is not a rate‐limiting step in diclofenac elimination in vivo, or whether the effect of the CYP2C9 polymorphisms is substrate‐dependent, needs further investigation.
NAT2 and CYP1A2 phenotyping with caffeine: head‐to‐head comparison of AFMU vs. AAMU in the urine metabolite ratios
Authors: Nyéki Á , Buclin T. , Biollaz J. , Decosterd L. A.
Published: March 2003

Aims  (i) To compare the phenotyping of healthy subjects for NAT2 and CYP1A2 activities with caffeine, by the simultaneous assay of the urinary metabolites AFMU and AAMU, and (ii) to ascertain whether NAT2 and CYP1A2 phenotyping is influenced by the use of AFMU or AAMU in the metabolite ratio.

Methods  Thirty‐five healthy subjects (16 men, 19 women) participated to the study. Caffeine metabolite concentrations were measured in urine collected 8 h after 2.5 mg kg−1 caffeine intake using a new validated h.p.l.c. method. The metabolite ratios AFMU/1X, AFMU/(AFMU+1X+1U), AAMU/1X, AAMU/(AAMU+1X+1 U), and (AFMU+1U+1X)/17U, (AAMU+1U+1X)/17U were determined as indices of NAT2 and CYP1A2 activity, respectively.

Results  Slow and rapid acetylators were similarly identified using the four NAT2 metabolite ratios in 139 out of 140 measurements. An appreciable amount of AAMU was present in urine that was immediately acidified and analysed. Consequently, the ratio using AFMU was lower than that using total AAMU following transformation of AFMU in basic conditions. The proportion of AFMU in urine analysed immediately expressed as AFMU/(AFMU+AAMU) ratio did not correlate with urine pH, but was a function of the acetylation phenotype, with a low intergroup variability (64 ± 3% and 32 ± 5%, for rapid and slow acetylators, respectively; P < 0.00001, anova). Regarding CYP1A2 activity, a good correlation (r = 0.99) was observed between the metabolite ratios calculated from AFMU and AAMU, although the ratios calculated from AFMU were proportionately and systematically lower P < 0.00001, paired t‐test, slope 1.2).

Conclusions  This study demonstrates that both AFMU and AAMU can be used for NAT2 and CYP1A2 metabolite ratio determinations. The reported conversion of AFMU into AAMU is unlikely to explain the large amount of AAMU in urine that was acidified and analysed immediately after voiding. The results suggest that AAMU is formed not solely through a nonenzymatic hydrolysis in urine, but in vivo by a NAT2 phenotype‐dependent pathway.
Safety profile of rofecoxib as used in general practice in England: results of a prescription‐event monitoring study
Authors: Layton Deborah , Riley Jane , Wilton Lynda V. , Shakir Saad A. W.
Published: March 2003

Aims A postmarketing Prescription‐Event Monitoring study was undertaken to monitor the safety of rofecoxib, a cyclo‐oxygenase (COX)‐2 selective inhibitor prescribed in primary care in England.

Methods Questionnaires requesting clinical event data were sent to prescribing physicians between February and November 2000, and the data analysed for all events.

Results There were 15 268 patients identified, mean age 62 years, 67% female. The commonest specified indication was osteoarthritis (24%). Dyspepsia and nausea were the most frequently reported adverse events. A history of dyspeptic or upper gastrointestinal (GI) conditions, recent use of other nonsteroidal anti‐inflammatory drugs (NSAIDs), use of selected concomitant gastroirritant drugs (NSAIDs, aspirin, anticoagulants, antiplatelet drugs), or gastroprotective drugs (misoprostol, antacids, proton‐pump inhibitors, histamine‐2 antagonists), and age (≥ 65 years) modified the risk of having minor GI events. During treatment or within 1 month of stopping, 110 serious GI events were reported (including 76 upper GI bleeds/peptic ulcers, one perforated colon), 101 thromboembolic events, three reports of acute renal failure, one each of Stevens–Johnson syndrome, severe anaphylaxis and angio‐oedema.

Conclusions Doctors should continue to prescribe NSAIDs including COX‐2 selective inhibitors with caution.
Mast cell mediators other than histamine induce pruritus in atopic dermatitis patients: a dermal microdialysis study
Authors: Rukwied R. , Lischetzki G. , Mcglone F. , Heyer G. , Schmelz M.
Published: March 2003

While histamine is the crucial mediator of pruritus in type 1 allergic reactions, its role in atopic dermatitis (AD) is unclear. In this study, the role of mast cell mediators in protein extravasation and pruritus was evaluated using intradermal microdialysis. The microdialysis capillaries were used to apply the mast cell degranulating substance compound 48/80 (C48/80; 0·05%) or histamine (0·01%) and also to deliver H1-blockers (cetirizine, 200 μg mL−1) in nine AD patients and nine controls. Large pore size membranes (3000 kDa) enabled simultaneous analysis of protein extravasation. Itch sensation was measured psychophysically and weal and flare reaction were evaluated planimetrically. Protein extravasation induced by histamine and C48/80 was significantly reduced in AD patients. Blockade of H1-receptors by cetirizine significantly reduced C48/80-induced protein extravasation in AD patients and controls to an identical level. C48/80-induced pruritus was abolished by cetirizine in controls, whereas pruritus in AD patients was unchanged after H1 blockade. We conclude that mast cell mediators others than histamine are involved in C48/80-induced pruritus in AD patients. Whether the reduced capacity of AD patients to induce protein extravasation is of pathophysiological relevance for pruritus remains to be established.
Cost evaluation of the medical management of neurofibromatosis 1: a prospective study on 201 patients
Authors: Wolkenstein P. , Durand-Zaleski I. , Moreno J.C. , Zeller J. , Hemery F. , Revuz J.
Published: March 2003

Neurofibromatosis 1 (NF1) is associated with many internal complications as well as skin manifestations, and patients may require a variety of medical and surgical interventions. We aimed to assess the medical needs of NF1 patients, and to evaluate the financial cost of the resources used for them in relation to the severity of the disease. We conducted a prospective analysis on a cohort of 201 patients in our referral centre for adults. Severity of the disease was assessed. Therapeutic management was considered as multidisciplinary if it required more than three different specialists. Plastic and dermatological surgery procedures performed were recorded. Hospital costs were computed over a 3-year period and included all hospitalization days, clinic visits and procedures performed in all departments where the patients were admitted. One hundred and thirty-seven patients had at least one out-patient procedure or one hospitalization during the follow-up period. The mean cost per patient per year was £810 (median 240; range 0–13,860). Multidisciplinary procedures were more frequent in moderately and severely affected NF1 patients than in milder cases (P < 0·0001); hence, the costs for moderate and severe cases were higher than for less severe groups (P = 0·005). Plastic and/or dermatological surgery was performed with the same frequency in the different severity groups (71%). Regardless of the presence of serious intractable complications, the patients’ priority is for treatment of the disfigurement due to the disease. The management of these patients can be considered relatively inexpensive from the viewpoint of the healthcare system.
Beta‐thalassaemia intermedia: is it possible consistently to predict phenotype from genotype?
Authors: Ho  , Hall  , Luo  , Weatherall  , Thein
Published: March 2003

Eighty‐seven patients with β thalassaemia of intermediate severity were investigated in our Unit to determine whether it is possible to consistently predict phenotypic severity from genotypic factors. The subjects were from the following ethnic backgrounds: Asian Indian (35.1%), Middle Eastern (24.3%), Mediterranean (21.6%), Northern European (14.9%) and South‐East Asian/Chinese (4.1%). There was a wide spectrum of phenotypic severity; 49 had mild disease, 22 moderate and 16 severe disease. 22/87 patients had inherited only a single copy of a β‐thalassaemia allele, of whom 11 had also co‐inherited triplicated α genes (ααα/αα or ααα/ααα) and seven had dominantly inherited β thalassaemia. In four of the heterozygotes no explanation was found for the thalassaemia‐intermedia phenotype. 65/87 patients were homozygous or compound heterozygous for 26 mutations (40 genotypes) which ranged from very mild β+to β° thalassaemia alleles. All patients with two mild or very mild β+thalassaemia alleles had mild to moderate disease. Although concurrent inheritance of extra α genes with heterozygous β thalassaemia results in thalassaemia intermedia, the disease is mild. Co‐inheritance of α thalassaemia as a modulating factor was not evident in this cohort of patients. Presence of thein‐cis XmnI‐Gγ site was a modulating factor but insufficient to explain the high fetal haemoglobin levels encountered. In conclusion, apart from the two categories of triplicated α genes with heterozygous β thalassaemia and inheritance of mild β+thalassaemia alleles, it was not possible to consistently predict phenotype from α and β genotypes alone, due to the influence of modulating factors, some implicated (such as inheritance of HPFH determinants) and others as yet unidentified.
A lymphohistiocytic variant of anaplastic large cell lymphoma with demonstration of the t(2;5)(p23;q35) chromosome translocation
Authors: Ott German , Bastian Boris C. , Katzenberger Tiemo , Decoteau John F. , Kalla Jörg , Rosenwald Andreas , Weissinger Florian , Ott M. Michaela , Kadin Marshall E. , Müller‐Hermelink Hans K.
Published: March 2003

Cytogenetic investigations were performed in a case of a nodal malignant non‐Hodgkin's lymphoma. Histopathological analysis from an involved lymph node as well as from a skin biopsy revealed a lymphohistiocytic variant of CD30‐positive anaplastic large cell lymphoma (ALCL). A t(2;5)(p23;q35) chromosome translocation could be observed in all metaphases analysed. This finding was confirmed both by RT‐PCR analysis of the NPM/ALK fusion protein and by positive staining with the p80NPM/ALK antibody. To the best of our knowledge, this is the first report of a t(2;5) documented by classic cytogenetics in the lymphohistiocytic variant of ALCL.
Hypermethylation of the M27β (DXS255) locus in chronic B‐cell leukaemia
Authors: Crossen Peter E. , Morrison Mary J.
Published: March 2003

We have investigated the methylation status of the M27β (DXS255) locus in 21 female patients with chronic B‐cell leukaemia and in 20 normal controls. DNA was digested with PstI and then with the methylation sensitive enzyme HpaII and probed with the M27β probe. Eight patients (38%) showed hypermethylation of the M27β locus which was not seen in any of the normal controls. Hypermethylation of the M27β locus has also been found in acute myeloid leukaemia, acute lymphocytic leukaemia and lymphoma, suggesting that hypermethylation of the M27β locus is associated with the leukaemic process.
Serum thrombopoietin levels in Kawasaki disease
Authors: Miura Nobuyuki , Terai Masaru , Meng Y. Gloria , Sato Takeyuki , Niimi Hiroo
Published: March 2003

Kawasaki disease (KD) is an acute febrile vasculitic syndrome with thrombocytosis occurring in childhood. Transient thrombocytosis of KD is sometimes the cause of complications such as coronary aneurysmal thrombosis and myocardial infarction. We have analysed blood TPO levels in KD and found that 26/31 acute‐phase KD patients had detectable blood TPO levels (mean 173 pg/ml; range 89–294 pg/ml), which decreased immediately with the elevation of platelet counts in 5/12 patients studied. Elevated serum level of TPO may have an important role for this ill‐defined disease.
Multidrug resistance analysis in lymphoproliferative disease of large granular lymphocytes
Authors: Lamy Thierry , Drenou Bernard , Fardel Olivier , Amiot Laurence , Grulois Isabelle , Le Prise Pierre‐Yves , Loughran Thomas P. , Fauchet Renee
Published: March 2003

Multi‐drug resistance (MDR) phenotype contributes to the ineffectiveness of chemotherapy. P‐glycoprotein (PgP) and lung resistance protein (LRP) are proteins implicated in chemoresistance. We analysed the expression of PgP and LRP respectively in 17 and 15 cases of lymphoproliferative disease of granular lymphocytes (LDGL) including 10 cases of clonal large granular lymphocytic (LGL) leukaemia, six cases of oligoclonal (n = 5) and polyclonal (n = 1) CD3+ lymphoproliferation and one case of CD3− NK lymphocytosis. Functional PgP activity, as determined by Rh123 dye efflux assay, was found in all the patients. The mean percentage of effluxing cells was 47 ± 22%, compared to 35 ± 8% on normal lymphocytes (P < 0.04). The efflux was blocked in the presence of verapamil, a PgP revertant agent. A high proportion of CD57+ cells (66 ± 10%) from these patients expelled Rh123. Functional PgP activity was associated with expression of MDR1 mRNA. By using immunocytochemistry, LRP expression was detected in 11/15 patients (73%). 7/10 LGL leukaemia patients presented a LRP+/Efflux+ phenotype and 5/7 had LRP+/Efflux+/MDR1 mRNA+ phenotype. These findings suggest that the PgP+/LRP+ phenotype is frequently observed in LDGL. Its clinical relevance in aggressive cases remains to be determined.
Familial acute myeloid leukaemia: four male members of a single family over three consecutive generations exhibiting anticipation
Authors: De Lord  , Powles  , Mehta  , Wilson  , Treleaven  , Meller  , Catovsky
Published: March 2003

A kindred with familial acute myeloid leukaemia (AML) is described displaying the concept of anticipation which refers to increased disease severity and/or earlier age at onset with succeeding generations. The affected individual in the first generation was the father (aged 34 years at diagnosis) of two affected members of a single sibship (aged 24 and 25 years). The fourth patient is the son (aged 4 years) of one of these brothers in the second generation. The AML was of different subtypes and a karyotypic abnormality (del[6q]) was detected in one of three patients. The first‐generation patient died during chemotherapy; the remaining three patients are in complete remission after chemotherapy and autologous bone marrow transplantation.
Gastric cancer associated with acute disseminated intravascular coagulation: successful initial treatment with weekly 24‐hour infusion of high‐dose 5‐fluorouracil and leucovorin
Authors: Yeh Kun‐Huei , Cheng Ann‐Lii
Published: March 2003

Acute disseminated intravascular coagulation (DIC) is a severe complication of gastric adenocarcinoma, and most of the patients die within 1–3 weeks. We have treated five such patients with an empirical non‐ myelosuppressive HDFL regimen (weekly 24 h infusion of high‐dose 5‐fluorouracil 2600 mg/m2 and leucovorin 300 mg/m2). Within 2 weeks of starting the treatment the clinical and laboratory evidence of acute DIC quickly resolved in all five patients. HDFL not only caused no further myelosuppression, but also resulted in normalization of the patient's haemogram within a few weeks. Other anti‐cancer drugs could then be safely added. Three patients had a survival time of more than 6 months. We suggest that HDFL is an ideal initial treatment for gastric cancer complicated by acute DIC.
Late‐onset noninfectious pulmonary complications after allogeneic bone marrow transplantation
Authors: Palmas Angelo , Tefferi Ayalew , Myers Jeffrey L. , Scott John P. , Swensen Stephen J. , Chen Michael G. , Gastineau Dennis A. , Gertz Morie A. , Inwards David J. , Lacy Martha Q. , Litzow Mark R.
Published: March 2003

We examined the incidence and clinical outcome of late‐onset noninfectious pulmonary complications (LONIPC) in a series of 234 patients who underwent allogeneic bone marrow transplantation at our institution between April 1982 and October 1996. The 179 patients who survived 3 months or more were evaluated. Clinical, radiologic, pulmonary function, and pathologic tests were reviewed to identify 18 patients (10%) who fulfilled the diagnostic criteria of LONIPC. Accordingly, the pulmonary processes included bronchiolitis obliterans (BO, five patients), bronchiolitis obliterans with organizing pneumonia (BOOP, three patients), diffuse alveolar damage (DAD, one patient), lymphocytic interstitial pneumonia (LIP, one patient), and nonclassifiable interstitial pneumonia (NCIP, eight patients). Various methods of enhanced immunosuppressive therapy resulted in marked durable remission in nine patients (50%) (3/3 with BOOP, 3/8 with NCIP, 1/1 with DAD, 1/1 with LIP, 1/5 with BO). The presence of chronic graft‐versus‐host disease (cGVHD) and prophylaxis for GVHD with cyclosporine and prednisone were the only variables significantly associated with the development of LONIPC (P = 0.0001 and 0.008, respectively). Regardless of histology, a reduction in the forced expiratory volume to < 45% of the predicted range was associated with poor response to treatment. These findings suggest a strong association between cGVHD and LONIPC and that the risk of LONIPC development may be influenced by the particular method of GVHD prophylaxis. Most patients with BOOP or mild airflow limitation at diagnosis achieved durable remissions.
Androgen effects on factor IX expression: in‐vitro and in‐vivo studies in mice
Authors: Brady Jolene N. , Notley Colleen , Cameron Cherie , Lillicrap David
Published: March 2003

A role for steroid hormones has been proposed for the post‐pubertal factor IX increment of ∼25% seen in both normal males and females, as well as in the post‐pubertal phenotypic recovery seen in haemophilia B Leyden. We have evaluated androgen receptor binding to the factor IX promoter and have assessed transcriptional activation of the factor IX gene in hepatocytes through transient transfection studies and through expression of factor IX in a murine model of androgen insensitivity. Whereas transfection of the androgen receptor alone did not activate expression from the factor IX promoter, co‐transfection with the CCAAT enhancer binding protein resulted in a synergistic 17‐fold enhancement of transcriptional activity. Using liver nuclear extracts and recombinant androgen receptor protein we have confirmed binding of this protein to the factor IX proximal promoter and disruption of binding with a mutation at nucleotide −26. Finally, studies in normal and testicular feminized male mice showed different developmental patterns of factor IX expression. In normal mice, expression recapitulates that seen in humans, with early post‐natal levels being ∼50% of the adult values and with a post‐pubertal increment of ∼25%. In contrast, testicular feminized animals did not show a significant post‐pubertal increment of factor IX. These studies provide further support for the role of androgen receptor binding to the factor IX promoter in regulating the developmental expression of factor IX.
Inverse immunostaining pattern for synthesized versus endocytosed alpha‐granule proteins in human bone marrow megakaryocytes
Authors: De Larouzière Vanina , Brouland Jean‐Philippe , Souni Fadila , Drouet Ludovic , Cramer Elisabeth
Published: March 2003

The time of appearance and pattern of expression of several α‐granule proteins, von Willebrand factor (VWF), fibrinogen and immunoglobulins (Ig) were examined and compared in human bone marrow megakaryocytes (MK) using an immunocytochemical approach. VWF is synthesized by immature MK, whereas it has been shown that fibrinogen is incorporated from the plasma into α‐granules. The present study was undertaken in order to determine whether there are chronological and morphological differences in the expression of VWF and fibrinogen in vivo in human MK. Seven paraffin‐embedded biopsies of normal human bone marrow were labelled with specific antibodies for VWF and for fibrinogen, detected by the alkaline phosphatase anti‐alkaline phosphatase (APAAP) method, and analysed by immunomorphometry. We found a clear, statistically significant, difference in the labelling pattern of VWF and fibrinogen. The expression of other endocytosed α‐granule proteins, immunoglobulins G and A, was therefore studied in bone marrow MK from two patients with multiple myeloma, one with monoclonal IgG and one with monoclonal IgA. The immunostaining pattern was similar to that of fibrinogen and different from VWF, and characteristic of endocytosed α‐granule proteins.

This study demonstrates that: (i) immunohistochemical staining of MK α‐granules proteins distinguishes the peripheral cockade distribution pattern of endocytosed protein from the perinuclear pattern of endogenously synthesized proteins; (ii) VWF is present in human bone marrow MK when fibrinogen is not yet detectable; (iii) VWF synthesis ceases while fibrinogen is still being incorporated; (iv) immunoglobulins can be detected in MK cytoplasm, with a staining pattern resembling that of fibrinogen.
Inter‐individual variability in Ca2+ signalling in platelets from healthy volunteers: effects of aspirin and relationship with expression of endomembrane Ca2+‐ATPases
Authors: Feijge Marion A. H. , Van Pampus Elisabeth C. M. , Lacabaratz‐Porret Christine , HamulyÀk Karly , Lévy‐Toledano Sylviane , Enouf Jocelyne , Heemskerk Johan W. M.
Published: March 2003

Increased Ca2+ signal generation may lead to hyperactivity of platelets and contribute to thrombotic complications. Using fura‐2‐loaded platelets from 51 healthy volunteers, high variability was detected in the Ca2+ responses evoked by the receptor agonists, thrombin and collagen, and the inhibitor of sarco/endoplasmic reticulum Ca2+‐ATPases (SERCA), thapsigargin (Tg). Oral intake of 500 mg aspirin reduced the magnitude of the Ca2+ responses, and lowered the intra‐individual coefficients of variance of the responses by 50%. However, the corresponding inter‐individual variance coefficients were only a little influenced by aspirin intake, pointing to subject‐dependent factors in Ca2+ handling that are unrelated to thromboxane formation. With each agonist, 6–9% of the subjects had platelets with relatively high Ca2+ responses (> mean + SD) both before and after aspirin intake. In 90% (9/10) of these cases the high responsiveness was confirmed in platelets obtained 6–12 months later. The Tg‐ but not thrombin‐induced Ca2+ responses correlated inversely with the expression levels of SERCA PL/IM 430 (SERCA‐3b) in platelets. After aspirin intake, the Ca2+ responses with collagen but not thrombin correlated inversely with SERCA‐2b expression. These results suggest that, in the absence of potentiating effects of thromboxane, (i) the amount of PL/IM 430‐recognizable SERCA may control the Ca2+ signal when SERCA‐2b is specifically inhibited (with Tg), and (ii) the expression of SERCA‐2b determine the collagen‐ but not the thrombin‐evoked Ca2+ signal. Accordingly, limited Ca2+‐pumping activity by low expression of one of the SERCA isoforms is likely to be one of the factors resulting in increased platelet activity towards collagen or thapsigargin but not thrombin.