bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of the vasoconstrictor effects of the selective 5‐HT1D‐receptor agonist L‐775,606 with the mixed 5‐HT1B/1D‐receptor agonist sumatriptan and 5‐HT in human isolated coronary artery
Authors: Longmore J. , Maguire J. J. , MacLeod A. , Street L. , Schofield W. N. , Hill R. G.
Published: March 2003

Aims Vasoconstriction in human coronary artery can be mediated via activation of both 5‐HT2 and 5‐HT1B‐receptors. Coronary vasoconstriction is a rare, but potential adverse effect of the antimigraine drug sumatriptan. In order to investigate the receptor population involved we compared the vasoconstrictor effects of sumatriptan (a mixed 5‐HT1B/1D‐receptor agonist) with those of L‐775,606 (a selective 5‐HT1D‐receptor agonist) and 5‐HT (the endogenous ligand) in human isolated coronary arteries.

Methods Coronary arteries were obtained from human hearts removed prior to transplant surgery. Several endothelium denuded ring segments (4 mm in length) were obtained from each artery and mounted for isometric tension recording. Each segment was first exposed to 45 mm KCl and then to 5‐HT (1 nm‐100 μm ). Concentration‐effect curves to L‐775,606 (1‐(3‐(5‐(1,2,4‐triazol‐4‐yl)‐1H‐indol‐3‐yl)propyl)‐4‐(2‐(3‐fluorophenyl)ethyl)piperazine) and sumatriptan were then performed in a consecutive and random manner. The response to repeated application of 5‐HT was obtained in separate segments.

Results Twenty‐five segments from seven different coronary arteries were studied. Concentration‐effect curves were fitted to the data using nonlinear regression analysis. The maximum contraction for L‐775,606 was significantly less than that for sumatriptan with Emax values (% relative to 45 mm KCl=100%) of 30.1±4.22 and 41.5±2.7, respectively. L‐775,606 was significantly (30‐fold) less potent than sumatriptan in causing contraction compared with sumatriptan (EC50 values were 6.0 μm and 0.2 μm, respectively). For comparison the Emax value for 5‐HT was 77.2% and the EC50 value was 0.2 μm.

Conclusions The selective 5‐HT1D‐receptor agonist L‐775,606 has less propensity towards vasoconstriction in human isolated coronary artery (endothelium‐denuded) than was mixed 5‐HT1B/1D‐receptor agonist sumatriptan. The contractions produced were at concentrations where L‐775,606 would be expected to occupy 5‐HT1B‐receptors.
Effects of the 5‐HT3 antagonist cilansetron vs placebo on phasic sigmoid colonic motility in healthy man: a double‐blind crossover trial
Authors: Stacher Georg , Weber Ute , Stacher‐janotta Giselheid , Bauer Peter , Huber Kurt , Holzäpfel Antje , Krause Günter , Steinborn Claus
Published: March 2003

Aims  5‐hydroxytryptamine3 receptor antagonists act antiemetically and slow colonic transit. This study evaluated effects of the high‐affinity 5‐HT3 antagonist, cilansetron, on fasting, meal‐and anticholinesterase‐stimulated phasic contractile activity of the human sigmoid colon as well as on bowel habits and stool consistency.

Methods  Five female and seven male healthy volunteers received, during three 7 day periods separated by 7 day wash‐out periods, 4 mg cilansetron, 8 mg cilansetron or placebo three times daily orally under random, double‐blind, crossover conditions. On day 8 of each treatment period, motility 20–40 cm from the anal verge was recorded using five pressure sensors spaced at 5 cm intervals. After a basal 30 min, subjects swallowed a further dose of the scheduled treatment; 60 min later, blood was taken for the determination of plasma cilansetron levels. Thereafter, subjects ingested a 4200 kJ meal and 250 ml sweetened mallow tea (166 kJ); 90 min after meal onset, 1 mg neostigmine was administered intramuscularly and motility recording was continued for 60 min

Results  Phasic contractile activity and intraluminal base‐line pressure increased postprandially and more so after neostigmine. With cilansetron, the area under the pressure curve as the primary outcome variable and the number of contractions were significantly greater than with placebo (P = 0.005), amplitude and duration of contractions and base‐line pressure were not affected. The effects of the two cilansetron dosages did not differ. With cilansetron, stool tended to become firmer. No adverse effects were observed. Plasma levels were highest with 8 mg cilansetron.

Conclusions  Cilansetron slightly augments meal‐stimulated and markedly neostigmine‐stimulated phasic motility of the sigmoid colon. When administered over 7 days, it tends to increase stool consistency and is well tolerated.
Topical application of doxepin hydrochloride, capsaicin and a combination of both produces analgesia in chronic human neuropathic pain: a randomized, double‐blind, placebo‐controlled study
Authors: McCleane Gary
Published: March 2003

Aims
To assess the analgesic efficacy of topical administration of 3.3% doxepin hydrochloride, 0.025% capsaicin and a combination of 3.3% doxepin and 0.025% capsaicin in human chronic neuropathic pain.

Methods
A randomized, double‐blind, placebo‐controlled study of 200 consenting adult patients. Patients applied placebo, doxepin, capsaicin or doxepin/capsaicin cream daily for 4 weeks. Patients recorded on a daily basis overall pain, shooting, burning, paraesthesia and numbness using a 0–10 visual analogue scale during the week prior to cream application (baseline levels) and for the 4 week study period. Side‐effects and desire to continue treatment were also recorded.

Results
Overall pain was significantly reduced by doxepin, capsaicin and doxepin/capsaicin to a similar extent. The analgesia with doxepin/capsaicin was of more rapid onset. Capsaicin significantly reduced sensitivity and shooting pain. Burning pain was increased by doxepin and by capsaicin and to a lesser extent by doxepin/capsaicin. Side‐effects were minor. One patient requested to continue placebo cream, 17 doxepin cream, 13 capsaicin and 9 the combination of doxepin and capsaicin.

Conclusions
Topical application of 3.3% doxepin, 0.025% capsaicin and 3.3% doxepin/0.025% capsaicin produces analgesia of similar magnitude. The combination produces more rapid analgesia.
Ovarian hormones in man: their effects on resting vascular tone, angiotensin converting enzyme activity and angiotensin II‐induced vasoconstriction
Authors: Lee Alison F. C. , McFarlane Lesley C. , Struthers Allan D.
Published: March 2003

Aims
Oestrogens in women have been shown to cause vasodilation which may reflect alterations in the activity of vascular angiotensin converting enzyme (ACE) and/or sensitivity to angiotensin II. The aim of this study was to assess the effect of ovarian hormones on vascular tone, vascular ACE activity and vasoconstriction to angiotensin II in males.

Methods
Eight volunteers were randomised in a crossover design to oestradiol, medroxy‐progesterone, and placebo. Vasoconstriction to angiotensin I and angiotensin II was assessed by forearm plethysmography.

Results
Although baseline forearm flow was increased with oestradiol, suggesting generalized vasodilation, there were no changes in the vasoconstrictor responses to angiotensin I or angiotensin II. Medroxy‐progesterone affected neither baseline flow nor vasoconstrictor responses. The results expressed as percentage reduction in flow (mean ± s.d.) were: angiotensin I 48 pmol ml−1: placebo −48 ± 14%; oestradiol −42 ± 16%; medroxyprogesterone −43 ± 8% and for angiotensin II 16 pmol ml−1: placebo −42 ± 10%; oestradiol −39 ± 11%; medroxyprogesterone −46 ± 13%.

Conclusions
Acute administration of oestradiol caused vasodilation in males, the effect was not due to alterations in vascular ACE activity or to altered sensitivity to angiotensin II.
Parkinson's disease and CYP1A2 activity
Authors: Forsyth, J. T. , Grünewald, R. A. , Rostami‐Hodjegan, A. , Lennard, M. S. , Sagar H. J. , Tucker G. T.
Published: March 2003

Aims
MPTP, a neurotoxin which induces parkinsonism is partially metabolized by the enzyme CYP1A2. Smoking appears to protect against Parkinson's disease (PD) and cigarette smoke induces CYP1A2 activity. Thus, we investigated the hypothesis that idiopathic PD is associated with lower CYP1A2 activity using caffeine as a probe compound.

Methods
CYP1A2 activity was assessed using saliva paraxanthine (PX) to caffeine (CA) ratios. Caffeine half‐life was also estimated from salivary concentrations of caffeine at 2 and 5 h post dose. 117 treated and 40 untreated patients with PD and 105 healthy control subjects were studied.

Results
PX/CA ratios were 0.57, 0.93 and 0.77 in treated patients, untreated patients and healthy control subjects, respectively, with no significant differences between study groups (95% CI: treated patients vs controls −0.24, 0.57; untreated patients vs controls −0.75, 0.35). However, patients with PD (treated or untreated) had caffeine half‐lives shorter than that in controls (treated patients: 262 min, untreated patients: 244 min, controls: 345 min; 95% CI: controls vs treated patients 23, 143 (P = 0.003); controls vs untreated patients 19, 184 (P = 0.011)). Amongst the patients with PD, caffeine half‐life was also inversely related to the age of onset of disease (P = 0.012); gender and concomitant drugs did not influence this significantly.

Conclusions
Based on PX/CA ratio, there was no evidence of decreased CYP1A2 activity in patients compared with control subjects. The observed decrease in the elimination half‐life of caffeine in PD may be caused by increased CYP2E1 activity, an enzyme that also contributes to the metabolism of caffeine. The latter warrants further investigation.
Drug use of children in the community assessed through pharmacy dispensing data
Authors: Schirm Eric , Van Den Berg Paul , Gebben Henkjan , Sauer Pieter , De Jong‐van den Berg Lolkje
Published: March 2003

Aims
To determine the extent of drug use in children and the types of drugs that children use.

Methods
Cross‐sectional study and cohort study, using computerized pharmacy dispensing records for all children aged 0–16 years in the northern part of The Netherlands in 1998. The main outcome measures were proportion of children that used drugs (per sex and age group), mean number of drugs per child, 10 most widely used drug groups and cumulative proportions of drugs users (per drug group) during the first 2 years of life.

Results
Drug use was the highest among infants, decreased till adolescence and increased from there. Overall, approximately 60% of all children used at least one drug in 1998. At younger ages, boys used more drugs than girls and at older ages girls used more drugs than boys. Systemic antibiotics were used by 21% of the children and were by far the most widely used drugs. Other frequently used drugs were analgesics (10%), corticosteroids for dermatologic use (9%), anthistamines (8%) and antiasthmatics (7%). Approximately 10% of the children had used at least one drug at the age of 1 month and at the age of 2 years this proportion was 81%.

Conclusions
The majority of children was exposed to one or more drugs and this exposure started at very young age. This shows the importance of good guidelines for drug use in children and emphasizes the necessity of research of pharmacokinetic and pharmacodynamic properties in children to obtain safety, efficacy and quality evidence of these drugs.
Design issues for drug epidemiology
Authors: McMahon Alex D. , MacDonald Tom M.
Published: March 2003

Despite the difficulties involved in designing drug epidemiology studies, these studies are invaluable for investigating the unexpected adverse effects of drugs. The aim of this paper is to discuss various aspects of study design, particularly those issues that are not easily found in either textbooks or review papers. We have also compared and contrasted drug epidemiology with the randomized controlled trial (RCT) wherever possible. Drug epidemiology is especially useful in the many situations where the RCT is not suitable, or even possible. The study base has to be defined before the appropriate cohort of subjects is assembled. If all of the cases are identified, then a referent sample of controls may be assembled by random sampling of the study base. If all of the cases cannot be assembled, a hypothetical secondary base may need to be created. Preferably, only new‐users of the drug should be included, and the risk‐ratio will be different for acute users and chronic users. Studies will usually only be possible when researching the unintended effects of drugs. It is difficult to study efficacy because of confounding by indication. In occasional circumstances it may be possible to study efficacy (examples are given). Discussion of the dangers of designing with generalisability in mind is provided. Additionally, the similarities in study design between drug epidemiology and the RCT are discussed in detail, as well as the design‐characteristics that cannot be shared between the two methods.
Stereoselective halofantrine disposition and effect:concentration‐related QTc prolongation
Authors: Abernethy Darrell R. , Wesche David L. , Barbey Jean T. , Ohrt Colin , Mohanty Sarina , Pezzullo John C. , Schuster Brian G.
Published: March 2003

Aims  1) To characterize the variability of multiple‐dose halofantrine pharmacokinetics over time in healthy adults, 2) to correlate the pharmacodynamic measure electrocardiographic (ECG) QT interval with (+)‐ and (−)‐halofantrine plasma concentration and 3) to evaluate the safety and tolerance of halofantrine hydrochloride given over time to healthy adults.

Methods  Twenty‐one healthy subjects were enrolled and 13 completed the study (180 days). Subjects received either 500 mg of racemic halofantrine once daily in the fasted state for 42 days, or placebo, and then halofantrine washout was documented for the following 138 days. Pharmacokinetic and pharmacodynamic (ECG QTc) measurements were obtained.

Results  Mean accumulation half‐times (days) for halofantrine were: 7.0 ± 4.8 [(+)‐halofantrine] and 7.3 ± 4.8 [(−)‐halofantrine]. Mean steady‐state concentrations were: 97.6 ± 52.0 ng ml−1[(+)‐halofantrine] and 48.5 ± 20.8 [(−)‐halofantrine]. Steady‐state oral clearance was: 139 ± 73 l h−1[(+)‐halofantrine] and 265 ± 135 l h−1[(−)‐halofantrine]. Peak plasma concentrations of both (+)‐ and (−)‐halofantrine were attained at 6 h and maximal ECG QTc prolongation was at 4–8 h following drug administration. Fourteen of 16 subjects who received active drug had ECG QTc prolongation that was positively correlated with both (+)‐ and (−)‐halofantrine concentration. The five subjects who received placebo had no demonstrable change in ECG QTc throughout the study.

Conclusions 
Halofantrine accumulates extensively and shows high intersubject pharmacokinetic variability, is associated with concentration‐related ECG QTc prolongation in healthy subjects, and is clinically well tolerated in this subject group.
Non‐sedating antihistamine drugs and cardiac arrhythmias – biased risk estimates from spontaneous reporting systems?
Authors: De Bruin M. L. , Van Puijenbroek E. P. , Egberts A. C. G. , Hoes A. W. , Leufkens H. G. M.
Published: March 2003

Aims  This study used spontaneous reports of adverse events to estimate the risk for developing cardiac arrhythmias due to the systemic use of nonsedating antihistamine drugs and compared the risk estimate before and after the regulatory action to recall the over‐the‐counter status of some of these drugs.

Methods  All suspected adverse drug reactions (ADRs) reported until July 1999 to the Netherlands Pharmacovigilance Foundation Lareb were used to calculate the ADR reporting odds ratio, defined as the ratio of exposure odds among reported arrhythmia cases, to the exposure odds of other ADRs (noncases), adjusted for gender, age, reporter, year of reporting and comedication, stratified for the periods before and after the governmental decision in the Netherlands.

Results  Seven‐hundred and thirty‐seven cases of arrhythmia were reported, out of which there were 43 instances where the patients were using nonsedating antihistamines. In general nonsedating antihistamines are associated with cardiac arrhythmia to a higher extent in comparison with other drugs (ADR reporting odds ratio 2.05 [95% CI: 1.45, 2.89]). The association between arrhythmias and nonsedating antihistamine drugs calculated before 1998 was not significantly higher than 1 (OR 1.37 [95% CI: 0.85, 2.23]), whereas the risk estimate calculated after the governmental decision did significantly differ from 1 (OR 4.19 [95% CI: 2.49, 7.05]).

Conclusions  Our data suggest that nonsedating antihistamines might have an increased risk for inducing arrhythmias. Our findings, however, strongly suggest that the increased risk identified can at least partly be explained by reporting bias as a result of publications about and mass media attention for antihistamine induced arrhythmias.
Pharmacokinetics and pharmacodynamics of combination chemotherapy with paclitaxel and epirubicin in breast cancer patients
Authors: Danesi Romano , Innocenti Federico , Fogli Stefano , Gennari Alessandra , Baldini Editta , Di Paolo Antonello , Salvadori Barbara , Bocci Guido , Conte Pier Franco , Del Tacca Mario
Published: March 2003

Aims  To investigate the pharmacokinetics and pharmacodynamics of epirubicin and paclitaxel in combination, as well as the effects of paclitaxel and its vehicle Cremophor EL on epirubicin metabolism.

Methods  Twenty‐seven female patients with metastatic breast cancer received epirubicin 90 mg m−2 i.v. followed 15 min or 30 h later by a 3 h i.v. infusion of paclitaxel 175, 200 and 225 mg m−2. Plasma concentrations of paclitaxel, epirubicin and epirubicinol were measured and the relationship between neutropenia and drug pharmacokinetics was evaluated using a sigmoid maximum effect (Emax) model. Finally, the influence of paclitaxel and Cremophor EL on epirubicin metabolism by whole blood was examined.

Results  An increase in epirubicinol plasma concentrations occurred after the start of the paclitaxel infusion, resulting in a significant increase in the area under the plasma concentration‐time curve (AUC) of epirubicinol (+0.5 µmol l−1 h [95% CI for the difference: 0.29, 0.71],+0.66 µmol l−1 h [95% CI for the difference: 0.47, 0.85] and +0.82 µmol l−1 h [95% CI for the difference: 0.53, 1.11] at paclitaxel doses of 175, 200 and 225 mg m−2, respectively), compared with epirubicin followed by paclitaxel 30 h later (0.61±0.1 µmol l−1 h). A significant increase in epirubicin AUC (+0.74 µmol l−1 h [95% CI for the difference: 0.14, 1.34] and +1.09 µmol l−1 h [95% CI for the difference: 0.44, 1.74]) and decrease in drug clearance (CLTB) (−25.35 l h−1 m−2[95% CI for the difference: −50.18, −0.52] and −35.9 l h−1 m−2[95% CI for the difference −63,4,−8,36]) occurred in combination with paclitaxel 200 and 225 mg m−2 with respect to the AUC (3.16±0.6 µmol l−1 h) and CLTB (74.4±28.4 l h−1 m−2) of epirubicin followed by paclitaxel 30 h later. An Emax relationship was observed between neutropaenia and the time over which paclitaxel plasma concentrations were equal to or greater than 0.1 µmol l−1 (tC0.1). The tC0.1 value predicted to yield a 50% decrease in neutrophil count was 7.7 h. Finally, Cremophor EL markedly inhibited the metabolism of epirubicin to epirubicinol in whole blood.

Conclusions  Paclitaxel/Cremophor EL affects the disposition of epirubicinol and epirubicin. Furthermore, the slope factor of the Emax relationship between neutropenia and tC0.1 of paclitaxel suggests that the drugs might also interact at the pharmacodynamic level.
Caffeine in apnoeic Asian neonates: a sparse data analysis
Authors: Lee How Sung , Khoo Yok Moi , Chirino‐Barcelo Yazmin , Tan Kim Leong , Ong Dorothy
Published: March 2003

Aims  To monitor plasma caffeine concentrations and adverse effects and to study the pharmacokinetics of caffeine in neonatal apnoea in the local Asian population after intravenous administration of caffeine.

Methods  Eighteen neonates with apnoea were treated with caffeine citrate at a loading dose of 10 mg caffeine base kg−1 and a maintenance dose of 2.5 mg kg−1 day−1. Blood samples, three after loading and two after the maintenance dose on day 2, 3, 7, 14 and 21 were taken and analysed for caffeine and its main metabolites using solid phase extraction and h.p.l.c. Adverse effects were monitored. Sparse data pharmacokinetic analysis was performed using P‐Pharm.

Results  Mean caffeine concentrations varied from 10 to 20 mg l−1 throughout treatment (range 3.6–28.4 mg l−1). These concentrations were efficacious; less so in those with lower concentrations. Adverse effects included gastrointestinal disturbances, diuresis and hyperglycaemia. Pharmacokinetic parameter estimates [mean (coefficient of variation%)] were CL=0.00628 (17.5%) l h−1 and V=0.961 (20.3%) l. CL (l h−1)=0.004248 * wt(kg)+0.00154; r=0.8, P<0.01, explained 64% of the variation. V (l)=0.6299 * wt(kg)+0.259; r=0.67, P<0.01, explained 45% of variation. Model‐predicted compared with observed plasma concentrations in a separate group of 10 neonates were unbiased and of good precision.

Conclusions  The dosing regimen studied was suitable for our local Asian neonates as it resulted in therapeutic caffeine concentrations for adequate treatment of apnoea. Adverse effects were tolerable. Therefore, to avoid a higher incidence of adverse effects, this regimen should be retained and not increased as proposed by other workers. CL and V were within values of those reported for neonatal apnoea. Sparse data analysis showed that weight alone was adequate as the influential variable for the accurate prediction of individual pharmacokinetic parameters, plasma concentrations and for dosage adjustment if required.
Involvement of CYP2C9 and UGT2B7 in the metabolism of zaltoprofen, a nonsteroidal anti‐inflammatory drug, and its lack of clinically significant CYP inhibition potential
Authors: Furuta Shigeru , Akagawa Nobuyoshi , Kamada Emiko , Hiyama Akio , Kawabata Yoshihiro , Kowata Nobuhiko , Inaba Atsuhiro , Matthews Anne , Hall Michael , Kurimoto Tadashi
Published: March 2003

Aims
To identify the cytochrome P450 (CYP) and UDP‐glucuronosyltransferase (UGT) isoforms responsible for the formation of the primary metabolite(s) of zaltoprofen, and to predict possible drug interactions by investigating the inhibition of CYP isoforms in vitro.

Methods
The metabolism of zaltoprofen was studied in vitro using recombinant CYP and UGT isoform cDNA‐expression systems. The effects of selective isoform inhibitors on zaltoprofen metabolism were studied using human liver microsomes. The inhibitory effects of zaltoprofen on the metabolism of selective probe substrates for CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4 were also determined in human liver microsomes.

Results
Zaltoprofen was extensively metabolized by CYP2C9 and UGT2B7. CYP2C9 catalysed sulphoxidation but not hydroxylation of zaltoprofen. In the human liver microsomal metabolism study, zaltoprofen metabolism was markedly inhibited by sulphaphenazole, a selective inhibitor of CYP2C9. In the drug interaction study, negligible inhibition (< 15%) of the activities of CYP1A2, CYP2C19, CYP2D6, CYP2E1 and CYP3A4 was apparent at 5 µg ml−1, the maximum plasma concentration   observed   in   humans   after   oral   administration   of   an   80 mg   zaltoprofen  tablet. However, zaltoprofen inhibited CYP2C9 by 26% at 5 µg ml−1. At higher concentrations, zaltoprofen produced some inhibition of CYP2C9 (IC50 = 19.2 µg ml−1; 64.4 µm) and CYP3A4 (IC50 = 53.9 µg ml−1; 181 µm). The free drug concentrations in plasma (0.02 µg ml−1, 67.0 nm) at the Cmax of the clinically effective doses are much lower than the IC50 values corrected for the nonspecific binding ratio of zaltoprofen to microsomal protein (15.5 µg ml−1 for CYP3A4, 49.5 µg ml−1 for CYP3A4). Furthermore, the maximum free drug concentrations in the hepatic intracellular was calculated to be 0.068 µg ml−1 and the increase in the AUC in the presence of zaltoprofen was estimated to be only 0.4% for CYP2C9 substrates and 0.1% for CYP3A4 substrates, respectively.

Conclusions
Zaltoprofen is predominantly metabolized by CYP2C9 and UGT2B7, and is considered unlikely to cause significant drug interactions in vivo when coadministered with CYP substrates at clinically effective doses.
Central and peripheral effects of sustained caffeine use: tolerance is incomplete
Authors: Watson Joanne , Deary Ian , Kerr David
Published: March 2003

Aims
It is widely held that tolerance develops to the effects of sustained caffeine consumption. This study was designed to investigate the effects of chronic, staggered caffeine ingestion on the responses of an acute caffeine challenge, during ­euglycaemia.

Methods
Twelve healthy volunteers were randomized using a double‐blind, cross‐over design to take either 200 mg caffeine (C‐replete) or placebo (C‐naïve) twice daily for 1 week. Following baseline measurements being made, the responses to 200 mg caffeine (blood‐pressure, middle cerebral artery velocity, mood and cognitive performance) were examined over the subsequent 120 min. Blood glucose was not allowed to fall below 4.0 mmol l−1.

Results
After the caffeine challenge, middle cerebral artery blood velocity decreased in both conditions but was greater in the C‐naïve condition (−8.0 [‐10.0, −6.1] cm s−1vs−4.9 [‐6.8, −2.9] cm s−1 C‐replete, P < 0.02). Systolic blood pressure rise was not significantly different in C‐naïve, although this rise was more sustained over time (P < 0.04). Mood was adversely affected by regular caffeine consumption with tense aspect of mood significantly higher at baseline in C‐replete 11.6 ± 0.6 C‐naïve vs 16.3 ± 1.6 C‐replete, P < 0.01). Cognitive performance was not affected by previous caffeine exposure.

Conclusions
Overall these results suggest that tolerance is incomplete with respect to both peripheral or central effects of caffeine.
Preliminary evidence of an association of tumour necrosis factor microsatellites with increased risk of multiple basal cell carcinomas
Authors: Hajeer A.H. , Lear J.T. , Ollier W.E.R. , Naves M. , Worthington J. , Bell D.A. , Smith A.G. , Bowers W.P. , Jones P.W. , Strange R.C. , Fryer A.A.
Published: March 2003

Tumour necrosis factor alpha (TNF-α) appears important in ultraviolet-induced immunosuppression, suggesting that it is a susceptibility candidate for cutaneous basal cell carcinoma (BCC). We now describe data on the association between TNF microsatellite polymorphisms, first on susceptibility in 202 controls and 133 cases each having two to 30 BCCs, and secondly, within the cases, on BCC numbers. The data show that the proportions of individuals with TNF a1- and a7-containing genotypes were significantly different (P = 0·0271, P = 0·0393, respectively) between cases and controls. Secondly, within the cases, TNF alleles d4 (P = 0·023) and d6 (P = 0·006) alone, and the TNF a2–b4–d5 haplotype (P = 0·007), were significantly associated with the number of BCC lesions. These preliminary data provide the first evidence that TNF microsatellite polymorphism may influence the pathogenesis of multiple BCC.
Angiomatoid (malignant) fibrous histiocytoma as a second tumour in a child with neuroblastoma
Authors: Gambini C. , Haupt R. , Rongioletti F.
Published: March 2003

Neuroblastoma occurring as a disseminated disease in children has a poor prognosis. Haematogenous metastases usually involve the marrow, bone, liver and skin. A second neoplasm may also develop. We describe a child with retroperitoneal neuroblastoma (stage 3) who developed a nodular mass in the inguinal area which was suspected to be a metastasis. Histopathology disclosed an angiomatoid (malignant) fibrous histiocytoma, and excision was curative. The occurrence of angiomatoid (malignant) fibrous histiocytoma as a second tumour in a patient with neuroblastoma has not previously been reported.
c-Myc expression in human anagen hair follicles
Authors: Rumio C. , Donetti E. , Imberti A. , Barajon I. , Prosperi E. , Brivio M.F. , Boselli A. , Lavezzari E. , Veraldi S. , Bignotto M. , Castano P.
Published: March 2003

The hair follicle represents a very attractive organ system for studying the precise balance between cell proliferation, growth, differentiation, and death of cells, because it periodically and regularly regenerates, retaining its morphogenetic signals throughout its life. One of the most intriguing oncogenes which is able to induce both cell growth and apoptosis, depending upon the environmental conditions, is c-myc. The aim of the present study was to investigate its presence and localization in human hair follicles by immunohistochemistry and immunofluorescence. Our observations demonstrated the consistent presence of two clusters of c-Myc-expressing cells in anagen follicles, located in two annular regions of the inner root sheath, at the border between cells characterized by putative trichohyalin granules and cells which are keratinized. The lower group belongs to Henle’s layer, while the upper group belongs to Huxley’s layer. c-Myc oncoprotein seems to favour apoptosis/differentiation and may be a marker for terminal differentiation of trichocytes, at least in the inner root sheath. Our findings agree with the interpretation that the complex morphology of the hair follicle reflects its complex function; the extrusion of a highly organized multicellular structure, the hair shaft, driven by another highly organized multicellular structure, the inner root sheath.
A novel PTEN mutation in a Japanese patient with Cowden disease
Authors: Kubo Y. , Urano Y. , Hida Y. , Ikeuchi T. , Nomoto M. , Kunitomo K. , Arase S.
Published: March 2003

Cowden disease (CD) is an autosomal dominant syndrome characterized by multiple hamartomatous lesions and an increased risk for malignancies. Recent evidence has indicated that the PTEN gene, encoding a protein tyrosine phosphatase, is the CD susceptibility gene. However, another line of evidence has suggested that CD might be genetically heterogeneous. Clinical features of CD are variable, and there are interfamilial differences in the expression of skin lesions . Therefore, information on PTEN mutations in CD patients should be accumulated to clarify the genotype–phenotype correlation. In the present study, we found heterozygous germline mutations of PTEN in all of three Japanese patients with CD examined, indicating no genetic heterogeneity among our patients. The mutations included two non-sense mutations of R335X and R130X, and a mis-sense mutation of C136R. To the best of our knowledge, the C136R mutation has not previously been reported in CD patients. This novel mutation was located outside the core motif of the phosphatase domain of PTEN protein, where most of the missense mutations previously reported in CD patients were clustered. Mucocutaneous manifestations were far fewer in the patient with this mutation than in the patients with nonsense mutations. Whether the phenotypic difference in mucocutaneous features was due to the different mutations remains unclear.
Role for tumour necrosis factor-α receptors in ultraviolet-induced skin tumours
Authors: Starcher B.
Published: March 2003

The biological effects of tumour necrosis factor (TNF)-α are mediated through either the TNFR1 or the TNFR2 receptor. In the present study, the effects of ultraviolet (UV) irradiation on skin pathology and tumour promotion were studied in hairless mice deficient in either the TNFR1 or the TNFR2 receptor. SKH-1 hairless mice were crossed with either TNFR1 knockout (KO) mice or TNFR2 KO mice to develop a strain of hairless mice deficient in either of these receptors. Elastosis and other pathological indications of UVB irradiation were not affected by the loss of either receptor. The absence of either receptor, however, resulted in a highly significant reduction in skin tumours in response to UVB irradiation. Inflammatory cell influx following chronic UV irradiation was virtually eliminated in the TNFR1 KO mice, while the TNFR2 KO mice responded to UV irradiation with the normal increase in inflammatory cells throughout the lower and upper dermis. Contact hypersensitivity responses were eliminated in the TNFR2 KO mice, whereas the TNFR1 KO mice retained normal contact hypersensitivity reactions. These studies suggest that TNF-α plays no part in the accumulation of excessive elastin in the skin during chronic UVB exposure. However, there appears to be an important role for TNF-α in mediating tumorigenesis, distinct from its role in initiating cutaneous immune responses.
Two cases of vulval pigmented extramammary Paget’s disease: histochemical and immunohistochemical studies
Authors: Chiba H. , Kazama T. , Takenouchi T. , Nomoto S. , Yamada S. , Tago O. , Ito M.
Published: March 2003

We describe two Japanese female patients with pigmented extramammary Paget’s disease (EMPD); one patient had a dark brown plaque and the other had a reddish patch with a pigmented area, both affecting the vulval region. Histochemical and immunohistochemical examinations confirmed EMPD with melanocyte colonization; plump tumour cells with a large nucleus and pale cytoplasm that were positive for CAM 5.2 and CEA proliferated singly or in nests in the epidermis, and scattered among the tumour cells were many dendritic cells with a large amount of melanin that were positive for HMB-45 and S-100 protein. Fontana–Masson (FM) stain showed many positive cells with well-developed dendritic processes within and around tumour nests. Histochemical and immunohistochemical studies of non-pigmented EMPD cases on the same region showed that HMB-45 positive cells were sparse or not detected at all, and that also FM staining-positive cells were decreased or not detected, and their dendritic processes were poorly formed. The present study suggests that there might be heterogeneity in EMPD in terms of relationships between Paget’s cells and melanocytes.
Granulocyte colony‐stimulating factor administration increases serum concentrations of soluble selectins
Authors: Ohsaka Akimichi , Saionji Katsu , Igari Jun
Published: March 2003

In order to explore the possible role of granulocyte colony‐stimulating factor (G‐CSF) in the inflammatory process, we examined the serum concentrations of soluble selectins (sL‐selectin, sE‐selectin and sP‐selectin) following in vivo administration of G‐CSF to five healthy volunteers and 12 neutropenic patients with haematological malignancies. The serum concentrations of both sL‐selectin and sE‐selectin were slightly but significantly increased after G‐CSF administration in the healthy volunteers. The serum concentrations of all three selectins were significantly increased after G‐CSF administration in the neutropenic patients concomitant with an increase in their neutrophil counts. These findings suggest that G‐CSF may participate in the leucocyte–endothelial cell interactions in vivo.
IL‐6 and SCF exert different effects on megakaryocyte maturation
Authors: Tajika Kenji , Ikebuchi Kenji , Inokuchi Koichi , Hasegawa Setuo , Dan Kazuo , Sekiguchi Sadayoshi , Nakahata Tatsutoshi , Asano Shigetaka
Published: March 2003

Megakaryocytic maturation has two aspects, nuclear maturation and cytoplasmic maturation, both of which are affected by several thrombopoietic cytokines. We cultured murine bone marrow cells separated by BSA velocity sedimentation which contained more immature megakaryocytes with lower ploidy in order to study the effects of rhIL‐6 and rmSCF on the two aspects of megakaryocyte maturation. During culture with rhIL‐6 or rmSCF for 4 d, the megakaryocytes underwent endomitosis, resulting in DNA polyploidization. These two cytokines had different effects on cytoplasmic maturation. Flow cytometry showed that the megakaryocytes cultured with rhIL‐6 became larger and had a more complex intracellular structure than those cultured with rmSCF. Morphological studies showed that the megakaryocytes cultured with rhIL‐6 were larger and more granular, whereas those cultured with rmSCF were smaller and more basophilic. rhIL‐6 promoted cytoplasmic process formation by the megakaryocytes, but rmSCF did not. Analysis of transcripts of PF‐4 and GPllb by RT‐PCR suggested that rmSCF kept more megakaryocytes at the protein synthesis stage than did rhIL‐6. Based on our findings, these cytokines exert different effects on megakaryocytic cytoplasmic maturational events.
t(3;17)(q21;q25) in Epstein‐Barr virus associated peripheral T‐cell lymphoma: a paediatric case
Authors: Millot Frédéric , Brizard Françoise , Babin Philippe , Canioni Danielle , Macintyre Elisabeth , Levard Guillaume , Gambert Catherine , Ricour Claude , Guilhot François
Published: March 2003

We describe an immunocompetent 8‐year‐old boy with a serological profile indicating chronic infection by Epstein‐Barr virus (EBV) who developed subcutaneous and pulmonary lesions related to a peripheral T‐cell proliferation. No clonal rearrangement of T‐cell receptor or immunoglobulin genes was seen. However, the finding of a t(3;17)(q21;q25) in 44 metaphases from one skin lesion demonstrated a clonal origin. We also showed that the proliferative T cells contained EBV genome leading to the diagnosis of EBV‐associated peripheral T‐cell lymphoma. Further cytogenetic and molecular studies are needed to identify genes implicated in the pathogenesis of this haematological malignancy.
Detection of monoclonal plasma cells in the peripheral blood of patients with primary amyloidosis
Authors: MCElroy, JR Edwin A. , Witzig Thomas E. , Gertz Morie A. , Greipp Philip R. , Kyle Robert A.
Published: March 2003

We evaluated the blood from 150 patients with primary AL‐amyloidosis for circulating monoclonal plasma cells using a sensitive slide‐based immunofluorescence technique. The percentage of monoclonal blood plasma cells (BPC) that were in S‐phase was determined by the bromodeoxyuridine labelling index (BLI). Monoclonal BPC were detected in 16% (24/150) of patients. The median number of monoclonal BPC was 1 × 106/l and 4.6% (7/150) of patients had a high number. The BLI was zero in all but three patients. This study demonstrates that monoclonal plasma cells circulate in the peripheral blood of patients with AL‐amyloidosis.
Analysis of the two microsatellite repeat polymorphisms of the factor VIII gene in the Turkish population
Authors: Jarjanazi˙ Hamdi˙ , Timur A. Anil , El‐Maarri˙ Osman , Çağlayan S. Hande
Published: March 2003

DNA‐based diagnosis of haemophilia A has previously been carried out by linkage analysis using two highly informative markers, Hind III RFLP and St14 VNTR, for affected Turkish families. In the present study the number and frequency of the microsatellite alleles at introns 13 and 22 in the factor VIII (FVIII) gene were analysed in order to increase the rate of informative females and accuracy of linkage analysis. Six alleles were observed at both loci. The two most frequent alleles of each locus were the same as the two common alleles found in Anglo‐Americans. The comparison of heterozygosity of both microsatellite loci showed that the Turkish population is slightly less polymorphic than Anglo‐Americans but more polymorphic than Chinese, Slavs and Uzbekians. The additional use of the two microsatellite repeat polymorphisms with the previously established informative markers has been accepted as the most effective strategy in DNA diagnosis by linkage analysis for the assessment of haemophilia A carriers and affected fetuses in the Turkish population. The modifications adopted in this study for the multiplex PCR analysis of the microsatellite repeat polymorphism eliminated the use of radioactivity and sequencing gels, reducing cost and labour.
Electrospray ionization mass spectrometry identification of fibrinogen Banks Peninsula (γ280Tyr→Cys): a new variant with defective polymerization
Authors: Fellowes Andrew P. , Brennan Stephen O. , Ridgway Hayley J. , Heaton David C. , George Peter M.
Published: March 2003

Fibrinogen Banks Peninsula was identified in the mother of a patient referred for investigation following recurrent epistaxis. Coagulation tests revealed prolonged thrombin and reptilase times and a decreased functional fibrinogen level. Thrombin‐catalysed release of fibrinopeptides A and B was normal, and no abnormalities were detected by DNA sequencing of the regions encoding the thrombin cleavage sites in the Aα and Bβ genes. Reducing SDS‐PAGE and reverse‐phase HPLC analysis of purified fibrinogen chains were normal, as was electrospray ionization mass spectrometry (ESI‐MS) analysis of isolated Aα and Bβ chains. However ESI‐MS revealed a mass of 48 345 D for the isolated γ chains, 31 D less than the measured mass of control chains (48 376 D). Since normal and abnormal γ chains were not resolved, this implies a 60–62 D mass decrease in 50% of the molecules. A 60 D decrease was confirmed when DNA sequencing indicated heterozygosity for a mutation of Tyr→Cys at codon 280 of the γ chain gene. Fibrin monomer polymerization revealed a delayed lag phase and reduced final turbidity and although factor XIIIa crosslinking of fibrinogen was normal, it is likely that this delay is due to impaired D:D self association. Recent crystallographic studies show residues γ280 and γ275 make contact across the D:D interface, suggesting a similar mechanism for the polymerization defects in fibrinogens Banks Peninsula and Tokyo II (γ275Arg→Cys).
A phase II trial of interleukin‐2 in myelodysplastic syndromes
Authors: Nand Sucha , Stock Wendy , Stiff Patrick , Sosman Jeffrey , Martone Brenda , Radvany Ruta
Published: March 2003

Patients with myelodysplastic syndromes (MDS) show a decrease in the number and function of natural killer (NK) cells, including lymphokine activated killer (LAK) cell activity. Interleukin‐2 (IL‐2) stimulates the proliferation and activity of these lymphocytes. Anecdotal clinical experience has shown haematological and cytogenetic improvement in myelodysplasia by low‐dose IL‐2 treatment.

A total of 10 patients with MDS were treated with 1 million units of IL‐2 subcutaneously daily for 12 weeks. Even though improvement in CD16+/CD56+ cell numbers was seen in a majority of the patients, the haematological status and transfusion requirements remained unchanged. There was minimal toxicity from this therapy.
Frequent de novo monoallelic expression of β‐spectrin gene (SPTB) in children with hereditary spherocytosis and isolated spectrin deficiency
Authors: Miraglia Del Giudice Emanuele , Lombardi Cinzia , Francese Matteo , Nobili Bruno , Conte Maria Luisa , Amendola Giovanni , Cutillo Stefano , Iolascon Achille , Perrotta Silverio
Published: March 2003

This report represents an attempt to define the rate of β‐spectrin de novo mutations affecting mRNA accumulation in patients with hereditary spherocytosis (HS). 19 HS children with haematologically normal parents and varying degrees of spectrin deficiency were studied. 13 of the 19 cases who were heterozygous at the genomic level for polymorphisms in the β‐spectrin coding region were further studied. However, in an analysis of reverse‐transcripted amplified cDNA from the regions of the polymorphisms, seven patients appeared to be homozygous, suggesting the occurrence of de novo mutational events affecting expression of one β‐spectrin allele. We conclude that in HS patients with isolated spectrin reduction and normal parents the apparently recessive pattern of inheritance may frequently be associated with de novo monoallelic expression of β‐spectrin.
Multiple myeloma and the translocation t(11;14)(q13;q32): a report on 13 cases
Authors: Fonseca Rafael , Witzig Thomas E. , Gertz Morie A. , Kyle Robert A. , Hoyer James D. , Jalal Syed M. , Greipp Philip R.
Published: March 2003

Complex cytogenetic abnormalities have been described in patients with multiple myeloma (MM). To better understand the significance of the most frequent translocation observed in MM, we studied the clinical characteristics of patients with MM and the t(11;14)(q13;q32) abnormality. A search of the cytogenetic database at the Mayo Clinic identified patients with MM and t(11;14)(q13;q32). The medical records were reviewed for the clinical characteristics of these patients. We identified 13 patients with MM and t(11;14)(q13;q32) determined by standard cytogenetic analysis; in 10 patients the abnormality was detected at the time of relapse (three with previously normal results of cytogenetic examination). At the time the translocation was detected, plasma cell (PC) leukaemia was clinically diagnosed in two patients. The median number of circulating PCs, as determined by the cytoplasmic immunofluorescence of T‐cell‐depleted peripheral blood mononuclear cells, was 1.1 × 109/l (mean 1.74; range 0.0017–6.26 × 109/l). On linear regression analysis there was a strong correlation between the number of circulating PCs and the number of bone marrow PCs. The median survival after demonstration of the translocation was 8.1 months. Of all patients, 10 died of disease progression and three were alive. Patients with MM who have t(11;14)(q13;q32) seem to have an aggressive clinical course, even when the abnormality is detected at the time of diagnosis, with evidence of many circulating PCs.
Clinical characteristics of patients with de novo acute myeloid leukaemia and isolated trisomy 11: a Cancer and Leukemia Group B study
Authors: Heinonen Kristiina , MRÓzek Krzysztof , Lawrence David , Arthur Diane C. , Pettenati Mark J. , Stamberg Judith , Qumsiyeh Mazin B. , Verma Ram S. , Maccallum Jane , Schiffer Charles A. , Bloomfield Clara D.
Published: March 2003

Isolated trisomy 11 is the third most common sole trisomy in de novo acute myeloid leukaemia (AML). However, only 49 cases have been published, and for only a fraction of these cases has full description of clinical and haematological features been provided. As a result, little is known about the clinical characteristics of de novo AML patients with solitary trisomy 11. We have identified 13 patients (0.9%) with isolated trisomy 11 among a total of 1496 consecutive adult patients successfully karyotyped as part of a prospective Cancer and Leukemia Group B (CALGB) cytogenetic study (CALGB 8461). Nine patients (69%) were over the age of 60 (range 29–73 years). Eight patients (62%) were diagnosed with AML of FAB M2 subtype, three patients (23%) had FAB M1 AML and one patient each had AML of FAB M0 and M7, respectively. Seven patients (54%) had high, >100 × 109/l, platelet counts (median 102 × 109/l; range 17–207 × 109/l). All patients received CALGB induction therapy with standard doses of cytarabine and daunorubicin. Six patients (46%) achieved a complete remission (CR). The median CR duration was 17.5 months (range 8.7–49.8). Only one patient, who underwent bone marrow transplantation in first CR, continues in initial CR. The median survival was 14.3 months (range 0.5–50.7); only one patient survives. We conclude that de novo AML with isolated trisomy 11 is predominantly associated with older age, M2 and M1 FAB subtypes, high platelet count and few long‐term disease‐free survivals, although it is currently unknown whether isolated trisomy 11 constitutes an independent prognostic factor.