bluuebunny/crossref_metadata_2025 huggingface dataset:
Gastric function measurements in drug development
Authors: Pohle Thorsten , Domschke Wolfram
Published: December 2004

The function of the stomach includes initiation of digestion by exocrine secretions such as acid and pepsin, which are under the control of the endocrine secretion of hormones that also coordinate intestinal motility. The stomach also stores and mechanically disrupts ingested food. Various techniques have been developed to assess gastric physiology, the most important of which is assessment of acid secretion, as well as gastric motility and gastric emptying. The influence of drugs on gastric function and the effect of gastric secretion and mechanical actions on the bioavailability of novel compounds are of critical importance in drug development and hence to clinical pharmacologists. The control of acid secretion is essential in the treatment of peptic ulcer disease as well as gastrooesophageal reflux disease (GORD); pH‐metry can be used to determine the necessary dose of an acid suppressant to heal mucosal damage. Disturbed gastric myoelectric activity leading to gastroparesis can cause delayed gastric emptying, often found in patients with diabetes mellitus. Electrogastrography (EGG) may be used to evaluate the influence of prokinetics and other drugs on this condition and aid in determining effective therapy.
Effects of passive smoking on blood pressure and aortic pressure waveform in healthy young adults – influence of gender
Authors: Mahmud Azra , Feely John
Published: December 2004

Aims  Passive smoking impairs the elasticity of the aorta in patients with coronary heart disease. We therefore studied the effect of passive smoking on wave reflection in the aorta, a marker of arterial stiffness, in healthy subjects.

Method  We examined the effects of acute exposure to passive smoking on blood pressure and the aortic pressure waveform in healthy young men (n = 10) and women (n = 11), aged 26 ± 5 years (mean ± SEM) compared with 12 healthy controls, aged 24 ± 2 years (six female) who were exposed to room air. The aortic pressure waveform was derived with radial applanation tonometry (SphygmoCor, AtCor Medical, version 6.2) and the augmentation index, a measure of arterial wave reflection in the aorta, calculated. Blood pressure (Omron Model HEM‐705 CP, Omron Corporation, Tokyo, Japan) and augmentation index were measured at baseline, 15, 30 and 60 min after exposure to environmental tobacco smoke (carbon monoxide 25–30 p.p.m. for 60 min) or room air.

Results  Passive smoking was associated with an increase in brachial (124 ± 4–137 ± 3 mmHg, P < 0.01) and aortic systolic blood pressure (110 ± 3–123 ± 4 mmHg, P < 0.01) at 60 min in the male subjects only. The augmentation index increased from −1.7 ± 5 to 14 ± 5 at 60 min (P < 0.001) only in the male subjects. The transit time of the pulse did not change significantly. The change in augmentation index was independent of the increase in blood pressure. Brachial and aortic diastolic blood pressure and heart rate did not change significantly in either male or female subjects. No haemodynamic changes were observed in the control group.

Conclusions  Acute exposure to passive smoking has a deleterious effect on the arterial pressure waveform in healthy young males but not in females, suggesting a possible protection of female gender from functional changes in arteries.
Sildenafil does not improve nitric oxide‐mediated endothelium‐dependent vascular responses in smokers
Authors: Dishy Victor , Harris Paul A. , Pierce Rosanna , Prasad Harish C. , Sofowora Gbenga , Bonar Holly L. , Wood Alastair J. J. , Stein C. Michael
Published: December 2004

Aims  To examine the hypothesis that sildenafil, a phosphodiesterase type 5 inhibitor that inhibits cGMP breakdown, could enhance nitric oxide‐mediated vasodilation and reverse endothelial dysfunction in chronic smokers.

Methods  Flow‐mediated dilation of the brachial artery and forearm postischemic reactive hyperemia (both nitric oxide‐mediated responses) were measured before and after sildenafil 50 mg and placebo in a double‐blind, randomized, crossover study in 9 men who were chronic smokers (21 ± 3 pack years).

Results  There was no significant change in flow‐mediated dilation after either sildenafil (0.18%, 95%CI −1.7–2%) or placebo (0.24%, 95%CI −2.8–3.3%) (P = 0.88 and 0.8, respectively). Sildenafil had no significant effect on resting forearm blood flow or postischemic reactive hyperemia (P = 0.39 and 0.7, respectively). Resting heart rate and blood pressure were unaffected by sildenafil.

Conclusions  Acute sildenafil administration did not improve endothelial function in chronic smoking men.
Protective role of glutathione S‐transferase P1 (GSTP1) Val105Val genotype in patients with bronchial asthma
Authors: Aynacioglu A. Sükrü , Nacak Muradiye , Filiz Ayten , Ekinci Erhan , Roots Ivar
Published: December 2004

Background  GlutathioneS‐transferase P1 (GSTP1), the abundant isoform of glutathione S‐transferases (GSTs) in lung epithelium, plays an important role in cellular protection against oxidative stress and toxic foreign chemicals. It has been suggested that polymorphisms in the GSTP1 gene are associated with asthma and related phenotypes. As significant interindividual and interethnic differences exist in the distribution of xenobiotic‐metabolizing enzymes, we have studied the GSTP1 Ile105Val polymorphism in patients with asthma in a Turkish sample.

Methods  GSTP1 Ile105Val polymorphism in exon 5 was determined in 210 patients with asthma (112 extrinsic and 108 intrinsic) and 265 control individuals without lung diseases and without history of allergy or atopy, using polymerase chain reaction (PCR)‐restriction fragment length polymorphism (RFLP) techniques.

Results  The proportion of GSTP1 Val105 homozygotes was significantly lower in the patients with asthma than in the control individuals (3.8% vs 12.1%). The odds ratio for GSTP1 Val105 homozygotes vs all other genotypes was 0.29 (95%CL 0.13–0.64, p = 0.01) for asthmatics. The distribution of GSTP1 Ile105Val genotypes and the frequency of GSTP1 Val105Val homozygotes (3.7% vs 3.9%) was not significantly different between extrinsic and intrinsic asthmatics.

Conclusion  These results suggest a significant association between GSTP1 Ile105Val polymorphism and susceptibility to asthma and that the GSTP1 Val105Val genotype may be protective against developing this disease.
Effect of food on the relative bioavailability of two oral formulations of posaconazole in healthy adults
Authors: Courtney Rachel , Wexler David , Radwanski Elaine , Lim Josephine , Laughlin Mark
Published: December 2004

Aims   This randomized, crossover, single‐dose study evaluated the relative oral bioavailability of posaconazole suspension and coprecipitate tablet formulations. Additionally, the study determined whether systemic exposure to posaconazole was affected by prandial status or by the fat content of a meal.

Methods   This was a randomized, open‐label, four‐way crossover, single‐dose study in 20 healthy men. Posaconazole pharmacokinetics were evaluated over 72 h following a single oral dose of posaconazole suspension (200 mg/5 ml) administered with a high‐fat meal, a nonfat breakfast, or after a 10 h fast, or posaconazole tablets (2 × 100 mg) administered with a high‐fat meal.

Results   The posaconazole suspension showed a significant increase in bioavailability compared with the tablet (increase in AUC(0,72 h) = 137% (90% confidence interval (CI) 119%, 156% and Cmax = 123% (90% CI 104%, 146%). The mean increases in AUC(0,72 h) and Cmax values were about 400% when administered with a high‐fat meal compared with administration of the suspension in the fasting state (AUC(0,72 h) 90% CI 343%, 448%; Cmax 90% CI 352%, 493%). Administration of the suspension with a nonfat meal enhanced exposure, resulting in an increase in AUC(0,72 h) of 264% (90% CI 231%, 302%) and in Cmax of 296% (90% CI 250%, 350%) relative to the fasted state.

Conclusions   The suspension formulation of posaconazole was associated with enhanced systemic exposure and increased relative bioavailability compared with the tablet. Food substantially enhanced the rate and extent of posaconazole absorption in healthy subjects.
Metabolic inertia in contracting skeletal muscle: a novel approach for pharmacological intervention in peripheral vascular disease
Authors: Greenhaff P. L. , Campbell‐O’Sullivan S. P. , Constantin‐Teodosiu D. , Poucher S. M. , Roberts P. A. , Timmons J. A.
Published: December 2004

Peripheral vascular disease (PVD) is generally accepted to result in the failure of skeletal muscle blood flow to increase adequately at the onset of muscular work. There are currently no routine pharmacological interventions towards the treatment of PVD, however, recent Phase III trials in the USA have demonstrated the clinical potential of the phosphodiesterase III inhibitor Cilostazol for pain‐free and maximal walking distances in patients with intermittent claudication. PVD is characterized by a marked reliance on oxygen‐independent routes of ATP regeneration (phosphocreatine hydrolysis and glycolysis) in skeletal muscle during contraction and the rapid onset of muscular pain and fatigue. The accumulation of metabolic by‐products of oxygen‐independent ATP production (hydrogen and lactate ions and inorganic phosphate) has long been associated with an inhibition in contractile function in both healthy volunteers and PVD patients. Therefore, any strategy that could reduce the reliance upon ATP re‐synthesis from oxygen‐independent routes, and increase the contribution of oxygen‐dependent (mitochondrial) ATP re‐synthesis, particularly at the onset of exercise, might be expected to improve functional capacity and be of considerable therapeutic value. Historically, the increased contribution of oxygen‐independent ATP re‐synthesis to total ATP generation at the onset of exercise has been attributed to a lag in muscle blood flow limiting oxygen delivery during this period. However, recent evidence suggests that limited inertia is present at the level of oxygen delivery, whilst considerable inertia exists at the level of mitochondrial enzyme activation and substrate supply. In support of this latter hypothesis, we have reported on a number of occasions that activation of the pyruvate dehydrogenase complex, using pharmacological interventions, can markedly reduce the dependence on ATP re‐synthesis from oxygen‐independent routes at the onset of muscle contraction. This review will focus on these findings and will highlight the pyruvate dehydrogenase complex as a novel therapeutic target towards the treatment of peripheral vascular disease, or any other disease state where premature muscular fatigue is prevalent due to metabolite accumulation.
Population pharmacokinetics of piperaquine in adults and children with uncomplicated falciparum or vivax malaria
Authors: Hung Te‐Yu , Davis Timothy M. E. , Ilett Kenneth F. , Karunajeewa Harin , Hewitt Sean , Denis Mey Bouth , Lim Chiv , Socheat Doung
Published: December 2004

Aims To study the population pharmacokinetics of piperaquine after co‐administration with dihydroartemisinin in uncomplicated malaria.

Methods The disposition of piperaquine was studied in 85 Cambodian patients with uncomplicated falciparum or vivax malaria treated with the piperaquine‐dihydroartemisinin coformulation Artekin®. All patients were given Artekin® orally at 0, 6, 24 and 32 h with a total piperaquine dose of 32–35 mg base kg−1. Adults were given tablets while children received either tablets or a dispersible granule formulation. Patients underwent either intensive (17–19 samples) or sparse (2–5 samples) blood sampling schedules over 35 days and clinical/parasitological follow‐up over > 28 days. Piperaquine in plasma was quantified by high performance liquid chromatography.

Results All patients achieved fever clearance within 24 h and parasite clearance within 72 h. The 28‐day cure rate was 97% in adults and 98% in children. A covariate‐free two‐compartment population model with first‐order absorption and elimination gave the most robust representation of the plasma concentration‐time data in both adults and children. In adults (n = 38), the median (interquartile range) derived pharmacokinetic descriptors CL/F, Vss/F and t1/2,z were 0.9 l h−1 kg−1 (0.79–1.02 l h−1 kg−1), 574 l kg−1(371–711 l kg−1) and 23 days (19–28 days), respectively. In children (n = 47), corresponding values were 1.8 l h−1 kg−1 (1.29–2.3 l h−1 kg−1), 614 l kg−1 (332–1205 l kg−1) and 14 days (10–18 days), respectively.

Conclusions Piperaquine is a highly lipid‐soluble drug with a large Vss/F, long t1/2,z and a clearance that is markedly higher in children than in adults.
Retrospective population pharmacokinetic analysis of cetirizine in children aged 6 months to 12 years
Authors: Pitsiu Maria , Hussein Ziad , Majid Oneeb , Aarons Leon , De Longueville Marc , Stockis Armel
Published: December 2004

Aims  To evaluate retrospectively the population pharmacokinetics of cetirizine, a second‐generation antihistamine, in children.

Methods  Data were pooled from six clinical trials, in which cetirizine was administered orally in various formulations and in single and multiple dosage regimens. The population consisted of 112 children (33 female and 79 male) aged 6 months to 12 years. A one‐compartment open model with first‐order absorption and elimination was fitted to the plasma concentration‐time profiles using nonlinear mixed effects modelling with first‐order estimation.

Results  A linear association was found between apparent clearance (CL/F) and age with the former increasing by 0.12 l h−1 per year. The intercept of the relationship was slightly lower for female children (0.59 vs. 0.77 l h−1 in male). The population estimate of CL/F for an average age of 7 years was 1.61 and 1.43 l h−1 for male and female children, respectively. A linear association was found between apparent volume of distribution (V/F) and age with the former increasing by 1.4 l year−1, with an intercept of 4.0 l. The population estimate of V/F for an average age of 7 years was 13.9 l. The magnitudes of interpatient variability were 35.6% for CL/F and 19.7% for V/F. The magnitude of residual variability in cetirizine concentrations was 26.9%.

Conclusions  Population  analysis  predicts  a  linear  increase  in  cetirizine  CL/F  and V/F with age, with CL/F being slightly lower in female children, relative to males of the same age. However, this gender difference probably has no clinical consequences. Since V/F increased more rapidly with age than CL/F, a nonlinear increase in half‐life was seen, from < 4 h in infants to near the adult value at 12 years of age. The current recommended dosing regimens that younger children should receive lower but more frequent doses, are confirmed by the present analysis.
Therapeutic drug monitoring of lopinavir/ritonavir given alone or with a non‐nucleoside reverse transcriptase inhibitor
Authors: Solas Caroline , Poizot‐Martin Isabelle , Drogoul Marie‐Pierre , Ravaux Isabelle , Dhiver Catherine , Lafeuillade Alain , Allegre Thierry , Mokhtari Malika , Moreau Jacques , Lepeu Gérard , Petit Nathalie , Durand Alain , Lacarelle Bruno
Published: December 2004

Aims

To evaluate the interindividual variability in the plasma concentrations of lopinavir in the context of routine monitoring with or without treatment with a non‐nucleoside reverse transcriptase inhibitor and to assess the interaction between the coformulation of lopinavir/ritonavir and efavirenz or nevirapine.

Methods

Plasma trough and peak concentrations (Ctrough, Cmax) of lopinavir from 182 HIV‐1‐infected patients were analysed by high‐performace liquid chromatography. Three lopinavir/ritonavir regimens were assessed, namely (A) 400 mg lopinavir/100 mg ritonavir twice daily given alone (n = 125), (B) 400/100 mg twice daily together with a non‐nucleoside reverse transcriptase inhibitor (n = 25), and (C) 533/133 mg twice daily together with a non‐nucleoside reverse transcriptase inhibitor (n = 32).

Results

Median (ng ml−1) Ctrough and Cmax lopinavir (interquartile range, CV) were: (A) 4852 (3198–6891, 56%) and 8501 (6333–11 584, 41%), (B) 2979 (1704–5186, 74%) and 5612 (3362–11 704, 76%) and (C) 5082 (2696–7226, 74%) and 9757 (4883–12 963, 60%). Median Ctrough of lopinavir was lower in patients taking both efavirenz [P = 0.01, 95% confidence interval (CI) for difference between medians 343, 2713] and nevirapine (P = 0.019, 95% CI for difference between medians 354, 3681) compared with those taking lopinavir/ritonavir alone. A higher interindividual variability was observed when lopinavir/ritonavir was given with a non‐nucleoside reverse transcriptase inhibitor. The risk of achieving a ‘suboptimal’Ctrough of lopinavir (below a threshold of 3000 ng ml−1) was statistically higher in patients treated with a non‐nucleoside reverse transcriptase inhibitor (P < 0.001, 95% CI for difference between percentages 8.8, 43.1%) compared with those receiving lopinavir/ritonavir alone.

Conclusions

Our results confirmed the interaction between lopinavir and efavirenz, and also demonstrated a significant interaction between the former drug and nevirapine, resulting in lower Ctrough of lopinavir. The wide interpatient variability in this interaction suggests that therapeutic drug monitoring may be useful in optimizing the dose of lopinavir.
Blood–brain barrier transport of morphine in patients with severe brain trauma
Authors: Ederoth Per , Tunblad Karin , Bouw René , Lundberg C. Johan F. , Ungerstedt Urban , Nordström Carl‐Henrik , Hammarlund‐Udenaes Margareta
Published: December 2004

Aims

In experimental studies, morphine pharmacokinetics is different in the brain compared with other tissues due to the properties of the blood–brain barrier, including action of efflux pumps. It was hypothesized in this clinical study that active efflux of morphine occurs also in human brain, and that brain injury would alter cerebral morphine pharmacokinetics.

Methods

Patients with traumatic brain injury, equipped with one to three microdialysis catheters in the brain and one in abdominal subcutaneous fat for metabolic monitoring, were studied. The cerebral catheter locations were classified as ‘better’ and ‘worse’ brain tissue, referring to the degree of injury. Morphine (10 mg) was infused intravenously over a 10‐min period in seven patients in the intensive care setting. Tissue and plasma morphine concentrations were obtained during the subsequent 3‐h period with microdialysis and regular blood sampling.

Results

The area under the concentration–time curve (AUC) ratio of unbound morphine in brain tissue to plasma was 0.64 (95% confidence interval 0.40, 0.87) in ‘better’ brain tissue (P < 0.05 vs. the subcutaneous fat/plasma ratio), 0.78 (0.49, 1.07) in ‘worse’ brain tissue and 1.00 (0.86, 1.13) in subcutaneous fat. The terminal half‐life and Tmax were longer in the brain vs. plasma and fat, respectively. The relative recovery for morphine was higher in ‘better’ than in ‘worse’ brain tissue. The Tmax value tended to be shorter in ‘worse’ brain tissue.

Conclusions

The unbound AUC ratio below unity in the ‘better’ human brain tissue demonstrates an active efflux of morphine across the blood–brain barrier. The ‘worse’ brain tissue shows a decrease in relative recovery for morphine and in some cases also an increase in permeability for morphine over the blood–brain barrier.
Clinical inhibition of CYP2D6‐catalysed metabolism by the antianginal agent perhexiline
Authors: Davies Benjamin J. L. , Coller Janet K. , James Heather M. , Gillis David , Somogyi Andrew A. , Horowitz John D. , Morris Raymond G. , Sallustio Benedetta C.
Published: December 2004

Aims

Perhexiline is an antianginal agent that displays both saturable and polymorphic metabolism via CYP2D6. The aim of this study was to determine whether perhexiline produces clinically significant inhibition of CYP2D6‐catalysed metabolism in angina patients.

Methods

The effects of perhexiline on CYP2D6‐catalysed metabolism were investigated by comparing urinary total dextrorphan/dextromethorphan metabolic ratios following a single dose of dextromethorphan (16.4 mg) in eight matched control patients not taking perhexiline and 24 patients taking perhexiline. All of the patients taking perhexiline had blood drawn for CYP2D6 genotyping as well as to measure plasma perhexiline and cis‐OH‐perhexiline concentrations.

Results

Median (range) dextrorphan/dextromethorphan metabolic ratios were significantly higher (P < 0.0001) in control patients, 271.1 (40.3–686.1), compared with perhexiline‐treated patients, 5.0 (0.3–107.9). In the perhexiline‐treated group 10/24 patients had metabolic ratios consistent with poor metabolizer phenotypes; however, none was a genotypic poor metabolizer. Interestingly, 89% of patients who had phenocopied to poor metabolizers had only one functional CYP2D6 gene. There was a significant negative linear correlation between the log of the dextrorphan/dextromethorphan metabolic ratio and plasma perhexiline concentrations (r2 = 0.69, P < 0.0001). Compared with patients with at least two functional CYP2D6 genes, those with one functional gene were on similar perhexiline dosage regimens but had significantly higher plasma perhexiline concentrations, 0.73 (0.21–1.00) vs. 0.36 (0.04–0.69) mg l−1 (P = 0.04), lower cis‐OH‐perhexiline/perhexiline ratios, 2.85 (0.35–6.10) vs. 6.51 (1.84–11.67) (P = 0.03), and lower dextrorphan/dextromethorphan metabolic ratios, 2.51 (0.33–39.56) vs. 11.80 (2.90–36.93) (P = 0.005).

Conclusions

Perhexiline significantly inhibits CYP2D6‐catalysed metabolism in angina patients. The plasma cis‐OH‐perhexiline/perhexiline ratio may help to both phenotype patients and predict those in whom perhexiline may be most likely to cause clinically significant metabolic inhibition.
In vivo coagulation activation following infusion of highly purified factor XI concentrate
Authors: RICHARDS E. M. , MAKRIS M. M. , COOPER P. , PRESTON F. E.
Published: December 2004

The use of factor XI concentrates has been associated with thrombosis. Plasma markers of coagulation activation were measured before and 30, 60, 120 and 240 min after six infusions of the BPL factor XI concentrate. Five studies were completed before surgical intervention, one was undertaken in a patient with an intracerebral haemorrhage. Significant elevation of levels of fibrinopeptide A (FpA) (P < 0.05) and thrombin–antithrombin (TAT) were demonstrated following six infusions and prothrombin fragment F1.2 following four. Levels of all three markers had risen 60 min following concentrate administration and FpA levels remained elevated throughout the study period. Levels of D‐dimer rose in four patients at 240 min. These results indicate significant thrombin generation by 60 min and subsequent plasmin generation consistent with coagulation activation by the factor XI concentrate. The greatest elevation of activation markers was seen in those subjects with pre‐existing coagulation activation. We advise caution in the use of these products and awareness of the risks in patients who may already have activated coagulation states.
Idiotype‐specific T lymphocytes in monoclonal gammopathies: evidence for the presence of CD4+ and CD8+ subsets
Authors: YI QING , ERIKSSON INGRID , HE WEN , HOLM GO¨RAN , MELLSTEDT HÅKAN , O¨STERBORG ANDERS
Published: December 2004

Tumour‐specific CD4+ T helper (Th) and CD8+ T cytotoxic (Tc) cells may participate in the control and eradication of tumour cells. In the present study, idiotype‐specific stimulation of CD4+ and CD8+ blood T cells from patients with monoclonal gammopathy of undetermined significance and patients with untreated multiple myeloma stage I was examined. Activation was measured in the CD4+ and CD8+ subsets enriched by magnetic microbeads as the incorporation of 3H‐thymidine and the secretion of interferon (IFN)‐γ, interleukin (IL)‐2 and IL‐4 by single cells using the enzyme‐linked immunospot assay. Idiotype‐specific T cells were found in four of seven patients. Stimulation was mainly confined to the CD4+ subset in three of the four responding patients. This type of response was major histocompatibility complex (MHC) class II restricted as it could be inhibited by monoclonal antibodies against MHC class II (HLA‐DR), but not against class I (HLA‐ABC) molecules. Idiotype‐specific CD8+ T cells were also demonstrated in these patients although at a lower frequency. One patient showed a strong and dominating activation of CD8+ T cells which could be blocked by antibodies against HLA‐ABC but not against HLA‐DR. Idiotype‐specific CD4+ or CD8+ T cells were mainly of the type‐1 subsets as judged by their secretion of IFN‐γ and IL‐2. Thus, this study provides evidence for the presence of idiotype‐specific and MHC‐restricted CD4+ and CD8+ T cells of the type‐1 subsets in patients with monoclonal gammopathies. Such T cells with the potential to control the growth of tumour B cells may be a suitable target for immunotherapeutic interventions in patients.
Long‐term follow‐up of leukaemia patients after related cryopreserved allogeneic bone marrow transplantation
Authors: STOCKSCHLA¨DER MARCUS , HASSAN HASSAN T. , KROG CORNELIA , KRU¨GER WILLIAM , LO¨LIGER CORNELIUS , HORSTMAN MARTIN , ALTNO¨DER M. , CLAUSEN JOHANNES , GRIMM JAN , KABISCH HARTMUT , ZANDER AXEL
Published: December 2004

We have previously shown that allogeneic bone marrow transplantation (BMT) with cryopreserved donor marrow cells can be used without prolonging the engraftment time or interfering with the reconstitution of haemopoiesis. In this report we extend our initial observations of the first 40 patients who underwent allogeneic bone marrow transplantation from related donors with cryopreserved donor bone marrow for haematological malignancies, including the long‐term follow‐up data of the previously reported patients. The outcome of these patients was compared with that of 40 related BMT recipients receiving fresh donor bone marrow (historic control group). Time until engraftment of all patients receiving cryopreserved bone marrow was not different from the control group (ANC > 0.5 × 109/l 17 d (range 11–24 d) versus 17.5 d (range 10–28 d); platelets > 20 × 109/l 21 d (range 11–85 d) versus 22 d (range 13–69 d), respectively). There was the same incidence of acute and chronic GvHD in patients receiving either cryopreserved bone marrow or fresh bone marrow (acute GvHD ≥ II 61% v 60% and chronic GvHD 56% v 52%, respectively). Chimaerism studies showed no difference between the patient groups. Furthermore, the two groups did not differ in day 100 survival (82% v 72%). With a median follow‐up of 520 d (range 47–1365 d) and 1289 d (range 48–1849 d), 60% of the patients receiving cryopreserved and 53% of the patients receiving fresh allogeneic donor bone marrow, respectively, are alive. We conclude that cryopreservation of allogeneic related donor bone marrow does not adversely affect engraftment, does not decrease the incidence of severe acute GvHD, and does not seem to affect the day 100 survival or long‐term haemopoiesis.
Persistence of residual tumour cells after cytokine‐mediated ex vivo expansion of mobilized CD34+ blood cells in multiple myeloma
Authors: VAN RIET IVAN , JUGE‐MORINEAU NADINE , SCHOTS RIK , DE WAELE MARC , DE GREEF CATHERINEE , THIELEMANS KRIS , VAN CAMP BEN , BAKKUS MARLEEN
Published: December 2004

Mobilized CD34+ blood cells were immunomagnetically enriched from leukapheresis products in five multiple myeloma (MM) patients. Thawed samples of selected CD34+ cells were cultured for up to 21 d in a liquid and stroma‐free culture system with different combinations of recombinant cytokines. The most successful cell expansion was obtained when a combination of rh‐IL‐1β, rh‐IL‐3, rh‐IL‐6, rh‐SCF, rh‐G‐CSF and rh‐GM‐CSF was used. After 14 d this mixture gave a 120–187‐fold overall increase of total nuclear cells and a 4–8‐fold overall increase of early CFU‐GM numbers. In four patients a very sensitive patient‐specific PCR analysis showed the presence of monoclonal cells in the initial leukapheresis products. After immunomagnetic separation a tumour cell depletion of 2–4 logs was observed, although all samples still contained malignant cells. Cell suspensions that were cultured with the most potent cytokine combination showed tumour contamination in two‐thirds of evaluable cases at the moment of maximal CFU‐GM output. Serial cDNA dilution experiments indicated that the positive PCR results at day 14 reflected the persistence of pre‐culture tumour cells rather than in vitro expansion of tumour cells in two cases. This study demonstrates that ex vivo expansion of myeloid precursor cells from mobilized CD34+ cells in MM patients does not always result in an effective purging of residual tumour cells. On the other hand, our culture conditions do not seem to favour in vitro expansion of malignant cells, despite the use of a cytokine cocktail that includes potential myeloma growth factors.
Human herpesvirus‐8/Kaposi's sarcoma‐associated herpesvirus in organ transplant patients with immunosupression
Authors: Alkan Serhan , Karcher Donald S. , Ortiz Antonia , Khalil Salem , Akhtar Mohammed , Ashraf Ali M.
Published: December 2004

Several recent studies have demonstrated Kaposi's sarcoma‐associated herpes virus (KSHV), also known as putative human herpes virus‐8 (HHV‐8), DNA in various epidemiologic forms of Kaposi's sarcoma (KS), including AIDS‐associated, classic, and endemic types. Risk of developing KS in non‐HIV‐infected immunosuppressed hosts, such as patients following solid organ transplantation, is also significantly higher compared to normal individuals. We have retrospectively evaluated 28 organ transplant patients with KS (23 cutaneous and five visceral) for the presence of KSHV genome by polymerase chain reaction (PCR) amplification of DNA isolated from formalin‐fixed, paraffin‐embedded archival tissue samples. 27/28 KS patients were positive for the presence of KSHV. In four KS patients, tissue samples with no histologic evidence of KS were also analysed for KSHV. No evidence of positivity in three samples was noted, but one patient had weak positive amplification products on DNA samples isolated from a gastric biopsy with chronic gastritis and lymph node with sinus histiocytosis. These data support the association of KSHV with KS developing in non‐HIV‐infected immunosuppressed patients, similar to other forms of KS, and suggest that KSHV may play a significant role in the development of all forms of KS.
Kinetics of myeloid and lymphocyte recovery and infectious complications after unrelated umbilical cord blood versus HLA‐matched unrelated donor allogeneic transplantation in adults
Authors: Hamza Nashaat S. , Lisgaris Michelle , Yadavalli Gopala , Nadeau Laura , Fox Robert , Fu Pingfu , Lazarus Hillard M. , Koc Omer N. , Salata Robert A. , Laughlin Mary J.
Published: December 2004

Sources for allogeneic stem cells for patients with haematological disorders lacking a histocompatible sibling donor include matched unrelated donor (MUD) and umbilical cord blood (UCB). A total of 51 patients with haematological disorders, treated with myeloablation and transplantation with either unrelated human leucocyte antigen (HLA) partially matched UCB (28 patients) or HLA‐matched MUD grafts (23 patients) during 1997–2003, were evaluated for life‐threatening infections, haematological reconstitution, graft versus host disease, relapse and event‐free survival (EFS). The median duration of neutropenia after transplantation was longer (29 d vs. 14 d) in the UCB group. The probability of donor‐derived neutrophil engraftment by day 42 was 0·86 [95% confidence interval (CI) 0·71–1·0] in UCB recipients versus 0·96 (95% CI 0·87–1·0) in MUD recipients surviving >28 d. Overall infection rates were higher in UCB recipients, particularly at the early time points (before day +50) after transplantation. Graft failure occurred in five UCB recipients and two MUD recipients and was associated with the occurrence of bacteraemia during neutropenia. The EFS at 3‐year follow‐up was 0·25 in UCB and 0·35 in MUD recipients. UCB transplantation in adults is associated with delayed neutrophil and lymphocyte recovery compared with MUD grafting, and higher rates of bacteraemia at early time points after transplantation.
Increased sensitivity to IL‐4 in cystic fibrosis patients with allergic bronchopulmonary aspergillosis
Authors: Knutsen A. P. , Hutchinson P. S. , Albers G. M. , Consolino J. , Smick J. , Kurup V. P.
Published: December 2004

Background:  Allergic bronchopulmonary aspergillosis (ABPA) is characterized by a heightened Th2 CD4+ T‐cell response to Aspergillus fumigatus allergens and a hyper‐immunoglobulin (Ig)E state compared with cystic fibrosis patients without ABPA. We hypothesize that one reason for this response is increased sensitivity to interleukin (IL)‐4 in ABPA resulting in increased expression of CD23 and CD86 and leading to a positive amplification mechanism that increases Th2 CD4+ T cell responses.

Methods:  Peripheral blood mononuclear cells (PBMC) were isolated from seven ABPA CF and 19 non‐ABPA CF patients and 16 nonatopic controls and stimulated with rIL‐4 (range 0.1–10 ng/ml) and rIL‐13 (range 1–10 ng/ml) for 48 h. The number of CD23 molecules and percentages of CD23+ B cells were quantified by flow cytometry. Both phorbol 12‐myristate 13‐acetate (PMA)/ionomycin (IO) and antigen stimulated, toxoid and Asp f2/f3/f4, PBMC were examined for cytoplasmic cytokine synthesis enumerated by cytokine staining using flow cytometry to measure Th2 and Th1 CD3+ T cells.

Results:  The numbers of CD23 molecules on B‐cells were significantly elevated at time 0 in ABPA CF patients compared with both non‐ABPA CF patients and nonatopic controls. Following IL‐4 stimulation in vitro, the numbers and percentages of CD23 expression on B cells were significantly up‐regulated in ABPA CF patients compared with non‐ABPA CF patients and controls. The IL‐13 stimulation up‐regulated CD23 expression; however, there was no significant difference in ABPA CF patients compared with non‐ABPA CF patients and controls. The percentages of interferon (IFN)‐γ+ CD3+T cells following PMA/IO stimulation were significantly decreased in both ABPA and non‐ABPA CF patients compared with controls. There were no significant differences of IL‐4+ and IL‐13+ CD3+ T cells between ABPA and non‐ABPA CF patients. When tetanus toxoid stimulated T cells were examined, both ABPA and non‐ABPA CF patients had significantly decreased IFN‐γ+ CD3+ T cells compared with controls. In Asp f2/f3/f4 stimulated T cells, ABPA CF patients had significantly increased IL‐4+ CD3+ T cells compared with non‐ABPA CF patients and controls.

Conclusions:  ABPA CF patients have increased sensitivity to IL‐4 but not to IL‐13 up‐regulation of CD23 molecules compared with non‐ABPA CF patients. There were decreased percentages of IFN‐γ+ and IL‐2+ Th1 T cells in CF patients compared with nonatopic controls but similar percentages of IL‐4+ Th2 T cells in all three groups. However, ABPA CF patients had increased frequency of Aspergillus‐stimulated Th2 T cells. This indicated that there is skewing of Th2 T cells in ABPA CF patients. Thus, in CF ABPA patients there is increased Th2 T cells and increased sensitivity to IL‐4.
Frequency of allergic rhinitis to laboratory animals in university employees as confirmed by chamber challenges
Authors: Ruoppi P. , Koistinen T. , Susitaival P. , Honkanen J. , Soininen H.
Published: December 2004

Background:  Occupational risk for laboratory animal allergy (LAA) is well known, but prevention is often insufficient and new cases keep appearing.

Methods:  A questionnaire on work‐related symptoms was sent to all laboratory animal workers in Kuopio University. Subjects with rhinitis were invited to an examination for LAA, which consisted of an otorhinolaryngological examination and skin prick testing (SPT) with 16 common environmental allergens and two to five relevant laboratory animals. In cases of occupational sensitization, a challenge test was performed.

Results:  The questionnaire was completed by 156 of 245 (64%) subjects. Rhinitis was reported by 65 of the respondents (42%) of whom 47 were examined for LAA. The duration of exposure before the onset of rhinitis varied from 1 month to 18 years. Twenty‐seven persons (57%) had a history of some previous atopic symptoms. Forty subjects (85%) showed positive reactions in SPT, 24 (51%) for laboratory animals. Fifteen of the 26 animal challenge tests performed were positive, confirming the diagnosis of occupational allergic rhinitis in three subjects, occupational dermatitis in five and both diseases in seven subjects. The frequency of occupational allergic rhinitis among all participants was 10 of 156 (6%).

Conclusions:  Atopic constitution and work‐related sensitization were common in rhinitic laboratory animal workers. Occupational allergic disease was diagnosed in nearly every third case.
Corticosteroid allergy in asthma
Authors: Kilpiö K. , Hannuksela M.
Published: December 2004

Background: Glucocorticosteroids form the basis of therapy for asthma and other allergic diseases. However, they frequently cause delayed contact allergy and occasionally immediate allergy. The purpose of this study was to investigate the occurrence of corticosteroid allergy among patients with asthma and with some complaints caused by inhaled corticosteroids.

Methods: Patch tests with corticosteroids were performed in 51 asthma patients with side‐effects from inhalant corticosteroids and in 50 symptom‐free asthma patients using the Finn Chamber® system. The corticosteroids and their vehicles were: betamethasone‐17‐valerate 1% in petrolatum, hydrocortisone‐17‐butyrate (Hc‐17‐B) 1% in ethanol, tixocortol‐21‐pivalate 1% in petrolatum, budesonide 0.1% in petrolatum, beclomethasone dipropionate 0.1 and 0.5% in petrolatum and as inhalant powder 200 μg, and fluticasone propionate 0.1 and 0.5% in petrolatum and as inhalant powder 250 μg. The results were read twice, on D4–5 and again on D10.

Results: Two patients in the symptomatic group reacted to corticosteroids in patch tests, one to betamethasone‐17‐valerate, Hc‐17‐B and budesonide, and the other to budesonide and Hc‐17‐B. The first patient suffered from widespread eczematous dermatitis when using beclomethasone. Fluticasone caused oropharyngeal irritation, hoarseness and shortness of breath. The second patient experienced a severe rash after the fourth budesonide inhalation. She had used various topical corticosteroids for her atopic dermatitis without any side‐effects. None of the symptom‐free patients showed positive results.

Conclusions: Delayed allergy to corticosteroids occurs occasionally in asthma, perhaps in the same frequency as in dermatitis. A positive patch test reaction usually means clinical allergy, i.e. the patient cannot use that particular steroid. Cross allergy between corticosteroids is common. However, such patients usually tolerate some other common corticosteroids.
Skin test evaluation in nonimmediate allergic reactions to penicillins
Authors: Torres M.‐J. , Sánchez‐Sabaté E. , Álvarez J. , Mayorga C. , Fernández J. , Padial A. , Cornejo‐García J.‐A. , Bellón T. , Blanca M.
Published: December 2004

Background:  Nonimmediate reactions (NIR) to aminopenicillins (AP) are frequent. Although patch testing (PT) and intradermal testing (IT) are used for diagnosis, comparative results have never been adequately performed. We compared PT and IT in subjects with NIR to AP.

Methods:  Twenty‐one subjects with NIR to AP and positive IT were re‐evaluated. Skin tests were performed with amoxicillin (AX) and ampicillin (AM) at different concentrations in petrolatum, 50, 25, and 5% w/w, for PT and in saline, 20, 2, and 0.2 mg/ml for both PT and IT. Skin biopsies from the site of the positive response were studied with haematoxylin–eosin and immunohistochemistry.

Results:  In the re‐evaluation, one case was IT and PT negative and was excluded; 20 were IT positive and 18 PT positive for both AX and AM. Decreasing concentrations of AP induced a reduction in positivity in both methods when diluted in saline, but not when mixed in petrolatum (PT only). With both PT and IT, immunohistochemical studies showed a perivascular mononuclear infiltrate with CD4 and CD8 memory cells expressing perforin and granzyme B.

Conclusions:  Both tests appear valuable for the diagnosis of NIR to AP. However, IT diagnosed more patients than PT. The vehicle (saline or petrolatum) had no influence on the response, although in the former the concentration was critical. The immunohistochemical analysis showed skin infiltrates compatible with a T‐cell drug reaction.
Allergenic pollen records (15 years) and sensitization in patients with respiratory allergy in Thessaloniki, Greece
Authors: Gioulekas D. , Papakosta D. , Damialis A. , Spieksma F. , Giouleka P. , Patakas D.
Published: December 2004

Background:  Very limited allergenic pollen records exist in Greece so far; moreover, there is a lack of investigation on patient sensitization. The above data are necessary for respiratory allergy diagnosis and treatment worldwide.

Methodology:  Daily records and identification of 16 airborne pollen species were made using a Burkard trap (1987–2001). Skin sensitivity to 13 most common pollen extracts was investigated, in a sample of 1311 asthmatics with atopy, admitted to the Out‐Patient Clinic for Asthma (1990–2001). Skin sensitivity to 55 allergens, including 13 pollen extracts, was detected by skin prick test.

Results:  The following pollen concentrations were recorded: cypress (24.9% of the total), oak (20.8%), wall pellitory (13.6%), olive (9.1%), pine (8.9%), grasses (6.3%), plane (5.4%), hazel (3%), goosefoot (2.5%) and poplar (1.4%). The respective percentages of birch, ragweed, mugwort, willow, alder and elm were lower than 1%. The highest counts of airborne pollen grains were detected from March to June. Regarding patient sensitization, sensitivity was detected to: grasses in 530 patients (40.4%), olive 417 (31.8%), goosefoot 240 (18.3%), wall pellitory 201 (15.3%), mugwort 198 (15.1%), plantain 194 (14.6%), cypress 166 (12.7%), hazel 126 (9.6%), pine 122 (9.3%), poplar 111 (8.4%), plane 107 (8,2%), oak 99 (7.6%) and to birch 89 patients (6.8%). The sensitivity to pollen grains displays preponderance (57.9%) to males.

Conclusions:  For the first time in Thessaloniki, Greece, 15‐year allergenic pollen records were conducted. Clinical observations confirm that the pollen mainly implicated in respiratory allergy symptoms are grasses, olive and wall pellitory.
Allergy markers in adults in relation to the timing of pet exposure: the EGEA* study
Authors: Oryszczyn M.‐P. , Annesi‐Maesano I. , Charpin D. , Kauffmann F.
Published: December 2004

Background: Studies suggest that early childhood exposure to pets may protect from the development of atopy, but limited information is available on adults. The association of allergy markers in adulthood with current and childhood exposure to pets was studied considering retrospectively the window of exposure.

Methods: Immunoglobulin E (IgE), skin prick tests (SPT), eosinophils were related to exposure to pets in 187 adult asthmatic cases and 243 controls from the Epidemiological Study on the Genetics and Environment of Asthma, bronchial hyperresponsiveness and atopy (EGEA) study. Analyses were redone after exclusion of subjects who removed pets or experienced symptoms to animals to take into account selection in that retrospective study.

Results: In asthmatic cases, current exposure to pets was unrelated to SPT positivity (+), whereas childhood exposure was significantly related to less SPT+ to any allergen, and to cat in particular, with an association restricted to those exposed before 2 years of age [OR = 0.30 (CI 0.12–0.76)]. Considering the relative timing of exposure in relation to asthma onset showed that the protective effect of exposure to pets occurs for pet exposure starting before asthma onset [OR for SPT+ = 0.19 (CI 0.08–0.48)].

Conclusion: Results support the hypothesis that exposure to pets in early life, and in particular before asthma onset, may protect against allergen sensitization in adulthood.
Mucosal T‐cell phenotypes in persistent atopic and nonatopic rhinitis show an association with mast cells
Authors: Powe D. G. , Huskisson R. S. , Carney A. S. , Jenkins D. , McEuen A. R. , Walls A. F. , Jones N. S.
Published: December 2004

Background:  Allergic rhinitis is characterized by selective expansion of T cell subsets with a CD4+ phenotype. Recently, we identified a subpopulation of nonallergic rhinitis subjects with increased epithelial mast cell and eosinophil populations, suggestive of local mucosal allergy. Previously, T cell subsets have not been characterized in this subselection of nonallergic subjects and furthermore, their relationship to mast cell and basophil effector cells remain unidentified.

Objective:  To determine if a subpopulation of nonallergic subjects with idiopathic rhinitis (IR) have localized allergy confined to their nasal mucosa by comparing the T cell subsets and major histocompatibility complex (MHC) II expressing cells to persistent allergic rhinitis (PAR). Furthermore, the relationship between T cell subsets and mast cells/basophils was investigated.

Methods:  None of the symptomatic patients in this study were clinically allergen‐challenged. Nasal turbinate mucosa was removed from patients with PAR, IR and normal controls. Morphometry was performed on immunostained sections for T cell subset populations including CD3+, CD4+, CD8+, CD25+, CD45RA+, CD45RO+, human leucocyte antigen (HLA)‐DRα (MHC class II), mast cell tryptase and for basophils.

Results:  Subjects with persistent allergic rhinitis differed to normal controls in showing significantly increased numbers of total (CD3+), activated (CD25+) and allergen‐naïve (CD45RA+) T lymphocytes in their nasal mucosa (P < 0.025). The naïve CD45RA+ memory T cells correlated to mucosal mast cells in PAR (P = 0.03). IR patients differ to allergic subjects in showing significantly reduced numbers of epithelial HLA‐DRα+ cells (P = 0.007), but increased numbers of CD8+ lymphocytes (P = 0.02). The CD8+ T cells correlated with mucosal mast cell numbers (P = 0.02). In both rhinitis groups, basophils were present in very low numbers obviating the need for statistical analysis.

Conclusion:  PAR is characterized by increased numbers of CD3+, CD25+ and CD45RA+ T lymphocytes compared with normal mucosa. Allergic and nonallergic rhinitis groups can be separated by significant differences in the number of epithelial antigen presenting cells (APCs) (HLA‐DRα+) and sub‐epithelial activated (CD25+) T cells. Moreover, IR patients do not significantly differ to their allergic counterparts with respect to total (CD3+) and naïve (CD45RA+) T cell numbers, or numbers of epithelial activated (CD25+) lymphocytes. IR subjects show significantly increased numbers of CD8+ lymphocytes compared with control mucosa and although our findings suggest that the initiating inflammatory events may differ, both rhinitis groups show a similarity in pathology involving mucosal mast cells with an association to infiltrating T cells.