bluuebunny/crossref_metadata_2025 huggingface dataset:
Pharmacodynamic interaction between the new selective cholesterol absorption inhibitor ezetimibe and simvastatin
Authors: Kosoglou Teddy , Meyer Ingo , Veltri Enrico P. , Statkevich Paul , Yang Bo , Zhu Yali , Mellars Lillian , Maxwell Stephen E. , Patrick James E. , Cutler David L. , Batra Vijay K. , Affrime Melton B.
Published: March 2003

Aims  The primary aims of these two single‐centre, randomized, evaluator‐blind, placebo/positive‐controlled, parallel‐group studies were to evaluate the potential for pharmacodynamic and pharmacokinetic interaction between ezetimibe 0.25, 1, or 10 mg and simvastatin 10 mg (Study 1), and a pharmacodynamic interaction between ezetimibe 10 mg and simvastatin 20 mg (Study 2). Evaluation of the tolerance of the coadministration of ezetimibe and simvastatin was a secondary objective.

Methods  Eighty‐two healthy men with low‐density lipoprotein cholesterol (LDL‐C) ≥130 mg dl−1 received study drug once daily in the morning for 14 days. In Study 1 (n=58), five groups of 11–12 subjects received simvastatin 10 mg alone, or with ezetimibe 0.25, 1, or 10 mg or placebo. In Study 2 (n=24), three groups of eight subjects received simvastatin 20 mg alone, ezetimibe 10 mg alone, or the combination. Blood samples were collected to measure serum lipids in both studies. Steady‐state pharmacokinetics of simvastatin and its β‐hydroxy metabolite were evaluated in Study 1 only.

Results  In both studies, reported side‐effects were generally mild, nonspecific, and similar among treatment groups. In Study 1, there were no indications of pharmacokinetic interactions between simvastatin and ezetimibe. All active treatments caused statistically significant (P<0.01) decreases in LDL‐C concentration vs placebo from baseline to day 14. The coadministration of ezetimibe and simvastatin caused a dose‐dependent reduction in LDL‐C and total cholesterol, with no apparent effect on high‐density lipoprotein cholesterol (HDL‐C) or triglycerides. The coadministration of ezetimibe 10 mg and simvastatin 10 mg or 20 mg caused a statistically (P<0.01) greater percentage reduction (mean −17%, 95% CI −27.7, −6.2, and −18%, −28.4, −7.4, respectively) in LDL‐C than simvastatin alone.

Conclusions  The coadministration of ezetimibe at doses up to 10 mg with simvastatin 10 or 20 mg daily was well tolerated and caused a significant additive reduction in LDL‐C compared with simvastatin alone. Additional clinical studies to assess the efficacy and safety of coadministration of ezetimibe and simvastatin are warranted.
Antagonism of long‐acting β2‐adrenoceptor agonism
Authors: Lipworth Brian J.
Published: March 2003

The established place of regular long‐acting β2‐adrenoceptor agonists at step 3 in asthma management guidelines has evolved as a consequence of evidence showing additive effects of salmeterol and formoterol on exacerbation rates, resulting in a putative inhaled corticosteroid sparing effect. There is however, evidence to show that although long‐acting β2‐adrenoceptor agonists facilitate using a lower dose of inhaled corticosteroid, this may occur at the expense of suboptimal anti‐inflammatory control. This is likely to be the case especially with fixed dose combination inhalers where it is not possible to properly titrate anti‐inflammatory therapy with inhaled corticosteroids without also inadvertently overtreating with unnecessarily high doses of long‐acting β2‐adrenoceptor agonists. Most patients with mild to moderate persistent asthma can be adequately controlled on monotherapy with inhaled corticosteroid in low or medium dosage, which is considerably cheaper than concomitant   use   of   a   long‐acting   β2‐adrenoceptor   agonist.   Subsensitivity   to   long‐acting β2‐adrenoceptor  agonists  is  a  predictable  pharmacological  phenomenon  which occurs despite concomitant inhaled corticosteroid therapy and occurs more readily for bronchoprotective  than  bronchodilator  effects.  Subsensitivity  of  salbutamol  protection against bronchoconstrictor stimuli occurs in patients receiving concomitant long‐acting β2‐adrenoceptor agonists, which may be due to β2‐adrenoceptor down‐regulation or prolonged receptor occupancy. Prospective large scale long‐term studies are required to further define the clinical relevance of β2‐adrenoceptor polymorphisms, to look at clinical control outcomes as well as propensity for subsensitivity. It would therefore make more sense to first of all optimize the dose of anti‐inflammatory therapy with inhaled corticosteroid and to then consider adding a long‐acting β2‐adrenoceptor agonist for patients who are poorly controlled.
Population pharmacokinetics of nevirapine in an unselected cohort of HIV‐1‐infected individuals
Authors: De Maat Monique M. R. , Huitema Alwin D. R. , Mulder Jan W. , Meenhorst Pieter L. , Van Gorp Eric C. M. , Beijnen Jos H.
Published: March 2003

Aims
To study the population pharmacokinetics of nevirapine and to identify relationships between patient characteristics and pharmacokinetics in an unselected population of patients attending our outpatient clinic.

Methods
Ambulatory HIV‐1‐infected patients from the outpatient clinic of the Slotervaart Hospital who were being treated with a nevirapine‐containing regimen were included. During each visit, blood samples were collected for the determination of nevirapine plasma concentrations and clinical chemistry parameters. Variables that were collected at baseline were serology for hepatitis B (HBV) and C (HCV) viruses, liver enzymes, and total bilirubin (TBR). In addition, information about concomitant use of St John's wort and patient demographics were included. The pharmacokinetics of nevirapine were described by first‐order absorption and elimination using nonlinear mixed effect modelling (NONMEM V1.1). Population pharmacokinetic parameters (apparent clearance (CL/F), volume of distribution (V/F), absorption rate constant (ka)) were estimated, as were interindividual, interoccasion, and residual variability in the pharmacokinetics. The influence of patient characteristics on the pharmacokinetics of nevirapine was determined.

Results
From 173 outpatients a total number of 757 nevirapine plasma concentrations at a single random time point and full pharmacokinetic curves for 13 patients were available resulting in a database of 1329 nevirapine plasma concentrations. Mean CL/F, V/F, and ka were 3.27 l h−1, 106 l, and 01.66 h−1, respectively. CL/F of nevirapine was correlated with weight, chronic HCV infection, and baseline aspartate aminotransferase (ASAT). Chronic HCV and baseline ASAT> 1.5 × upper limit of normal (ULN) decreased CL/F by 27.4% and 13.2%, respectively, whereas an increase in body weight of 10 kg increased CL/F by 0.14 l h−1. A trend towards a lower CL/F in patients of the Negroid race was observed. No significant covariates were found for V/F.

Conclusions
The pharmacokinetics of nevirapine were adequately described by our population pharmacokinetic model. Weight, chronic HCV infection, and baseline ASAT were found to be significant covariates for CL/F of nevirapine. The model incorporating these significant covariates may be an important aid in further optimizing nevirapine‐containing therapy.
The unbound percentage of saquinavir and indinavir remains constant throughout the dosing interval in HIV positive subjects
Authors: Boffito Marta , Hoggard Patrick G. , Reynolds Helen E. , Bonora Stefano , Meaden E Rhiannon , Sinicco Alessandro , Di Perri Giovanni , Back David J.
Published: March 2003

Aims
To measure the unbound plasma concentrations of saquinavir (SQV) and indinavir (IDV) and to relate them to the total plasma concentrations in order to establish the unbound percentage of protease inhibitors in vivo during a full dosage interval profile.

Methods
HIV‐infected   subjects   (n = 35;   median   CD4   cell   count = 340 × 106 cells l−1, range: 120–825; viral load < 50 copies ml−1 in 22/35) treated with SQV or IDV containing regimens were studied. Plasma drug samples were collected at 0, 2, 4, 8 and 12 h postdose for the twice daily regimens and 0, 1, 2, 4 and 8 h for the three times daily regimens. Ultra‐filtration was used to separate unbound IDV and SQV in plasma and their respective concentrations were measured by a fully validated method using high performance liquid chromatography‐mass spectometry (h.p.l.c.‐MS/MS).

Results
Based on the ratio AUCunbound/AUCtotal, the median unbound percentage (95% CI for differences) of SQV and IDV from all the samples studied was 1.19% (0.99, 1.58%) and 36.3% (35.1, 44.2%), respectively. No significant difference was seen in the percentage binding of SQV between patients receiving SQV alone (median = 1.49%) or with ritonavir (median = 1.09%; P = 0.141; 95% CI for difference between medians = −0.145, 0.937) over the pharmacokinetic profile. Similarly, no significant difference was seen in the percentage binding of IDV in patients receiving IDV alone (median 35.2%) or with ritonavir (median = 41.3%; P = 0.069; 95% CI for difference between medians = −0.09, 15.4). The unbound concentrations of SQV (P < 0.0001; 95% CI for r  2 = 0.634, 0.815) and IDV (P < 0.0001; 95% CI for r  2 = 0.830, 0.925) remained constant as a proportion of total concentration over the full dosing profile.

Conclusions
These in vivo data confirm previously published in vitro measurements of SQV and IDV protein binding. The unbound percentage of both protease inhibitors remained constant over the dosing interval.
Pharmacokinetic and pharmacodynamic profile following oral administration of the phosphodiesterase (PDE)4 inhibitor V11294A in healthy volunteers
Authors: Gale Donna Donigi , Landells Linda J. , Spina Domenico , Miller Alan J. , Smith Kate , Nichols Terry , Rotshteyn Yakov , Tonelli Alfred , Lacouture Peter , Burch Ronald M. , Page Clive P. , O'Connor Brian J.
Published: March 2003

Aims  To assess the pharmacokinetic and pharmacodynamic profile of the novel PDE4 inhibitor V11294A (3‐(3‐cyclopentyloxy‐4‐methoxybenzyl)‐6‐ethylamino‐8‐isopropyl‐3H purine hydrochloride) in healthy male volunteers.

Methods  This was a double‐blind, single dose, randomized crossover study in eight healthy volunteers who received a single oral, fasting dose of V11294A (300 mg) or placebo. Blood samples were taken before and 0.5, 1, 2, 2.5, 3, 4, 6, 9, 12, 18 and 24 h after oral dosing for determination of plasma concentrations of V11294A. Blood samples were also taken before and 3 and 24 h after dosing for the assessment of the effect of V11294A on mononuclear cell proliferation and tumour necrosis factor (TNF) release in whole blood.

Results  Following a single oral dose of 300 mg V11294A, plasma concentrations of V11294A and its active metabolite V10332 reached Cmax (ng ml−1; mean ± s.d.; 1398 ± 298, 1000 ± 400, respectively) after 2.63 ± 0.79 and 5.9 ± 2.3 h, respectively. For V11294A and V10332, t1/2 were 9.7 ± 3.9 and 9.5 ± 1.7 h, and AUC(0,∞) were 18100 ± 6100 and 18600 ± 8500 ng ml−1 h, respectively. At 3 h dosing, plasma concentrations of V11294A and V10332 (3‐(3‐cyclopentyloxy‐4‐methoxy‐benzyl)‐8‐isopropyl‐3H‐purin‐6‐ylamine) were 1300 ± 330 and 860 ± 300 ng ml−1, 7 and 3 times their in vitro IC50s for inhibition of TNF release and proliferation, respectively. Treatment with V11294A resulted in a significant reduction of lipopolysaccharide (LPS)‐induced TNF release at 3 h (P < 0.001) and at 24 h (P < 0.05) post ingestion. The amount of TNF released (pmol ml−1) in response to a submaximal concentration of LPS (4 ng ml−1) was not significantly altered following placebo treatment (before 681 ± 68 vs 3 h postdose 773 ± 109, P = 0.27). In contrast, there was a significant reduction in the amount of TNF released following treatment with V11294A (before 778 ± 87 vs 3 h postdose 566 ± 72, P = 0.02). Phytohaemagluttinin (PHA) stimulated the incorporation of [3H]‐thymidine in whole blood prior to drug administration. V11294A inhibited the PHA‐induced proliferation at 3 h (P < 0.05). No adverse reactions were noted following single oral administration of V11294A.

Conclusions  A single oral 300 mg dose of V11294A administered to healthy volunteers results in plasma concentrations adequate to inhibit activation of inflammatory cells ex vivo, which persists for at least 24 h without any adverse reactions.
Bisphosphonate (olpadronate) retention and its determinants in Paget's disease of bone
Authors: Cremers S. , Eekhoff E. , Dam S. v. , Den Hartigh J. , Hamdy H. , Vermeij P. , Papapoulos S.
Published: March 2003

Bisphosphonates (BPs) are the treatment of choice for Paget's disease of bone. They concentrate selectively in the skeleton at active sites and suppress osteoclastic bone resorption. Understanding the pharmacokinetics and pharmacodynamics can lead to improvements in the management of patients. Because of the specific pharmacology of BPs, the amount of BP delivered to the skeleton is likely to determine its effect. This amount can be estimated by measuring the whole body retention (WBR) of the BP [1].

In the present study BP WBR was investigated in 75 patients with active Paget's disease of bone, and PK/PD relationships were explored.

The BP WBR (WBR=dose BP−amount excreted in urine) was investigated during 5 days of intravenous treatment with olpadronate 8 mg day−1. Relationships between WBR, body weight, renal function and pretreatment disease activity (serum alkaline phosphatase (sAP), urinary hydroxyproline and urinary cross‐linked N‐telopeptides of type 1 collagen, both corrected for renal function (uOHP/Cr, uNTx/Cr)) were investigated. The relationships between change in biochemical parameters of bone metabolism (to day 5 of treatment and to nadir) and pretreatment disease activity and WBR corrected for body weight (WBR/BW) were investigated (Pearson correlation test). Statistical significance was regarded as P<0.05.

The mean WBR during the 5 day treatment period was 47 ± 14% of the dose administered (range 13% to 91%). The WBR and WBR/BW were significantly related to renal function (r=−0.43 and −0.56, respectively). Correlation with the activity of the disease was examined after logarithmic transformation of the bone turnover data. All pretreatment biochemical parameters of bone turnover measured were significantly correlated with WBR and WBR/BW of olpadronate (r=0.50, 0.61 and 0.57, respectively for WBR/BW), even after adjustment for renal function.

The decreases in biochemical parameters of bone turnover were significantly related to the pretreatment values of these parameters (e.g. r=0.96 for the change of sAP from pretreatment to nadir and 0.99 for uNTx/Cr). They were also significantly related to olpadronate WBR/BW (e.g. r=0.57 (uOHP/Cr), r=0.50 (uNTx/Cr) and r=0.35 (uSAP) for the change to nadir).

The decrease in bone turnover by intravenous olpadronate in patients with Paget's disease of bone strongly depends on the pretreatment rate of bone turnover. This may be explained, at least partially, by the amount of BP delivered to the skeleton, which is related to renal function and rate of bone turnover.

This is the first clinical study on PK/PD relationships of a BP, where the specific pharmacology of BPs is taken into account. Future studies and more longitudinally obtained data can help in better understanding the PK/PD properties of BPs in patients with Paget's disease of bone.
Hand dermatosis in upper secondary school pupils: 2-year comparison and follow-up
Authors: Yngveson M. , Svensson Å. , Johannisson A. , Isacsson Å.
Published: March 2003

The purpose of this comparative study was to record the prevalence and to estimate the incidence of self-reported hand dermatosis in a cohort of upper secondary school pupils on two different occasions, in 1995 and 1997. A previously validated questionnaire was used. Of the 1273 pupils invited to participate, 1136 (89·2%) responded to the questionnaire. The 1-year prevalence increased, although not significantly, from 9·6% in 1995 to 10·5% in 1997. For girls, the 1-year prevalence increased significantly, from 12·4% to 15·7%, while for boys the 1-year prevalence decreased, but not significantly, from 6·7% to 5·1%. The cumulative incidence was significantly higher (P < 0·001) among the girls (10·5%) than among the boys (2·8%). Multiple logistic regressions showed that the highest risk factors for 1-year prevalence in 1995 were having self-reported childhood eczema, and a family history of atopic eczema or asthma. In 1997, the highest risk factors were self-reported childhood eczema, female gender and a family history of atopic eczema. This study has identified the point at which the genders diverge with regard to hand dermatosis, with boys tending to recover, while girls show a steady or even increasing prevalence.
Use of mycophenolate mofetil in the treatment of paraneoplastic pemphigus
Authors: Williams J.V. , Marks JR. J.G. , Billingsley E.M.
Published: March 2003

Paraneoplastic pemphigus (PNP) is a rare autoimmune blistering disease with circulating antibodies that bind the cell surface of the epidermis and other non-stratifying epithelia, and immunoprecipitate a complex of four or five proteins (250 kDa, 230 kDa, 210 kDa, 190 kDa and occasionally 170 kDa). 1, 2 Combinations of immunosuppressive agents are usually required to obtain even partial control of the skin lesions. 3 Mucous membrane lesions are refractory to treatment. We describe a patient with PNP whose skin and oral lesions are quiescent following treatment with oral mycophenolate mofetil.
Two cases of Merkel cell tumour arising in patients with chronic lymphocytic leukaemia
Authors: Ziprin P. , Smith S. , Salerno G. , Rosin R.D.
Published: March 2003

Chronic lymphocytic leukaemia (CLL) has been associated with an increased incidence of second neoplasms, especially skin cancers such as basal and squamous cell carcinomas. No association with the rarer skin cancer, the Merkel cell tumour (MCT), has previously been reported. Two patients with MCT had a previous diagnosis of CLL. MCT is an aggressive skin cancer, as up to 45% of patients have lymph node involvement at presentation and 5-year disease-free survival is as low as 30%. It is most commonly found on sun-exposed areas of the body, and ultraviolet radiation together with drug-induced or CLL-induced immunosuppression may be the underlying mechanism in the observed relationship between CLL and other skin cancers.
Viral folliculitis on the face
Authors: Jang K-A. , Kim S-H. , Choi J-H. , Sung K-J. , Moon K-C. , Koh J-K.
Published: March 2003

The common clinical presentations of herpes simplex virus (HSV) and molluscum contagiosum (MC) are well known to dermatologists. However, folliculitis due to these viruses is an infrequently reported entity and might be considered a sign of immunosuppression [such as infection with human immunodeficiency virus, (HIV)], especially in cases of folliculitis due to MC. The purpose of this study was to describe the clinical and histopathological characteristics of viral folliculitis due to HSV and MC. We retrospectively collected all our cases with histologically proven folliculitis due to HSV and MC between 1994 and 1999. A total of seven patients aged 7–54 years was identified. Prior to establishment of the diagnosis of folliculitis due to HSV and MC, they were treated with topical antibiotics or topical steroids, without improvement. Tentative diagnoses were bacterial folliculitis, syringoma, perifollicular fibrosis, contact dermatitis or pseudolymphoma. Biopsy of the lesions revealed multiple molluscum bodies in the follicular epithelium with sparing of the epidermis in four patients, and ballooning degeneration and intranuclear viral inclusions in the follicular epithelium in the other three. Three patients had evidence of underlying immune suppression, such as pregnancy, chronic viral hepatitis B and nasopharyngeal carcinoma. One patient had suffered from epilepsy for 4 years. Testing for HIV by enzyme-linked immunosorbent assay was negative in the four patients in whom this was performed, and T-cell subsets were normal in the three patients in whom these were quantified. In cases of molluscum folliculitis treated with simple curettage, the lesions cleared without scarring or recurrence. In cases of herpetic folliculitis, the lesions improved with antihistamines or aciclovir. MC or HSV should be considered in cases that present with folliculitis-like dermatoses but which are refractory to anti-infective and anti-inflammatory treatment.
Expression of Fc receptors for IgG during acute and chronic cutaneous inflammation in atopic dermatitis
Authors: Kiekens R.C.M. , Thepen T. , Bihari I.C. , Knol E.F. , Van De Winkel J.G.J. , Bruijnzeel-Koomen C.A.F.M.
Published: March 2003

Atopic dermatitis is an allergic skin disease characterized by elevated total and antigen-specific serum IgE and IgG4 levels. In acute and chronic cutaneous inflammation, large cellular infiltrates including T cells, dendritic cells and macrophages are found, especially in the dermis. These cells play an important part in the regulation of local inflammatory reactions. Receptors binding IgG (FcγR) are involved in dendritic cell and macrophage function. In this study, we examined the in vivo distribution and cellular expression of the three classes of leucocyte FcγR in human skin during acute and chronic cutaneous inflammation in atopic dermatitis. Atopy patch test skin was used as a model for acute inflammation in atopic dermatitis, while chronic lesional skin was used to investigate FcγR expression in chronically inflamed skin. In atopy patch test sites no increase in the number of CD1a+ dendritic cells and a slight increase in macrophages compared with non-lesional skin was observed. Our results showed increased expression of FcγRI (CD64) and FcγRIII (CD16) in acutely inflamed skin as well as in chronically inflamed lesional skin, compared with healthy and non-lesional atopic dermatitis skin. FcγRI was expressed by RFD1+, RFD7+ and CD68+, but not by CD1a+ dermal dendritic cells. RFD1+ dendritic cells and CD68+ macrophages were the main FcγRIII-expressing cells during the acute inflammatory reaction. The significant increase in expression of FcγRIII (CD16) and FcγRI (CD64) probably results from upregulation of the receptors on resident cells. Insight into the presence of FcγR+ cells in human skin during inflammation is important both for our understanding of skin immune reactions and the development of new therapeutic concepts.
The use of two substrates to improve the sensitivity of indirect immunofluorescence in the diagnosis of pemphigus
Authors: Harman K.E. , Gratian M.J. , Bhogal B.S. , Challacombe S.J. , Black M.M.
Published: March 2003

The aim of this study was to re-evaluate indirect immunofluorescence (IIF) comparing two substrates, normal human skin (HS) and monkey oesophagus (MO) using serum from 29 pemphigus patients classified according to the presence of serum autoantibodies to either desmoglein (Dsg) 1 or Dsg3 detected by enzyme-linked immunosorbent assay (ELISA). Overall, the sensitivity of IIF was 83% on HS and 90% on MO. When data from both substrates were combined, the sensitivity increased to 100%. When sera from pemphigus foliaceus (PF) patients were studied, which contained Dsg1 antibodies only, the sensitivity of IIF was greatest on HS and titres were on average 4·8 doubling dilutions higher than on MO. In contrast, when sera containing autoantibodies only to Dsg3 from pemphigus vulgaris (PV) patients was studied, the sensitivity was greatest on MO and titres were on average 4·4 doubling dilutions higher than on HS. There was a significant correlation between Dsg1 antibody levels and IIF titres on HS and between Dsg3 antibody levels and IIF titres on MO. The investigation of immunobullous disorders in the future is likely to move towards antigen-specific techniques such as the Dsg ELISAs used in this study. However, in laboratories which currently rely on IIF for detecting pemphigus autoantibodies, the data presented in this study strongly suggest that two substrates should be used for IIF screening: one rich in Dsg1, such as HS, and the other rich in Dsg3, such as MO. This combination of substrates should not only increase the sensitivity of detecting pemphigus antibodies, but will aid in the differentiation of PV from PF. It is also possible that the data might be more useful for disease monitoring.
B‐lymphocyte suppression in multiple myeloma is a reversible phenomenon specific to normal B‐cell progenitors and plasma cell precursors
Authors: Rawstron Andrew C. , Davies Faith E. , Owen Roger G. , English Anne , Pratt Guy , Child J. Anthony , Jack Andrew S. , Morgan Gareth J.
Published: March 2003

The reduced levels of normal immunoglobulin in patients with myeloma may be due to suppression of normal B‐cell differentiation. However, reports on the numbers of B cells vary, with some finding decreases consistent with immunoparesis, and others reporting expansions of phenotypically aberrant cells. We have therefore assessed the phenotype and levels of B lymphocytes in patients at presentation (n = 23), in plateau or complete remission (PB n = 42, BM n = 18), and in relapse (PB n = 17, BM n = 14), in comparison to normal individuals (n = 10). Phenotypic analysis was performed using five‐parameter flow cytometry, with CD14 used to exclude monocytes where necessary.

We found no evidence of a phenotypically distinctive blood or marrow B‐cell population in patients with myeloma, nor of an increase in the levels of any B‐cell subset. Numbers of blood CD19+38+ normal plasma cell precursors were significantly reduced in presentation/relapse patients, but not in patients in plateau/remission. Total CD19+ cells were significantly reduced only in patients with circulating myeloma cells, detected by IgH‐PCR. In the marrow, CD19+ B cells expressing CD5, CD10, CD34, CD38, CD45low and Syndecan‐1 were significantly decreased at presentation/relapse, but not in patients in plateau/remission. The majority of these antigens are expressed by normal B‐cell progenitors, indicating that myeloma also affects the early stages of B‐cell development. The suppression of progenitor cells was not restricted to B‐lymphoid differentiation, as total CD34+ cells were also significantly reduced in the marrow of myeloma patients at presentation.

These results indicate that, if neoplastic B cells are present in myeloma, they are low in number and have a phenotype similar to their normal counterparts. Furthermore, there is a reversible suppression of CD19+ B lymphocytes that correlates inversely with disease stage, and specifically affects the early and late stages of normal B‐cell differentiation.
The role of rosetting in the multiplication of Plasmodium falciparum: rosette formation neither enhances nor targets parasite invasion into uninfected red cells
Authors: Clough Barbara , Atilola F. Abiola , Pasvol Geoffrey
Published: March 2003

The effect of rosette formation on the multiplication in vitro of Plasmodium falciparum was studied in order to establish whether rosetting acts as a major virulence factor in the pathogenesis of severe malaria by facilitating invasion of uninfected red cells. Invasion rates for rosetting (R+) and non‐rosetting (R−) parasites selected from the same clone, PA1, of P. falciparum were similar over a range of starting parasite concentrations when assayed in both static cultures and conditions of shear stress comparable with microvascular flow. However, incubation of both R+ and R− parasites under simulated conditions of flow led to decreased invasion and fewer multiply‐infected red cells as we have previously observed. Studies using fluorescently labelled red cells or reticulocytes demonstrated that rosetting did not alter the rates of invasion or target merozoites into the uninfected cells comprising a rosette. Preferential invasion of reticulocytes occurred regardless of rosetting or conditions of flow. Although the role of rosetting in the pathogenesis of malaria might relate to microvascular obstruction or perhaps the restriction of phagocytosis, our data suggest that rosetting does not play a role in the invasion or targeting of parasites into uninfected cells, eliminating this mechanism to explain the association of virulence with the rosetting parasite phenotype.
Bcr‐Abl kinase promotes cell cycle entry of primary myeloid CML cells in the absence of growth factors
Authors: Jonuleit Tarja , Peschel Christian , Schwab Renate , Kuip Heiko van der , Buchdunger Elisabeth , Fischer Thomas , Huber Christoph , Aulitzky Walter E.
Published: March 2003

Cell cycle control of both immature and differentiated primary myeloid normal and chronic‐phase chronic myeloid leukaemia (CML) cells to growth factor deprivation was studied. CD34+ cells were cultured in liquid culture. After removal of growth factors for 48 h normal cells were very efficiently arrested with the fraction of cells in S phase reduced by 70.8 ± 6.5% in CD34+ and 50.5 ± 4.2% in CD34− cells. In contrast, a significantly higher proportion of leukaemic cells remained in S phase. The fraction of S‐phase cells was reduced by only 29.3 ± 5.7% in CD34+ CML cells and 21.2 ± 3.8% in CD34− cells. This abnormal negative cell cycle control in leukaemic cells was specific for growth factor deprivation. Reaction to IFN‐α and TNF‐α treatment was identical both in normal and CML cells. Equal quantities of the cytokines TNF‐α, IL‐1α, IL‐1RA and IL‐6 were produced by CML and normal cells. However, production of GM‐CSF, with a median of 11 ± 5 pg/ml, was found only in the supernatants of CML cells. But antibodies to GM‐CSF did not restore growth factor dependence of the leukaemic cells. The defect was completely corrected by the abl‐specific tyrosine kinase inhibitor CGP 57148 without effecting cell cycle control of normal cells. Our results demonstrate a directly Bcr‐Abl‐dependent defective response of both immature and differentiated primary myeloid CML cells to growth factor deprivation.
Stimulation of leukaemic cells from adult T‐cell leukaemia patients with bacterial superantigens
Authors: Ogata Masao , Kikuchi Hiroshi , Ohtsuka Eiichi , Kohno Kazuhiro , Ito Morio , Nasu Masaru
Published: March 2003

Bacterial superantigens stimulate T cells in a manner that is restricted to the Vβ of the T‐cell receptor. We examined the ability of adult T‐cell leukaemia (ATL) cells to respond to these superantigens. Mononuclear cells from 10 patients were cultured with staphylococcal enterotoxin A (SEA), staphylococcal enterotoxin B (SEB) or toxic shock syndrome toxin‐1 (TSST‐1), and their response was determined by MTT assay and 3H‐thymidine incorporation assay. Cells from six patients showed a specific response to a single superantigen. In two cases the cells responded to TSST‐1 and bore Vβ2, the known target of TSST‐1. In three cases the cells responded to SEA with one bearing Vβ9, a target of SEA, and one bearing Vβ16. In one case the cells responded to SEB. Most of the cells which proliferated in response to superantigens were determined genetically to be leukaemic. The response to TSST‐1 was inhibited by anti‐Vβ2 antibody. The responding cells showed a strongly enhancement expression of interleukin‐2 receptor. These findings indicate that leukaemic cells from a proportion of ATL patients have an ability to respond to T‐cell receptor‐dependent superantigens. This suggests that bacterial infection in such patients may contribute to the expansion of ATL cells.
Anti‐idiotypic T‐cell activation in multiple myeloma induced by M‐component fragments presented by dendritic cells
Authors: Dabadghao Sunil , Bergenbrant Susanne , Anton Doina , HE Wen , Holm GÖran , YI Qing
Published: March 2003

The monoclonal immunoglobulin (Ig) (M‐component) secreted by the tumour plasma cells in multiple myeloma (MM) has specific antigenic determinants (idiotypes; id) that can serve as tumour‐specific antigens. The intact Ig molecule is a weak antigen, and small fragments of id protein might be more immunogenic for the induction of id‐specific immunity. Dendritic cells (DC) have attracted attention as the most efficient antigen‐presenting cells and promising adjuvants for immunotherapy in tumours. In this study the in vitro T‐cell response against F(ab′)2 and Fab fragments, heavy and light chains of the M‐component was examined in five patients with MM clinical stage I. All fragments were able to stimulate T cells but F(ab′)2 or Fab fragments and heavy chains induced a stronger response than light chains. DC induced a significantly stronger id‐specific immune response than monocytes. Moreover, with DC as antigen‐presenting cells, a predominant interferon (IFN)‐γ (type‐1 T‐cell) response was seen in all patients. Both IFN‐γ and interleukin (IL)‐4 (type‐1 and type‐2 T‐cell) responses were noted when monocytes were used. Our study suggests that DC pulsed with idiotypic fragments such as F(ab′)2 fragment and heavy chain can be used for the induction of type‐1 anti‐idiotypic T‐cell response for immunotherapy in MM.
MRC trial of α2b‐interferon maintenance therapy in first plateau phase of multiple myeloma
Authors: Drayson Mark T. , Chapman Catherine E. , Dunn Janet A. , Olujohungbe Ade B. , MacLennan Ian C. M. ,   for the MRC Working Party on Leukaemia in Adults
Published: March 2003

Plateau phase has been achieved in 64% of all newly diagnosed patients with multiple myeloma treated with the ABCM (adriamycin, BiCNU, cyclophosphamide and melphalan) regimen in the Medical Research Council (MRC) trials; this stable clinical stage of the disease is associated with no more than minimal symptoms. Several studies have found that α‐interferon (α‐IFN) maintenance therapy increases the duration of plateau phase, but it is less clear if this translates into prolonged survival. We report the effect of α‐IFN on the duration of plateau phase and overall survival in a trial with 284 patients who were randomized to receive α2b‐IFN (Intron‐A) or no maintenance therapy during first plateau phase. The minimum follow‐up after randomization was 21 months. There was no significant difference in the overall survival between the two treatment groups (χ2 = 0.32, P = 0.57). There was a trend towards longer relapse‐free survival in the patients allocated α‐IFN, but this trend to longer plateau phase was not statistically significant (χ2 = 1.62, P = 0.2). Disease progression at relapse on α‐IFN appears to be more severe with greater elevations from plateau levels of serum paraprotein (P = 0.06) and β2‐microglobulin (P = 0.03) levels. Physicians tended to start chemotherapy sooner after diagnosis of relapse when patients had received α‐IFN (P = 0.16). Although, in common with most other studies, there is a trend for patients treated with α‐IFN to have a longer plateau phase, this is counteracted by morbidity attributable to the treatment and a somewhat shortened survival post relapse. Meta‐analysis of interferon trials is required to assess whether the minor trend for longer survival in patients maintained on α‐IFN found in some studies is significant and, if so, the extent of this advantage.
GM‐CSF promotes differentiation of a precursor cell of monocytes and Langerhans‐type dendritic cells from CD34+ haemopoietic progenitor cells
Authors: Herbst Birgit , KÖhler Gabriele , Mackensen Andreas , Veelken Hendrik , Lindemann Albrecht
Published: March 2003

Epithelia‐associated dendritic cells (DC) including Langerhans cells in the skin (LC) are precursors of lymph node located interdigitating DC (iDC). CD1a+ LC are known to be derived from CD34+ haemopoietic progenitor cells (HPC); however, cells of an intermediate differentiation state that are CD34− and CD1a− have not been identified. Monitoring the differentiation pathway of HPC in the presence of GM‐CSF + IL‐4, we observed the emergence of a distinct LC precursor population that was CD33+ CD13+ CD4+ CD38+ CD44+ CD34− CD14− CD1a−. The cells could be separated by FACS due to a unique CD44/CD38 expression pattern or by CD44 expression in conjunction with the SSC profile. It was found that they were similarly generated in the presence of GM‐CSF alone and were detectable in culture for at least a week. Irrespective of being generated in the presence of GM‐CSF + IL‐4 or GM‐CSF alone, CD44/SSC‐sorted precursor cells matured to MHC class II compartments (MIIC) and Birbeck granules (BG) expressing LC, when subsequently cultured in the presence of GM‐CSF + IL‐4. When IL‐4 was omitted, however, the same cells matured to phagocytically active adherent macrophages (MΦ). These culture conditions were associated with a > 4‐fold increase in the concentration of IL‐6 when compared to those used for LC differentiation. The identification of a distinct oligopotent precursor cell population that can deliberately be induced to give rise to BG+ MIIC+ CD1a+ CD14− LC or to adherent CD14+ MΦ further substantiates the close relationship of monocytes and DC and may help to identify its in vivo equivalent.
The effect of monoclonal or ion‐exchange purified factor VIII concentrate on HIV disease progression: a prospective cohort comparison
Authors: Hay  , Ludlam  , Lowe  , Mayne  , Lee  , Prescott  , Lee
Published: March 2003

The CD4 count has been reported to decline less rapidly in HIV‐infected haemophiliacs treated with monoclonally purified factor VIII concentrates than in those using intermediate‐purity concentrates. No survival advantage has been demonstrated for this effect, and it is unclear whether this effect occurs with all high‐purity concentrates. Two cohorts of patients with severe haemophilia A and HIV treated with either ion‐exchange‐purified or monoclonally‐purified concentrates were compared. The CD4 count, survival, AIDS‐defining illnesses, CDC category and anti‐retroviral therapy were recorded at 6‐monthly intervals for 3 years following the change from intermediate to high‐purity factor VIII. 116 patients were recruited, 37 of whom were treated with an ion‐exchange purified factor VIII concentrate at three centres, mean (SD) age 31.1 (12.2) years, and 79 were treated with monoclonally purified factor VIII concentrate at two centres, mean (SD) age 29.8 (11.2) years. At the start of the study the median CD4 count was (monoclonal v ion‐exchange) 0.30 v 0.16 × 109/l.  The CD4 count declined in both arms to a median of (monoclonal v ion‐exchange) 0.16 v 0.08 × 109/l at the final visit. Analysis of the (CD4 count)1/2 over time, using a random coefficients model, found that the mean (SE) rates of decline were not statistically significantly different in the two treatment groups (monoclonal v ion exchange: −0.050 (0.008) v−0.034 (0.011) (CD4 count)1/2 per year, P = 0.24). No statistically significant difference in survival (log‐rank test: P = 0.33) was found. There was no difference in the proportion of individuals experiencing one or more AIDS‐defining illnesses (P = 0.32) or in the proportion progressing to CDC category IV (P = 0.28) during the study. The CD4 count declined during the study at a rate similar to that previously reported in patients treated with intermediate‐purity factor VIII concentrate, and there was no evidence of any difference between the two treatment groups.
Down‐regulation of the CD62L antigen as a possible mechanism for neutrophilia during inflammation
Authors: Rogowski Ori , Sasson Yifaat , Kassirer Micha , Zeltser David , Maharshak Nitsan , Arber Nadir , Halperin Pinchas , Serrov Jacob , Sorkin Patrick , Eldor Amiram , Berliner Shlomo
Published: March 2003

The expressions of both the CD11b/CD18 and CD62L ( L‐selectin) antigens as well as the absolute number of white blood cells in the peripheral circulating pool of blood in 116 patients with various inflammatory conditions were determined. A highly significant (P < 0.0001) negative correlation (r = −0.57) was noted between the absolute number of peripheral blood polymorphonuclear leucocytes and the expression of the CD62L antigen on their surface. In view of the role of CD62L antigen in the rolling of leucocytes along endothelial cells in the microvasculature, its down‐regulation might explain, at least in part, the demargination and appearance of neutrophilia during inflammation.
Differential effect of unfractionated heparin and low molecular weight heparin on intravascular tissue factor pathway inhibitor: evidence for a difference in antithrombotic action
Authors: Hansen John‐Bjarne , Sandset Per Morten , Huseby Kirsten Raanaas , Huseby Nils‐Erik , Bendz Bjørn , Østergaard Per , Nordøy Arne
Published: March 2003

Tissue factor pathway inhibitor (TFPI) is a potent inhibitor of tissue factor (TF)‐induced blood coagulation, which is increased several‐fold in post‐heparin plasma and thought to contribute significantly to the antithrombotic action of heparin. In the present study we investigated whether subcutaneous (s.c.) administration of a low molecular weight heparin (LMWH), enoxaparin, had a different effect on intravascular pools of TFPI compared with continuous i.v. infusion of unfractionated heparin (UFH). 18 healthy male volunteers were randomly assigned to continuous i.v. infusion with UFH (initially 450 U/kg/24 h, n = 6) or to s.c. treatment with LMWH once daily (enoxaparin, 1.5 mg/kg, n = 12) for 72 h. A bolus injection of 5000 IU UFH i.v. caused an 8–13‐fold increase in plasma‐free TFPI antigen (TFPI Ag), followed by a progressive decrease (81 ± 4%, P < 0.001) during the 72 h infusion with UFH. 4 h after discontinuation of the infusion, basal free TFPI Ag and heparin‐releasable TFPI were significantly decreased compared with the concentrations before the infusion (30 ± 9% and 27 ± 7%, respectively). In contrast, LMWH treatment did not reduce either basal or heparin‐releasable TFPI Ag. The changes in plasma TFPI Ag by UFH and LMWH were statistically different between groups both in pre‐ (P < 0.001) and post‐heparin (P < 0.0001) plasma. The differential effect of UFH and LMWH on intravascular pools of TFPI may contribute to the understanding of the apparent superior efficacy of LMWHs in the treatment of both arterial and venous thrombosis.
Unrelated donor marrow transplantation for chronic myelogenous leukaemia
Authors: Dini  , Lamparelli  , Rondelli  , Lanino  , Barbanti  , Costa  , Manfredini  , Guidi  , Rosti  , Alessandrino  , Locatelli  , Marenco  , Soligo  , DI Bartolomeo  , Aversa  , LA Nasa  , Busca  , Majolino  , DE Laurenzi  , Bacigalupo
Published: March 2003

Between January 1989 and July 1995 the search for an unrelated donor (UD) was started for 379 consecutive Italian patients with Philadelphia positive (Ph+) chronic myelogenous leukaemia (CML). 89 (23%) were transplanted. The overall probability of transplant before and after December 1991 was 16% and 49%, respectively (P = 0.0001), and average interval between search activation and graft was 23 months and 13 months, respectively (P = 0.0001). Disease‐free survival (DFS) following 60 consecutive transplants performed before February 1996 was 41.5% at 48 months and was 64% for patients grafted after January 1993. In univariate analysis, five variables had a favourable effect on DFS: year of bone marrow transplantation (BMT) after 1993 (P = 0.0002), HLA‐DRB1 donor/recipient (D/R) match (P = 0.0006), total body irradiation (TBI) containing regimen (P = 0.0006), graft‐versus‐host disease (GvHD) prophylaxis including ‘early’ cyclosporin before the transplant, and a marrow cell dose > 3 × 108/kg of recipient body weight (P = 0.04). Multivariate analysis confirmed that HLA identity (P = 0.006), TBI‐containing regimen (P = 0.0001) and ‘early cyclosporin’ (P = 0.04) were associated with higher DFS. Transplant‐related mortality (TRM) was 67% in patients grafted before January 1993 and 30% in patients grafted subsequently (P = 0.002). Multivariate analysis confirmed DRB1 identity (P = 0.03) and TBI‐containing regimen (P = 0.0005) to be independent factors predictive of low TRM.

This suggests that the outcome of patients transplanted from an HLA DRB1 matched donor, after a TBI‐containing preparative regimen, is similar to results recently reported in patients transplanted from geno‐identical siblings.

These results indicate that the search should be initiated at diagnosis for patients < 45 years of age and UD BMT should be considered early in the disease course for those with an available DRB1‐matched unrelated donor.
Prolonged disease‐free survival in relapsed/refractory low‐grade lymphoma treated with fludarabine: a report of four cases
Authors: Sternberg  , Matutes  , Killick  , Wothersponn  , Catovsky
Published: March 2003

We report four patients with relapsed or refractory follicular (three) and lymphoplasmacytic (one) lymphoma who achieved complete remission (CR) with fludarabine (FDR) lasting from 3.2 to 6 years. One had stage III and three stage IV and were resistant to chlorambucil and/or anthracycline. FDR was well tolerated, the only complication being herpes zoster infection in three patients, which was controlled with aciclovir. One patient developed a gastric MALT lymphoma from a different clone than the follicular lymphoma. In conclusion, some patients with refractory low‐grade lymphoma achieve long‐term CR with FDR; therefore this agent may be considered as alternative to stem‐cell transplantation.
Haemopoietic transformation by the TEL/ABL oncogene
Authors: Hannemann JÜrgen R. , Mcmanus Deborah M. , Kabarowski Janusz H. S. , Wiedemann Leanne M.
Published: March 2003

Rare, novel forms of activated ABL kinase, the result of a fusion between TEL (or ETV6, a member of the ETS transcription factor family), and the non‐receptor tyrosine kinase ABL, have been identified. We have analysed the TEL/ABL fusion protein (type A) cloned from an acute lymphoblastic leukaemia patient. In contrast to a second TEL/ABL fusion (type B) identified in two cases of myeloid leukaemia, the portion of TEL contained in the type A TEL/ABL fusion was smaller and did not contain a potential Grb2 binding site. The type A TEL/ABL cDNA we used in this study encoded a 155 kD protein with elevated tyrosine kinase activity and was responsible for the phosphorylation of a number of proteins in vivo. Its expression in factor‐dependent murine haemopoietic precursor cells efficiently converted these cells to factor independence for both survival and growth. These cells continued to express high levels of myc mRNA after growth factor depletion. We also demonstrated that type A TEL/ABL self‐associated in stably expressing haemopoietic cells. Although the TEL portion of the TEL/ABL fusion protein has no sequence similarity to that of BCR in the BCR/ABL protein, all forms of these fusion proteins contain a structure implicated in oligomerization. Our results support the conclusion that the protein interaction domain of BCR and TEL, but not the Grb2 binding site, are the important functional components in the activation of ABL kinase in haemopoietic disease.
Associations between TGF‐β1 receptors in human bone marrow stromal cells
Authors: Robledo Mar M. , Ursa Ma Angeles , Sánchez‐Madrid Francisco , TeixidÓ JoaquÍn
Published: March 2003

Bone marrow stromal cells are required for sustained haemopoiesis. Transforming growth factor‐β (TGF‐β) is a multifunctional cytokine present in the bone marrow microenvironment which regulates the expression of several cytokines, cytokine receptors and cell adhesion elements. The TGF‐β receptors type I and II, and endoglin, mediate TGF‐β1 binding to the membrane of human bone marrow stromal cells. [125I]TGF‐β1‐affinity labelling experiments showed that three different anti‐endoglin monoclonal antibodies co‐immunoprecipitated a 68 kD TGF‐β1‐labelled polypeptide together with TGF‐β1/endoglin complexes. Here, we have shown that the 68 kD receptor corresponds to the type I receptor, indicating that endoglin and the type I receptor associate on the membrane of these cells upon ligand binding. The expression of endoglin by stromal cells was found to be up‐regulated by TGF‐β1, but not by IL‐1β. The association of endoglin with signalling components of the TGF‐β receptor system on the membrane of bone marrow stromal cells might modulate TGF‐β1 access to the signalling pathways, and therefore it could regulate TGF‐β1‐mediated stromal cellular responses.