bluuebunny/crossref_metadata_2025 huggingface dataset:
Repeatability of local forearm vasoconstriction to endothelin‐1 measured by venous occlusion plethysmography
Authors: Strachan Fiona E. , Newby David E. , Sciberras David G. , McCrea Jacqueline B. , Goldberg Michael R. , Webb David J.
Published: March 2003

Aims
 We  investigated  the  repeatability  of  the  forearm  blood  flow  response  to intra‐arterial infusion of endothelin‐1 (ET‐1), assessed by venous occlusion ­plethysmography.

Methods
In eight healthy men (aged 18–50 years), on four separate occasions, ET‐1 (2.5 or 10 pmol min−1) was infused for 120 min via a 27 SWG cannula sited in the brachial artery of the nondominant arm. Each dose level was administered twice on consecutive visits. The dose order was randomized. Results are expressed as percentage change from baseline at 120 min (mean ± s.e. mean).

Results
ET‐1 caused significant vasoconstriction (P < 0.0001 anova) at both doses (38 ± 3%, 2.5 pmol min−1 and 62 ± 3%, 10 pmol min−1; mean visit 1 and 2). There was no difference in the response to either dose on repeated challenge. Responses appeared to be less variable when expressed as percentage change in the ratio of blood flow (infused:noninfused) in both arms than as percentage change in blood flow in the infused arm alone, as indicated by repeatability coefficients (15% vs 21%, 2.5 pmol min−1 and 11% vs 13%, 10 pmol min−1; ratio vs infused arm alone).

Conclusions
We have shown dose‐dependent vasoconstriction in the forearm vascular bed to intra‐arterial infusion of ET‐1 and that this response is less variable when expressed as percentage change in the ratio of forearm blood flow than percentage change in the infused arm. These data should also provide useful information to determine the power of early clinical pharmacology studies investigating the activity of endothelin receptor antagonists.
Effect of severe renal impairment on the pharmacokinetics of azimilide following single dose oral administration
Authors: Corey Alfred E. , Agnew Jeffrey R. , Valentine Suzanne N. , Parekh Nikhil J. , Powell James H. , Thompson Gary A.
Published: March 2003

Aims To assess the influence of severe renal impairment on azimilide pharmacokinetics.

Methods A single oral dose of 125 mg azimilide dihydrochloride was administered to subjects with normal and severely impaired renal function. Blood and urine samples were collected for 22–28 and 10 days, respectively.

Results Azimilide renal clearance decreased in subjects with renal impairment (mean 14 vs 4.8 ml h−1 kg−1, 95% confidence interval on the ratio 0.23, 0.50). However, no change in any other pharmacokinetic parameter including oral clearance (mean 109 vs 104 ml h−1 kg−1, 95% confidence interval on the ratio 0.67, 1.36) was observed.

Conclusions Since azimilide blood concentrations are essentially unaffected by renal function, an a priori dosage regimen adjustment is not required in patients with renal impairment.
Biomarkers for the effects of benzodiazepines in healthy volunteers
Authors: De Visser S. J. , Van Der Post J. P. , De Waal P. P. , Cornet F. , Cohen A. F. , Van Gerven J. M. A.
Published: March 2003

Studies of novel centrally acting drugs in healthy volunteers are traditionally concerned with kinetics and tolerability, but useful information may also be obtained from biomarkers of clinical endpoints. A useful biomarker should meet the following requirements: a consistent response across studies and drugs; a clear response of the biomarker to a therapeutic dose; a dose–response relationship; a plausible relationship between biomarker, pharmacology and pathogenesis. In the current review, all individual tests found in studies of benzodiazepine agonists registered for anxiety in healthy volunteers since 1966 were progressively evaluated for compliance with these requirements. A MedLine search yielded 56 different studies, investigating the effects of 16 different benzodiazepines on 73 different (variants of ) neuropsychological tests, which could be clustered into seven neuropsychological domains. Subjective and objective measures of alertness were most sensitive to benzodiazepines. The most consistent effects were observed on saccadic peak velocity (SPV) and visual analogue scores ( VAS) of alertness, where 100% and 79% of all studies respectively showed statistically significant effects. A dose–response relationship could be constructed for temazepam and SPV, which was used to determine dose equivalencies relative to temazepam, for seven different benzodiazepines. These dose equivalencies correlated with the lowest recommended daily maintenance dose (r2 = 0.737, P < 0.05). This relationship between SPV reduction and clinical efficacy could reflect the clinical practice of aiming for maximum tolerated levels, or it could represent a common basis behind SPV reduction and anxiolytic activity for benzodiazepines (probably sedation). The number of tests used in human psychopharmacology appears to be excessive and their sensitivity and reproducibility low.
Comparative study of analgesic efficacy and morphine‐sparing effect of intramuscular dexketoprofen trometamol with ketoprofen or placebo after major orthopaedic surgery
Authors: Hanna M. H. , Elliott K. M. , Stuart‐Taylor M. E. , Roberts D. R. , Buggy D. , Arthurs G. J.
Published: March 2003

Aims Multimodal analgesia is thought to produce balanced and effective postoperative pain control. A combined therapy with nonsteroidal anti‐inflammatory drugs (NSAIDs) and opiates could result in synergistic analgesia by acting through different mechanisms. Currently there are very few parenterally administered NSAIDs suitable for the immediate postoperative period. Therefore, this study was undertaken to assess the analgesic efficacy, relative potency, and safety of parenteral dexketoprofen trometamol following major orthopaedic surgery.

Methods One hundred and seventy‐two patients elected for prosthetic surgery, were randomized to receive two intramuscular injections (12 hourly) of either dexketoprofen 50 mg, ketoprofen 100 mg or placebo in a double‐blind fashion. Postoperatively, the patient's pain was stabilized, then they were connected to a patient‐ controlled analgesia system (PCA) of morphine for 24 h (1 mg with 5 min lockout).

Results The mean cumulative amount of morphine (CAM) used was of 39 mg in the dexketoprofen group and 45 mg in the ketoprofen group vs 64 mg in the placebo group. (Reduction in morphine use was approximately one‐third between the active compounds compared with placebo (adjusted mean difference of −25 mg between dexketoprofen and placebo and −23 mg between ketoprofen and placebo. These differences were statistically significant: P ≤ 0.0003; 95% CI −35, −14. Pain‐intensity scores were consistently lower with the active compounds, the lowest corresponded to the dexketoprofen‐treated patients. Regarding sedation, there were statistically significant differences between the two active compounds and placebo only at the 2nd and 13th hours. Wound bleeding was specifically measured with no statistically significant differences found between all the groups.

Conclusions Intramuscular administration of dexketoprofen trometamol 50 mg has good analgesic efficacy both in terms of opioid‐sparing effect and control of pain after major orthopaedic surgery.
A population‐based study of maternal use of amoxicillin and pregnancy outcome in Denmark
Authors: Jepsen Peter , Skriver Mette V. , Floyd Andrea , Lipworth Loren , Schønheyder Henrik C. , Sørensen Henrik T.
Published: March 2003

Aims Amoxicillin is a widely used penicillin, but data on its safety in pregnancy are limited. We examined the association between amoxicillin exposure during pregnancy and birth weight, preterm delivery, congenital malformations, perinatal death, and spontaneous abortion.

Methods We identified all primiparous women with a live birth, or a stillbirth after the 28th gestational week, from 1 January 1991–31 December 2000 in the County of North Jutland, Denmark. Data on prescriptions for amoxicillin and outcome were obtained from population‐based registries. Using a follow‐up and a case–control design, we compared pregnancy outcomes between women who had been prescribed amoxicillin during pregnancy and those who had not, adjusting for available potentially confounding factors.

Results We identified 401 primiparous women who redeemed a prescription for amoxicillin during their pregnancy. The control group consisted of 10 237 primiparous women who did not redeem any prescriptions from 3 months before pregnancy until the end of pregnancy. The adjusted mean birth weight of children born to amoxicillin‐exposed mothers was 57 g [95% confidence interval (CI) 9, 105] higher than that of children born to controls. Odds ratios among amoxicillin‐exposed relative to controls were: low birth weight 0.63 (95% CI 0.26, 1.53), preterm delivery 0.77 (95% CI 0.49, 1.21), congenital malformation 1.16 (95% CI 0.54, 2.50), and spontaneous abortion 0.89 (95% CI 0.66, 1.18). We did not observe any cases of perinatal death in the amoxicillin‐exposed women.

Conclusions We did not find any increased risk of adverse pregnancy outcome associated with amoxicillin exposure during pregnancy, but additional studies are warranted.
Differential effects of carvedilol and atenolol on plasma noradrenaline during exercise in humans
Authors: Herman Rahmatina B. , Jesudason Peter J. , Mustafa Ali M. , Husain Ruby , Choy Anna Maria J. , Lang Chim C.
Published: March 2003

Aims Evidence of long‐term beneficial effects of β‐blockers on mortality and morbidity in patients with heart failure has been demonstrated in recent randomized trials. However, not all β‐blockers are identical. Carvedilol, a nonselective β‐ and α‐adrenergic blocker, can potentially blunt the release of noradrenaline by blocking presynaptic β2‐adrenergic receptors. To test this hypothesis, we have compared the effects of carvedilol and atenolol on plasma noradrenaline during exercise in healthy young volunteers.

Methods This study investigated the differential effects of 2 weeks pretreatment with carvedilol 25 mg day−1 and atenolol 50 mg day−1 on plasma noradrenaline at rest and during exercise on a treadmill in a double‐blind randomized crossover study, involving 12 healthy male volunteers (mean age 21.6 ± 0.3 years).

Results Haemodynamic parameters at rest and during exercise were not significantly different in either carvedilol or atenolol pretreatment groups. However, carvedilol pretreatment significantly blunted the increase in plasma noradrenaline during exercise [393.8 ± 51.7 pg ml−1 (pretreatment) to 259.7 ± 21.2 pg ml−1 (post‐treatment)], when compared with atenolol [340.4 ± 54.6 pg ml−1 (pretreatment) to 396.2 ± 32.0 pg ml−1 (post‐treatment)]. The difference between carvedilol and atenolol (95% confidence interval) was −145.2, −351.0, P < 0.05.

Conclusions We have demonstrated that carvedilol but not atenolol significantly blunted the increase in plasma noradrenaline during exercise. These findings may suggest a sympathoinhibitory effect of carvedilol that may enhance its ability to attenuate the cardiotoxicity associated with adrenergic stimulation in patients with heart failure.
Regressing Merkel cell carcinoma—a case showing replacement of tumour cells by foamy cells
Authors: Maruo K. , Kayashima K-I. , Ono T.
Published: March 2003

Merkel cell carcinoma (MCC) is a malignant neuroendocrine tumour with a high rate of recurrence and metastasis. However, some cases of spontaneous regression have recently been reported. We describe the clinical course of an 80-year-old Japanese woman with regressing MCC. We also report histological findings of the regressing tumour for the first time. After the patient’s first visit to our hospital, the lesion was a rapidly progressive tumour, but suddenly began decreasing in size, and rapidly regressed. The surface of the tumour flattened, the colour changed from red to dark red, and finally the lesion appeared as a small yellowish plaque. Histopathological analysis of the completely regressed tumour revealed that the tumour cells were completely replaced by numerous foamy cells. This is the first report demonstrating the histopathological features of regressing MCC.
Consequences of factor IX mutations in 26 families with haemophilia B
Authors: Weinmann Ann F. , Murphy Michael E. P. , Thompson Arthur R.
Published: March 2003

Haemophilia B is due to a variety of mutations within the factor IX gene. In the Seattle series, 26 additional unrelated families have had a mutation identified within the past 2 years. Of these, 11 were common recurrent point mutations identifiable by rapid restriction digest screening; eight of these probably represent founder mutations. 15 others were identified by sequencing amplified coding region fragments; eight are novel. Two each had frameshift and donor splice mutations and 11 had missense mutations. Five of these mutations associated with normal levels of circulating dysfunctional factor IX were computer modelled into coordinates for factor IXa.
Acute myeloid leukaemia expressing the leucocyte integrin CD11b — a new leukaemic syndrome with poor prognosis: result of an ECOG database analysis
Authors: Paietta Elisabeth , Andersen Janet , Yunis Jorge , Rowe Jacob M. , Cassileth Peter A. , Tallman Martin S. , Bennett John M. , Wiernik Peter H. ,   and the Eastern Cooperative Oncology Group, Boston, Massachusetts, U.S.A.
Published: March 2003

While assessing the prognostic implications of immunophenotyping in 382 patients enrolled in treatment protocols of the Eastern Cooperative Oncology Group (ECOG) for de novo adult acute myeloid leukaemia, we identified 95 patients with a unique antigen profile characterized by high expression of the leucocyte integrin CD11b (CD11b+ AML). High expression of CD11b was defined as 32% positive blasts based on the retrospectively established prognostic cut‐off point for this antigen. Although CD11b is normally expressed by mature monocytes, natural killer cells and granulocytes, leukaemic blasts in CD11b+ AML lacked other immunologic monocytic features (e.g. CD14 and CD122, the interleukin‐2 receptor β chain) and demonstrated a high degree of immaturity, as reflected by a high incidence of blasts expressing the stem cell factor receptor, CD117, and few blasts positive for the myeloid differentiation antigen CD15. Furthermore, by FAB criteria, only 41% of CD11b+ AML cases were classified as M4/M5. Patients with CD11b+ AML had a low response rate (54%) when compared with acute monocytic leukaemia (AMOL; 82%, P = 0.006) or AML overall (68%, P = 0.031), independent of age, cytogenetic abnormalities and P‐glycoprotein expression. Because of its poor prognosis, recognition of CD11b+ AML is clinically warranted and, given its morphologic and cytogenetic ambiguity, must be based on the unique antigen profile.
A randomized trial of the effect of clodronate on skeletal morbidity in multiple myeloma
Authors: McCloskey  , MacLennan  , Drayson  , Chapman  , Dunn  , Kanis  ,   for the MRC Working Party on Leukaemia in Adults
Published: March 2003

In patients with multiple myeloma, despite a major reduction of bone pain achieved with chemotherapy, skeletal disease continues to progress. The effects of clodronate, an inhibitor of osteoclastic bone resorption, are evaluated on the natural history of skeletal disease in patients with newly diagnosed multiple myeloma. Within the framework of the VIth MRC Multiple Myeloma Trial, 536 patients (218 women, 318 men) with recently diagnosed multiple myeloma were randomized to receive either clodronate 1600 mg daily (n=264) or an outwardly identical placebo (n=272) in addition to chemotherapy. Treatment with clodronate was associated with a 50% decrease in the proportion of patients with severe hypercalcaemia (5.1% v 10.1%, P=0.06) and a similar reduction in reported non‐vertebral fractures (6.8% v 13.2%, P=0.04). Fewer patients receiving clodronate sustained vertebral fractures after entry to the trial (38% v 55%, P=0.01) and patients also lost less height over 3 years compared to those receiving placebo (2.0 v 3.4 cm, P=0.01). Biochemical indices of bone turnover were significantly lower in patients receiving concomitant clodronate, both at plateau and at disease relapse. The frequencies of back pain and poor performance status were significantly lower at 24 months in clodronate than in placebo‐treated patients (10.9% v 19.9%, P=0.05, and 18.3% v 30.5% P=0.03 respectively.) There was no statistically significant difference in survival between the clodronate and placebo treated patients. The study indicates that long‐term oral clodronate slows the progression of skeletal disease in multiple myeloma and decreases the associated morbidity. Patients without overt skeletal disease at diagnosis were also found to benefit from clodronate, indicating that this treatment should be initiated as early in the course of the disease as possible.
Thrombopoietin serum levels in patients with aplastic anaemia: correlation with platelet count and persistent elevation in remission
Authors: Schrezenmeier Hubert , Griesshammer Martin , Hornkohl Alex , Nichol Janet L. , Hecht Thomas , Heimpel Herrmann , Kubanek Bernhard , Raghavachar Aruna
Published: March 2003

In an attempt to evaluate the role of thrombopoietin (TPO) in the pathobiology of aplastic anaemia (AA), we have examined TPO levels in sera from 54 AA patients and 119 healthy controls. A total of 92 samples were collected from AA patients: 43 samples were harvested at diagnosis, 23 samples in the cytopenic period after treatment, and 26 samples when patients were in partial (n = 10) or complete remission (n = 16) following immunosuppressive treatment. TPO serum levels were assessed by a sandwich‐antibody ELISA that utilized a polyclonal rabbit antiserum for both capture and signal. Serum samples from normal donors revealed a mean TPO level of 95.3 ± 54.0 pg/ml (standard deviation). Mean TPO levels in AA sera collected at diagnosis and before onset of treatment were 2728 ± 1074 pg/ml (P < 0.001 compared to normal controls; mean platelet count at that time: 27 × 109/l). TPO serum levels of AA patients in partial or complete remission after immunosuppressive treatment were significantly lower than TPO levels at diagnosis (P < 0.001). However, despite normal platelet counts (mean 167 × 109/l), TPO levels remained significantly elevated in complete remission (mean TPO 1009 ± 590 pg/ml, P < 0.001 compared to normal controls). There was a significant inverse correlation between serum TPO levels and platelet counts in AA patients who were not transfused for at least 2 weeks prior to sample collection (coefficient of correlation (r) = −0.70, P < 0.0001).

In summary, TPO levels were highly elevated in sera of patients with AA. Thus there is no evidence to suggest an impaired TPO response contributing to thrombocytopenia in AA. Thrombopoietin did not return to normal levels in remission, indicating a persisting haemopoietic defect in remission of AA. We hypothesize that elevated levels of TPO may be required to maintain normal or near normal platelet counts in remission of AA.
Immunogenicity and cross‐reactivity with idiotypic IgA of VH CDR3 peptide in multiple myeloma
Authors: Wen Yue‐Jin , Ling Min , Lim Seah H.
Published: March 2003

Multiple myeloma idiotypic protein is clone‐specific and therefore represents an ideal tumour antigen for immune targeting. In this study we determined whether a synthetic peptide corresponding to the autologous idiotypic VH CDR3 sequence could elicit peptide‐specific immune responses in a patient with IgA myeloma. Not unlike B‐cell lymphoma, the immune repertoire of the patient contained T cells capable of mounting proliferative and cytotoxic responses to antigen‐presenting cells loaded with the CDR3 peptide. Furthermore, the T cells were also able to secrete interferon‐γ upon peptide rechallenge. Antigen recognition by peptide‐primed T cells was MHC dependent and could be blocked by antibodies to both monomorphic MHC class I and class II molecules. These results therefore indicate the presence of T‐cell epitopes on the VH CDR3 sequence. In addition, CDR3 peptide‐primed T cells were also able to mount similar immune responses when rechallenged with the intact IgA idiotypic protein, suggesting that functional T‐cell epitopes had been derived from the CDR3 sequence of the idiotypic protein. Our results therefore provide a new perspective to the immunogenicity of the idiotypic protein in myeloma.
Secondary haematological neoplasm after treatment of adult acute lymphoblastic leukaemia: analysis of 1170 adult ALL patients enrolled in the GIMEMA trials
Authors: Pagano Livio , Annino Luciana , Ferrari Antonella , Camera Andrea , Martino Bruno , Montillo Marco , Tosti Maria Elena , Mele Alfonso , Pulsoni Alessandro , Vegna Maria Luce , Leone Giuseppe , Mandelli Franco ,   for the GIMEMA (Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto) Group
Published: March 2003

Between 1983 and 1994 the incidence of secondary haematological neoplasms (SHM) was evaluated in 1170 new cases of ALL enrolled in the GIMEMA trials. Of the 942 patients who achieved complete remission (CR); seven developed a SHM: four AMLs and three NHLs.

The median latency from onset of ALL and of secondary haematological neoplasm was 69 months for AML and 61 months for NHL. Three out of four patients with secondary AML were unresponsive to the new chemotherapy and died, whereas the fourth patient achieved a new CR. Among the three NHL cases, two patients are presently alive in CR, whereas the third patient was refractory to chemotherapy and died.

The relative risk of haematological malignancy among the GIMEMA trials population, as compared to that of the Italian Cancer Registries, was 15.25‐fold higher, and the actuarial estimated cumulative proportion of ALL patients with a secondary haematological neoplasm at 5 and 10 years were 0.59% and 3.63% respectively.

The incidence of adult ALL who developed a SHM, although apparently lower than in the paediatric ALL series, was higher when compared to the normal population. The difference between paediatric and adult ALL is probably due to the lack of craniospinal radiotherapy and to the lower doses of epipodoxiphyllotoxins used in adult trials. The higher percentage of childhood ALL with a prolonged event‐free survival could result in an increase of secondary neoplasms in these cases, which suggests that secondary haematological neoplasms in adult ALL patients are real, although rare, events.
The Platelet Function Analyzer (PFA‐100®): a novel in‐vitro system for evaluation of primary haemostasis in children
Authors: Carcao  , Blanchette  , Dean  , He  , Kern  , Stain  , Sparling  , Stephens  , Ryan  , Freedman  , Rand
Published: March 2003

The PFA‐100® system provides an in‐vitro method of assessing primary platelet‐related haemostasis by measuring the time (the closure time, or CT) taken for a platelet plug to occlude a microscopic aperture cut into a membrane coated with collagen and either epinephrine or ADP. We used the system to establish normal ranges for CTs in healthy children, adults and neonates. Mean CTs of healthy children were independent of the needle gauge used (21G or 23G) for blood sampling; they were very similar to the mean CTs of healthy adults, but longer than mean CTs of healthy neonates. Although children with haemophilia had normal CTs, the PFA‐100 system was found to be potentially useful in screening for von Willebrand disease in children.
Trisomy 12 is seen within a specific subtype of B‐cell chronic lymphoproliferative disease affecting the peripheral blood/bone marrow and co‐segregates with elevated expression of CD11a
Authors: Su'ut  , O'Connor  , Richards  , Jones  , Roberts  , Davies  , Fegan  , Jack  , Morgan
Published: March 2003

In order to delineate the specific morphological and immunophenotypic features of B‐cell lymphoproliferative disorders associated with trisomy 12, 172 sequential unselected cases of CD19+CD5+ B‐cell disorders, primarily affecting the peripheral blood and bone marrow, were studied. Trisomy 12 was found in 24 cases (13.9%), with all cases morphologically classified as either CLL‐PL or CLL‐mixed by FAB criteria. Trisomy 12 was not found in any cases of typical CLL. Trisomy 12 cases demonstrated a significant higher expression of CD11a (P<0.0001) and CD20 (P<0.0006) when compared to cases with the equivalent morphology and immunophenotype, but without the chromosomal abnormality. Trisomy 12 cases also demonstrated a higher frequency of FMC7, CD38 expression and moderate to strong surface immunoglobulin staining. However, no correlation was detected between the percentages of trisomy 12 cells and cells expressing CD11a, CD38, FMC7 or sIg mean fluorescent intensity. Cells from trisomy 12 positive cases were sorted according to their CD11a expression using fluorescent activated cell sorting. There was a significant increase in the percentage of trisomy 12 cells within the CD11a+ sorted fraction compared to the unsorted population (P<0.05), implying that trisomy 12 is associated with increased expression of CD11a.

With the highly specific morphological and immunophenotypic features demonstrated by trisomy 12 cases in this study, it is highly likely that these cases constitute a specific group of B‐cell lymphoproliferative disorders.
Evaluation of immunotoxins containing single‐chain ribosome‐inactivating proteins and an anti‐CD22 monoclonal antibody (OM124):in vitroandin vivostudies
Authors: Bolognesi Andrea , Tazzari Pier Luigi , Olivieri Fabiola , Polito Letizia , Lemoli Roberto , Terenzi Adelmo , Pasqualucci Laura , Falini Brunangelo , Stirpe Fiorenzo
Published: March 2003

Immunotoxins were prepared with three ribosome‐inactivating proteins (RIP), momordin, pokeweed antiviral protein from seeds (PAP‐S) and saporin‐S6, linked to the anti‐CD22 monoclonal antibody OM124. These immunotoxins inhibited protein synthesis by CD22‐expressing cell lines Daudi, EHM, BJAB, Raji and BM21 with IC50(concentration causing 50% inhibition) ranging from < 5 × 10−15to 7.6 × 10−11 Mas RIP, and IC90(concentration causing 90% inhibition) ranging from 5 × 10−14to 5 × 10−8 M, with no effect on a CD22‐negative HL60 cell line at the highest concentration tested (5 × 10−8 M). Apoptosis was induced in sensitive cells. The formation of bone marrow colonies was inhibited by no more than 40% by the immunotoxins at concentrations up to 10−9 M. Treatment with the immunotoxins, alone or in combination, significantly extended the survival time of mice bearing transplanted Daudi cells. A treatment with cyclophosphamide and OM124/saporin immunotoxin was particularly effective in SCID mice transplanted with a low number of cells (3 × 10−6), when 60% of the animals remained tumour‐free.
Comparison of bone marrow transplant and chemotherapy for relapsed childhood acute lymphoblastic leukaemia: the MRC UKALL X experience
Authors: Wheeler Kate , Richards Sue , Bailey Clifford , Chessells Judith ,   for the Medical Research Council Working Party on Childhood Leukaemia
Published: March 2003

We examined the outcome of the 489 children with acute lymphoblastic leukaemia (ALL) who relapsed in the UKALL X trial, and produced graphical displays of adjusted comparisons of event‐free survival (EFS) for chemotherapy versus bone marrow transplantation (BMT) from a sibling or volunteer unrelated donor, and autologous BMT (ABMT).

EFS at 5 years was only 3% (95% CI 0–6%) for children who relapsed in the bone marrow (BM) within 2 years of diagnosis, irrespective of type of post‐relapse treatment, whereas for those with late extramedullary relapse it was 66% (95% CI 48–85%). Comparison of the types of treatment did not show benefit for ABMT. For allogeneic BMT the overall reduction in the odds of an event was 26% (95% CI 1–51%) (2P = 0.05), resulting in an absolute increase in 5‐year event‐free survival of 14% (from 26.4% to 40.7%).

New approaches are needed for children with early BM relapses whose prognosis is virtually hopeless with current therapy; however, a conventional chemotherapy approach may be justifiable for late extramedullary relapses. For the remaining patients (71%), with later BM or early extramedullary relapses, the optimal treatment is still not clear. This uncertainty warrants a formal randomized comparison of BMT and chemotherapy, to avoid the biases due to unmeasurable selection factors.
Marked improvements in outcome with chemotherapy alone in paediatric acute myeloid leukaemia: results of the United Kingdom Medical Research Council's 10th AML trial
Authors: Stevens Richard F. , Hann Ian M. , Wheatley Keith , Gray Richard G. ,   on behalf of the MRC Childhood Leukaemia Working Party
Published: March 2003

359 eligible children with acute myeloid leukaemia (AML) entered the MRC AML 10 trial between May 1988 and March 1995. Patients received four courses of intensive induction and consolidation chemotherapy, with or without subsequent autologous (A‐BMT) or allogeneic (allo‐BMT) bone marrow transplant. There were randomized comparisons of thioguanine versus etoposide in induction and of A‐BMT versus not. Allo‐BMT was recommended for patients with a HLA‐matched sibling and was evaluated by donor versus no donor comparison. The complete remission rate was 92%. In first remission there were 20 deaths during consolidation chemotherapy and 11 after BMT (8/61 allo‐BMTs, 1/60 A‐BMTs and 2/4 matched unrelated donor transplants). The relapse rate was low, decreasing from 26% in the first year to 2% in the fourth. Long‐term outcome was excellent with survival at 7 years from entry of 56% and event‐free survival of 48%. There were no significant differences between thioguanine and etoposide, whereas both A‐BMT and allo‐BMT reduced relapse risk but did not produce a significant survival benefit. It appears that over half the children entered into AML 10 are cured, a result which compares favourably with other reported series. We conclude that four courses of intensive chemotherapy are an effective approach to the treatment of paediatric AML, which avoids the acute toxicity and long‐term side‐effects of BMT and also avoids the need for prolonged maintenance therapy or cranial irradiation.
Association between hyperflexibility of the thumb and an unexplained bleeding tendency: is it a rule of thumb?
Authors: Kaplinsky Chaim , Kenet Gili , Seligsohn Uri , Rechavi Gideon
Published: March 2003

A bleeding tendency manifested by petechiae and ecchymoses is one of the most common causes for referral of patients to haematology clinics. Vessel wall pathology is not usually considered to be a cause for deranged haemostasis, although coexistence of increased capillary fragility and joint hypermobility have been reported.

We determined the frequency of thumb hyperextensibility and scored the findings in a series of 44 patients referred because of ecchymoses and petechiae, as well as 261 control children and their mothers. All 44 patients had normal coagulation studies. Thumb flexibility score was +4 in 30 patients, +3 in eight patients, +2 in five patients and +1 in one of the index patients. In the control group, only one of 261 had a +4, and three had a +3 score, and two of 260 mothers had a +4 score. Ecchymoses were not observed in any of these subjects, nor in the +1 patients.

Based on clinical presentation and normal coagulation studies, we suggest that our patients had an underlying subtype of Ehlers‐Danlos syndrome. In view of the dramatically high occurrence of thumb hyperextensibility in patients with unexplained mild bleeding tendency, costly haemostatic and coagulation studies on such patients may not be necessary.
Microsatellite instability in adult T‐cell leukaemia
Authors: Hatta Yoshihiro , Yamada Yasuaki , Tomonaga Masao , Miyoshi Isao , Said Jonathan W. , Koeffler H. Phillip
Published: March 2003

Microsatellite instability (MSI) is thought to represent a defect of the DNA mismatch repair system which has been implicated in the tumourigenesis of several human malignancies. We investigated MSI in acute/ lymphomatous adult T‐cell leukaemia (ATL; n=22) using 54 highly polymorphic dinucleotide short‐tandem repeat sequences. The corresponding control DNA from each individual was obtained from the peripheral blood in either chronic phase (n=5) or when complete remission was achieved (n=17). 10/22 (41%) patients had MSI, six of whom showed MSI in multiple loci; four loci had MSI in multiple samples. The incidence of MSI in ATL was found to be higher than in other haematological malignancies, indicating MSI as a feature of ATL, which may be involved in the progression of the disease.
Differential constitutive activation between STAT‐related proteins and MAP kinase in primary acute myelogenous leukaemia
Authors: Hayakawa Fumihiko , Towatari Masayuki , Iida Hiroatsu , Wakao Hiroshi , Kiyoi Hitoshi , Naoe Tomoki , Saito Hidehiko
Published: March 2003

Many cytokines and growth factors stimulate multiple signal transduction pathways essential for proliferation in human acute leukaemia cells, including a mitogen‐activated protein (MAP) kinase pathway and a Janus kinase (JAK)‐STAT (signal transducers and activators of transcription) pathway. We have previously shown constitutive activation of MAP kinase in approximately 50% of acute myelogenous leukaemia (AML) samples. Recently, STAT proteins have been reported to be constitutively activated in 10–20% of AML cases. STAT3 and STAT5 are the main STAT proteins activated in haemopoietic progenitors in response to cytokines such as IL‐3, GM‐CSF, erythropoietin and thrombopoietin. Although the possibility of STAT1 protein as a substrate for MAP kinase at a serine residue has been suggested, the cross‐talk between STATs and MAP kinase pathways in vivo, especially in leukaemia cells, remains unknown.

We examined the phosphorylation of STAT3 and STAT5 at the tyrosine residues in AML samples in which MAP kinase activity had already been found. 40/50 primary AML cases (80%) exhibited constitutive tyrosine phosphorylation of STAT5. Electrophoretic mobility shift assay showed DNA binding activity of STAT5 correlated with tyrosine phosphorylation of STAT5. Similarly, with respect to STAT3, 17/23 cases examined (74%) showed constitutive tyrosine phosphorylation of STAT3. In addition, we examined the tyrosyl‐phosphorylation of STAT5 isoforms, STAT5A and STAT5B, in 20 AML cases, and found selective STAT5B phosphorylation in the absence of STAT5A phosphorylation in three cases. Furthermore, in certain AML cases, constitutive activation of MAP kinase and STAT proteins occurred independently. No significant correlation of MAP kinase activation was observed with either tyrosine phosphorylation of STAT3/STAT5 or positive DNA binding of STAT proteins. These results suggest that constitutive activation of STAT proteins occurs commonly and that the causes of constitutive activation of these two major cascades are heterogenous in AML.
A novel mature B‐cell line (DOBIL‐6) producing both parathyroid hormone‐related protein and interleukin‐6 from a myeloma patient presenting with hypercalcaemia
Authors: Ohmori Minoru , Nagai Masami , Fujita Mitsuhiro , Dobashi Hiroaki , Tasaka Taizo , Yamaoka Genji , Kawanishi Koichi , Taniwaki Masafumi , Takahara Jiro
Published: March 2003

A novel human EBV‐negative B‐cell line, designated DOBIL‐6, was established from a patient with non‐secretary myeloma. The DOBIL‐6 cell has cytoplasmic γ protein and expresses CD19, 20, 38, 45RO, VLA‐4 and PCA‐1 antigens, but lacks CD10, 45RA and VLA5 antigens. Chromosome analysis showed that DOBIL‐6 cells had many complex structural abnormalities, including t(11;14) (q13;q32), which were consistent with that of the fresh tumour cells. Interestingly, abundant interleukin‐6 (IL‐6) and parathyroid hormone‐related protein (PTHrP) accumulated in the culture supernatant of DOBIL‐6 cells. Hypercalcaemia and splenomegaly associated with plasma cell proliferations which resulted in the expansion of the light zones in the follicles were observed in DOBIL‐6 transplanted nude mice. RT‐PCR analysis detected mRNA for PTHrP, and IL‐6 as well as its receptor (GP80) in DOBIL‐6 cells. Treatment of the DOBIL‐6 cells with neutralizing anti‐IL‐6 antibody inhibited their growth in a dose‐dependent manner, whereas the addition of exogenous IL‐6 stimulated it in serum‐depleted conditions. These findings suggest that both IL‐6 and PTHrP are produced in DOBIL‐6 cells, and that IL‐6 promotes its growth by an autocrine mechanism. Since IL‐6 is known to stimulate not only the growth of B‐cell neoplasms but also osteoclastic bone resorption by cooperating with PTHrP, this simultaneous production of IL‐6 and PTHrP might be synergistically linked and play a role in the development of hypercalcaemia of the patient. The DOBIL‐6 cell is a useful tool to clarify the mechanism of hypercalcaemia associated with mature B‐cell neoplasms.
Chemotaxis of macrophages is abolished in the Wiskott‐Aldrich syndrome
Authors: Zicha  , Allen  , Brickell  , Kinnon  , Dunn  , Jones  , Thrasher
Published: March 2003

Wiskott‐Aldrich syndrome (WAS) is a rare disease characterized by microthrombocytopenia, eczema and immune deficiency. In this study a direct‐viewing chemotaxis chamber was used to analyse chemotactic responses of WAS neutrophils and macrophages in stable linear concentration gradients. In five patients with classic WAS, chemotaxis of macrophages but not of neutrophils was found to be abolished, whereas the speed of random motility of both cell types was found to be indistinguishable from control cells. This supports the existence of an essential functional link, previously suggested by biochemical studies, between Cdc42, WAS protein (WASp) and the actin cytoskeleton in primary human macrophages. Moreover, these data suggest that Cdc42‐WASp‐mediated filopodial extension is a requirement for chemotaxis but not for chemokinesis in these cells. Abnormal directional cell motility of macrophages and related antigen‐presenting cells may play a significant part in the immune deficiency and eczema of WAS.
Generation and functional characterization of human dendritic cells derived from CD34+ cells mobilized into peripheral blood: comparison with bone marrow CD34+ cells
Authors: Ratta Marina , Rondelli Damiano , Fortuna Alessandra , Curti Antonio , Fogli Miriam , Fagnoni Francesco , Martinelli Giovanni , Terragna Carolina , Tura Sante , Lemoli Roberto M.
Published: March 2003

Dendritic cells (DCs) are the most powerful professional antigen‐presenting cells (APC), specializing in capturing antigens and stimulating T‐cell‐dependent immunity. In this study we report the generation and characterization of functional DCs derived from both steady‐state bone marrow (BM) and circulating haemopoietic CD34+ cells from 14 individuals undergoing granulocyte colony‐stimulating factor (G‐CSF) treatment for peripheral blood stem cells (PBSC) mobilization and transplantation. Clonogenic assays in methylcellulose showed an increased frequency and proliferation of colony‐forming unit‐dendritic cells (CFU‐DC) in circulating CD34+ cells, compared to that of BM CD34+ precursors in response to GM‐CSF and TNF‐α with or without SCF and FLT‐3L. Moreover, peripheral blood (PB) CD34+ cells generated a significantly higher number of fully functional DCs, as determined by conventional mixed lymphocyte reactions (MLR), than their BM counterparts upon different culture conditions. DCs derived from mobilized stem cells were also capable of processing and presenting soluble antigens to autologous T cells for both primary and secondary immune response. Replacement of the early‐acting growth factors SCF and FLT‐3L with IL‐4 at day 7 of culture of PB CD34+ cells enhanced both the percentage of total CD1a+ cells and CD1a+CD14− cells and the yield of DCs after 14 d of incubation. In addition, the alloreactivity of IL‐4‐stimulated DCs was significantly higher than those generated in the absence of IL‐4. Furthermore, autologous serum collected during G‐CSF treatment was more efficient than fetal calf serum (FCS) or two different serum‐free media for large‐scale production of DCs. Thus, our comparative studies indicate that G‐CSF mobilizes CD34+ DC precursors into PB and circulating CD34+ cells represent the optimal source for the massive generation of DCs. The sequential use of early‐acting and intermediate‐late‐acting colony‐stimulating factors (CSFs) as well as the use of autologous serum greatly enhanced the growth of DCs. These data may provide new insights for manipulating immunocompetent cells for cancer therapy.
Patients with high‐risk myelodysplastic syndrome can have polyclonal or clonal haemopoiesis in complete haematological remission
Authors: Delforge Michel , Demuynck Hilde , Verhoef Gregor , Vandenberghe Peter , ZachÉE Pierre , Maertens Johan , Van Duppen Viktor , Boogaerts Marc A.
Published: March 2003

The clonality of mature peripheral blood‐derived myeloid and lymphoid cells and bone marrow haemopoietic progenitors from 18 females with myelodysplasia (MDS) (five refractory anaemia, RA; one RA with ringed sideroblasts, RARS; three chronic myelomonocytic leukaemia, CMML; four RA with excess of blasts, RAEB; five RAEB in transformation, RAEB‐t) was studied by X‐chromosome inactivation analysis. Using the human androgen‐receptor (HUMARA) assay, we analysed the clonal patterns of highly purified immature CD34+38− and committed CD34+38+ marrow‐derived progenitors, and CD16+14− granulocytes, CD14+ monocytes, CD3+ T and CD19+ B lymphocytes from peripheral blood. In high‐risk patients (RAEB, RAEB‐t), clonality analysis was performed before and after intensive remission‐induction treatment. All patients, except one with RA, had predominance of a single clone in their granulocytes and monocytes. The same clonal pattern was found in CD34+ progenitor cells. In contrast, CD3+ T lymphocytes were polyclonal or oligoclonal in 14/18 patients. X‐chromosome inactivation patterns of CD19+B cells were highly concordant with CD3+ T cells except for two patients (one RA, one CMML) with monoclonal B and polyclonal T lymphocytes, therefore suggesting a clonal mutation in a progenitor common to the myeloid and B‐lymphoid lineages or the coexistence of MDS and a B‐cell disorder in these particular patients. After high‐dose non‐myeloablative chemotherapy, polyclonal haemopoiesis was reinstalled in the mature myeloid cells and immature and committed marrow progenitors in three of four patients achieving complete haematological remission. Therefore we conclude that most haematological remissions in MDS are associated with restoration of polyclonal haemopoiesis.
Monocyte tissue factor‐like activity in post myocardial infarction patients
Authors: Freeburn  , Wallace  , Strain  , Sinnamon  , Craig  , Johnson  , Gilmore
Published: March 2003

It is widely recognized that thrombosis is the major event in the evolution of stable vascular disease to unstable ischaemic syndromes including myocardial infarction and stroke. The purpose of this case–control study was to establish clinical and laboratory data on the possible relationship between specific components of the haemostatic system and coronary heart disease. The procoagulant activity (PCA) of peripheral monocytes and polymorphonuclear neutrophils was assessed in 21 males who had suffered a myocardial infarction (MI) and in age‐matched controls. In addition, total factor VII activity, fibrinogen, tissue factor pathway inhibitor (TFPI), D‐dimers, tissue plasminogen activator (t‐PA), plasminogen activator inhibitor (PAI‐1), tumour necrosis factor‐α (TNF‐α) and full blood counts were measured. Post MI patients had significantly higher monocyte PCA, higher plasma concentrations of TFPI, fibrinogen, t‐PA, T/P100 and also higher total white blood cell and neutrophil counts compared to age‐matched controls. This elevated procoagulant state in post MI patients could further exacerbate the disease process and increase the risk of subsequent acute ischaemic events.
Randomized clinical study comparing aggressive chemotherapy with or without G‐CSF support for high‐risk myelodysplastic syndromes or secondary acute myeloid leukaemia evolving from MDS
Authors: Bernasconi Carlo , Alessandrino E. Paolo , Bernasconi Paolo , Bonfichi Maurizio , Lazzarino Mario , Canevari Angelo , Castelli Guglielmo , Brusamolino Ercole , Pagnucco Guido , Castagnola Carlo
Published: March 2003

One hundred and five consecutive primary high‐risk myelodysplastic syndromes (MDS) or secondary acute myeloid leukaemia (sAML) evolving from MDS (performance status 0–3, ECOG) entered this study. Induction chemotherapy (CT) consisted of idarubicine 12 mg/m2 i.v. on days 1 and 2, etoposide 60 mg/m2/12 h i.v. for 5 d, Ara‐C 120 mg/m2/12 h i.v. for 5 d (one or two courses). Patients were randomized to receive or not G‐CSF (5 μg/kg/d subcutaneously 48 h after the end of CT). 52 cases underwent CT alone and 53 CT+G‐CSF. The CT + G‐CSF patients had a significantly shorter duration of neutropenia (8 v 16 d) with a lower incidence of infections and significantly better responses (CR+PR: 74% v 52%, P < 0.05). 40 patients entered CR: 17 with CT and 23 with CT+G‐CSF. Responders underwent two consolidation courses with the same CT, followed by high‐dose Ara‐C (2 g/m2 every 12 h for 3 d). Most CRs were clonal. At present 21 responders have relapsed (median relapse‐free survival 4.5 months). Eight responders received an allo‐BMT, six are alive in CR 7–57 months post‐transplant. Therefore allo‐BMT only increases the chance of a long survival and possible cure. In conclusion, CT+G‐CSF did not prolong either CR duration or survival; the growth factor support, however, increased the number of allo‐transplantable cases by inducing higher remission rates and improving clinical conditions.
Chronic myelogenous leukaemia CD34+ cells exit G0/G1 phases of cell cycle more rapidly than normal marrow CD34+ cells
Authors: Traycoff Christie M. , Halstead Bartley , Rice Susan , Mcmahel Jon , Srour Edward F. , Cornetta Kenneth
Published: March 2003

To investigate the mechanisms behind the leukaemic expansion of chronic myelogenous leukaemia (CML), we examined the cell cycle status and activation kinetics of purified subpopulations of CD34+ cells from normal and CML bone marrow (BM). Propidium iodide staining was used to assess cell cycle status of fresh cells or those stimulated with cytokines. Although the cell cycle status of fresh low‐density cells from CML and normal BM was similar, a larger percentage of CML CD34+ cells were cycling than those from normal BM. The HLA‐DR− compartment of CML CD34+ cells, a fraction enriched for normal, non‐leukaemic progenitors, contained a higher percentage of quiescent cells than the CD34+ HLA‐DR+ fraction. When the activation of CD34+ cells was examined in response to SCF or IL‐3 alone, or SCF+IL‐3+IL‐6, CML CD34+ cells exited G0/G1 more rapidly than normal CD34+ cells. Interestingly, although normal BM CD34+ cells failed to cycle in response to IL‐6 alone, or in the absence of exogenous cytokines, 30% of CML cells cycled under these conditions. No differences in the degree of apoptosis were documented among CML and normal CD34+ cells in these cultures. These data suggest that enhanced cell cycle activation of CML CD34+ cells, by either autocrine stimuli or via enhanced sensitivity to exogenous stimuli, may be partially responsible for the pronounced cellular expansion characteristic of CML.