bluuebunny/crossref_metadata_2025 huggingface dataset:
Direct measurement of alcohol and its metabolites
Authors: Swift Robert
Published: November 2003

Background  Ethanol can be easily and accurately measured in body fluids or vapours by several chemical and enzymatic methods. However, the persistence of ethanol in the body is short lived and ethanol's metabolism and distribution within the body are extremely complex.

Aim  This article discusses the complex pharmacokinetics of ethanol and how pharmacokinetics can affect measured ethanol concentrations when ethanol is sampled across different individuals or from different body fluids or vapors, such as whole blood, plasma, breath, urine, saliva, or transdermal ethanol.

Findings  Several physical methods for the quantification of ethanol concentrations are described, including gas chromatography, electrochemical detection, infrared detection and enzymatic methods, along with their advantages and limitations. Also discussed, is the utility of the measurement of several immediate metabolites of ethanol, including acetaldehyde, acetate, and fatty acid ethyl esters.

Conclusions  The complex distribution and metabolism of ethanol and its metabolites can obscure the relationship between the ethanol concentration that is measured in body fluids or vapours and the amount of ethanol that is actually taken into the body.
Evaluation of a new quality of life questionnaire for patients with irritable bowel syndrome
Authors: Hahn B. A. , Kirchdoerfer L. J. , Fullerton S. , Mayer E.
Published: November 2003

Background:

We describe the development and evaluation of a new disease‐specific instrument, the Irritable Bowel Syndrome Quality of Life Questionnaire (IBSQOL), which was designed for use in patients with irritable bowel syndrome. The IBSQOL measures 10 domains found to be relevant to patients with irritable bowel syndrome: emotional health, mental health, health belief, sleep, energy, physical functioning, diet, social role, physical role, and sexual relations.

Methods:

During its development and evaluation, the IBSQOL was administered to over 500 patients with irritable bowel syndrome—two groups of patients from tertiary care centres, three focus groups of 8–12 patients each, and 287 patients in a national irritable bowel syndrome support network. As a control, the IBSQOL was also administered to 37 patients who did not have irritable bowel syndrome but had other gastrointestinal disorders. Statistical analyses to test the reliability and validity of the IBSQOL were performed using Cronbach's α coefficient.

Results:

Responses from the focus groups indicated that the IBSQOL was easy to complete and did not require too much time to fill out (≈ 25 min). Statistical analyses of the final 30‐item version of the IBSQOL demonstrated that it had both adequate validity and reliability (α ≥ 0.60). A comparison of mean IBSQOL scores of persons with and without irritable bowel syndrome (but with other gastrointestinal conditions) showed no difference between the two groups with irritable bowel syndrome; however, scores for both irritable bowel syndrome groups were considerably lower than for the non‐irritable bowel syndrome group, suggesting better health‐related quality of life in patients who do not have irritable bowel syndrome. This further demonstrated the validity of the IBSQOL in targeting questions and domains specific to patients with irritable bowel syndrome.

Conclusions:

Evaluation of the IBSQOL included testing the questionnaire in a large number of patients, which resulted in a revised and well‐constructed instrument that demonstrated both adequate validity and reliability. The IBSQOL is currently being used in large‐scale clinical trials to measure changes in quality of life in patients with irritable bowel syndrome following treatment intervention.
Doubling the omeprazole dose (40 mg b.d. vs. 20 mg b.d.) in dual therapy with amoxycillin increases the cure rate of Helicobacter pylori infection in duodenal ulcer patients
Authors: Labenz J. , Beker J.A. , Dekkers C.P.M. , Farley A. , Klör H.‐U. , Jönsson A.
Published: November 2003

Background:

Several studies have shown that dual therapy with omeprazole and amoxycillin may cure Helicobacter pylori infection. However, the optimum dose of omeprazole has still to be established.

Methods:

An international, randomized, double‐blind multicentre trial was conducted in patients with duodenal ulcers to compare the H. pylori cure rates obtained by dual therapy consisting of either omeprazole 20 mg b.d. plus amoxycillin 750 mg b.d. or omeprazole 40 mg b.d. plus amoxycillin 750 mg b.d. for 2 weeks. Dual therapy was followed by omeprazole 20 mg once daily for 2 weeks. Before entering the trial and 4 weeks after cessation of treatment H. pylori infection was assessed by histology and a 13C‐urea breath test.

Results:

381 patients were randomized into the study, of whom 345 were evaluable for the all‐patients‐treated analysis of efficacy and 378 were valid for the evaluation of safety. Histology results showed that H. pylori infection was cured in 64 out of 174 patients treated with omeprazole 20 mg b.d. plus amoxycillin and in 102 out of 171 patients treated with omeprazole 40 mg b.d. plus amoxycillin (37% vs. 60%; P <  0.001). Both treatment regimens were well tolerated, with adverse events reported by 29 (15.2%) and 35 patients (18.7%), respectively.

Conclusions:

This study has shown that dual therapy with amoxycillin 750 mg b.d. and omeprazole 40 mg b.d. is superior to dual therapy with amoxycillin and omeprazole 20 mg b.d. in patients with H. pylori‐positive duodenal ulcers. Thus, a true dose–response relationship exists between omeprazole and treatment success. However, a combination of omeprazole with two of amoxycillin, clarithromycin and a nitroimidazole is a preferable alternative for routine clinical use.
Effect of acute pentoxifylline treatment in an experimental model of colitis
Authors: Peterson T. C. , Davey K.
Published: November 2003

Background:

The effect of acute pentoxifylline treatment in an experimental model of colitis was assessed using the trinitrobenzene sulphonic acid (TNBS)‐induced rat model of colitis.

Methods:

Animals were treated with intracolonic injection (250 μL) of TNBS (50 mg in 50% ethanol) to induce inflammation and ulcers. Animals received pentoxifilline (100 mg/kg intracolonically) or saline 24 and 48 h following TNBS treatment. Five days following TNBS treatment, colons were dissected and scored according to the morphology damage score. The colons were then rolled longitudinally, fixed in formalin and embedded in paraffin. The collagen content of colonic sections was determined by a Sirius red–Fast green technique.

Results:

Animals treated with TNBS alone had significantly higher gross morphology damage scores compared to animals treated with saline. Pentoxifylline significantly reduced the gross morphology damage score in animals receiving TNBS. Colonic collagen levels were significantly elevated in TNBS‐treated animals compared to animals receiving saline. Pentoxifylline treatment did not alter the collagen content of colons from TNBS‐treated animals.

Conclusion:

TNBS treatment significantly elevates morphology damage score compared to controls. The results also suggest that colonic collagen was significantly elevated in animals treated with TNBS compared to controls. Pentoxifylline treatment was not sufficient to reduce the elevation in colonic collagen, although pentoxifylline treatment was sufficient to reduce the pathological changes due to TNBS, thus bringing the morphology damage score down to control levels.
Mucosal interleukin‐1β production and acid secretion in enlarged fold gastritis
Authors: YASUNAGA Y. , SHINOMURA Y. , KANAYAMA S. , HIGASHIMOTO Y. , YABU M. , MIYAZAKI Y. , MURAYAMA Y. , NISHIBAYASHI H. , KITAMURA S. , MATSUZAWA Y.
Published: November 2003

Background: We have previously shown that eradication of Helicobacter pylori increases acid secretion in H. pylori‐associated enlarged fold gastritis.

Aim: To investigate whether locally produced interleukin‐1β is possibly involved in the inhibition of acid secretion in H. pylori gastritis.

Methods: IL‐1β release from the gastric body mucosa was determined by short‐term culture of biopsy specimens in 13 patients with enlarged fold gastritis (all H. pylori‐positive), five H. pylori‐positive and 10 H. pylori‐negative patients without enlarged folds. The acid‐inhibitory effect of locally produced IL‐1β was examined by []>
14C]‐aminopyrine uptake assay using isolated rabbit gastric glands.

Results: IL‐1β release was significantly greater in patients with enlarged fold gastritis, significantly correlated with both basal and tetragastrin‐stimulated acid outputs in the H. pylori‐positive patients (r = −0.591 and r = −0.641, respectively; P < 0.01), and significantly decreased with concomitant increases in acid secretions after eradication of H. pylori. [14C]‐aminopyrine uptake was inhibited by IL‐1β in a dose‐dependent manner.

Conclusions: Increased production of IL‐1β caused by H. pylori infection is possibly involved in the inhibition of acid secretion in enlarged fold gastritis.
The vascular and glandular organoprotective properties of metronidazole in the rodent stomach
Authors: KO J. K. S. , CHING C. K. , CHOW J. Y. C. , ZHANG S. T. , LAM S. K. , CHO C. H.
Published: November 2003

Background: The gastroprotective action of metronidazole, an antimicrobial used in the therapy against Helicobacter pylori infection, is unclear. Thus, the aim of the present investigation was to study the organoprotective action and antiulcer mechanisms of this drug in rodents.

Methods and results: Metronidazole (10 mg/kg), given either per os or intraperitoneally, 30 min beforehand, reduced ethanol (40%, 10 mL/kg, p.o.)‐induced gastric mucosal damage in male rats. Likewise, oral administration of metronidazole dose‐dependently attenuated the indomethacin (30 mg/kg, p.o.)‐induced gastric lesion formation and the concurrent depletion of mucosal mucus. However, metronidazole did not affect the basal mucosal prostaglandin E2 content. In an ex vivo gastric chamber preparation, 40% ethanol incubation markedly lowered transmucosal potential difference and increased mucosal vascular permeability in rat stomachs. Incubation with all doses of metronidazole did not modulate gastric mucosal blood flow nor transmucosal potential difference, either before or after ethanol treatment. Nevertheless, the increase in vascular permeability by 40% ethanol was significantly alleviated by either p.o. or i.p. metronidazole pre‐treatment. In addition, exposure of the isolated rabbit gastric gland preparation to metronidazole (10−5 and 10−4 m) significantly attenuated the damaging action of 10% ethanol.

Conclusion: It is concluded that metronidazole possesses a direct vascular and glandular organoprotective property in the rodent stomach. However, the anti‐ulcer action does not appear to involve prostaglandins nor act through the improvement of gastric mucosal blood flow. Preservation of intramucosal mucus may partly contribute to the prevention of indomethacin‐induced ulceration in rats.
A randomized controlled study comparing elemental diet and steroid treatment in Crohn's disease
Authors: ZOLI G. , CARÈ M. , PARAZZA M. , SPANÒ C. , BIAGI P. L. , BERNARDI M. , GASBARRINI G.
Published: November 2003

Background: Elemental diet is considered an effective primary treatment for active Crohn's disease, but it is usually given by a feeding tube.

Methods: Twenty‐two patients (12 males, median age 30 years, range 18–60) with moderately active Crohn's disease were enrolled in a randomized study in which the efficacy of an elemental diet administered orally was compared to high‐dose corticosteroids in achieving clinical and laboratory remission. Ten patients were treated by oral elemental diet (Peptamen, Clintec, USA) and 10 received corticosteroids. Both treatment regimens lasted 2 weeks. The two groups did not differ with respect to age, sex, body weight, location of disease, treatment or disease activity prior to the study. In all patients studied, simple Crohn's disease activity index, nutritional status (expressed as body mass index), percentage of ideal body weight, fat mass, fat free mass, erythrocyte sedimentation rate, interleukin‐6, intestinal permeability (expressed as permeability index), prealbumin, retinol binding protein and multiskin test were evaluated before and after treatment.

Results: After 2 weeks of treatment, there were significant improvements in simple Crohn's disease activity index, erythrocyte sedimentation rate, permeability index, body mass index, prealbumin, retinol binding protein and multiskin test in the elemental diet group. There were significant improvements in simple Crohn's disease activity index and fat free mass in the corticosteroid group.

Conclusions: These data suggest that, in the short term, an oral elemental diet is at least as effective as steroids in inducing remission of mild‐moderately active Crohn's disease, but it may be more effective in improving the nutritional status of these patients, probably through a more rapid restoration of normal intestinal permeability.
Effect of a novel 5‐HT3 receptor agonist MKC‐733 on upper gastrointestinal motility in humans
Authors: Coleman N. S. , Marciani L. , Blackshaw E. , Wright J. , Parker M. , Yano T. , Yamazaki S. , Chan P. Q. , Wilde K. , Gowland P. A. , Perkins A. C. , Spiller R. C.
Published: November 2003

Background : Although 5‐HT3 antagonists have been used to treat chemotherapy‐induced emesis and diarrhoea‐predominant irritable bowel syndrome, the effects of 5‐HT3 agonists in humans are unknown.

Aim : To determine the effect of MKC‐733, a selective 5‐HT3 receptor agonist, on upper gastrointestinal motility.

Methods : Oral MKC‐733 (0.2, 1 and 4 mg) was compared with placebo in three randomized, double‐blind, cross‐over studies in healthy males. Antroduodenal manometry was recorded for 8 h during fasting and 3 h post‐prandially (n = 12). Gastric emptying and small intestinal transit were determined by gamma‐scintigraphy (n = 16). Gastric emptying, accommodation and antral motility were determined by echoplanar magnetic resonance imaging (n = 12).

Results : MKC‐733 (4 mg) increased the number of migrating motor complexes recorded in the antrum and duodenum (P < 0.001), but had no effect on post‐prandial motility. MKC‐733 delayed scintigraphically assessed liquid gastric emptying (P = 0.005) and accelerated small intestinal transit (P = 0.038). Echoplanar magnetic resonance imaging confirmed the delayed gastric emptying (P < 0.001) and demonstrated a significant increase in cross‐sectional area of the proximal stomach (P < 0.01).

Conclusions : MKC‐733 delays liquid gastric emptying in association with relaxation of the proximal stomach, stimulates fasting antroduodenal migrating motor complex activity and accelerates small intestinal transit.
The population pharmacokinetics of long‐term methotrexate in rheumatoid arthritis
Authors: Godfrey C. , Sweeney K. , Miller K. , Hamilton R. , Kremer J.
Published: November 2003

Aims Methotrexate is considered by many practitioners to be the agent of choice in the treatment of rheumatoid arthritis. The pharmacokinetics of methotrexate have been reported to exhibit significant intersubject variability. Therefore, this study was undertaken to evaluate the population pharmacokinetics of methotrexate during long‐term administration in adults with rheumatoid arthritis.

Methods Methotrexate pharmacokinetics were evaluated in a 36 month study of 62 adults with rheumatoid arthritis. Patients received oral or intramuscular doses of methotrexate weekly with pharmacokinetic studies performed every 6 months. Data were analyzed with nonlinear mixed effects modeling.

Results Three thousand two hundred and sixty post oral or intramuscular dose serum methotrexate concentrations comprising 425 individual concentrationvstime profiles were modeled using NONMEM. Covariates that significantly (P<0.005) influenced the disposition of methotrexate were age (AGE, years), body weight (BW, kg), creatinine clearance (CLCR, l h−1 ), gender (GEN; 0=male, 1=female), dose (DOSE, μmol), and fedvsfasted state (FED; 0=fasted, 1=fed). The final model describing the biexponential disposition of methotrexate was clearance(CL, l h−1 )=(0.0810*BW+0.257*CLCR)*(1–0.167*GEN); central volume (V c, l)=0.311*BW; peripheral volume (V p, l)=0.469*BW‐0.169*AGE; intercompartmental clearance (Q, l h−1 )=4.27*(1–0.355*GEN); oral absorption rate constant (kapo, h−1 )=4.70–0.0439*DOSE*(1–0.507*FED); intramuscular absorption rate constant (kaim, h−1 )=0.122*DOSE; relative bioavailability (F )=93.4%; and oral absorption lag time (LAGpo, min)=13.5. Pharmacokinetic parameters (%CV) for a typical fasted male subject in this study were CL, 7.34 l h−1(27%);V c, 23.5 l (28%);V p, 25.3 l (31%); Q, 4.25 l h−1(41%);kapo, 3.67 h−1(77%); andkaim, 3.09 h−1(44%).

Conclusions The population pharmacokinetics of methotrexate in adults with rheumatoid arthritis were well described by this investigation. Substantial interpatient variability was explained by incorporating patient specific data into regression equations predicting pharmacokinetic parameters.
Haemostatic and rheological factors in intermittent claudication: the influence of smoking and extent of arterial disease
Authors: Lee Amanda , Fowkes F. , Rattray Anna , Rumley Ann , Lowe Gordon
Published: November 2003

Patients with intermittent claudication have been reported to have disturbances in blood rheology and haemostasis. Whether these disturbances are a result of, or largely independent of, smoking history and arterial narrowing has not yet been established. The levels of whole blood and plasma viscosity, haematocrit, von Willebrand factor antigen, fibrin D‐dimer antigen and urinary fibrinopeptide A antigen were compared in 617 claudicants and 722 controls from two epidemiological studies in Edinburgh. After adjustment for age and sex, all factors, except whole blood viscosity and haematocrit, were significantly higher in the claudicants compared to controls (P≤0.001). The risk of intermittent claudication was significantly raised for unit change in each factor, except for whole blood viscosity and haematocrit. Adjustment for lifetime smoking had little effect on the odds ratios. After further adjustment for the ankle brachial pressure index (as a measure of the extent of peripheral arterial disease), haematocrit, von Willebrand factor and urinary fibrinopeptide A showed a significant independent relationship with the risk of intermittent claudication. We conclude that the association between selected rheological and haemostatic factors and leg ischaemia is largely independent of both smoking history and the extent of arterial narrowing, and may be directly related to microvascular ischaemia.
Interleukin‐10 mRNA expression in B‐cell chronic lymphocytic leukaemia inversely correlates with progression of disease
Authors: Sjöberg Jan , Aguilar ‐Santelises Miguel , Sjögren Ann‐Marie , Pisa Eva K. , Ljungdahl ÅKe , Björkholm Magnus , Jondal Mikael , Mellstedt HÅkan , Pisa Pavel
Published: November 2003

Interleukin‐10 (IL‐10) has been shown in vitro to inhibit survival and spontaneous DNA synthesis in B‐cell chronic lymphocytic leukaemia (B‐CLL) cells by induction of programmed cell death. We have analysed the presence of mRNA transcripts for IL‐10 in purified B‐CLL cells from 35 patients by RT‐PCR. Transcripts for IL‐10 were detected in 11/20 patients with non‐progressive disease. In cell preparations from patients with progressive B‐CLL IL‐10 mRNA were detected in only 2/15 samples (P ≤ 0.01). The Epstein‐Barr virus status of the cells did not account for the difference in IL‐10 mRNA expression observed between the two groups of patients. Thus, IL‐10 mRNA expression in leukaemic cells from patients with B‐CLL was strongly associated with non‐progressive disease. This finding may support other observations suggesting that IL‐10 might be a candidate for immune therapy of progressive B‐CLL.
Generation of human natural killer cells from peripheral blood CD34+ cells mobilized by granulocyte colony‐stimulating factor
Authors: Takenaka Katsuto , Mizuno Shin‐ichi , Harada Mine , Nagafuji Koji , Miyamoto Toshihiro , Iwasaki Hiromi , Fujisaki Tomoaki , Kubota Akira , Ohno Yuju , Arima Fumitou , Shigematsu Hirokazu , Gondo Hisashi , Okamura Takashi , Okamura Seiichi , Inaba Shoichi , Niho Yoshiyuki
Published: November 2003

We studied the generation of human natural killer (NK) cells from CD34+ cells that were isolated from peripheral blood stem cells (PBSC) mobilized by granulocyte colony‐stimulating factor (G‐CSF). The isolated CD34+ cells were cultured in the presence of a combination of interleukin‐1 (IL‐1α), IL‐2, and stem cell factor for 5 weeks without marrow stroma. We found that the CD34+ cells isolated from G‐CSF‐mobilized PBSC (G‐CSF/PBSC) could differentiate into a population of NK cells which were CD56+(bright)/CD3− and showed morphologic characteristics of large granular lymphocytes. Immunophenotypic analysis of the NK cells thus generated showed that a small proportion of them expressed CD2, CD8 and CD16 surface markers and approximately half of them coexpressed CD7. This NK population exhibited cytotoxic activity against a NK‐sensitive cell line, K562. These observations suggest that CD34+ cells from G‐CSF/PBSC contain precursors of NK cells that can differentiate into functional NK cells.
Inhibition of platelet adhesion to collagen by monoclonal anti‐CD36 antibodies
Authors: MATSUNO KAZUHIKO , DIAZ‐RICART MARIBEL , MONTGOMERY ROBERT R. , ASTER RICHARD H. , JAMIESON G. A. , TANDON NARENDRA N.
Published: November 2003

Monoclonal anti CD36 antibodies capable of inhibiting platelet adhesion to collagen have not previously been identified. We have now prepared two groups of monoclonal antibodies. One group was prepared using, as immunogen, highly purified (99+%) CD36 prepared by a denaturing procedure. These antibodies (Mo series) reacted strongly with CD36 on protein blots but did not immunoprecipitate native CD36 from platelet lysates nor inhibit platelet adhesion to collagen. The second group of monoclonal antibodies (131 series) was prepared using CD36 purified to >95% by a non‐denaturing procedure. These antibodies reacted with control platelets, but not Naka‐negative platelets which lack CD36, as measured by flow cytometry and by immunoprecipitation. Three monoclonal antibodies of this latter group (131.4, 131.5 and 131.7) inhibited platelet adhesion to collagen in static systems under Mg2+‐independent conditions but had little effect in the presence of Mg2+. 131.4 and 131.7 also inhibited adhesion to collagen using citrated whole blood in a parallel plate flow chamber at physiological shear rates (800 s−1), whereas 131.5 was without effect. These are the first anti‐CD36 monoclonal antibodies shown to be capable of inhibiting platelet adhesion to collagen and provide further evidence that CD36 plays a role in platelet–collagen interaction.
Bencyclane as an anti‐sickling agent
Authors: KOÇAK RIKKAT , BAŞLAMIŞLI FIKRI , GÜVENÇ BIROL , TAMER LÜLÜFER , AIKIMBAEV KAIRGUELDY S. , ISBIR TURGAY
Published: November 2003

A vasodilating Ca2+ channel blocker, bencyclane, was used in 18 patients with homozygous sickle cell anaemia (SCD) to test the possible anti‐sickling effect. With bencylane intervention the Na+‐K+ ATPase activity increased from 256±29 to 331±37 nmol Pi/mg protein/h (P<0.0001) and the Ca2+‐Mg2+ ATPase level increased from 172±12 to 222±44 nmol Pi/mg protein/h (P<0.0001). The intracytoplasmic Ca2+ concentration reduced from 3.5±0.6 to 2.7±0.25 μmol/l (P<0.0001). The patient’s blood contained fewer irreversibly sickled cells (ISCs) (a reduction from 21.4% to 14.4%) (P<0.05). At the same time MCHC of the erythrocytes decreased from 34.5 to 33.0 g/dl (P<0.05). Bencyclane appears to be a promising anti‐sickling agent that can be used orally in SCD.
t(3;21)(q26;q22) with AML1 rearrangement in a de novo childhood acute monoblastic leukaemia
Authors: Johansson Bertil , Fioretos Thoas , Garwicz Stanislaw , Heim Sverre , Mitelman Felix
Published: November 2003

t(3;21)(q26;q22) is a recurrent chromosomal abnormality in Philadelphia‐positive chronic myeloid leukaemia in blast crisis and in treatment‐related myelodysplastic syndrome and acute myeloid leukaemia. The molecular consequences of the t(3;21) are presently being unravelled; various transcripts between the AML1 gene in 21q22 and several unrelated genes, i.e. EAPEVI1 and MDS1, in 3q26 are generated, resulting in the formation of a chimaeric transcription factor. The t(3;21) has only rarely been described in de novo leukaemias and never before in an acute leukaemia in a child. We here present the clinical, cytogenetic and molecular genetic findings in a boy with a de novo acute monoblastic leukaemia with t(3;21)(q26;q22) and AML1 rearrangement.
Recombinant human G‐CSF‐mobilized peripheral blood stem cells for second allogeneic transplant after bone marrow graft rejection in children
Authors: Zecca Marco , Perotti Cesare , Marradi Pierluigi , Montagna Daniela , Giorgiani Giovanna , BALTER RITA , Prete Luisella , Locatell i Franco
Published: November 2003

Two children affected by severe aplastic anaemia and sickle cell anaemia rejected the allogeneic bone marrow transplantation from an HLA‐matched unrelated volunteer and an HLA‐identical sibling, respectively. In both cases a second transplant using granulocyte‐colony stimulating factor (G‐CSF) mobilized peripheral blood stem cells (PBSC) was performed. Donors were the HLA‐haploidentical mother and the same HLA‐identical sibling who was employed for the first marrow allograft, respectively. Treatment with G‐CSF and PBSC collection were well tolerated. Both patients had engraftment of donor haemopoiesis and did not experience severe graft‐versus‐host disease. These cases confirm that PBSC transplant should be considered as a feasible treatment to reverse graft failure in paediatric patients.
The interaction of α° thalassaemia with Hb Icaria: three unusual cases of haemoglobinopathy H
Authors: Kanavakis E. , Traeger‐Synodinos J. , Papasotiriou I. , Vrettou C. , Metaxotou‐Mavromati A. , Stamoulakatou A. , Lagona E. , Kattamis C.
Published: November 2003

The clinical, haematological, biosynthetic and molecular data of three Greek haemoglobin H (HbH) disease patients with a distinctive clinical phenotype are described. During infancy all three patients had unusually severe clinical manifestations for HbH disease, with anaemia necessitating blood transfusions, signs of bone changes, growth impairment, and splenomegaly. Molecular analysis identified a rare α‐thalassaemia genotype (− −Med/α Icα) . Splenectomy resulted in marked amelioration of the clinical signs; post splenectomy all three patients preserve adequate haemoglobin levels (9–10 g/dl) with growth restored to normal. Despite the initial severe clinical phenotype in these patients, our experience indicates that splenectomy modifies the clinical course to that of mild thalassaemia intermedia. This observation should be considered carefully when giving genetic counselling to families carrying the rare Hb Icaria mutation and an α° thalassaemia mutation.
A −793 G to A transition in the factor IX gene promoter is polymorphic in the caucasian population
Authors: Rai Harjeet , Winship Peter
Published: November 2003

We report the incidence of a prevalent polymorphism at position −793 in the promoter region of the factor IX gene in caucasian individuals. This DNA change was originally reported as one of two changes in the factor IX gene of a severely affected haemophilia B patient from Japan. We confirm the neutral nature of this change and demonstrate that, despite showing linkage disequilibrium with the previously reported MseI RFLP <100 bp distant, the use of these two loci together in a carrier screening strategy significantly increases the level of informativity over that achieved using either polymorphism alone.
Minimal residual disease after allogeneic bone marrow transplantation for chronic myeloid leukaemia: a metaphase‐FISH study
Authors: El‐Rifai Wa'el , Ruutu Tapani , Vettenrant a Kim , Temtam y Samia , Knuutila Sakari
Published: November 2003

Metaphase‐FISH was adopted for the detection of proliferating Philadelphia‐positive (Ph+) residual leukaemic cells in 25 patients with chronic myeloid leukaemia treated with allogeneic bone marrow transplantation (BMT). Patients were followed up during their clinical remission for 4–50 months (median 17 months) after BMT. 80 bone marrow samples were studied. For most of the cases no fewer than 1000 metaphases were analysed. Six patients (24%) showed residual Ph+ cells during the first 6 months and two others by the end of the first year after BMT. Three patients relapsed during the study and in two of them residual Ph+ cells were detected during the first 6 months after BMT. In 17 patients no Ph+ cells were detected at any stage of follow‐up and 16 (94.1%) of them continue in complete clinical and haematological remission. Our results indicate that metaphase‐FISH is a reliable tool in the quantitation of proliferating residual leukaemic cells. We suggest that consecutive findings of equal amounts of residual leukaemic cells do not necessarily predict a relapse. However, their presence calls for follow‐up at shorter intervals where an increasing number of these cells predicts an ensuing relapse.
Neutralizing anti‐interferon‐α antibodies and response to treatment in patients with Ph+ chronic myeloid leukaemia sequentially treated with recombinant (α2a) and lymphoblastoid interferon‐α
Authors: Russo Domenico , Candoni Anna , Zuffa Eliana , Minisini Rosalba , Silvestri Federico , Fanin Renato , Zaja Francesco , Martinelli Giovanni , Tura Sante , Botta Giuseppe , Baccarani Michele
Published: November 2003

Neutralizing anti‐IFNα antibodies (nIFNα Abs) occur in a significant proportion of patients with hairy cell leukaemia, hepatitis or solid tumours treated with recombinant IFNα (IFNα2a or IFNα2b), but information on their incidence in chronic myeloid leukaemia (CML) is scanty and their clinical relevance is not yet completely defined. By using an IFNα antiviral neutralization bioassay, the frequency of nIFNα2a Abs was evaluated in 67 Ph+ CML patients during IFNα2a therapy at doses ranging from 6 to 9 MU/d. 15 patients (22%) developed nIFNα2a Abs (titre ranging from 1:40 to 1:20480) and 11/15 (73%) were haematologically and/or karyotypically unresponsive to treatment. 52 patients did not develop antibodies and 11 of them (21%) were unresponsive. The negative relationship between the positivity for nIFNα2a Abs and the response to treatment was highly significant (P = 0.0001). In nine nIFNα2a Abs positive patients, treatment was changed from recombinant IFNα2a to lymphoblastoid IFNα (IFNα‐ly), at the same dose and schedule. After 9 months of IFNα‐ly treatment a haematological response was achieved in 4/7 cases who were non‐responsive to prior IFNα2a therapy and was maintained in the other two patients previously responsive to IFNα2a. However, no karyotypic response was observed. This data shows that a significant proportion of Ph+ CML patients receiving treatment with IFNα2a can develop neutralizing antibodies and that these antibodies are associated with a loss of IFNα2a efficacy. Changing the patients to treatment with lymphoblastoid IFNα may restore haematological response but it is not likely to induce a karyotypic response.
Acute arterial thrombosis due to platelet aggregation in a patient receiving granulocyte colony‐stimulating factor
Authors: Kawachi Yasunori , Watanabe Ayako , Uchida Tatsumi , Yoshizawa Kiyoshi , Kurooka Nobuaki , Setsu Kojun
Published: November 2003

We describe a 44‐year‐old man with non‐Hodgkin's lymphoma receiving granulocyte colony‐stimulating factor (G‐CSF) who developed an acute arterial thrombosis. The removed thrombus contained large amounts of platelet aggregation. A rapid increase of platelets and increased adenosine diphosphate (ADP)‐ and collagen‐induced platelet aggregation were observed at the time of the thrombotic event. A challenge test of G‐CSF showed an increase in the platelet count and an augmentation of ADP‐ and collagen‐induced platelet aggregation. In the use of G‐CSF, patients who produce a rapid increase in platelet levels could be at greater risk for thrombotic events and need to be followed‐up carefully.
Denaturing gradient gel electrophoresis: a novel method for determining Rh phenotype from genomic DNA
Authors: Steers Fiona , Wallace Maura , Johnson Philip , Carritt Ben , Daniels Geoff
Published: November 2003

Denaturing gradient gel electrophoresis (DGGE) was carried out on PCR products amplified from exons 2 and 5 of RHD and RHCE. Exon 2 of RHD and exon 2 of the C allele of RHCE have an identical sequence, which differs from that of the c allele of RHCE. One band representing D and/or C, and another representing c, could be distinguished by DGGE of exon 2 amplifications of genomic DNA from individuals with the appropriate Rh phenotype. C and c could only be distinguished in D‐negative samples. Exon 5 of RHD and exon 5 of the E and e alleles of RHCE all have different nucleotide sequences. Bands representing D, E and e could be distinguished following DGGE of the products of exon 5 amplification of genomic DNA from individuals with red cells of the appropriate Rh phenotype. In samples from individuals with VS+ red cells (V+ or V−) there was a shift of the band representing e. Sequencing demonstrated that VS is associated with a RHCE e sequence with a single base change predicting a Leu245 → Val substitution in the Rh polypeptide. This substitution may be responsible for the VS and es antigens.
A plasmocyte selective monoclonal antibody (B‐B4) recognizes syndecan‐1
Authors: Wijdenes John , Vooijs Wim C. , Clément Claude , Post Jacob , Morard Florence , Vita Natalio , Laurent Patrick , Sun Ren‐Xiao , Klein Bernard , Dore Jean‐Michel
Published: November 2003

We developed a new monoclonal antibody, B‐B4, which specifically identifies human plasma cells. It strongly reacts with all multiple myeloma cell lines and with malignant plasma cells of all tumour samples of the multiple myeloma patients tested. B‐B4 does not react with any peripheral blood, bone marrow or tonsil cells. Cloning of the B‐B4 antigen reveals that the monoclonal antibody recognizes syndecan‐1. It appears that the monoclonal antibody B‐B4 is a suitable marker for human plasmocyte identification among haemopoietic cells and a useful probe for the diagnosis of haematological malignancies. Furthermore, this monoclonal antibody can be used for depletions prior to CD34 grafting.
Combination therapy with interferon alpha and ribavirin for chronic hepatitis C virus infection in thalassaemic patients
Authors: Telfer P. T. , Garson J. A. , Whitby K. , Grant P. R. , Yardumian A. , Hoffbrand A. V. , Wonke B.
Published: November 2003

Hepatitis C virus (HCV) infection is common in multi‐transfused thalassaemic patients, and, in combination with transfusional iron overload, can result in progressive liver disease. Therapy with interferon‐alpha causes a sustained loss of HCV in only 15–25% of patients, and there is as yet no established effective therapy for those who fail to respond. We have conducted a pilot study of combination anti‐viral therapy for patients who failed to respond, or relapsed after an initial response to single‐agent interferon‐alpha. Patients were treated for 6 months with interferon‐alpha 2b, given subcutaneously three mega units thrice weekly, together with ribavirin, orally 1 g daily. 11 patients were enrolled, their median age was 24.9 years. 8/10 evaluable patients had cirrhosis on biopsy, five were infected with HCV type 1 and all but one had initial HCV RNA titres > 106 genomes/ml. Five patients (45.5%) had a sustained virological response with loss of serum HCV RNA for > 6 months after finishing therapy. There was no clear association between response to therapy and age, histology, HCV genotype, or HCV RNA titre. Transfusion requirements were significantly increased during the treatment phase, probably due to ribavirin‐induced haemolysis, and this necessitated intensification of iron chelation therapy. Serum ferritin levels decreased significantly in those who responded. These results suggest that combination therapy is potent in clearing HCV infection, and may provide effective second‐line therapy for thalassaemic patients who have failed to respond to interferon‐alpha monotherapy.
Constitutive and inducible expression of megakaryocyte‐specific genes in Friend erythroleukaemia cells
Authors: Vannucchi Alessandro M. , Linari Silvia , Cellai Cristina , Koury Mark J. , Paoletti Francesco
Published: November 2003

Friend murine erythroleukaemia cells (MELC) were analysed by semiquantitative RT‐PCR for the constitutive and inducible expression of megakaryocyte‐specific genes. Uninduced MELC expressed detectable levels of mRNAs for acethylcholinesterase (AChE), platelet factor‐4 (PF4), glycoprotein IIb (GPIIb) and von Willebrand factor (VWF), whereas the erythroid α‐ and β‐globin genes were not transcribed appreciably. However, MELC exposed to 5 mm hexamethylene bisacetamide (HMBA) or 1.5% dimethyl sulphoxide (DMSO) seemed to be channelled towards a mixed erythroid/megakaryocytic phenotype characterized by unaltered levels of VWF mRNA, increased levels of AChE, GPIIb and PF4 mRNA, and simultaneous induction of the globin genes. Megakaryocyte‐related genes were expressed, in the absence of globin gene transcription, by MELC treated with either phorbol‐12‐myristate acetate (PMA; 100 ng/ml) or colcemid (40 nm), an antimicrotubule agent capable of promoting polyploidization in this model. Moreover, PMA and colcemid induced also de novo expression of the thrombopoietin receptor c‐mpl. PMA and colcemid did not affect high basal c‐myb mRNA levels which, in turn, were down‐regulated upon HMBA or DMSO induction. Additionally, both uninduced and induced MELC exhibited significant levels of Epo‐R and IL‐3R mRNAs, whereas no expression of granulocyte/macrophage‐related genes was detected. Megakaryocyte gene expression of MELC was also compared to that of other haemopoietic cell populations from normal mice and mice infected with the anaemic strain of the Friend virus. According to our results, MELC should be seen as an unique erythro‐megakaryocytic model of differentiation, potentially useful for studying molecular events governing lineage commitment as well as some steps of megakaryocytopoiesis.
T cells selectively infiltrate bone marrow areas with residual haemopoiesis of patients with acquired aplastic anaemia
Authors: Mlenhorst J. Joseph , Van Krieken Johan H. J. M. , Dreef Enno , Landegent James E. , Willemze Roel , Fibbe Willem E.
Published: November 2003

Aplastic anaemia (AA) is characterized by pancytopenia and bone marrow (BM) hypocellularity. In some patients AA may be mediated by T cells. To localize inflammatory cell infiltrates, we carried out a quantitative immunohistochemical analysis of BM biopsies of AA patients. In five out of eight biopsies, significantly higher numbers T cells were found in the areas with residual haemopoiesis (RH). The significantly increased numbers of CD3+ T cells in areas with RH supports the hypothesis of a site‐directed infiltration and/or a local proliferation of T cells in the BM of patients with AA.
Reduction of heat‐induced haemotoxicity in a hyperthermic purging protocol of murine acute myeloid leukaemic stem cells by AcSDKP
Authors: Wierenga Pieter K. , Dillingh Jan H. , Konings Antonius W. T.
Published: November 2003

The tetrapeptide AcSDKP (Goralatide) is a cytokine with known inhibitory effects on cell proliferation. Many purging agents used in autologous bone marrow transplantation protocols, including hyperthermia, preferentially kill cycling cells. A pretreatment with Goralatide offers a possibility to reduce the haemotoxicity in many purging settings. The impact of Goralatide on the hyperthermic purging protocol was investigated in normal and myeloid leukaemic (SA8) murine cells. The median survival time after transplantation (i.e. leukaemia incidences) was used as an in vivo parameter to determine the effects on leukaemic cells. The hyperthermic effect on normal and leukaemic cells was also investigated in vitro using the cobblestone area‐forming cell (CAFC) assay. A heat treatment of 90 min at 43°C resulted in a 4‐log depletion of leukaemic stem cells. For normal progenitor cells (CFU‐GM) a 2‐log cell kill was shown. The reduction in proliferative activity of the CFU‐GM after an 8 h incubation with 10−9 m Goralatide resulted in a decrease in the heat sensitivity of the progenitor subset to approximately a 1‐log cell kill. The leukaemic precursor cells seem insensitive to Goralatide inhibition, implicating an increase in the therapeutic window of the hyperthermic purging protocol. Finally, simulated remission bone marrow (5% leukaemic blasts) was incubated with Goralatide followed by a heat treatment of 90 min at 43°C. Lethally irradiated (10 Gy) mice transplanted with heat‐treated remission bone marrow (106 normal bone marrow cells versus 5 × 104 leukaemic cells) died of aplasia while Goralatide‐pretreated remission bone marrow could rescue the irradiated mice without revealing leukaemic engraftment. These findings confirmed the enhanced protection against hyperthermia of the normal haemopoietic subsets by Goralatide and thus increased the success of the hyperthermic purging protocol.
A comparison of outcome from febrile neutropenic episodes in children compared with adults: results from four EORTC studies
Authors: Hann I. , Viscoli C. , Paesmans M. , Gaya H. , Glauser M. ,   the International Antimicrobial Therapy Cooperative Group (IATCG) of the European Organization for Research and Treatment of Cancer (EORTC)
Published: November 2003

The object of this study was to determine whether there were any differences between the ‘typical’ child with fever and neutropenia and their adult counterpart with regard to infection type and outcome, by analysis of 3080 patients, including 759 children < 18 years of age and 2321 adults. These represented patients randomized in previous trials, between 1986 and 1994, which compared empirical antibiotic regimens for fever in neutropenic patients. There were fewer childhood acute myeloid leukaemia patients than adults but more acute lymphoblastic leukaemia cases and more with solid tumours undergoing intensive myelosuppressive therapy. The children were less likely to be undergoing first induction therapy but the relative incidence of patients receiving relapse schedules or maintenance therapies were not significantly different in the two age groups. Children less frequently had a defined site of infection than adults and where they had a defined site there were more upper respiratory tract but fewer lung infections. There was a similar low incidence of shock at presentation in the two groups but the children's median neutrophil count was lower, and their median duration of granulocytopenia before the trial was shorter. The incidence of bacteraemia was similar, but clinically documented infection was less frequent and fever of unknown origin consequently more common in children. Children developed more streptococcal bacteraemias and fewer staphylococcal bacteraemias than adults (P = 0.003) but the relative incidence of various gram‐negative species was similar (P = 0.57). In general, the children had a better overall success rate and lower mortality than adults. Death from infection was only 1% in children versus 4% in adults (P = 0.001), and time to defervescence was shorter in children. In the younger age group, univariate logistic regression models showed high temperature, prolonged neutropenia before the trial and shock as prognostic indicators for the presence of bacteraemia. Solid tumour patients were significantly less likely to have a bacteraemia. Multivariate analysis confirmed the independent prognostic value of these indicators. Using the logistic equation of the selected model, the overall discriminant ability was poor. However, it was possible to identify a small subgroup without shock or high fever and with a short prior duration of neutropenia which carries a particularly low risk of bacteraemia, who could be considered for early discharge, monotherapy and shortened courses of antibodies, in prospective trials.
Dexamethasone, cyclophosphamide, idarubicin and etoposide (DC‐IE): a novel, intensive induction chemotherapy regimen for patients with high‐risk multiple myeloma
Authors: BALLESTER O. F. , MOSCINSKI L. C. , FIELDS K. K. , HIEMENZ J. W. , ZORSKY P. E. , GOLDSTEIN S. C. , SABA H. I. , SPIERS A. S. D. , KRONISH L. , SULLIVAN P. , ELFENBEIN G. J.
Published: November 2003

We evaluated toxicities and responses to a novel, dose intensive and time sequenced, chemotherapy programme (DC‐IE) in 45 patients with high‐risk myeloma. DC‐IE consisted of: dexamethasone (days 1–4); cyclophosphamide (day 5); idarubicin and etoposide (days 8–10). Complete response (CR) was achieved in four patients, six patients achieved near complete responses (nCR) and 21 patients achieved a partial remission (PR). Overall response rate was 76% (CI 56–94%) for newly diagnosed patients (n =21) and 62% (CI 36–81%) for relapsed/refractory patients (n =24). Toxicities were limited to myelosupression; two patients died of sepsis during neutropenia (4%). DC‐IE is active and tolerable for high‐risk multiple myeloma, including patients with relapsed or refractory disease to anthracycline containing regimens.
Frequency of CBFβ/MYH11 fusion transcripts in patients entered into the U.K. MRC AML trials
Authors: LANGABEER S. E. , WALKER H. , GALE R. E. , WHEATLEY K. , BURNETT A. K. , GOLDSTONE A. H. , LINCH D. C.
Published: November 2003

It has been established that cytogenetic findings at diagnosis of acute myeloid leukaemia (AML) are a powerful prognostic indicator. Patients who have the inv(16)(p13q22), closely associated with the FAB subtype M4Eo, are deemed to have good‐risk disease. This subtle translocation may be difficult to detect in poor‐quality metaphase preparations and if missed could lead to the incorrect assignment of risk group and influence further treatment strategies.

We studied 321 patients with AML at diagnosis for the presence of inv(16)(p13q22) and CBFβ/MYH11 fusion transcripts by cytogenetic and RT‐PCR techniques respectively. Karyotypic analysis detected 21 cases of inv(16) (p13q22), all of which were PCR positive. A further 12 cases were detected at the molecular level only, in FAB types other than M4Eo. The observed frequencies of CBFβ/MYH11 fusion transcripts in our study have been adjusted for the reported incidence of each FAB subtype and we calculate that 10.1% of all new cases of AMLs have molecular evidence of inv(16)(p13q22), only half of which are of the M4Eo subtype.

We conclude that molecular screening for the presence of CBFβ/MYH11 fusion transcripts should be mandatory in all cases of AML at diagnosis.
The role of compliance as a cause of instability in oral anticoagulant therapy
Authors: Van der Meer Felix J. M. , BriëT Eenest , Vandenbroucke Jan P. , ŠRámek Dagmar I. , Versluijs Myrthe H. P. M. , Rosendaal Frits R.
Published: November 2003

To assess the role of non‐compliance as a cause of instability in patients on oral anticoagulant therapy, a follow‐up study of stably and instably anticoagulated patients and of patients beginning oral anticoagulant therapy was performed. Compliance was assessed by pill counting and with the use of pill bottles in the cap of which a microprocessor registered the exact date and time of opening of the bottle. (In)stability of oral anticoagulant therapy was expressed as the number of INRs and as the time spent within the target range and by squared sigma. Subsequently, as a pilot study, a randomized intervention study in instable, non‐compliant patients was performed in which these patients were or were not informed about the real nature of the cap of the pill bottle.

Nineteen stable and 19 unstable patients and 30 patients beginning therapy were followed for 3 months or less until therapy stopped. Compliance was better in the previously stably anticoagulated patients, although differences were small. Poor compliance was not a major cause of instability in patients starting therapy. Although the pilot intervention study was too small to assess the role of the special pill bottle, it was shown that compliance can be positively affected.