bluuebunny/crossref_metadata_2025_split huggingface dataset:
A comparison of relative abundance, activity factor and inhibitory monoclonal antibody approaches in the characterization of human CYP enzymology
Authors: Soars Matthew G. , Gelboin Harry V. , Krausz Kristopher W. , Riley Robert J. 
Published: March 2003

Aims The objective of this study was to evaluate the potential uses of relative abundance, relative activity approaches and inhibitory monoclonal antibodies (mAbs) in the characterization of CYP enzymology in early drug discovery.Methods Intrinsic clearance estimates for the oxidation of ethoxyresorufin (a selective probe of CYP1A2 activity), tolbutamide (CYP2C9), S‐mephenytoin (CYPC19), dextromethorphan (CYP2D6) and testosterone (CYP3A4) were used to determine relative activity factors (RAFs). CLint values were determined for the metabolism of 14 drugs in human liver microsomes (HLM) and for these major CYPs. The relative contribution of each individual CYP to the oxidation of each drug was then assessed using relative abundance and activity techniques in addition to inhibitory mAbs.Results Relative abundance and activity methods as well as inhibitory mAbs qualitatively assigned the same CYP isoform as predominantly responsible for the clearance of each drug by HLM. Metabolism catalysed by CYP1A2, 2C9, 2D6 and 3A4 was also predicted to be quantitatively similar using both abundance and activity techniques. However, the relative contribution of the polymorphic CYP2C19 appeared to be over‐estimated approximately two‐fold using recombinant CYP compared with that from the HLM and mAb approach.Conclusions All three methods investigated in this study appear suitable for use in the characterization of the CYP metabolism of new chemical entities produced during early drug discovery.
Blood pressure response to conventional and low‐dose enalapril in chronic renal failure
Authors: Elung‐Jensen Thomas , Heisterberg Jens , Kamper Anne‐Lise , Sonne Jesper , Strandgaard Svend 
Published: March 2003

Aims In chronic renal failure, the clearance of most ACE inhibitors including enalapril is reduced. Hence, with conventional dosage, plasma enalaprilat may be markedly elevated. It is unclear whether this excess of drug exposure affords an improved control of blood pressure. The aim of the present study was to evaluate short‐term blood pressure response to two different plasma levels of enalaprilat.Methods As part of an open, randomized, controlled trial of the effect of high and low dosage of enalapril on the progression of renal failure, short‐term blood pressure response was evaluated. Data were analysed in all patients completing 3 months of follow‐up. The patients were allocated to two trough plasma concentrations of enalaprilat, either above 50 ng ml−1 (high) (n = 17) or below 10 ng ml−1 (low) (n = 18), and the daily dose of enalapril titrated accordingly.Results Median (range) glomerular filtration rate (GFR) at baseline was 18 (7.9) in the high enalaprilat concentration group and 17 (7.3) ml min−1 1.73 m2 in the low concentration group (NS). Nine patients in each group were on treatment with enalapril at baseline with a median daily dose of 5 mg in both the high (5–10) and low (2.5–20) concentration group. At 3 months’ follow‐up, the dose was 10 (2.5–30) and 1.9 (1.25–5) mg (P < 0.0001), respectively. After 3 months median trough concentrations of enalaprilat were 82.5 (22–244) ng ml−1 and 9.1 (2.5–74.8) ng ml−1 (P < 0.002). At baseline the median systolic blood pressures in the two groups were 140 (110–200) and 133 (110–165), in the high and low enalaprilat concentration groups, respectively, and after 3 months they were 135 (105–170) and 130 (105–170) mmHg (NS). Median diastolic blood pressure was 80 mmHg in each group both at baseline (65–100) and at follow‐up (60–95) (NS). There was no difference between the groups in concomitant antihypertensive treatment (number of patients treated, mean daily dose) during the observation period. Proteinuria remained stable during the study period in both groups; patients in the high concentration group had higher plasma potassium concentrations at day 90 and patients in the low group experienced a slight increase in GFR.Conclusions  In moderate to severe chronic renal insufficiency the same degree of blood pressure control was achieved on low as well as moderate daily doses of enalapril. This was irrespective of concomitant antihypertensive treatment.
Low‐dose atorvastatin therapy does not augment endothelial function in active hypercholesterolaemic males
Authors: Parnell Melinda M. , Chin‐Dusting Jaye P. F. , Starr Jennifer , Kaye David M. 
Published: March 2003

Aims As statin therapy has been demonstrated to augment endothelial function in sedentary hypercholesterolaemia (HC), we aimed to investigate the effects of atorvastatin therapy on endothelial function in physically active, HC men.Method and results Eleven physically active, HC males were recruited. Endothelial function [forearm blood flow response to brachial artery infusion of acetylcholine (Ach)] was assessed twice in each subject following atorvastatin or no therapy in a randomized crossover design. In addition, endothelial function was compared with an active, normolipidaemic control group (C). Atorvastatin therapy reduced total and LDL cholesterol, but had no effect on basal blood flow or endothelial function (peak ACh mean difference ± standard error 0.75 ± 1.75 ml min−1 per 100 ml tissue) [95% confidence interval (CI) −3.1, 4.6]. In addition, there was no difference in endothelial function between the HC and C groups (−1.14 ± 2.60 ml min−1 per 100 ml tissue; CI −6.53, 4.25).Conclusion Statin therapy in HC patients with normal endothelial function does not augment endothelial function.
Changes in collagen I and collagen III metabolism in patients with generalized atopic eczema undergoing medium-dose ultraviolet A1 phototherapy
Authors: Mempel M. , Schmidt T. , Boeck K. , Brockow K. , Stachowitz S. , Fesq H. , Schäfer T. , Thomsen S. , Schnopp C. , Ring J. , Probst R. , Luppa P. , Abeck D. 
Published: March 2003

AbstractFourteen patients suffering from acute, exacerbated atopic eczema were screened for changes in collagen I and collagen III metabolism in serum (n = 11), urine (n = 11) and skin biopsies (n = 9) before and after medium-dose ultraviolet (UV) A1 phototherapy (15 exposures of 50 J/cm2 over a 3-week period, total dose 750 J/cm2). Mature collagen I and, to a lesser extent, mature collagen III were found to be decreased after the therapy in skin samples from the irradiated patients. As markers of collagen I degradation, the cross-links pyridoline and deoxypyridoline were analysed in urine using high-performance liquid chromatography. Both cross-links were found to be mildly increased after UVA1 phototherapy, without reaching statistical significance. As markers of de novo collagen synthesis we screened for the procollagen I-carboxyterminal peptide (PICP) and procollagen III-aminoterminal peptide (PIIINP) levels in serum and skin. The ratio of PICP to PIIINP in serum dropped significantly after the UVA1 phototherapy, suggesting a different impact of UVA1 on the two collagens. These findings were paralleled by a diminished ratio of PICP to PIIINP in tissue samples. Staining for matrix metalloproteinase 1 (MMP-1) and its specific counterpart, tissue inhibitor of MMP-1 (TIMP-1), showed slight increases for both proteins by therapeutic UVA1; this was also seen in serum for TIMP-1 but not MMP-1. In our study, high-energy UVA1 doses induced changes of the skin collagens in patients with atopic eczema which are measurable by their metabolites in serum and urine.
The treatment of atopic dermatitis with adjunctive high-dose intravenous immunoglobulin: a report of three patients and review of the literature
Authors: Jolles S. , Hughes J. , Rustin M. 
Published: March 2003

Abstract
There are few reports of the use of high-dose intravenous immunoglobulin (hdIVIg) in the treatment of atopic dermatitis (AD). We describe our experience using this therapy in three patients with severe AD, all of whom had steroid-related side-effects. These patients received either Alphaglobin® or Sandoglobulin® 2 g/kg monthly: all had improved skin scores, allowing reduction of their steroid dose. Total IgE fell in one of three patients. We discuss the side-effects of hdIVIg and their management, and detail the differences between the available immunoglobulin products available in the U.K. There are several proposed mechanisms of action of this therapy which may be operative, and those most important in AD are discussed. In view of the time and expense involved in the treatment of patients with hdIVIg, careful patient assessment is vital. We describe dose reduction strategies and methods for cost containment. In addition, one of the patients has embarked on IVIg home therapy training. This will be the first time this has been attempted for a dermatological indication. Training of this type may be available through an immunotherapy service such as exists for patients with primary immunodeficiencies.
Regulation of the uPAR/uPA system expressed on monocytes by the deactivating cytokines, IL‐4, IL‐10 and IL‐13: consequences on cell adhesion to vitronectin and fibrinogen
Authors: Paysant Jérôme , Vasse Marc , Soria Jeannette , Lenormand Bernard , Pourtau Jérôme , Vannier Jean‐Pierre , Soria Claudine 
Published: March 2003

Urokinase (uPA) and its receptor (uPAR) have been proposed to be involved in monocyte migration by inducing degradation of matrix proteins. In addition, uPAR is also implicated in cell adhesion to the vascular wall. The adhesive function of uPAR depends on a direct interaction with vitronectin which is increased by uPA and by modification of cell surface integrin (such as CD11b–CD18) when associated to uPAR. In this study we analysed the role of three deactivating cytokines, IL‐4, IL‐10 and IL‐13, on the surface expression of uPA, uPAR and CD11b by monocytes and their consequences on monocyte adhesion to immobilized fibrinogen and vitronectin.IL‐10 induced a decrease in uPA and CD11b after 18 h incubation and a delayed decrease in uPAR which was only significant after 48 h incubation. These results may explain the decrease in monocyte adhesion, which was observed after an 18 h incubation with IL‐10, on immobilized vitronectin and fibrinogen. In contrast, IL‐4 and IL‐13 induced a decrease in uPAR after 18 h and a significant increase in uPA both in the cell lysates and at the cell surface, as well as an increase in cell surface associated CD11b. These cytokines did not modify cell adhesiveness to vitronectin or fibrinogen despite the increase in CD11b–CD18. This could be due to the decrease in uPAR because CD11b–CD18/uPAR forms a cell adhesion complex. In addition, the increase in uPA induced by IL‐4 could counterbalance the direct interaction of uPAR with vitronectin. The increase in uPA suggests that IL‐4 and IL‐13 could induce plaque fissuring by monocytes, whereas IL‐10 may induce protection against matrix protein degradation by decreasing uPA.
Comparative adhesion of human haemopoietic cell lines to extracellular matrix components, bone marrow stromal and endothelial cultures
Authors: Turner Marc L. , Masek Lisa C. , Hardy Cheryl L. , Parker Alistair C. , Sweetenham John W. 
Published: March 2003

We used flow cytometry to characterize cell adhesion molecule expression of the human haemopoietic cell lines KG1a, K562, HL‐60, NALM‐6 and CEM. A 51chromium labelling assay was used to study the adhesion of these cell lines to extracellular matrix components and to bone marrow stromal and endothelial cultures. Both adhesion molecule expression and functional binding behaviour varied between cell lines. All five cell lines expressed the integrins α4β1 and α5β1 and all adhered to fibronectin. However, differences in intensity of expression of these integrins failed to correlate with extent of fibronectin adhesion. Inhibition experiments demonstrated that adhesion of KG1a to fibronectin was completely inhibited by divalent cation chelation and partially inhibited by RGDS peptides and chondroitinase ABC, suggesting that both α4β1 and α5β1 as well as CD44 were responsible for this interaction. Adhesion to bone marrow stromal and endothelial layers was superior to that to purified extracellular matrix components and was partially inhibited by divalent cation chelation. RGD peptides and anti‐α4 monoclonal antibody also partially inhibited KG1a adhesion to bone marrow endothelium.Discordance between cell adhesion molecule expression and adhesive behaviour suggest that current phenotypic descriptions remain incomplete and reinforce the need for complementary functional binding studies.
Minimal residual disease analysis for the prediction of relapse in children with standard‐risk acute lymphoblastic leukaemia
Authors: Goulden Nicholas J. , Knechtli Christopher J. C. , Garland Russell J. , Langlands Kenneth , Hancock Jeremy P. , Potter Michael N. , Steward Colin G. , Oakhill Anthony 
Published: March 2003

We report a largely retrospective analysis of minimal residual disease (MRD) in a cohort of 66 children suffering from acute lymphoblastic leukaemia (ALL). All patients lacked high‐risk features at diagnosis, i.e. the presenting white cell count was <50 × 109/l, age 1–16 years and translocations t(9;22) and t(4;11) were not present. All were treated according to either the MRC protocols UKALL X or XI. PCR of IgH, TCRδ and TCRγ gene rearrangements and allele‐specific oligoprobing were employed for the detection of MRD. Sensitivity was at least 10−4 in 78/82 (93%) probes examined.A total of 33 patients relapsed (seven on therapy and 26 off) and 33 remain in continuing complete remission (CCR) (median follow‐up 69 months from diagnosis). Of those who remain in CCR, MRD was present in the bone marrow in 32%, 10% and 0% at 1, 3 and 5 months into therapy respectively. This is in marked contrast to the presence of MRD at these times in 82%, 60% and 41% of patients who relapsed (P<0.001, P<0.005 and P<0.005). These results provide further evidence of a strong correlation between clearance of MRD early in therapy and clinical outcome in childhood ALL.
Factors which affect the CFU‐GM content of the peripheral blood haemopoietic progenitor cell harvests in patients with acute myeloid leukaemia
Authors: Jowitt  , Chang  , Morgenstern  , Howe  , Ryder  , Testa  , Scarffe  
Published: March 2003

Autologous peripheral blood haemopoietic stem cells (PBSC) were harvested from 30 patients withde novoacute leukaemia, 29 of whom had entered remission following standard chemotherapy. Correlation of CD34+cells/kg to CFU‐GM/kg in the harvests was good (correlation coefficient=0.72,P<0.001). We demonstrated significant associations between the CFU‐GM content of the harvest and the following: time to platelets >50×109/l post final induction course (P<0.0001), days to harvest from day 1 of intensification/mobilization (correlation coefficient=−0.73,P<0.001), platelets >20×109/l at time of harvest (P=0.02), time to WBC >1.0×109/l post intensification/mobilization (correlation coefficient=−0.70,P<0.001), and WBC on day of harvest (correlation coefficient=0.60,P<0.001). In contrast, we found no relationship between the CFU‐GM content of the harvest and patient age up to 65 years, presence of absence of coexistent features of trilineage myelodysplasia at diagnosis, number of induction courses to remission or total number of courses of chemotherapy prior to intensification/mobilization. Haemopoietic recovery after reinfusion of PBSC was highly correlated to the number of CFU‐GM infused (neutrophils >0.5×109/lrs=−0.72,P=0.001; platelets >20×109/l unsupportedrs=−0.71,P=0.001).Our results show that the number of induction courses received, and thus exposure to cytotoxic agents received, made no significant difference to subsequent CFU‐GM harvest content. We collected superior harvests from those patients with faster platelet recovery following mobilization therapy. We also found that faster platelet recovery following the final induction therapy was a better predictor of the CFU‐GM harvest following mobilization than was the neutrophil recovery following final induction.
Cytotoxic effectors activated by low‐dose IL‐2 plus IL‐12 lyse IL‐2‐resistant autologous acute myeloid leukaemia blasts
Authors: Vitale Antonella , Guarini Anna , Latagliata Roberto , Cignetti Alessandro , Foa Robert 
Published: March 2003

We have investigated the susceptibility of primary acute myeloid leukaemia (AML) samples to the lytic action of normal peripheral blood mononuclear cells (PBMC), as well as of the patients' autologous and allogeneic PBMC collected at the time of remission following stimulation with different concentrations of interleukin (IL)‐2 and IL‐12, both alone and in variable combinations. Primary AML blasts were resistant to IL‐2‐activated PBMC effectors generated from normal individuals, allogeneic and autologous patients in five, six and eight of the 10 AML samples tested, respectively. IL‐12 alone proved ineffective in generating anti‐leukaemic activity, whereas, in combination, the two cytokines induced anti‐leukaemic killing in all five cases resistant to normal PBMC, in 4/6 samples resistant to allogeneic AML effectors and in 5/8 cases resistant to autologous effectors. In each case the lytic effect was maintained at the lowest cytokine combination (10 + 10 IU/ml) utilized. The suggestion of a synergistic effect was further strengthened by the evidence that at the lowest doses of IL‐2 and IL‐12 the degree of killing was greater than that promoted by each cytokine independently.The results of this study suggest that two major limitations associated with the administration of IL‐2 to AML patients, the resistance of the blasts to IL‐2‐generated killing and the toxicity associated with high‐dose IL‐2, may be overcome by the combined use of very low doses of IL‐2 and IL‐12. As IL‐2‐resistant blasts may be lysed by a low‐dose combination of IL‐2/IL‐12, feasibility studies with this cytokine combination are worthy of exploration in vivo.
Mutation analysis by a non‐radioactive single‐strand conformation polymorphism assay in nine families with X‐linked severe combined immunodeficiency (SCIDX1)
Authors: Wengler Georg S. , Giliani Silvia , Fiorini Maurilia , Mella Patrizia , Mantuano Elide , Zanola Alessandra , Pollonini Gabriella , Eibl Martha M. , Ugazio Alberto G. , Notarangelo Luigi D. , Parolini Ornella 
Published: March 2003

X‐linked severe combined immunodeficiency (SCIDX1) is an inherited disease characterized by profound abnormalities of cell‐mediated and humoral immunity. Patients with SCIDX1 have defects in the common cytokine receptor gamma chain gene (IL2RG) that encodes a shared, essential component of the receptors for interleukin‐2 (IL‐2), IL‐4, IL‐7, IL‐9 and IL‐15. We have characterized nine SCIDX1 families by using a DNA‐based, non‐radioactive screening method and DNA sequencing. Nine different mutations were found, scattered from exon 1 to exon 5 of the IL2RG gene. Two of these mutations have been previously identified in other unrelated patients; the other seven are novel mutations that differ from all of the 95 already reported in the IL2RG mutation data base.In addition to describing novel mutations in the IL2RG gene, this study shows that the knowledge of the genetic defect and the use of an efficient, non‐radioactive, and rapid screening approach have important implications for prenatal and postnatal diagnosis, carrier female identification, and possibly prenatal therapy.
Haemopoietic defect and decreased expansion potential of bone marrow autografts from patients with acute myeloid leukaemia in first remission
Authors: Straetmans  , Herman  , Van Bockstaele  , Michaux  , Hagemeijer  , Ferrant  
Published: March 2003

Autologous bone marrow (BM) transplantation for acute myeloid leukaemia (AML) in complete remission (CR) is frequently followed by a slow haemopoietic recovery. We assessed the haemopoietic capacity of purified BM stem cell (CD34+DR−) and progenitor cell (CD34+DR+) populations from patients with AML in CR, and compared these data with those of normal BM. The feasibility of ex vivo expansion in stroma‐conditioned medium supplemented with cytokines was also investigated.The number of CFU‐GM produced by initial patient CD34+DR− cells was decreased compared to normal, whereas these values were similar to normal for CD34+DR+ cells. BFU‐E, HPP‐CFC and LTC‐IC were reduced for both patient CD34+DR− and CD34+DR+ subpopulations. In contrast to normal, the patient CD34+DR− fraction was not enriched in LTC‐IC.CFU‐GM expansion from patient CD34+DR− cells was poor and decreased after 14 d of culture. No HPP‐CFC expansion could be observed for patient cells. LTC‐IC were below the level of detection after 14–21 d of expansion culture of CD34+DR− patient cells, whereas they were variably maintained or expanded for normal cells. After expansion culture, cytogenetic and/or FISH analyses did not reveal the anomalies present at diagnosis, regardless of the cell subpopulation analysed.In conclusion, BM cells of patients with AML in CR show a profound defect at the level of a stem cell enriched population. No meaningful ex vivo expansion could be obtained in culture conditions allowing for a significant expansion from a normal stem cell population.
Tumourigenicity of MTG8, a leukaemia‐related gene, in concert with v‐Ha‐ras gene in BALB/3T3 cells
Authors: Sueoka Eisaburo , Sueoka Naoko , Okabe Sachiko , Komori Atsumasa , Suganuma Masami , Kozu Tomoko , Fujiki Hirota 
Published: March 2003

The MTG8 (ETO) gene has been identified as the translocation partner of AML1 (PEBP2αB or CBFα2) gene in the AML1/MTG8 (ETO) fused gene caused by t(8;21) translocation in human acute myeloid leukaemia, M2 type. Although AML1/MTG8 chimaeric protein is known to inhibit the functioning of AML1 protein, the precise function of MTG8 gene itself is not known yet. We studied the significance of MTG8 gene in the oncogenicity of AML1/MTG8 fused gene, by introducing full‐length MTG8 cDNA into both BALB/3T3 cells containing v‐Ha‐ras gene (Bhas 42 cells) and BALB/3T3 cells without v‐Ha‐ras gene.Irrespective of the overexpression of MTG8 gene in both groups of cells, Bhas‐MTG8 clones which contained v‐Ha‐ras gene and expressed the MTG8 gene at a level more than twice that of parental Bhas 42 cells induced cell transformation, whereas BALB‐MTG8 clones without v‐Ha‐ras gene did not. Furthermore, injection of the transformed Bhas‐MTG8 clones into the subcutaneous tissue of nude mice induced tumours, whereas that of BALB‐MTG8 clones did not. These results suggest that MTG8 gene product, in cooperation with viral Ras protein, resulted in tumour formation. We provide the first evidence that MTG8 gene by itself has a carcinogenic property within the AML1/MTG8 (ETO) fused gene.