bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of plasma DNA in detection of loss of heterozygosity in patients with multiple myeloma
Authors: Ahmed Maha , Giles Francis , Joe Youngson , Weber Donna M. , Jilani Iman , Manshouri Taghi , Giralt Sergio , Lima Marcos De , Keating Michael , Albitar Maher
Published: August 2003

Background: Deletions or structural abnormalities in chromosomes 11 and 13 have been shown to be important in predicting clinical behavior in patients with multiple myeloma (MM). However, cytogenetic analysis in MM is frequently difficult because of poor yield of informative metaphases and the disease is frequently patchy, which complicates fluorescent in situ hybridization studies.

Objectives: The purpose of this study was to explore the potential of using peripheral plasma DNA for the detection of loss of heterozygosity (LOH) in chromosomes 11 and 13 in patients with MM.

Methods: Peripheral blood (PB) plasma of 81 patients with MM, was used as a source of DNA for the detection of LOH at chromosomes 13q14 (D13S319 and D13AFMaw301wb5), and 11q21 (D11S2179) using polymerase chain reaction.

Results: Only 62 of the studied patients were informative for the two 13q microsatellite markers and 16 (26%) of these patients showed LOH. Only seven (11%) of 61 patients with informative D11S2179 microsatellite maker showed LOH. Purified plasma cells (PCs) from six bone marrow (BM) samples using anti‐CD138‐coated magnetic beads showed identical results to those detected in DNA isolated from PB plasma. Three patients with LOH underwent autologous BM transplantation, and two of three reverted to a normal state (no LOH) after transplantation. Seven of the patients with 13q LOH in PB plasma had <10% PCs (PCs) in their BM at the time of testing.

Conclusion: PB plasma appears to be enriched by tumor‐specific DNA and can be used to detect chromosomal abnormalities in patients with MM. Further studies are needed to establish the clinical relevance of this approach in comparison with other techniques.
A clinical and molecular study of a Bedouin family with dysmegakaryopoiesis, mild anemia, and neutropenia cured by bone marrow transplantation
Authors: Tamary H. , Yaniv I. , Stein J. , Dgany O. , Shalev Z. , Shechter T. , Resnitzky P. , Shaft D. , Zoldan M. , Kornreich L. , Levy R. , Cohen A. , Moser R. A. , Kapelushnik J. , Shalev H.
Published: August 2003

Objectives: Familial thrombocytopenia is a relatively rare and heterogeneous group of clinical and genetic syndromes of unknown etiology. Recently, mutations in a few hematopoietic transcription factors were implicated in dysmegakaryopoiesis with and without dyserythropoietic anemia. The aim of the present study was to describe the clinical and hematologic picture of members of a Bedouin family with severe congenital thrombocytopenia associated with neutropenia and anemia and to determine the possible involvement of hematopoietic transcription factor genes in their disease.

Patients and methods: Four members of a Bedouin family presented with severe bleeding tendency, including intracranial hemorrhage in three. Three of the four were successfully treated with allogenic human leukocyte antigen (HLA)‐matched bone marrow transplants. Measurements of serum erythropoietin and thrombopoietin levels, bone marrow electron microscopy, and megakaryocytic colony were grown for each patient in addition to DNA amplification and single‐strand conformation polymorphism of each exon of the NF‐E2, Fli‐1, FOG‐1, and Gfi‐1b in genes.

Results: Bone marrow studies revealed dysmegakaryopoiesis and mild dyserythropoiesis. A low number of bone marrow megakaryocyte colony‐forming units was found, as well as a slightly elevated serum thrombopoietin level. No mutation was identified in any of the transcription factor genes examined.

Conclusions: A unique autosomal recessive bone marrow disorder with prominent involvement of megakaryocytes is described. Defects were not identified in transcription factors affecting the common myeloid progenitor.
The angioregulatory phenotype of native human acute myelogenous leukemia cells: influence of karyotype, Flt3 abnormalities and differentiation status
Authors: Glenjen Nils , Hovland Randi , Wergeland Line , Wendelbo Øystein , Ernst Peter , Bruserud Øystein
Published: August 2003

Introduction: The cytogenetic abnormalities and the response to induction therapy have been regarded as the most important prognostic parameters in acute myelogenous leukemia (AML) patients. Recent studies have demonstrated that internal tandem duplications and specific D‐835 point mutations of the Flt3 gene, as well as the angioregulatory phenotype represent additional adverse prognostic factors. The aim of the study was to investigate possible associations between genetic abnormalities, differentiation status and angioregulatory phenotype in native human AML blasts.

Method: Native AML blasts derived from consecutive patients were cultured in vitro and concentrations of angioregulatory molecules determined in the supernatants.

Results: Most patients released at least two different angioregulatory mediators. Pro‐angiogenic interleukin 8 (IL8) was released at relatively high levels for most patients, many of these patients showed additional release of pro‐angiogenic vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). High release of anti‐angiogenic IL12 was associated with high release of pro‐angiogenic IL8 and VEGF. Furthermore, patients with D‐835 mutations showed increased IL12 release, whereas patients with normal karyotype had decreased HGF release. Myelomonocytic differentiation was associated with IL18 release and CD34 expression with low IL12 release.

Conclusion: Our results suggest that native human AML blasts have a pro‐angiogenic phenotype. Although the investigated genetic abnormalities are associated with variation in the in vitro release of angioregulators, these differences are relatively small and do not quantitatively involve the most important IL8 release. It therefore seems unlikely that this phenotypic variation can explain the prognostic impact of the genetic abnormalities.
Core binding factor (CBF) acute myeloid leukemia: is molecular monitoring by RT–PCR useful clinically?
Authors: Marcucci Guido , Caligiuri Michael A. , Bloomfield Clara D.
Published: August 2003

Abstract: Clonal chromosomal abnormalities are the most important prognostic indicators in acute myeloid leukemia (AML). Two of the most prevalent cytogenetic subtypes of adult primary AML, t(8;21)(q22;q22) and inv(16)(p13q22)/t(16;16)(p13;q22), are characterized by disruption of the AML1(CBFA2, RUNX1) and CBFβ genes, respectively, which encode subunits of core binding factor (CBF), a regulator of normal hematopoiesis. At the molecular level, t(8;21) and inv(16)/t(16;16) result in the creation of novel fusion genes, AML1/ETO and CBFβ/MYH11, respectively, which encode fusion transcripts readily detectable by the reverse transcription–polymerase chain reaction (RT–PCR). Although the detection of t(8;21) or inv(16)/t(16;16) in adult patients with primary AML represents a favorable independent prognostic indicator for achievement of cure following intensive chemotherapy or stem cell transplantation, a substantial number of these patients (i.e. 40–50%) relapse and eventually die of their disease. Therefore, timely identification and therapeutic stratification of those patients deemed at high risk for disease relapse could ultimately result in a further improvement of clinical outcome within these cytogenetic subgroups of AML. As relapse is likely to occur as the result of failure of treatment to completely eradicate leukemic blasts, the detection of the AML1/ETO and CBFβ/MYH11 fusion transcripts using sensitive RT–PCR assays has been utilized as a surrogate marker for resistant disease and, in turn, to predict disease recurrence during remission. The purpose of this paper is to review the applicability of this strategy to the clinical management of t(8;21) and inv(16)/t(16;16) primary AML, here collectively referred to as CBF AML.
Review article: the role of rebamipide in the management of inflammatory disease of the gastrointestinal tract
Authors: Genta R. M.
Published: August 2003

Rebamipide stimulates the generation of endogenous prostaglandins in the gastric mucosa and is reported to accelerate ulcer healing. This review discusses whether rebamipide can prevent Helicobacter pylori infection, reduce inflammation, accelerate healing after eradication, promote ulcer healing, and prevent progression of preneoplastic lesions. Furthermore, we evaluate its usefulness in other inflammatory conditions of the gastrointestinal tract. We conclude that rebamipide is an important candidate for long‐term suppression of gastro‐intestinal inflammation, praticularly if reducing the complications of H. pylori infection without eradicating the organism becomes accepted. If its ability to accelerate mucosal normalization is confirmed, rebamipide could be added to eradication regimens. Little information exists on whether such therapy could help limit the development of pre‐neoplastic lesions. In light of the dearth of effective drugs to control inflammation in idiopathic inflammatory bowel disease, the potential of any promising new and safe compound deserves to be fully explored. The next step is to devise a targeted plan of translational research, so that results from the bench may be used to design rigorously controlled international clinical trials.
Review article: rebamipide and the digestive epithelial barrier
Authors: Matysiak‐Budnik T. , Heyman M. , Mégraud F.
Published: August 2003

Rebamipide exerts a positive effect on the digestive epithelial barrier by reinforcing its integrity in normal and in inflammatory conditions, and by normalizing the macromolecular transport across this barrier, increased by Helicobacter infection. Moreover, in mice, rebamipide is capable of diminishing allergic sensitization and of counteracting the inhibitory effect of Helicobacter pylori on oral tolerance to dietary antigens. These properties of rebamipide could explain its anti‐inflammatory activity with respect to the digestive mucosa and could provide protection against allergic sensitization to foreign antigens in susceptible individuals.
Early Middle Miocene broken foreland development in the southern Central Andes: evidence for extension prior to regional shortening
Authors: Dávila Federico M. , Astini Ricardo A.
Published: August 2003

Most authors suggest that the main contraction phases in the southern Central Andes started in the Late Miocene. Along the flat‐slab segment, deformation has progressively involved basement and a broken foreland has developed. Recent work suggests that construction of the Andes by late Neogene shortening may have been controlled by lithospheric thinning and crustal structure generated during mid‐Tertiary times in the Southern and Central Andes. Exposures at the eastern border of the Famatina Ranges in western Argentina in the flat‐slab segment document basement involved extension of approximately this age. The Del Abra Formation, the lower unit of a major Andean synorogenic cycle (Angulos Group), reveals a distinct and previously unrecognized early Middle Miocene tectonic event. This is suggested by a 505‐m‐thick thinning‐ and fining‐upward megasequence. Dominantly conglomeratic facies record a continuous progression from fault‐scarp‐related high‐gradient colluvium to relatively distal terminal‐fan facies. The fining–thinning upward megasequence characterizes progressive scarp backstepping and decreasing relief after active extension. Interpretation of the stratigraphic fill and the associated structure (high‐angle hinterland‐dipping fault) favours tectonic inversion of an originally normal fault. This allows reappraisal and new understanding of the early‐stage architecture of the Central Andean foreland. Early Middle Miocene extension may have had an important bearing on the later evolution of the broken foreland.
Synclinal‐horst basins: examples from the southern Rio Grande rift and southern transition zone of southwestern New Mexico, USA
Authors: Mack Greg H. , Seager William R. , Leeder Mike R.
Published: August 2003

In areas of broadly distributed extensional strain, the back‐tilted edges of a wider than normal horst block may create a synclinal‐horst basin. Three Neogene synclinal‐horst basins are described from the southern Rio Grande rift and southern Transition Zone of southwestern New Mexico, USA. The late Miocene–Quaternary Uvas Valley basin developed between two fault blocks that dip 6–8° toward one another. Containing a maximum of 200 m of sediment, the Uvas Valley basin has a nearly symmetrical distribution of sediment thickness and appears to have been hydrologically closed throughout its history. The Miocene Gila Wilderness synclinal‐horst basin is bordered on three sides by gently tilted (10°, 15°, 20°) fault blocks. Despite evidence of an axial drainage that may have exited the northern edge of the basin, 200–300 m of sediment accumulated in the basin, probably as a result of high sediment yields from the large, high‐relief catchments. The Jornada del Muerto synclinal‐horst basin is positioned between the east‐tilted Caballo and west‐tilted San Andres fault blocks. Despite uplift and probable tilting of the adjacent fault blocks in the latest Oligocene and Miocene time, sediment was transported off the horst and deposited in an adjacent basin to the south. Sediment only began to accumulate in the Jornada del Muerto basin in Pliocene and Quaternary time, when an east‐dipping normal fault along the axis of the syncline created a small half graben. Overall, synclinal‐horst basins are rare, because horsts wide enough to develop broad synclines are uncommon in extensional terrains. Synclinal‐horst basins may be most common along the margins of extensional terrains, where thicker, colder crust results in wider fault spacing.
Stratigraphic evidence for a Late Devonian possible back‐bulge basin in the Appalachian basin, United States
Authors: Filer Jonathan K.
Published: August 2003

Isopach and sedimentary facies maps of Upper Devonian (upper Frasnian and lower Famennian) strata deposited in a part of the central Appalachian foreland basin (eastern United States) during the Acadian orogeny show a significant change in depositional style over time. Maps of two successive upper Frasnian intervals show steady thickening to the east towards the hinterland. Coarser‐grained sediment was deposited in distinct tongues in front of the Augusta lobe, a previously recognized locus of sediment input in the central Appalachian basin. Maps of two subsequent lower Famennian stratigraphic intervals show distinct depocentres in the study area. Famennian strata thin eastward (by about 50%) over a distance of about 90 km from these depocentres to the limit of mapping at the Allegheny structural front. This is towards the Acadian sediment source and in contrast to general Upper Devonian thickening in that direction. The axes of these lower Famennian depocentres are stacked on top of each other. Also, coarser‐grained lower Famennian sediment is concentrated in strike trends just east of the axes of the depocentres, and no coarser tongues exist in front of the Augusta lobe, in contrast to the underlying (upper Frasnian) strata.

The duration of each of the four study intervals is estimated to be between 0.5 and 3.0 Myr. The early Famennian depocentres may be in a back‐bulge basin, with a forebulge uplifted to the east of the study area. Earlier deposition may have occurred in a basin with a subtle, subdued, and longer wavelength forebulge (perhaps located west of the study area). Previously published regional isopachs of Upper Devonian strata suggest that the main axis of subsidence of the Acadian foreland basin (foredeep depozone) at this time was over 350 km east of the study area. Examination of published quantitative flexural models of other foreland basins with flexural rigidities close to published rigidities calculated for the Appalachian basin suggests that the proposed back‐bulge basin is in the correct location, relative to the suggested position of the foredeep at that time. Several previously recognized structural features of the northern Appalachian basin support the interpretations presented herein. Much of the Acadian foreland basin may be eroded in the central Appalachian basin. The present study demonstrates the difficulties in recognizing foreland basin depozones in partially preserved orogens.
Insights into extramedullary tumour cell growth revealed by expression profiling of human plasmacytomas and multiple myeloma
Authors: Hedvat Cyrus V. , Comenzo Raymond L. , Teruya‐Feldstein Julie , Olshen Adam B. , Ely Scott A. , Osman Keren , Zhang Yana , Kalakonda Nagesh , Nimer Stephen D.
Published: August 2003

Summary. Malignant plasma cells generally grow within the bone marrow microenvironment; however, they can also grow at extramedullary sites. To identify the tumour‐specific alterations required for extramedullary growth, we analysed the expression profiles of a series of plasma cell neoplasms including primary multiple myeloma (MM), plasma cell leukaemia (PCL) and extramedullary plasmacytoma (EPC). Hierarchical clustering analysis segregated the EPCs from the remaining samples, and revealed an expression pattern associated with angiogenesis in the EPCs, involving higher expression of the genes TIE2, NOTCH3, CD31 and endoglin. Direct comparison of EPC samples with the MM samples identified 156 genes significantly upregulated and 85 genes significantly downregulated (P < 0·005, t‐test) in the EPCs, including several genes involved in angiogenesis and adhesion that were upregulated (including angiopoietin 1, SPARC, Notch3 and fibronectin 1). Immunohistochemical staining demonstrated CD31 and endoglin protein expression in the EPC tumour cells, which are both angiogenesis related and could confer malignant plasma cells with the ability to grow outside the normal bone marrow environment. Defining how malignant plasma cell growth is regulated in the bone marrow versus at extramedullary sites will help to delineate the mechanisms underlying the dependence of tumour cell growth on angiogenesis and cell adhesion.
Chromosome 11 rearrangements and specific MLL amplification revealed by spectral karyotyping in a patient with refractory anaemia with excess of blasts (RAEB)
Authors: Calabrese Giuseppe , Fantasia Donatella , Morizio Elisena , Toro Patrizia M. , Franchi Paolo Guanciali , Fornaro Antonella , Spadano Antonio , Stuppia Liborio , Palka Giandomenico
Published: August 2003

Summary. A patient with refractory anaemia with excess of blasts (RAEB) had a complex karyotype with multiple markers. Spectral karyotyping (SKY) showed rearrangements including three different der(11), containing a very high number of MLL gene copies, shown by fluorescence in situ hybridization (FISH) analysis. Fibre‐FISH experiments disclosed the presence of chromatin fibres with multiple MLL copies with a head‐to‐tail pattern. Apparently, no other region flanking the MLL site was present in the three der(11). MLL amplification was confirmed by the reverse transcription polymerase chain reaction (RT‐PCR). The patient died 6 months after diagnosis, supporting the severe prognosis of sole MLL amplification.
Patient self‐management of oral anticoagulation and external quality assessment procedures
Authors: Murray E. T. , Kitchen D. P. , Kitchen S. , Jennings I. , Woods T. A. L. , Preston F. E. , Fitzmaurice D. A.
Published: August 2003

Summary. The role of external quality assessment (EQA) is a contentious issue for patient self‐management (PSM) of oral anticoagulation. Patients from general practices in the West Midlands undertaking PSM were recruited to compare efficacy of patients' and health professionals' EQA procedure using the UK National External Quality Assessment Scheme (NEQAS). Patients using Coaguchek (Roche Diagnostics) were trained to perform EQA as part of their PSM training. They undertook PSM for 26 weeks and were asked to perform EQA using material provided by the UK NEQAS twice at home without supervision and twice at the practice with supervision. Patients' results were compared with health care professional users of Coaguchek S. Twenty‐three PSM patients were compared with 75 health care professional users of the NEQAS scheme. The PSM group international normalized ratio (INR) percentage time in range was 74%. There was no significant difference in the median results on NEQAS samples obtained by the patients and those obtained by professionals. Three patients were outwith consensus (results > 15% from the median INR) on more than one occasion. Patients were able to perform the EQA tests competently. The data show that good agreement can be achieved between patients analysing the same EQA samples, with coefficients of variation ranging from 22·3% to as low as 5·4%. Further study is required to determine how precision within these EQA schemes relates to the stability of treatment in patients' management of their own anticoagulation.
Long‐term administration of pegylated recombinant human megakaryocyte growth and development factor dramatically improved cytopenias in a patient with myelodysplastic syndrome
Authors: Kizaki Masahiro , Miyakawa Yoshitaka , Ikeda Yasuo
Published: August 2003

Summary. To date, there are no curative therapeutic options for patients with myelodysplastic syndromes (MDS) other than allogeneic stem cell transplantation. We treated an MDS patient with 10 µg/kg pegylated recombinant human megakaryocyte growth and development factor (rHuMGDF) for more than 450 d. The patient's platelet counts increased from < 10 × 109/l to 50 × 109/l. Interestingly, haemoglobin levels increased dramatically and reached over 13 g/dl without additional transfusion. Adverse events and neutralizing antibodies were not observed during treatment, suggesting that the long‐term administration of rHuMGDF might be of clinical benefit to patients with MDS.
Novel 112 kb (εGγAγ) δβ‐thalassaemia deletion in a Dutch family
Authors: Harteveld Cornelis L. , Osborne Cameron S. , Peters Marjolein , Van Der Werf Steffie , Plug Rob , Fraser Peter , Giordano Piero C.
Published: August 2003

Summary. An adult autochthonous Dutch patient who had exhibited severe perinatal anaemia, with partial recovery a few months after birth, was studied for the presence of β‐thalassaemia. Southern blotting showed that the patient was heterozygous for a novel deletion in the β‐globin gene cluster, leaving the β‐gene intact. Inverse polymerase chain reaction was used to determine the breakpoint sequence. The deletion removed 112 kb starting upstream of the HOR5′b6 gene to the second intron of the Aγ‐globin gene, including the locus control region. The breakpoint fragment identified a 13‐bp orphan sequence not present at either side of the breakpoint.
The IgG subclasses of platelet‐associated autoantibodies directed against platelet glycoproteins IIb/IIIa in patients with idiopathic thrombocytopenic purpura
Authors: Chan Howard , Moore Jane C. , Finch Clara N. , Warkentin Theodore E. , Kelton John G.
Published: August 2003

Summary. The majority of patients with idiopathic thrombocytopenic purpura (ITP) have antiplatelet autoantibodies that are most frequently directed against platelet glycoproteins IIb/IIIa or Ib/IX/V. However, there is some debate whether the immune response is oligoclonal or polyclonal in nature. We investigated the subclass distribution of anti‐IIb/IIIa IgG autoantibodies in 59 prospectively studied patients with ITP. We also tested patients with a variety of thrombocytopenic disorders (n = 31) and healthy controls (n = 30). Platelet lysates were tested for IgG anti‐IIb/IIIa autoantibodies, and the specific IgG subclass distribution was measured using antigen capture assays. All testing was done blinded to diagnosis and other assay results. After unblinding, we found that 43 of the 59 ITP patients had anti‐IIb/IIIa autoantibodies (sensitivity = 73%). Anti‐IIb/IIIa autoantibodies were also detected in five of the 31 non‐ITP patients, but in none of the 30 healthy controls (specificity = 91%). The IgG subclass assay was positive in 39 of the 43 ITP patients with anti‐IIb/IIIa antibodies (sensitivity = 92%) and in 12 samples that had no detectable anti‐IIb/IIIa antibodies including two ITP patients (specificity = 83%). The most common subclass in the ITP patient samples was IgG1 (77%), either alone (n = 14) or with other IgG subclass antibodies (n = 19). However, there were also patients with only IgG2 (n = 2), IgG3 (n = 3) or IgG4 (n = 3) antibodies. Our results are consistent with the hypothesis that ITP is a heterogeneous disorder and that some patients have evidence of oligoclonality, whereas other patients have polyclonal autoantibodies.
Defective function of Fas in T cells from paediatric patients with autoimmune thyroid diseases
Authors: BONA G , DEFRANCO S , CHIOCCHETTI A , INDELICATO M , BIAVA A , DIFRANCO D , DIANZANI I , RAMENGHI U , CORRIAS A , WEBER G , DE SANCTIS V , IUGHETTI L , RADETTI G , DIANZANI U
Published: August 2003

Triggering of the Fas receptor induces T cell apoptosis and is involved in shutting-off the immune response. Inherited defects impairing Fas function cause the autoimmune lymphoproliferative syndrome, and may play a role in other autoimmune diseases. The aim of this work was to analyse the Fas function in paediatric patients with thyroid autoimmunities. We found that T cells from 24/28 patients with Graves’ disease (GD) and 12/35 patients with Hashimoto's thyroiditis (HT) displayed defective Fas function. In HT, the defect was more frequent in patients requiring replacement therapy (11/20) than in those not requiring (1/15); moreover, in untreated HT the highest defect was displayed by patients with the highest levels of autoantibodies. Fas was always expressed at normal levels and no Fas mutations were detected. Analysis of the healthy parents of seven Fas-resistant patients showed that several of them were Fas-resistant, which suggests a genetic component. Fusion of Fas-resistant T cells with the Fas-sensitive HUT78 T cell line generated Fas-resistant hybrid cells, which suggests the presence of molecules exerting a dominant negative effect on Fas function. Analysis of Fas-induced activation of caspase-8 and -9 showed decreased activity of both caspases in HT, whereas activity of caspase-9 was increased and that of caspase-8 was decreased in GD. These data suggest that heterogeneous inherited defects impairing Fas function favour the development of thyroid autoimmunities.
Binding of live conidia of Aspergillus fumigatus activates in vitro-generated human Langerhans cells via a lectin of galactomannan specificity
Authors: PERSAT F , NOIREY N , DIANA J , GARIAZZO M-J , SCHMITT D , PICOT S , VINCENT C
Published: August 2003

Aspergillus fumigatus is the most common aetiological fungus responsible for human pulmonary aspergilloses. This study investigated the primary contact between Langerhans cells (LC), corresponding to dendritic cells present in pulmonary mucosa and live conidia of A. fumigatus. LC play a key role in antigen presentation for initiation of the primary T cell response. In vitro-generated LC (iLC) were differentiated from cultured human cord blood CD34+ cells and incubated at 4°C or 37°C with fluorescein-isothiocyanate (FITC)-stained conidia or control latex beads. In vitro, conidia were shown by microscopy and cytometry to adhere to iLC in a dose- and time-dependent manner. This adhesion was not limited to iLC because interstitial dendritic and other cells also fluoresced in the presence of conidia-FITC. A lectin other than mannose receptor-type lectin was demonstrated to be responsible of conidial binding. Inhibition of binding was observed with heterologous galactomannan and EDTA, indicating a C-lectin-like receptor with galactomannan structure specificity. After binding only a few conidia were internalized in acidic vesicles, as indicated by the cessation of conidial fluorescence. Conidial binding was followed by activation and maturation of iLC, suggesting that LC present in the lung may play a role in cellular host defence against aspergilloses.
Marked increase in vascular endothelial growth factor concentrations during Escherichia coli endotoxin‐induced acute inflammation in humans
Authors: Mittermayer F. , Pleiner J. , Schaller G. , Weltermann A. , Kapiotis S. , Jilma B. , Wolzt M.
Published: August 2003

Background  Bacterial endotoxins can induce the synthesis and release of vascular endothelial growth factor (VEGF), which may alter vascular permeability and cause vascular leakage.

Materials and methods  The effect of acute systemic inflammation on VEGF concentration was measured in healthy males after an intravenous bolus infusion of Escherichia coli endotoxin (lipopolysaccharide, LPS, 20 IU kg−1) in a double‐blind, placebo‐controlled parallel group study. LPS administration was followed by an infusion of lepirudin (bolus 0·1 mg kg−1, continuous infusion of 0·1 mg kg−1 h−1, n= 12) or saline (n = 12).

Results  Plasma VEGF increased from a mean of 15·1 pg mL−1 to 74·6 pg mL−1 5 h after LPS (P < 0·003). Body temperature, pulse rate, leukcytes, prothrombin fragment 1 + 2 (F1 + 2) and lactoferrin increased and platelets decreased after LPS (P < 0·05). The LPS‐induced increase in VEGF was paralleled by the neutrophil cell degranulation marker lactoferrin but not by F1 + 2, and was not affected by lepirudin, which blunted F1 + 2 formation (P < 0·05).

Conclusions  Inflammation‐induced activation of leukcytes rather than platelets plays a role in the marked increase in VEGF, which cannot be abrogated by antithrombotic therapy.
Are elevated homocysteine plasma levels related to peripheral arterial disease? Results from a cross‐sectional study of 6880 primary care patients
Authors: Darius H. , Pittrow D. , Haberl R. , Trampisch H. J. , Schuster A. , Lange S. , Tepohl H. G. , Allenberg J. R. , Diehm C.
Published: August 2003

Background  It is still unclear whether the strength of the association between elevated plasma homocysteine (HC) levels and peripheral arterial disease (PAD), coronary artery disease (CAD) and cerebrovascular disease (CVD) is similar.

Methods  Fasting homocysteine plasma levels were measured in 6880 unselected primary care patients aged 65 years or older. Presence of PAD was determined with the ankle brachial index, and both CAD and CVD were recorded according to patient history.

Results  Median homocysteine levels in the total sample (58·0% females, mean age 72·5 years, mean body mass index 27·3 kg m−2) differed between patients with and without PAD: 15·2 µmol L−1 (95% confidence interval [CI] 14·8; 15·7, vs. 13·9 µmol L−1 (CI: 13·8; 14·1; P < 0·001). Peripheral arterial disease prevalence moderately increased from 13·0% in the lowest HC quintile to 24·3% in the highest quintile (crude odds ratio [OR], 2·1 [CI: 1·7; 2·6]). The frequency of atherothrombotic manifestations in the patients’ history increased nearly linearly across the homocysteine quintiles in the univariate analysis. However, the association diminished substantially after adjusting for known interfering variables: the ORs between the HC highest fifth vs. lowest fifth (adjusted for age, gender, smoking status, diabetes, hypertension lipid disorders, and estimated glomerular filtration rate levels) for PAD decreased to 1·4, for CAD to 1·0 (NS), and for CVD to 1·1. (NS).

Conclusions  Elevated HC is only slightly more related to PAD than to CAD and CVD. After adjustment for known risk factors, the effect size is small, and an association can no longer be observed between homocysteine and CAD and CVD.
On the mechanism of post‐splenectomy leukocytosis in mice
Authors: Djaldetti M. , Bergman M. , Salman H. , Cohen A. M. , Fibach E. , Bessler H.
Published: August 2003

Background  Increased number of peripheral white blood cells (PWBCs) has been noted after removal of the spleen.

Design  To clarify the possible mechanisms by which splenectomy affects the PWBC number, the percentage of apoptotic PWBCs, the number and migration rate of peritoneal cells, as well as the 3H‐TdR incorporation into PWBCs, were examined in splenectomized, sham‐operated and control mice. In addition, the effect of control plasma injected to splenectomized animals on the number of PWBCs was examined.

Results  One and two months after splenectomy the PWBC counts significantly increased, whereas the percentage of apoptotic PWBCs and the number of cells in the peritoneal cavity decreased in comparison with that of the control and sham‐operated mice. Seventeen days after injection of carboxy‐fluorescein diacetate succinimidyl ester (CFSE)‐labelled peritoneal cells into the peritoneal cavity of the animals, their number was significantly higher in the peripheral blood and lower in the peritoneal cavity of the splenectomized animals in comparison with that of the control and sham‐operated mice. Injection of control plasma into the splenectomized mice prevented the development of postsplenectomy leukocytosis. Finally, 3H‐TdR incorporation into nonstimulated and Con A stimulated PBMCs from the splenectomized mice was higher as compared with cells from the control and sham‐operated mice.

Conclusions  The results of the study present several mechanisms that may clarify the cause of postsplenectomy leukocytosis.
Enterohepatic cycling of bilirubin as a cause of ‘black’ pigment gallstones in adult life
Authors: Vítek L. , Carey M. C.
Published: August 2003

In contrast to bile salts, which undergo a highly efficient enterohepatic circulation with multiple regulatory and physiologic functions, glucuronic acid conjugates of bilirubin are biliary excretory molecules that in health do not have a continuing biologic life. Intestinal absorptive cells are devoid of recapture transporters for bilirubin conjugates, and their large size and polarity prevent absorption by passive diffusion. However, unconjugated bilirubin, the β‐glucuronidase hydrolysis product of bilirubin glucuronides can be absorbed passively from any part of the small and large intestines. This can occur only if unconjugated bilirubin is kept in solution and does not undergo rapid bacterial reduction to form urobilinoids. Here we collect, and in some cases reinterpret, experimental and clinical evidence to show that in addition to the well‐known occurrence in newborns, enterohepatic cycling of unconjugated bilirubin can reappear in adult life. This happens as a result of several common conditions, particularly associated with bile salt leakage from the small intestine, the most notable ileal dysfunction resulting from any medical or surgical cause. We propose that when present in excess, colonic bile salts solubilize unconjugated bilirubin, delay urobilinoid formation, prevent calcium complexing of unconjugated bilirubin and promote passive absorption of unconjugated bilirubin from the large intestine. Following uptake, reconjugation, and resecretion into bile, this source of ‘hyperbilirubinbilia’ may be the important pathophysiological risk factor for ‘black’ pigment gallstone formation in predisposed adult humans.
Low plasma aldosterone despite normal plasma renin activity in uncomplicated type 1 diabetes mellitus: effects of RAAS stimulation
Authors: Luik P. T. , Kerstens M. N. , Hoogenberg K. , Navis G. J. , Dullaart R. P. F.
Published: August 2003

Background  Data on levels and responsiveness of PRA and aldosterone in type 1 diabetes mellitus are conflicting. Earlier studies were not standardized with respect to the type of diabetes mellitus, the presence of diabetic complications or sodium intake. Therefore, we studied plasma renin activity and plasma aldosterone in uncomplicated type 1 diabetes mellitus by evaluating the effects of endogenous (sodium restriction) and exogenous (angiotensin I infusion) stimulation.

Design  Twenty‐four type 1 diabetic patients and 24 matched healthy subjects were studied after 1 week of liberal sodium diet (200 mmol 24 h−1) and 1 week of low sodium diet (50 mmol 24 h−1). Angiotensin (Ang)I was infused at 4 and 8 ng kg−1 min−1 during both study days.

Results  During liberal and low sodium intake, plasma aldosterone was lower in type 1 diabetic patients compared with healthy subjects both at 08:00 h (P < 0·05) and after a 2‐h euglycaemic clamp (P < 0·05), despite similar PRA levels. The correlations between changes in PRA and changes in plasma aldosterone when shifting sodium intake were similar in both groups. During liberal sodium intake, the aldosterone levels after AngI infusion were lower in type 1 diabetic patients, whereas during low sodium they were not different.

Conclusions  Plasma aldosterone was deceased relative to PRA in uncomplicated type 1 diabetic patients, irrespective sodium intake. The responsiveness to sodium restriction was adequate and sodium restriction was able to overcome the decreased plasma aldosterone response to exogenous AngI, which was observed during liberal sodium in diabetic patients. The lower aldosterone is not secondary to diabetic complications and does not depend on the level of sodium intake.
Cellular characterization of the tropism of recombinant adenovirus for the adrenal glands
Authors: Wang Y. , Groot‐Wassink T. , Lemoine N. R. , Vassaux G.
Published: August 2003

Background  Recombinant adenoviruses are widely used in gene therapy clinical trials. A particular tropism for the adrenal glands has been reported but the precise cellular base for this tropism has not been determined.

Materials and methods  Recombinant adenoviruses were injected intravenously into Balb/c nu/nu or C57BL/6 mice. Seventy‐two hours later, the animals were sacrificed and the adrenal glands and livers collected. The glands were sectioned and analyzed using immunohistochemical methods to detect adenoviral epitopes and transgene expression. Total RNA were extracted from the liver and adrenal glands of some animals and subjected to real‐time RT‐PCR.

Results  The only cell type infected in the adrenal glands of Balb/c nu/nu or C57BL/6 mice is the adrenocortical cells in the zona fasciculata. Quantitatively, the relative level of gene expression in the adrenal gland is comparable but lower than that measured in the liver.

Conclusions  Systemic injection of recombinant adenovirus could be used as a procedure to restore adrenal steroidogenesis in clinical gene therapy protocols. In addition, our study suggest that adrenal dysfunction should be considered when criteria are established to assess the safety of gene therapy formulations administered systemically.
Effects of bosentan on neointimal response following coronary angioplasty
Authors: Sanmartín M. , Fernández‐Ortiz A. , Fantidis P. , Aragoncillo P. , Fernández‐Durango R. , Rollín R. , Alfonso F. , Hernández R. , Escaned J. , Macaya C.
Published: August 2003

Background  Endothelins are important vasoconstrictors and cellular‐growth promoters. ETA‐specific antagonists have been shown to reduce neointimal response to injury in some experimental angioplasty models. However, there is little information on the effects of dual ETA/ETB receptor blockers, such as bosentan, on neointimal proliferation following experimental coronary angioplasty.

Materials and methods  Coronary injury was achieved by directional atherectomy in the left anterior descending artery of 20 pigs. Animals were randomly assigned to receive a daily dose of oral bosentan (30 mg kg−1) (group I, n= 10) or no treatment (group II, n= 10). At 4 weeks, arteries were processed for histomorphometry and endothelin characterization.

Results  Vessel injury score was similar among the two groups. Overall, a linear correlation was found between injury and neointimal development (r = 0·57, P= 0·01). This correlation had a lower slope in group I compared with group II (P < 0·001), indicating a smaller amount of neointimal development for a similar degree of injury in bosentan‐treated animals. Multivariate regression showed that neointimal response was reduced by oral treatment with bosentan (beta: –0·59 mm2, 95% CI: −1·11/−0·06 mm2). In addition, immunostaining revealed fewer positive endothelin areas in group I arteries.

Conclusions  Oral treatment with bosentan reduces neointimal development following coronary angioplasty in this experimental model.
Controlled, randomized trial in maintenance treatment of intravenous buprenorphine dependence with naltrexone, methadone or buprenorphine: a novel study
Authors: Ahmadi J. , Ahmadi K. , Ohaeri J.
Published: August 2003

Aims  To test the effect of a 50‐mg day−1 dose of naltrexone in the maintenance treatment of intravenous‐buprenorphine dependence in comparison with a 50‐mg day−1 dose of methadone and also a 5‐mg day−1 dose of buprenorphine over a 24‐week treatment period.

Participants  Two hundred and four intravenous‐buprenorphine‐dependent patients who met the DSM‐IV criteria for opioid dependence were randomized to three equal groups and were treated in a clinic in Iran where they were also offered a weekly 0·5‐h counselling session, in 2002.

Findings  The mean age was 31·2 years (range 17–53). The majority [63·2%] was between 20 and 34 years of age. The educational level of most of them [55·4%] was between 6 and 12 years of study. Completion rates by groups were 83·8% for the 50‐mg dose methadone group, 58·8% for the 5‐mg dose buprenorphine group and 20·6% for the 50‐mg dose naltrexone group (P = 0·000). Retention in the 50‐mg methadone group was significantly better than in the 5‐mg dose buprenorphine group (P = 0·001) and the 50‐mg dose naltrexone group (P = 0·000). Retention in the 5‐mg buprenorphine group was significantly better than in the 50‐mg naltrexone dose group (P = 0·000).

Conclusions  Evaluation of the Iranian experience on this subject may be useful for other countries.
Resveratrol modulates apoptosis and oxidation in human blood mononuclear cells
Authors: Losa G. A.
Published: August 2003

Background  We examined the effect of resveratrol (RS), a nonflavonoid polyphenolic phytoalexin found in grapes and red wine, and RS coincubated with the oxidant 2‐deoxy‐D‐ribose (dR), on apoptosis and on the oxidative metabolic status of normal human peripheral blood mononuclear cells (PBMNCs) isolated ex vivo from healthy donors.

Material and methods  Apoptosis was measured by changes of membrane permeability to propidium iodide (PI), plasma membrane exposure of phosphatidylserine (PS) and intracellular caspase activity. Oxidative status was assessed by recording the intracellular glutathione concentration (GSH), the activities of the enzymes y‐glutamyltransferase (y‐GT) and glutathione‐S‐transferase (GST), and intracellular lipid peroxidation (MDA).

Results  Neither apoptotic nor oxidative parameters were affected by culturing PBMNCs in medium containing RS up to 20 µM for 5 days, while the frequency of cells with intermediate permeability to PI (17% ± 5) increased at 50 µM of RS. Thus resveratrol was slightly toxic, but there was little apoptosis in these cells. Peripheral blood mononuclear cells were also grown first in medium plus RS for 24 h and then for 96 h in medium containing RS plus 10 mM of dR, an oxidant sugar that is apoptogenic for human lymphocytes. The apoptotic changes triggered by dR were counteracted by the phytoalexin in a dose‐dependent manner, but RS activity was absent at the lowest concentration (5 µM) and significantly reduced at the highest concentration used (50 µM). In PBMNCs coincubated with 20 µM of RS and 10 mM of dR the antioxidant effect of RS manifested with a significant reduction of caspases‐3, ‐8, y‐GT, GST activities and MDA content.

Conclusions  Peripheral blood mononuclear cells acquire antioxidant capacity when treated with RS. Grape resveratrol may make a useful dietary supplement for minimizing oxidative injury in immune‐perturbed states and human chronic degenerative diseases.
Experimental and interventional dietary study in humans on the role of HDL fatty acid composition in PGI2 release and Cox‐2 expression by VSMC
Authors: Escudero I. , Martínez‐González J. , Alonso R. , Mata P. , Badimon L.
Published: August 2003

Background  High‐density lipoproteins (HDLs) induce prostacyclin (PGI2) release in vascular smooth muscle cells (VSMCs) by up‐regulation of cyclooxygenase‐2 (Cox‐2). Our goal was to analyze the role of human HDL lipid moiety on Cox‐2‐dependent PGI2 synthesis in human VSMCs and to assess the impact that the intake of diets with different fatty acid composition exert on HDL‐induced PGI2 release.

Materials and methods  Human VSMCs were treated with HDL or fatty acids in the presence or absence of different cell signalling inhibitors and PGI2 (by enzyme immunoassay) and Cox‐2 protein levels (by Western blot) were analyzed. High‐density lipoproteins were obtained from a plasma pool or from plasma of 12 volunteers subjected to a longitudinal dietary interventional study of three consecutive diets periods enriched in monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids n‐6 (PUFA n‐6) or n‐3 (PUFA n‐3).

Results  High‐density lipoprotein delipidation attenuated the effect of HDL on both PGI2 synthesis and Cox‐2 up‐regulation, while arachidonic acid (AA) but not other fatty acids mimicked the effects of HDL. Arachidonic acid induced PGI2 synthesis and Cox‐2 expression through similar mechanisms to those activated by HDL [pertussis toxin‐sensitive G proteins, p42/44 mitogen‐activated protein kinase (MAPK), p38MAPK, and c‐Jun N‐terminal kinase‐1 (JNK‐1) pathways]. Finally, we observed that HDL from the PUFA n‐3 dietary period induced lower PGI2 release than that from the PUFA n‐6 period (64% vs. 100%).

Conclusions  Our results suggest that lipid moiety modulates HDL‐induced PGI2 release/Cox‐2 up‐regulation in human VSMCs, and that changes in fatty acids as accomplished with the diet can modulate vascular PGI2 homeostasis.
Mechanisms of renal hyporesponsiveness to ANP in heart failure
Authors: Charloux A. , Piquard F. , Doutreleau S. , Brandenberger G. , Geny B.
Published: August 2003

The atrial natriuretic peptide (ANP) plays an important role in chronic heart failure (CHF), delaying the progression of the disease. However, despite high ANP levels, natriuresis falls when CHF progresses from a compensated to a decompensated state, suggesting emergence of renal resistance to ANP.

Several mechanisms have been proposed to explain renal hyporesponsiveness, including decreased renal ANP availability, down‐regulation of natriuretic peptide receptors and altered ANP intracellular transduction signal. It has been demonstrated that the activity of neutral endopeptidase (NEP) is increased in CHF, and that its inhibition enhances renal cGMP production and renal sodium excretion. In vitro as well as in vivo studies have provided strong evidence of an increased degradation of intracellular cGMP by phosphodiesterase in CHF. In experimental models, ANP‐dependent natriuresis is improved by phosphodiesterase inhibitors, which may arise as new therapeutic agents in CHF. Sodium‐retaining systems likely contribute to renal hyporesponsiveness to ANP through different mechanisms. Among these systems, the renin‐angiotensin‐aldosterone system has received particular attention, as angiotensin II and ANP have renal actions at the same sites and inhibition of angiotensin‐converting enzyme and angiotensin‐receptor blockade improve ANP hyporesponsiveness. Less is known about the interactions between the sympathetic nervous system, endothelin or vasopressin and ANP, which may also blunt ANP‐induced natriuresis.

To summarize, renal hyporesponsiveness to ANP is probably multifactorial. New treatments designed to restore renal ANP efficiency should limit sodium retention in CHF patients and thus delay the progression to overt heart failure.