bluuebunny/crossref_metadata_2025 huggingface dataset:
A preliminary investigation into mining and smelting impacts on trace element concentrations in the soils and vegetation around Tharsis, SW Spain
Authors: Chopin E. I. B. , Black S. , Hodson M. E. , Coleman M. L. , Alloway B. J.
Published: May 2003

Toxic trace elements present an environmental hazard in the vicinity of mining and smelting activities. However, the processes of transfer of these elements to groundwater and to plants are not always clear. Tharsis mine, in the Iberian pyrite belt (SW Spain), has been exploited since 2500 BC, with extensive smelting taking place from the 1850s until the 1920s. Sixty four soil (mainly topsoils) and vegetation samples were collected in February 2001 and analysed by ICP-AES for 23 elements. Concentrations are 6—6300 mg kg-1 As and 14—24800 mg kg-1 Pb in soils, and 0.20—9 mg kg-1 As and 2—195 mg kg-1 Pb in vegetation. Trace element concentrations decrease rapidly away from the mine, with As and Pb concentrations in the range 6—1850 mg kg—1 (median 22 mg kg—1) and 14—31 mg kg—1 (median 43 mg kg—1), respectively, 1 km away from the mine. These concentrations are low when compared to other well-studied mining and smelting areas (e.g. 600 mg kg—1 As at 8 km from Yellowknife smelter, Canada; >100 mg kg—1 Pb over 270 km2 around the Pb-Zn Port Pirie smelter, South Australia; mean of 1419 mg kg—1 Pb around Aberystwyth smelter, Wales, UK). The high metal content of the vegetation and the low soil pH (mean pH 4.93) indicate the potential for trace element mobility which could explain the relatively low concentration of metals in Tharsis topsoils and cause threats to plans to redevelop the Tharsis area as an orange plantation.
Metal dispersion in sediments and waters of the River Conwy draining the Llanrwst Mining Field, North Wales
Authors: Brydie J. R. , Polya D. A.
Published: May 2003

The dispersion and sediment-water partitioning of Pb and Zn have been studied in the Conwy River, North Wales. Analysis included major and trace element water chemistry and concentrations of sediment-hosted Pb and Zn. In situ solution pH, Eh, temperature and conductivity were also measured. Sediments were characterized via SEM, XRD, nitric acid leaching and sequential chemical extraction to quantify metal distribution and sediment phase associations. Dissolved and sediment-bound Pb and Zn within river and estuary waters and sediments have been used to calculate whole sediment- and phase-specific apparent partition coefficients.

Weathering of galena and sphalerite, associated with the Llanrwst Mining Field, provide point sources of elevated dissolved and sedimentary Pb and Zn in the upper catchment. Dissolved Pb is actively adsorbed onto sedimentary surface coatings of Fe-Mn (hydr)oxides and organics whilst Fe-Mn (hydr)oxides, organics and carbonates were the main hosts for Zn. Systematic changes in metal sorption are evident between the estuary and the upper catchment, with organic matter becoming progressively more important upstream. This change is ascribed to the sorptive properties of sedimentary organic material. Solution pH, [Cl] and aqueous metal speciation are the dominant solution controls on metal partitioning. Laboratory sorption experiments parallel Pb sorption behaviour in the natural system.
Mineralogical controls on metal distribution in stream sediment derived from the Caledonides of the Scottish Southern Uplands and English Lake District
Authors: Stone P. , Breward N. , Merriman R. J.
Published: May 2003

Stream sediment geochemistry provides an innovative method of assessing the basinal history of the Caledonian slate belts. Despite glaciation, the stream sediment geochemical patterns spatially mimic the outcrop of underlying bedrock lithologies. However, erosion from rock to sediment by fluvial processes may either increase or reduce an element’s abundance depending on the nature of its mineral host. An element held in heavy, resistate minerals will be concentrated, whereas one residing in unstable ferromagnesian minerals, which readily break down to clays during weathering, may be preferentially removed. Examples are provided from the Cr-Ni-V-Mg, base metal and Rb-Sr element suites. Primary and secondary bedrock patterns are recognized in the stream sediments. Primary patterns follow the original composition of the source bedrock, with steep gradients in the element distribution coinciding with lithostratigraphical boundaries. Such patterns also reveal subtle divisions within the established geological units for which the main compositional control was the nature of the ancient sedimentary provenance. Secondary patterns reflect remobilization of elements within the bedrock and so may cut across lithostratigraphical boundaries. These patterns (or their absence) are influenced by the thermal histories of the Caledonian basins, and so are indicative of the geotectonic regime in which the sedimentary sequences were originally deposited.
A model for the origin of Al-rich efflorescences near fumaroles, Melos, Greece: enhanced weathering in a geothermal setting
Authors: Hall A. J. , Fallick A. E. , Perdikatsis V. , Photos-Jones E.
Published: May 2003

Efflorescences in the geothermal field of SE Melos, Greece, contain significant amounts of hydrated Al sulphate, alunogen, which could represent the Melian alumen exploited in Roman times and commended by Pliny. The efflorescences at subaerial fumaroles are explained as follows: Sulphur crystallizes on oxidation of H2S emanating from depth. Weathering produces sulphuric acid enhancing groundwater alteration of volcanic rocks. The high geothermal gradient and arid climate stimulate efflorescences. Salts are recycled during wet and dry weather leading to Al-enrichment on loss of Fe(II,III) and other cations. δ34S‰ V-CDT values for sulphur in fumarole sublimates, solfatara soils and ‘veins’ range from —0.3 to 6.4‰, mean 3.8‰ (n = 8) while Al, Ca and Mg-sulphates in diverse settings range from —4.1 to 6.8‰ (n = 16). The values for sulphur indicate that the initial H2S had an igneous source and the signature is largely inherited by the sulphates.

This study aims to underpin research into the exploitation of industrial minerals in the Roman period. When searching for early alumen workings, areas with evidence of acid sulphate alteration (white rocks) and sulphurous fumarole activity should be investigated.
Replacement and authigenic mineralogy of metal contaminants in stream and estuarine sediments at Newtownards, Northern Ireland
Authors: Moles N. R. , Betz S. M. , McCready A. J. , Murphy P. J.
Published: May 2003

Tidal mudflats are locally enriched in heavy metals at the head of Strangford Lough in Northern Ireland, where drainage from the hinterland enters the sea lough via a tidal canal in an urban area. To characterize the metallic contaminants and investigate their provenance, heavy particles separated from stream, canal and estuarine sediments were analysed by electron microprobe and laser Raman microspectroscopic methods. Potential metal sources are mineralization in the catchment area and industrial or domestic pollution. Anthropogenic particles include metallic grains, alloys and compounds of Pb, Zn, Cu, Fe, Cr and Sn. Alteration of metallic particles includes de-zincification of brass in freshwater sediment and replacement of Cu wire by covellite in brackish to marine sediment. Mobility of Cu, Fe and S in canal and estuarine sediments is indicated by the authigenic growth of framboidal Fe sulphide on oxide substrates and of chalcopyrite rims on covellite. Intricate colloform and platy crystalline textures suggest a cyclical deposition of covellite and chalcopyrite under conditions of varying redox and salinity. Lead and Cr mobility in the contaminated estuarine sediment is shown by the authigenic formation on Pb-rich substrates of heterogeneous Pb- and Cr-rich sulphate-phosphate compounds and Pb-oxychlorides.
Fe-sulphate-rich evaporative mineral precipitates from the Río Tinto, southwest Spain
Authors: Buckby T. , Black S. , Coleman M. L. , Hodson M. E.
Published: May 2003

The soluble metal sulphate salts melanterite, rozenite, rhomboclase, szomolnokite, copiapite, coquimbite, hexahydrite and halotrichite, together with gypsum, have been identified, some for the first time, on the banks of the Rio Tinto, SW Spain. Secondary Fe-sulphate minerals can form directly from evaporating, acid, sulphate-rich solutions as a result of pyrite oxidation. Chemical analyses of mixtures of these salt minerals indicate concentrations of Fe (up to 31 wt.%), Mg (up to 4 wt.%), Cu (up to 2 wt.%) and Zn (up to 1 wt.%). These minerals are shown to act as transient storage for metals and can store on average up to 10% (9.5 — 11%) and 22% (20—23%), Zn and Cu respectively, of the total discharge of the Rio Tinto during the summer period.

Melanterite and rozenite precipitates at Rio Tinto are only found in association with very acidic drainage waters (pH <1.0) draining directly from pyritic waste piles. Copiapite precipitates abundantly on the banks of the Rio Tinto by (1) direct evaporation of the river water; or (2) as part of a paragenetic sequence with the inclusion of minor halotrichite, indicating natural dehydration and decomposition. The natural occurrences are comparable with the process of paragenesis from the evaporation of Rio Tinto river water under laboratory experiments resulting in the formation of aluminocopiapite, halotrichite, coquimbite, voltaite and gypsum.
Mineralogical and S isotopic features of the supergene profile of the Zapadno-Ozernoe massive sulphide and Au-bearing gossan deposit, South Urals
Authors: Belogub E. V. , Novoselov C. A. , Spiro B. , Yakovleva B. A.
Published: May 2003

The profile of the supergene zone of the Zapadno-Ozernoe massive sulphide Cu-Zn deposit differs from the classic model (Emmons, 1917) in that it includes a prominent dark sooty subzone rich in secondary sulphides. This subzone is situated above residual pyrite sands, which overlie the massive sulphide body and below quartz-baryte leached sands. It contains a diverse mineral assemblage which consists of secondary sulphides such as galena, sphalerite, metacinnabar, Se-bearing pyrite—dhzarkenite series, tiemannite, native Au, native S and native Se, and unidentified sulphosalts of Ag and Hg. The very light S isotope composition of the secondary sulphides (lowest values δ34S = —17.2‰ (VCDt) in comparison with primary pyrite ∼0‰ and baryte +18.4‰ is indicative of bacterial sulphate reduction. The overlying oxidized part of the supergene column contains minerals of the jarosite-beudantite- segnitite series. The maximum concentrations of Au, up to 150 ppm, occur in the lower part of the profile. The atypical structure, mineral assemblage and S isotope composition of the secondary sulphides in the dark layer of the supergene profile are indicative of particular geochemical conditions due to the existence of a stagnant water body that gave rise to intense bacterial activity, in turn controlled by fluctuations in the redox boundary.
Supergene iron sulpho-selenides from the Zapadno-Ozernoe copper-zinc massive sulphide deposit, South Urals, Russia: a new solid-solution series between pyrite FeS2 and dzharkenite FeSe2
Authors: Yakovleva V. A. , Belogub E. V. , Novoselov K. A.
Published: May 2003

Supergene Fe sulpho-selenides (pyrite, dzharkenite and their intermediate compositions) were discovered in the oxidation zone of the Zapadno-Ozernoe copper-zinc massive sulphide deposit, South Urals, in association with other rare supergene sulphides, such as galena, tetrahedrite, metacinnabarite, tiemannite, Pb sulpharsenides of the jordanite group, undetermined sulphosalts of Hg and Ag, and native elements, such as S, Au and Se. This is the second known occurrence of dzharkenite. The results of the investigation of Fe sulpho-selenides using electron microprobe analysis, X-ray diffraction (XRD) and reflected light microscopy show the existence of a limited solid-solution series between pyrite and dzharkenite, which has not been described before.
Mineralogical, numerical and analytical studies of the coupled oxidation of pyrite and coal
Authors: Evans K. A. , Gandy C. J. , Banwart S. A.
Published: May 2003

Mineralogical, bulk and field leachate compositions are used to identify important processes governing the evolution of discharges from a coal spoil heap in County Durham. These processes are incorporated into a numerical one-dimensional advective-kinetic reactive transport model which reproduces field results, including gas compositions, to within an order of magnitude. Variation of input parameters allows the effects of incorrect initial assumptions on elemental profiles and discharge chemistry to be assessed. Analytical expressions for widths and speeds of kinetic reaction fronts are developed and used to predict long-term development of mineralogical distribution within the heap. Results are consistent with observations from the field site. Pyrite oxidation is expected to dominate O2 consumption in spoil heaps on the decadal timescale, although C oxidation may stabilize contaminants in effluents on the centennial scale.
The kinetics of O2(aq) reduction during oxidative weathering of naturally occurring fracture minerals in groundwater
Authors: Rivas-Perez J. , Tullborg E.-L. , Banwart S. A.
Published: May 2003

Aqueous chemistry methods assessed the kinetic reactivity and reduction capacity of fracture-filling minerals from a granitic groundwater environment at the Aspo Hard Rock Laboratory, a field station for research and development into the geological disposal of spent nuclear fuel. Naturally occurring fracture filling reacted with oxygenated test solutions of known composition in recirculating batch reactors.

The loss of O2(aq) with time was consistent with second-order reaction kinetics where O2(aq) is consumed through reduction by reaction with structural Fe(II) at the surface of the fracture minerals. Values of the second-order rate constant (k, l mole—1 h—1) varied between experiments within the range 1.7 < log k < 2.9 and values for the total concentration of reducing sites as a measure of the reduction capacity of the mineral (St, mole g—1) varied within the range 8.5 x 10—5 < St < 4.3 x 10—4. Values for the rate constant are somewhat less than those published previously for reaction between O2(aq) and Fe(II) surface complexes; i.e. structural Fe(II) appears to be less reactive than adsorbed Fe(II)(aq) but is significantly more reactive than Fe2+.

Values of the rate constant did not depend on release of network ions from the reacting minerals, and did not vary significantly with pH, mineral mass, specific surface area or mineral Fe(II) content. Oxidative weathering of sulphide minerals does not occur to any measurable extent. When corrected to rock/water ratios for granite aquifers, the rate constants correspond to a half-life for O2(aq) on the order of seconds indicating an essentially instantaneous reaction on repository time scales. Comparison of reducing capacity for the fracture fillings and oxidizing capacity for groundwater saturated with O2(aq) indicates that oxidizing fronts within the geological barrier would travel ~4000 times more slowly than the velocity of groundwater flow.
Altered erythrocyte endothelial adherence and membrane phospholipid asymmetry in hereditary hydrocytosis
Authors: Gallagher Patrick G. , Chang Seon Hee , Rettig Michael P. , Neely John E. , Hillery Cheryl A. , Smith Brian D. , Low Philip S.
Published: May 2003

The risk for thrombosis is increased in patients with hereditary hydrocytosis, an uncommon variant of hereditary stomatocytosis. Erythrocytes from 2 patients with hydrocytosis were studied to gain insight into the mechanism of thrombosis in this disorder. Erythrocytes demonstrated abnormal osmotic scan ektacytometry and decreased erythrocyte filtration rates. There was also a mild increase in adherence of erythrocytes to endothelial monolayers in a micropipette assay. Adhesion of erythrocytes to the subendothelial matrix proteins thrombospondin and laminin, however, was not significantly increased. Percentages of hydrocytosis erythrocytes and reticulocytes with phosphatidylserine exposed on the outer surfaces were increased in both patients compared with healthy controls, indicating altered membrane phospholipid asymmetry. Increased phosphatidylserine exposure accelerating thrombin-forming processes has been proposed as a mechanism for thrombosis in sickle cell disease and β-thalassemia and may play a similar role in hereditary hydrocytosis.
The in vivo profile of transcription factors during neutrophil differentiation in human bone marrow
Authors: Bjerregaard Malene Digmann , Jurlander Jesper , Klausen Pia , Borregaard Niels , Cowland Jack Bernard
Published: May 2003

In vivo distribution of myeloid transcription factors during granulopoiesis was investigated by Northern and Western blotting in 3 neutrophil precursor populations from human bone marrow: immature (myeloblasts [MBs] and promyelocytes [PMs]); intermediate mature (myelocytes [MCs] and metamyelocytes [MMs]); and mature neutrophil cells (band cells [BCs] and segmented neutrophil cells [SCs]). Nonneutrophil cells were removed with magnetic-bead–coupled antibodies against CD2, CD3, CD14, CD19, CD56, CD61, glycophorin-A, and CD49d (BCs/SCs) before RNA and protein extraction. Polymorphonuclear neutrophils (PMNs) from peripheral blood depleted with anti-CD49d antibodies were also included. Expression of acute myeloid leukemia 1b (AML-1b), c-myb, GATA-1, and CCAAT/enhancer binding protein γ (C/EBP-γ) was seen primarily in MBs/PMs, and little expression was found in more mature cells. The level of C/EBP-α was constant in the bone marrow–derived cells and decreased in PMNs. C/EBP-ϵ was found primarily in MCs/MMs and was almost absent in more mature cells. Expression of C/EBP-β, C/EBP-δ, and C/EBP-ζ was observed from the MC/MM stage onward, with peak levels in the most mature cells. The amount of PU.1 increased throughout maturation whereas the level of Elf-1 reached a nadir in MCs/MMs The PU.1 coactivator c-jun and c-jun's dimerization partner c-fos were both detectable in MCs/MMs and increased in amount with maturity. CCAAT displacement protein (CDP) was found at comparable levels at all stages of differentiation. This demonstrates a highly individualized expression of the transcription factors, which can form the basis for the heterogeneous expression of granule proteins during granulopoiesis and cell cycle arrest in metamyelocytes.
CCR5 deficiency decreases susceptibility to experimental cerebral malaria
Authors: Belnoue Elodie , Kayibanda Michèle , Deschemin Jean-Christophe , Viguier Mireille , Mack Matthias , Kuziel William A. , Rénia Laurent
Published: May 2003

Infection of susceptible mouse strains with Plasmodium berghei ANKA (PbA) is a valuable experimental model of cerebral malaria (CM). Two major pathologic features of CM are the intravascular sequestration of infected erythrocytes and leukocytes inside brain microvessels. We have recently shown that only the CD8+ T-cell subset of these brain-sequestered leukocytes is critical for progression to CM. Chemokine receptor–5 (CCR5) is an important regulator of leukocyte trafficking in the brain in response to fungal and viral infection. Therefore, we investigated whether CCR5 plays a role in the pathogenesis of experimental CM. Approximately 70% to 85% of wild-type and CCR5+/- mice infected with PbA developed CM, whereas only about 20% of PbA-infected CCR5-deficient mice exhibited the characteristic neurologic signs of CM. The brains of wild-type mice with CM showed significant increases in CCR5+ leukocytes, particularly CCR5+ CD8+ T cells, as well as increases in T-helper 1 (Th1) cytokine production. The few PbA-infected CCR5-deficient mice that developed CM exhibited a similar increase in CD8+ T cells. Significant leukocyte accumulation in the brain and Th1 cytokine production did not occur in PbA-infected CCR5-deficient mice that did not develop CM. Moreover, experiments using bone marrow (BM)–chimeric mice showed that a reduced but significant proportion of deficient mice grafted with CCR5+ BM develop CM, indicating that CCR5 expression on a radiation-resistant brain cell population is necessary for CM to occur. Taken together, these results suggest that CCR5 is an important factor in the development of experimental CM.
Regulation of the human endothelial cell protein C receptor gene promoter by multiple Sp1 binding sites
Authors: Rance James B. , Follows George A. , Cockerill Peter N. , Bonifer Constanze , Lane David A. , Simmonds Rachel E.
Published: May 2003

The human endothelial cell protein C receptor (hEPCR) is normally expressed by the endothelium of large blood vessels, but the molecular basis for its in vivo specificity is uncertain. In this study, DNaseI hypersensitive site mapping demonstrated the presence of a hypersensitive site in the 5′ flanking region of the hEPCR gene in endothelial cells and certain transformed cells (HeLa and U937) known to express hEPCR in vitro. Conversely, this site was only weakly hypersensitive in HepG2 cells, cells which do not express hEPCR mRNA. Functional analysis of this 5′ flanking region by in vivo dimethylsulfate footprinting in cultured endothelial cells identified multiple regions, containing high and low homology consensus Sp1 binding sequences, that were protected from methylation in endothelial cells. These sequences were not protected in HepG2 cells. Reporter gene analysis of this region in endothelial cells demonstrated the presence of promoter activity conferred by the proximal 572 bp but failed to identify a functional TATA-box. This promoter was inactive in HepG2 cells. Electrophoresis mobility shift assays using endothelial cell nuclear extracts identified Sp1 family proteins binding to sites that were protected during footprinting. Sp1 sites were identified in regions at –368, –232, –226, –201, –146, and –102 bp relative to the translation start site. With the exception of the site at –102 bp, each identified Sp1 binding site made a positive contribution to reporter gene expression, although no individual site was critically important. We conclude that transcription factor binding to Sp1 binding sites in the 5′ flanking region is critical for normal hEPCR gene expression in endothelial cells.
Nonmyeloablative conditioning is sufficient to allow engraftment of EGFP-expressing bone marrow and subsequent acceptance of EGFP-transgenic skin grafts in mice
Authors: Andersson Goran , Illigens Ben M. W. , Johnson Kevin W. , Calderhead David , LeGuern Christian , Benichou Gilles , White-Scharf Mary E. , Down Julian D.
Published: May 2003

Immunologic reactions against gene therapy products may prove to be a frequent problem in clinical gene therapy protocols. Enhanced green fluorescence protein (EGFP) is commonly used as a marker in gene transfer protocols, and immune responses against EGFP-expressing cells have been documented. The present study was designed to investigate the effect of a pharmacologic, nonmyeloablative, conditioning regimen on the development of EGFP+ donor/recipient mixed bone marrow chimerism and ensuing tolerance to EGFP-expressing transplants. To this end, C57BL/6J (B6) mice were treated with soluble formulations of either busulfan (Busulfex) or the closely related compound treosulfan, followed by transplantation of bone marrow cells from EGFP-transgenic (B6-EGFP.Tg) donor mice. Such conditioning regimens resulted in long-term persistence of donor EGFP+ cells among various hematopoietic lineages from blood, bone marrow, and thymus. Stable hematopoietic chimeras transplanted at 10 to 17 weeks after bone marrow transplantation (BMT) with B6-EGFP.Tg skin grafts all accepted their transplants, whereas non-EGFP chimeric B6 control animals were able to mount rejection of the EGFP+ B6 skin grafts. Control third-party grafts from major histocompatibility complex (MHC)–mismatched mice were rejected within 20 days, indicating that acceptance of EGFP-expressing skin grafts was the result of specific immune tolerance induction by the transplantation of EGFP-transgenic bone marrow. Long-term tolerance to EGFP in chimeric recipients was confirmed by the absence of anti-EGFP–reactive T cells and antibodies. These results broaden the therapeutic potential for using hematopoietic molecular chimerism in nonmyeloablated recipients as a means of preventing rejection of genetically modified cells.
The “stomatin” gene and protein in overhydrated hereditary stomatocytosis
Authors: Fricke Britta , Argent Annette C. , Chetty Margaret C. , Pizzey Arnold R. , Turner E. Jane , Ho Mei M. , Iolascon Achille , von Düring Monika , Stewart Gordon W.
Published: May 2003

In overhydrated hereditary stomatocytosis (OHSt), Coomassie- and silver-stained polyacrylamide gels show an apparently complete deficit of the 32-kDa membrane protein, stomatin. We have used an antistomatin antibody to examine peripheral blood films, bone marrow, splenic tissue, and hepatic tissue from these patients by immunocytochemistry. This technique revealed that, in fact, some red cells did show positive stomatin immunoreactivity; and consistent with this result, Western blot analysis of the red cell membranes confirmed that about one twentieth to one fiftieth of the normal amount of stomatin was in fact present. Flow cytometry, combining immunoreactive quantitation of stomatin expression with thiazole orange staining for reticulocytes, showed that in OHSt, it was the young cells that had more stomatin. Magnetic-activated cell separation studies, using beads to which an anti–transferrin receptor antibody was conjugated, confirmed that in OHSt there was a correspondence between expression of stomatin and the transferrin receptor. Immunocytochemistry and Western blotting revealed that in OHSt patients, the protein was present in spleen, liver, neutrophils, platelets, monocytes, and about 50% of the peripheral lymphocytes, with the same distribution as in healthy controls. Neither Southern blots, nor direct sequencing of multiple subclones of the cDNA, nor sequencing of amplicons from genomic DNA revealed any significant abnormality in stomatin gene sequence in these patients. The deficiency of stomatin from red cells appears to be due to a loss of stomatin from these red cells on maturation in the bone marrow and in the circulation.
L-selectin stimulation enhances functional expression of surface CXCR4 in lymphocytes: implications for cellular activation during adhesion and migration
Authors: Ding Ziqiang , Issekutz Thomas B. , Downey Gregory P. , Waddell Thomas K.
Published: May 2003

L-selectin mediates leukocyte tethering and rolling, the first step in a sequential process of leukocyte adhesion and migration. Additionally, L-selectin has important signaling roles perhaps contributing to leukocyte activation and integrin-mediated adhesion. Because chemokines are critically involved in leukocyte activation, we questioned whether L-selectin signaling affects chemokine receptor expression and function. We observed that whereas only 5% to 15% of freshly isolated lymphocytes expressed CXCR4 on the cell surface, intracellular CXCR4 was detectable in all cells. Engagement of L-selectin by antibody cross-linking or the L-selectin ligands fucoidan or sulfatide mobilized intracellular CXCR4 to significantly increase surface CXCR4 expression but did not affect CCR5, CCR7, or β2-integrin expression. L-selectin stimulation also inhibited stromal-derived factor 1 (SDF-1)–induced CXCR4 internalization. The combined effects of L-selectin on CXCR4 trafficking are likely important in markedly enhancing cell activation by SDF-1. Blockade of SDF-1–induced CXCR4 internalization resulted in enhanced actin polymerization on subsequent exposure to SDF-1. Physiologically more important, L-selectin stimulation increased SDF-1–induced lymphocyte adhesion and transendothelial migration, which were inhibited by anti–leukocyte function-associated antigen 1 antibodies, tyrosine kinase inhibitors, and pertussis toxin. To further corroborate the additive stimulating effects, L-selectin signaling and SDF-1 increased β2-integrin activation. Taken together, L-selectin–mediated signals specifically enhance CXCR4 expression and function, suggesting a novel mechanism for the modulation of lymphocyte activation during cell adhesion and transmigration.
Activation of pp125FAK by type 2B recombinant von Willebrand factor binding to platelet GPIb at a high shear rate occurs independently of αIIbβ3 engagement
Authors: Mekrache Médina , Bachelot-Loza Christilla , Ajzenberg Nadine , Saci Abdelhafid , Legendre Paulette , Baruch Dominique
Published: May 2003

Shear-induced platelet aggregation (SIPA) involves the sequential interaction of von Willebrand factor (VWF) with both glycoprotein Ib (GPIb) and αIIbβ3 receptors. Type 2B recombinant VWF (2B-rVWF), characterized by an increased affinity for GPIb, induces strong SIPA at a high shear rate (4000 s–1). Despite the increased affinity of 2B-rVWF for GPIb, patients with type 2B von Willebrand disease have a paradoxical bleeding disorder, which is not well understood. The purpose of this study was to determine if SIPA induced by 2B-rVWF was associated with αIIbβ3-dependent platelet activation. To this end, we have addressed the influence of 2B-rVWF (Val553Met substitution) on SIPA-dependent variations of tyrosine protein phosphorylation (P-Tyr) and the effect of αIIbβ3 blockers. At a high shear rate, 2B-rVWF induced a strong SIPA, as shown by a 92.7% ± 0.4% disappearance of single platelets (DSP) after 4.5 minutes. In these conditions, increased P-Tyr of proteins migrating at positions 64 kd, 72 kd, and 125 kd were observed. The band at 125 kd was identified as pp125FAK using anti–phospho-FAK antibody. This effect, which required a high level of SIPA (> 70% DSP), was observed at 4000 s–1 but not at 200 s–1. Monoclonal antibodies (MoAbs) 6D1 (anti-GPIb) and 328 (anti-VWF A1 domain), completely abolished SIPA and p125FAK phosphorylation mediated by 2B-rVWF. In contrast, neither RGDS peptide nor MoAb 7E3, both known to block αIIbβ3 engagement, had any effect on SIPA and pp125FAK. The size of aggregates formed at a high shear rate in the presence of 2B-rVWF was decreased by genistein, demonstrating the biologic relevance of pp125FAK. These findings provide a unique mechanism whereby the enhanced interaction of 2B-rVWF with GPIb, without engagement of αIIbβ3, is sufficient to induce SIPA but does not lead to stable thrombus formation.
Host stem cells can selectively reconstitute missing lymphoid lineages in irradiation bone marrow chimeras
Authors: Rivera Amariliz , Chen Chiann-Chyi , Dougherty Joseph P. , Ben-Nun Avraham , Ron Yacov
Published: May 2003

The regulatory elements governing the process of lymphopoiesis from pluripotential stem cells to mature lymphocytes are not well understood. In this study we found that in bone marrow chimeras made by reconstituting lethally irradiated normal mice with bone marrow taken from genetically B-cell–deficient animals (μMT.B6 → F1) the B-cell compartment is reconstituted with host-derived B cells. Similarly, in animals reconstituted with bone marrow taken from mice with genetic deficiencies in the development of T cells (TCR–/– → F1) or both B and T cells (RAG–/– → F1), the missing lymphocyte lineage(s) was specifically reconstituted from host-derived cells. In all chimeras, all other blood lineages were generated from donor-derived stem cells. Control chimeras (B6 → F1) had only donor-derived hematopoietic cells as expected. The reconstituted, host-derived lymphoid compartments contained normal functional cell populations as determined by the presence of T cells expressing all 16 common TCR Vβ families, and the presence of all antibody isotypes in the serum. Reconstituted TCR–/– → F1 chimeras were also able to mount T-cell proliferative responses to foreign antigens equal to those of control animals. This observation would seem to suggest that during lymphopoietic reconstitution, missing lymphoid lineages can dictate their own reconstitution.
Peripheral T-cell lymphomas unspecified presenting in the skin: analysis of prognostic factors in a group of 82 patients
Authors: Bekkenk Marcel W. , Vermeer Maarten H. , Jansen Patty M. , van Marion Ariënne M. W. , Canninga-van Dijk Marijke R. , Kluin Philip M. , Geerts Marie-Louise , Meijer Chris J. L. M. , Willemze Rein
Published: May 2003

In the present study the clinicopathologic and immunophenotypic features of 82 patients with a CD30– peripheral T-cell lymphoma, unspecified, presenting in the skin were evaluated. The purpose of this study was to find out whether subdivision of these lymphomas on the basis of cell size, phenotype, or presentation with only skin lesions is clinically relevant. The study group included 46 primary cutaneous CD30– large cell lymphomas and 17 small/medium-sized T-cell lymphomas as well as 17 peripheral T-cell lymphomas with both skin and extracutaneous disease at the time of diagnosis. Patients with primary cutaneous small- or medium-sized T-cell lymphomas had a significantly better prognosis (5-year-overall survival, 45%) than patients with primary cutaneous CD30– large T-cell lymphomas (12%) and patients presenting with concurrent extracutaneous disease (12%). The favorable prognosis in this group with primary cutaneous small- or medium-sized T-cell lymphomas was particularly found in patients presenting with localized skin lesions expressing a CD3+CD4+CD8– phenotype. In the primary cutaneous T-cell lymphoma (CTCL) group and in the concurrent group, neither extent of skin lesions nor phenotype had any effect on survival. Our results indicate that peripheral T-cell lymphomas, unspecified, presenting in the skin have an unfavorable prognosis, irrespective of the presence or absence of extracutaneous disease at the time of diagnosis, cell size, and expression of a CD4+ or CD8+ phenotype. The only exception was a group of primary cutaneous small- or medium-sized pleomorphic CTCLs with a CD3+CD4+CD8– phenotype and presenting with localized skin lesions.
Ectopic hTERT expression extends the life span of human CD4+ helper and regulatory T-cell clones and confers resistance to oxidative stress–induced apoptosis
Authors: Luiten Rosalie M. , Pène Jérôme , Yssel Hans , Spits Hergen
Published: May 2003

Human somatic cells have a limited life span in vitro. Upon aging and with each cell division, shortening of telomeres occurs, which eventually will lead to cell cycle arrest. Ectopic hTERT expression has been shown to extend the life span of human T cells by preventing this telomere erosion. In the present study, we have shown that ectopic hTERT expression extends the life span of CD4+ T helper type 1 or 2 and regulatory T-cell clones and affected neither the in vitro cytokine production profile nor their specificity for antigen. In mixed cell cultures, ectopic hTERT-expressing clones were found to expand in greater numbers than untransduced cells of the same replicative age. This ectopic hTERT-induced growth advantage was not due to an enhanced cell division rate or number of divisions following T-cell receptor–mediated activation, as determined in carboxyfluorescein diacetate succinimidyl ester (CFSE)–labeling experiments. Moreover, the susceptibility to activation-induced cell death of both cell types was similar. However, cultures of resting hTERT-transduced T cells contained higher frequencies of Bcl-2–expressing cells and lower active caspase-3–expressing cells, compared with wild-type cells. Furthermore, hTERT-transduced cells were more resistant to oxidative stress, which causes preferential DNA damage in telomeres. Taken together, these results show that ectopic hTERT expression not only protects proliferating T cells from replicative senescence but also confers resistance to apoptosis induced by oxidative stress.
In vitro identification of human pro-B cells that give rise to macrophages, natural killer cells, and T cells
Authors: Reynaud Damien , Lefort Nathalie , Manie Elodie , Coulombel Laure , Levy Yves
Published: May 2003

In this study we report the molecular and functional characterization of very early interleukin 7 receptor α (IL-7Rα)+-CD79a+CD19– B-cell progenitors, produced by human CD34+CD19–CD10– cord blood cells grown in the presence of stromal cells and cytokines. Purified IL-7Rα+CD79a+CD19– cells transcribed the B-lymphoid specific genes E2A, EBF, TdT, Rag-1, had initiated DJH rearrangements, but almost lacked Pax-5 mRNA. When exposed to appropriate environmental conditions, these cells repressed B-cell genes and completely differentiated into CD14+ macrophages, CD56+ natural killer cells, and CD4high T cells. Retention of the DJH rearranged genes in both CD14+ and CD56+ cells unambiguously demonstrates that early B-cell genes, expressed prior to Pax-5, can be activated in a multipotent human progenitor cell whose final fate, including in non-B lineages, is determined by external signals.
The carboxy-terminal region of the granulocyte colony-stimulating factor receptor transduces a phagocytic signal
Authors: Santini Valeria , Scappini Barbara , Indik Zena K. , Gozzini Antonella , Rossi Ferrini Pierluigi , Schreiber Alan D.
Published: May 2003

Granulocyte colony-stimulating factor (G-CSF) induces proliferation, maturation, and functional activities of myeloid progenitors and mature neutrophils through a specific receptor, the G-CSF-R. Different signals are mediated by distinct regions of the cytoplasmic domain of G-CSF-R, but the precise role of each region has not yet been fully clarified. We evaluated the involvement of Syk kinase, essential in mediating phagocytic signals by Fcγ receptors, in G-CSF–induced phagocytosis, using murine myeloid 32D cells transfected with wild-type (WT) human G-CSF-R (hG-CSF-R) or with a G-CSF-R mutant truncated at cytoplasmic amino acid 715. The G-CSF-R mutant lacks the immunoreceptor tyrosine-based activation motif (ITAM), putative binding site for Syk. Following treatment of WT hG-CSF-R transfectants with IgG-coated particles, there was a significant increase in phagocytosis in G-CSF–stimulated cells, in which Syk tyrosine phosphorylation occurred, paralleled by enhancement of its tyrosine kinase activity. In the mutant transfectants, no significant increase in phagocytosis or Syk tyrosine phosphorylation occurred after stimulation with G-CSF. We also demonstrated that tyrosine phosphorylation of the Src kinases Hck and Lyn occurs following G-CSF stimulation of cells expressing WT G-CSF-R, but that Hck is not phosphorylated in mutant G-CSF-R transfectants. The increase in phagocytosis following G-CSF stimulation cannot be attributed to a rapid de novo increase in expression of Fcγ receptors. G-CSF induced expression of Fcγ receptors only after prolonged stimulation. Our data provide evidence that the carboxy-terminal region of G-CSF-R plays a role in the phagocytosis of IgG-coated particles and that Syk and Hck kinase tyrosine phosphorylation is involved.
Differential response of human acute myeloid leukemia cells to gemtuzumab ozogamicin in vitro: role of Chk1 and Chk2 phosphorylation and caspase 3
Authors: Amico Donatella , Barbui Anna Maria , Erba Eugenio , Rambaldi Alessandro , Introna Martino , Golay Josée
Published: May 2003

Gemtuzumab ozogamicin (GO) is a humanized anti-CD33 antibody conjugated to the anticancer agent calicheamicin, approved for the treatment of CD33+-relapsed acute myeloid leukemia. We have investigated the effects of GO on 4 human myeloid leukemia lines of different French-American-British (FAB) types (KG-1, THP-1, HL-60, and NB-4), observing 3 different types of response. Exposure to GO (10-1000 ng/mL) induced G2 arrest (up to 80% of the cells) followed by apoptosis (45% of the cells) in HL-60 and NB-4 cells. By contrast, in THP-1 cells we observed a strong G2 arrest (up to 75% of the cells) with little apoptosis. Finally, the KG-1 line was completely resistant to the same concentrations of GO. These different responses did not correlate with the levels of expression of either CD33 or multiple-drug resistance proteins, although the higher cyclosporin A (CsA)–inhibitable efflux activity of KG-1 cells may play a role in the resistance of this line to the drug. We could show that Chk1 and Chk2 phosphorylation, but not p53 or p21 expression, correlated with G2 arrest, implicating the ataxia-telangiectasia mutated/ataxia-telangiectasia related (ATM/ATR)–Chk1/Chk2 pathway in the cell cycle response to GO. However, apoptosis was associated with caspase 3 activation. Freshly isolated acute myeloid leukemia (AML) cells showed patterns of response to GO in vitro similar to those observed with the cell lines, including phosphorylation of Chk2 and caspase 3 activation. Our results suggest that the different molecular pathways induced by the drug in vitro may reflect, at least in part, the variable response to GO obtained in vivo.
Estimating the rate of thrombin and fibrin generation in vivo during cardiopulmonary bypass
Authors: Chandler Wayne L. , Velan Tomas
Published: May 2003

Our objective was to estimate the in vivo rates of thrombin and fibrin generation to better understand how coagulation is regulated. Studied were 9 males undergoing cardiopulmonary bypass (CPB). The rates of thrombin, total fibrin, and soluble fibrin generation in vivo were based on measured levels of prothrombin activation peptide F1.2, thrombin-antithrombin complex, fibrinopeptide A, and soluble fibrin, combined with a computer model of the patient's vascular system that accounted for marker clearance, hemodilution, blood loss, and transfusion. Prior to surgery, the average thrombin generation rate was 0.24 ± 0.11 pmol/s. Each thrombin molecule in turn generated about 100 fibrin molecules, of which 1% was soluble fibrin. The thrombin generation rate did not change after sternotomy or administration of heparin, then rapidly increased 20-fold to 5.60 ± 6.65 pmol/s after 5 minutes of CPB (P = .000 05). Early in CPB each new thrombin generated only 4 fibrin molecules, of which 35% was soluble fibrin. The thrombin generation rate was 2.14 ± 1.88 pmol/s during the remainder of CPB, increasing again to 5.47 ± 4.08 pmol/s after reperfusion of the ischemic heart (P = .000 08). After heparin neutralization with protamine, thrombin generation remained high (5.34 ± 4.01 pmol/s, P = .0002) and total fibrin generation increased, while soluble fibrin generation decreased. By 2 hours after surgery, thrombin and fibrin generation rates were returning to baseline levels. We conclude that cardiopulmonary bypass and reperfusion of the ischemic heart results in bursts of nonhemostatic thrombin generation and dysregulated fibrin formation, not just a steady increase in thrombin generation as suggested by previous studies.
Identification of differentially expressed genes representing dendritic cell precursors and their progeny
Authors: Wilson Heather L. , O'Neill Helen C.
Published: May 2003

The development of dendritic cells (DCs) from hematopoietic progenitors is not well understood. Using a spleen-derived long-term culture (LTC) system, it has been possible to continuously generate DCs from progenitors maintained in culture. The nonadherent LTC-DC population is composed of 2 major subsets. These are the small LTC-DC or DC precursors and their progeny, the large LTC-DCs that phenotypically resemble immature DCs. In this study, subtracted cDNA libraries were generated containing sequences differentially expressed in small or large LTC-DCs. Differential screening was then used on plated library clones to select genes expressed in either the small or the large cell population. Real-time polymerase chain reaction (PCR) has been used to verify the selection procedure for several genes of particular interest. Known genes isolated from subtracted libraries were related to stages in DC development and supported previous findings regarding the function of small and large LTC-DCs. Large LTC-DCs expressed a number of immunologically important genes encoding CD86, CCR1, osteopontin, and lysozyme. Small LTC-DCs resembled progenitor DCs expressing genes related to the organization of the cytoskeleton, the regulation of antigen processing, and a number of mitochondrial and ribosomal proteins. Novel transcripts were isolated from small and large LTC-DC–subtracted libraries that could encode novel proteins important in DC development. This study describes changes in gene expression related to the development of CD11c+CD11b+ major histocompatibility complex 2 low (MHC2lo) CD8α– DCs from precursors in a stroma-dependent culture system in the absence of exogenous cytokines.
Induction of acute myeloid leukemia in mice by the human leukemia-specific fusion gene NUP98-HOXD13 in concert with Meis1
Authors: Pineault Nicolas , Buske Christian , Feuring-Buske Michaela , Abramovich Carolina , Rosten Patty , Hogge Donna E. , Aplan Peter D. , Humphries R. Keith
Published: May 2003

HOX genes, notably members of the HOXA cluster, and HOX cofactors have increasingly been linked to human leukemia. Intriguingly, HOXD13, a member of the HOXD cluster not normally expressed in hematopoietic cells, was recently identified as a partner of NUP98 in a t(2;11) translocation associated with t-AML/MDS. We have now tested directly the leukemogenic potential of the NUP98-HOXD13 t(2; 11) fusion gene in the murine hematopoietic model. NUP98-HOXD13 strongly promoted growth and impaired differentiation of early hematopoietic progenitor cells in vitro; this effect was dependent on the NUP98 portion and an intact HOXD13 homeodomain. Expression of the NUP98-HOXD13 fusion gene in vivo resulted in a partial impairment of lymphopoiesis but did not induce evident hematologic disease until late after transplantation (more than 5 months), when some mice developed a myeloproliferative-like disease. In contrast, mice transplanted with bone marrow (BM) cells cotransduced with NUP98-HOXD13 and the HOX cofactor Meis1 rapidly developed lethal and transplantable acute myeloid leukemia (AML), with a median disease onset of 75 days. In summary, this study demonstrates that NUP98-HOXD13 can be directly implicated in the molecular process leading to leukemic transformation, and it supports a model in which the transforming properties of NUP98-HOXD13 are mediated through HOX-dependent pathways.
Tetrocarcin-A—induced ER stress mediates apoptosis in B-CLL cells via a Bcl-2—independent pathway
Authors: Anether Gabriele , Tinhofer Inge , Senfter Monika , Greil Richard
Published: May 2003

Tetrocarcin-A (TC-A), an antibiotic agent isolated from actinomycetes, has recently been described to antagonize Bcl-2 functions, thereby sensitizing tumor cells to cell death signals under control of Bcl-2. In this study, we analyzed the direct proapoptotic effect of TC-A in the B-chronic lymphocytic leukemia (B-CLL) model. We focused on the signal cascade triggered by TC-A in B-CLL cells and identified activated mitochondrial as well as endoplasmatic reticulum (ER) stress signals. The expression levels of known effector molecules mediating mitochondrial signaling, such as Bax and Bid, and the antagonistic molecule Bcl-2 did not influence sensitivity of B-CLL cells to TC-A. Furthermore, the molecular chaperone and sensor of ER stress, HSP70, though significantly up-regulated in B-CLL cells undergoing TC-A—triggered apoptosis, was ineffective to exert its anti-apoptotic function described in multiple cell death pathways. Autologous T cells of B-CLL patients were significantly less sensitive to TC-A as were also T cells from healthy donors when compared with their normal B-cell fraction. Furthermore, sensitivity of B-CLL cells to TC-A treatment in vitro was dependent neither on the expression levels of CD38—a prognostic factor for survival of B-CLL patients as well as for their response to therapy—nor on the clinical stage or pretreatment status of patients. From our data showing that TC-A induced a cell death pathway via ER stress preferentially in B cells and that it acted independently of important markers of drug sensitivity and of clinical markers, we conclude that TC-A might represent an attractive candidate drug for further evaluation in preclinical trials.