bluuebunny/crossref_metadata_2025 huggingface dataset:
Diversity of the bacterial hyperparasite Pasteuria penetrans in relation to root-knot nematodes (Meloidogyne spp.) control on Acacia holosericea
Authors: Duponnois Robin , Thioulouse Jean , Fargette Mireille , Davies Keith , Fould Sabine
Published: July 2002

Pasteuria penetrans isolates sampled from different geographical areas were characterised both for the heterogeneity of the endospore surface using monoclonal antibodies and for the ability of spores to attach to different isolates of Meloidogyne spp. The efficacy of these different Pasteuria isolates as biological control agents was tested in a glasshouse experiment with M. incognita from Senegal on Acacia holosericea. The immunoprofiles divided the P. penetrans isolates broadly differently from the attachment tests. Isolate PP16 from Senegal was associated with better seedling development of M. incognita-inoculated A. holoceria than were other isolates. Substantial variation in root and shoot biomass was not related to the observed variation in spore attachment tests. The difficulties involved in obtaining consistent biological control with Pasteuria are discussed in relation to the high degree of variability of this bacterium. Diversité chez l'hyperparasite bactérien Pasteuria penetrans en relation avec le contrôle du nématode Meloidogyne spp. sur Acacia holosericea - Des isolats de Pasteuria penetrans provenant de différentes régions géographiques ont été caractérisés en ce qui concerne, d'une part l'hétérogénéité de la surface des endospores - par utilisation d'anticorps monoclonaux -, d'autre part la capacité des spores à s'attacher à différentes souches de Meloidogyne spp. L'efficacité de ces différents isolats de P.penetrans en tant qu'agent de contrôle a été testée lors d'une expérience en serre utilisant un M. incognita d'origine sénégalaise et Acacia holosericea. Les isolats de P.penetrans ont montré des différences dans les profils immunologiques, différences non corrélées avec celles observées lors des tests d'attachement. En comparaison avec d'autres isolats, l'isolat PP16 provenant du Sénégal était associé à une meilleure croissance des plants de A. holosericea infestés par M. incognita. La variabilité importante touchant les biomasses aérienne et souterraine ne montrait aucune relation avec les différences observées lors des tests d'attachement. Les difficultés rencontrées pour obtenir un contrôle valable à l'aide de P.penetrans sont discutées en tenant compte du degré important de variabilité chez cette bactérie.
Electron microscopic study of the illite–smectite transformation in the bentonites from Cerro del Aguila (Toledo, Spain)
Authors: de Santiago Buey C. , Suarez Barrios M. , Garcia Romero E. , Dominguez Diaz M. C. , Doval Montoya M.
Published: July 2002

A mineralogical and microstructural study of bentonites from 'Cerro del Aguila' located in the Tagus Basin (Toledo, Spain) was carried out using XRD, SEM and TEM observations and chemical data obtained by TEM-EDX. The bentonites are mainly composed of trioctahedral smectite with a unit-cell formula (Si3.76Al0.24)(Al0.44Fe3+0.26Mg1.81)Ca0.05K0.19O10(OH)2and small amounts of illite. The relationships between illite and smectite particles observed by SEM and TEM allowed the study of the weathering process of illite to form smectite through possible intermediate stages. The transformation begins as an exfoliation normal to the stacking direction and develops by opening of the interlayer spacing, the replacement of K+by hydrated interlayer cations and slight reorganization of the 2:1 layer structure. The so-called solid-state transformation process then proceeds by further dissolution and the formation of a colloidal phase acting as raw material in the growth of new phyllosilicates such as smectite.
Nb-Ta oxide minerals from miarolitic pegmatites of the Baveno pink granite, NW Italy
Authors: Aurisicchio C. , De Vito C. , Ferrini V. , Orlandi P.
Published: July 2002

Chemical composition and cell parameters were ascertained on new data from Ti-Nb-Ta complex oxides from the miarolitic pegmatites of the Baveno pink granite (Southern Alps, NW Italy). The crystals are tiny, single or aggregated in small sprays, prismatic or tabular, from yellow-orange to brownish in colour. Typical associated minerals include fluorite, zinnwaldite, gadolinite-group minerals and Sc-minerals. Cavity paragenesis is typical of NYF pegmatites, and shows two stages of crystallization developing in magmatic-pneumatolitic and hydrothermal conditions. X-ray data show that some oxides belong to the aeschynite mineral group; others are polycrase and fersmite. Aeschynite and polycrase are chemically heterogeneous and structurally disordered because of their metamict state. This disorder does not always seem to be related to radionuclide contents. Two main trends are indicated, considering the behaviour of Y. The high Y contents fit with very low Ca andLREEcontents in theAsite; theHREEcontents follow the Y trend. In theBsite, Ti is the dominant cation, followed by some Nb and very little Ta. Small quantities of Y fit with increasing Ca, U, Th andREEcontents. In theBsite, Nb cations often exceed those of Ti. The Th contents are often greater than those of U. Besides the already known aeschynite-(Y) and vigezzite, new varieties, ‘titano-vigezzite’ and ‘niobo-aeschynite-(Y)’, are identified here in the Baveno miarolitic cavities. Samples 14 (analysis a) and 2502 (analysis b) have Ca as the main occupant of the A site, followed by Y, Th andREE; in theBsite, Ti prevails over Nb. These compositions cannot be considered as pure vigezzite, but as a new variety called ‘titano-vigezzite’. In the same way, analysis a of sample 3 may be considered a new variety of aeschynite-(Y), with Nb prevailing over Ti in theBsite, and here called ‘niobo-aeschynite-(Y)’. Neither variety has ever been mentioned in the literature. Epitaxial growth of aeschynite on polycrase (sample 3194) allows some inferences on the crystallization sequence.
Mineralogy and40Ar/39Ar geochronology of orangeites (Group II kimberlites) from the Damodar Valley, eastern India
Authors: Kent Ray W. , Kelley Simon P. , Pringle Malcolm S.
Published: July 2002

A suite of ultramafic-mafic alkaline igneous rocks in the Damodar Valley, eastern India, contains carbonate, phosphate and titanate minerals that are not characteristic or common in minettes or lamproites, but are typical of orangeites (Group II kimberlite) from southern Africa. Phlogopite grains from the Damodar alkaline rocks yield mean40Ar/39Ar ages of 116.6±0.8 Ma, 113.5±0.5 Ma and 109.1±0.7 Ma (1σ errors) using laser dating techniques. These ages are similar to the Rb-Sr ages of African orangeites, which lie mostly in the range 121 to 114 Ma. Prior to this study, only one possible occurrence of orangeite (the ∼820 m.y.-old Aries pipe, Western Australia) was known outside the Kaapvaal craton and its environs. If the Damodar alkaline rocks are bona fide orangeites, it is likely that they were generated at depths of >150 km, within the stability field of diamond.
The basophil activation marker defined by antibody 97A6 is identical to the ectonucleotide pyrophosphatase/phosphodiesterase 3
Authors: Bühring Hans-Jörg , Seiffert Martina , Giesert Christina , Marxer Anke , Kanz Lothar , Valent Peter , Sano Kimihiko
Published: July 2002

It has recently been shown that monoclonal antibody (MoAb) 97A6 detects a surface antigen expressed on basophils and their CD34+ precursor cells, as well as the basophil cell line KU-812. In this report the partial amino acid sequence of affinity chromatography– and sodium dodecyl sulfate–polyacrylamide gel electrophoresis–separated 97A6 antigen(s) from KU-812 lysates was determined. Sequence alignment of high-performance liquid chromatography–selected tryptic peptides from the resulting 130- and 150-kd bands revealed a 100% identity with amino acids 393 to 405 of ectonucleotide pyrophosphatase/phosphodiesterase-3 (E-NPP3; CD203c) but not of the related ectoenzyme PC-1 (E-NPP1). Moreover, MoAb 97A6 selectively recognized 293 cells transfected with human E-NPP3, but did not react with cells transfected with PC-1 or parental 293 cells. In addition, E-NPP3 messenger RNA expression was detected in basophils but not other peripheral blood cells. Finally, MoAb 97A6 immunoprecipitated phosphodiesterase activity from KU-812 cells and peripheral blood basophils, but not from other cell populations. These data demonstrate that MoAb 97A6 recognizes the functionally active type II transmembrane ectoenzyme E-NPP3.
JC-1: a very sensitive fluorescent probe to test Pgp activity in adult acute myeloid leukemia
Authors: Legrand Ollivier , Perrot Jean-Yves , Simonin Ghislaine , Baudard Marion , Marie Jean-Pierre
Published: July 2002

One of the best-characterized resistance mechanisms in acute myeloid leukemia (AML) is the drug extrusion mediated by P-glycoprotein (Pgp). Recently the results of workshops organized by several groups concluded that accurate measurement of low activity of Pgp is a difficult goal in clinical samples. Therefore, highly sensitive and specific assays were developed to assess the functionality of Pgp using JC-1, a fluorescent molecule with the different emission wavelength (green and red fluorescence) according to its concentration in 129 AML samples. It was shown that JC-1 (green and red bands) may define 3 groups of patients: resistant (R) (29% of patients), intermediate (I) (36%), and sensitive (S) (35%). In contrast, rhodamine 123 assay detected only the R group defined by JC-1. Nevertheless, the I group has an intermediate expression of Pgp (0.39, 0.29, and 0.19 for the R, I, and S groups, respectively, P = .002), an intermediate biologic profile (percentage of CD34, 95%, 67%, and 44%, respectively, P < .0001; in vitro resistance to daunorubicin, 94 μM, 20 μM, and 12 μM, respectively,P = .02), and an intermediate prognosis (achievement of complete remission, 55%, 65%, and 87%, P = .006; 3-year disease-free survival, 11%, 16%, and 36%, respectively,P = .005; and 3-year overall survival, 0%, 20%, and 51%, respectively, P < .0001). Therefore, JC-1 appeared to be a more convenient and simple way to detect a functional Pgp in clinical AML samples than rhodamine 123.
Activated fibroblast growth factor receptor 3 is an oncogene that contributes to tumor progression in multiple myeloma
Authors: Chesi Marta , Brents Leslie A. , Ely Sarah A. , Bais Carlos , Robbiani Davide F. , Mesri Enrique A. , Kuehl W. Michael , Bergsagel P. Leif
Published: July 2002

The t(4;14) translocation occurs frequently in multiple myeloma (MM) and results in the simultaneous dysregulated expression of 2 potential oncogenes, FGFR3 (fibroblast growth factor receptor 3) from der(14) and multiple myeloma SET domain protein/Wolf-Hirschhorn syndrome candidate gene 1 from der(4). It is now shown that myeloma cells carrying a t(4;14) translocation express a functional FGFR3 that in some cases is constitutively activated by the same mutations that cause thanatophoric dysplasia. As with activating mutations of K-ras and N-ras, which are reported in approximately 40% of patients with MM, activating mutations ofFGFR3 occur during tumor progression. However, the constitutive activation of ras and FGFR3 does not occur in the same myeloma cells. Thus the activated forms of these proteins appear to share an overlapping role in tumor progression, suggesting that they also share the signaling cascade. Consistent with this prediction, it is shown that activated FGFR3—when expressed at levels similar to those seen in t(4;14) myeloma—is an oncogene that acts through the MAP kinase pathway to transform NIH 3T3 cells, which can then generate tumors in nude mice. Thus,FGFR3, when overexpressed in MM, may be not only oncogenic when stimulated by FGF ligands in the bone marrow microenvironment, but is also a target for activating mutations that enable FGFR3to play a ras-like role in tumor progression.
Cytogenetic, interphase, and multicolor fluorescence in situ hybridization analyses in primary plasma cell leukemia: a study of 40 patients at diagnosis, on behalf of the Intergroupe Francophone du Myélome and the Groupe Français de Cytogénétique Hématologique
Authors: Avet-Loiseau Hervé , Daviet Axelle , Brigaudeau Christophe , Callet-Bauchu Evelyne , Terré Christine , Lafage-Pochitaloff Marina , Désangles François , Ramond Sylvie , Talmant Pascaline , Bataille Régis
Published: July 2002

Primary plasma cell leukemia (PCL) is a rare plasma cell malignancy. Consequently, few large reports have been published. Presented is a cytogenetic analysis of 40 patients with primary PCL compared with 247 newly diagnosed patients with stage III multiple myeloma (MM). Cytogenetic abnormalities were observed in 23 of 34 patients, with usually complex hypodiploid or pseudodiploid karyotypes. Analysis of rearrangements of the 14q32 region revealed significant differences with high cell mass MM—a higher incidence of t(11;14) (33% vs 16%; P < .025) and of t(14;16) (13% vs 1%;P < .002) though incidences of t(4;14) were identical and a higher incidence of monosomy 13 (68% vs 42%;P = .005). Hypodiploid karyotypes and monosomy 13 may explain, at least in part, the poorer prognosis of primary PCL. In contrast, significantly longer survival was observed in patients displaying t(11;14) in comparison with those lacking this translocation (P = .001).
Interleukin-6 is a growth factor for nonmalignant human plasmablasts
Authors: Jego Gaëtan , Bataille Régis , Pellat-Deceunynck Catherine
Published: July 2002

Interleukin-6 (IL-6), although often regarded as a B-cell differentiation factor, was recently described as the essential survival factor for human plasmablasts in vivo in reactive plasmacytosis. The present study reinvestigated the roles of IL-6 and IL-2 in the generation of plasma cells from human memory B cells in vitro. The cells involved in this differentiation process were identified as preplasmablasts (CD20±CD38±CD138−), plasmablasts (CD20−CD38++CD138−), and early plasma cells (CD20−CD38+++CD138+++). IL-2 or IL-10 induced a strong generation of plasmablasts and early plasma cells (PCs). Compared to IL-2 or IL-10, IL-6 alone was inefficient at PC generation. However, when combined with IL-2 or IL-10, IL-6 enhanced generation of early PCs. Moreover, anti–IL-6 monoclonal antibody markedly reduced IL-2–induced generation of early plasma cells, but not of plasmablasts. These roles of IL-2 and IL-6 were consistent with the difference in the expression of their respective receptors (R). CD25 (IL-2Rα) was increased 72 ± 10-fold on activated B cells, but decreased and then disappeared on plasmablasts. Conversely, CD126 (IL-6Rα) was barely expressed on activated B cells, but increased 18 ± 2-fold on preplasmablasts. Finally, IL-6 enhanced the proliferation (2-fold increase) of IL-2–generated plasmablasts. In conclusion, the data indicate that IL-6 is a growth factor for nonmalignant human plasmablasts.
Antiplatelet agents in tissue factor–induced blood coagulation
Authors: Butenas Saulius , Cawthern Kevin M. , van't Veer Cornelis , DiLorenzo Maria E. , Lock Jennifer B. , Mann Kenneth G.
Published: July 2002

Several platelet inhibitors were examined in a tissue factor (TF)–initiated model of whole blood coagulation. In vitro coagulation of human blood from normal donors was initiated by 25 pM TF while contact pathway coagulation was suppressed using corn trypsin inhibitor. Products of the reaction were analyzed by immunoassay. Preactivation of platelets with the thrombin receptor activation peptide did not influence significantly the clotting time or thrombin–antithrombin III complex (TAT) formation. Addition of prostaglandin E1 (5 μM) caused a significant delay in clotting (10.0 minutes) versus control (4.3 minutes). The prolonged clotting time is correlated with delays in platelet activation, formation of TAT, and fibrinopeptide A (FPA) release. In blood from subjects receiving acetylsalicylic acid (ASA or aspirin), none of the measured products of coagulation were significantly affected. Similarly, no significant effect was observed when 5 μM dipyridamole (Persantine) was added to the blood. Antagonists of the platelet integrin receptor glycoprotein (gp) IIb/IIIa had intermediate effects on the reaction. A 1- to 2-minute delay in clot time and FPA formation was observed with addition of the antibodies 7E3 and Reopro (abciximab) (10 μg/mL), accompanied by a 40% to 70% reduction in the maximal rate of TAT formation and delay in platelet activation. The cyclic heptapetide, Integrilin (eptifibatide), at 5 μM concentration slightly prolonged clot time and significantly attenuated the maximum rate of TAT formation. The disruption of the gpIIb/IIIa-ligand interaction not only affects platelet aggregation, but also decreases the rate of TF-initiated thrombin generation in whole blood, demonstrating a potent antithrombotic effect superimposed on the antiaggregation characteristics.
Antithymocyte globulin for graft-versus-host disease prophylaxis in transplants from unrelated donors: 2 randomized studies from Gruppo Italiano Trapianti Midollo Osseo (GITMO)
Authors: Bacigalupo Andrea , Lamparelli Teresa , Bruzzi Paolo , Guidi Stefano , Alessandrino Paolo Emilio , di Bartolomeo Paolo , Oneto Rosa , Bruno Barbara , Barbanti Mario , Sacchi Nicoletta , Van Lint Maria Teresa , Bosi Alberto ,   for Gruppo Italiano Trapianti Midollo Osseo (GITMO)
Published: July 2002

One hundred nine patients with hematologic malignancies, undergoing bone marrow transplants (BMT) from unrelated donors, were randomized in 2 consecutive trials to receive or not to receive antithymocyte globulin (ATG) in the conditioning regimen, as follows: (A) 54 patients (median age, 28 years; 39% with advanced disease) were randomized to no ATG (n = 25) versus 7.5 mg/kg rabbit ATG (Thymoglobulin; Sangstat, Lyon, France) (n = 29) ; (B) 55 patients (median age, 31 years, 71% with advanced disease) were randomized to no ATG (n = 28) versus 15 mg/kg rabbit ATG (n = 27). Grade III-IV graft-versus-host disease (GVHD) was diagnosed in 36% versus 41% (P = .8) in the first and in 50% versus 11% (P = .001) in the second trial. Transplant-related mortality (TRM), relapse, and actuarial 3-year survival rates were comparable in both trials. In fact, despite the reduction of GVHD in the second trial, a higher risk for lethal infections (30% vs 7%; P = .02) was seen in the arm given 15 mg/kg ATG. Extensive chronic GVHD developed overall more frequently in patients given no ATG (62% vs 39%;P = .04), as confirmed by multivariate analysis (P = .03). Time to 50 × 109/L platelets was comparable in the first trial (21 vs 24 days; P = .3) and delayed in the ATG arm in the second trial (23 vs 38 days;P = .02). These trials suggest that (1) 15 mg/kg ATG before BMT significantly reduces the risk for grade III-IV acute GVHD, (2) this does not translate to a reduction in TRM because of the increased risk for infections, and (3) though survival is unchanged, extensive chronic GVHD is significantly reduced in patients receiving ATG.
Rearrangements of the c-myc oncogene are present in 15% of primary human multiple myeloma tumors
Authors: Avet-Loiseau Hervé , Gerson Fabienne , Magrangeas Florence , Minvielle Stéphane , Harousseau Jean-Luc , Bataille Régis
Published: July 2002

Rearrangements of the c-myc oncogene have been found in most plasmacytomas induced in mice and human myeloma cell lines (HMCLs) analyzed so far. However, neither induced mouse plasmacytomas nor HMCLs represent relevant models for human multiple myeloma (MM). To evaluate the incidence of c-mycrearrangements in human plasma cell dyscrasias, sets of probes were generated to allow direct assessment of c-myctranslocations on interphase plasma cells by using fluorescence in situ hybridization. After validation of these probes, a large cohort of patients with either newly diagnosed MM (n = 529), relapsed MM (n = 58), primary plasma cell leukemia (PCL; n = 23), monoclonal gammopathy of undetermined significance (n = 65), or smoldering MM (n = 24) were analyzed. C-myc rearrangements were identified in 15% of patients with MM or primary PCL, independently of the stage of the disease (ie, diagnosis or relapse and MM or primary PCL). Analysis of the 2 main translocations observed on karyotyping, ie, t(8;14) and t(8;22), revealed that these specific translocations represented only 25% (23 of 91) of c-mycrearrangements. c-myc rearrangements were then correlated with several other patients' characteristics: illegitimateIgH recombinations, chromosome 13 deletions, and serum β2-microglobulin levels. The only significant correlation was with a high β2-microglobulin level (P = .002), although a trend for association with t(4;14) was observed (P = .08). Thus, c-myc rearrangement analysis in patients with MM revealed a strikingly lower incidence than that in HMCLs and plasmacytomas induced in mice, indicating that data obtained with these models cannot be directly extrapolated to human MM.
Dermal and pulmonary inflammatory disease in E-selectin and P-selectin double-null mice is reduced in triple-selectin–null mice
Authors: Collins Robert G. , Jung Unsu , Ramirez Maricela , Bullard Daniel C. , Hicks M. John , Smith C. Wayne , Ley Klaus , Beaudet Arthur L.
Published: July 2002

In the initial phase of an inflammatory response, leukocytes marginate and roll along the endothelial surface as a result of adhesive interactions between molecules on the endothelial cells and leukocytes. To evaluate the role of the 3 selectins (E, L, and P) in leukocyte rolling and emigration, a null mutation for L-selectin was introduced into previously described embryonic stem cells with null mutations in the genes for both E-selectin and P-selectin (E/P double mutants) to produce triple-selectin–null mice (E-selectin, L-selectin, and P-selectin [E/L/P] triple mutants). Triple-selectin homozygous mutant mice are viable and fertile and only rarely develop the severe mucocutaneous infections or pulmonary inflammation characteristic of E/P double-mutant mice. Surface expression of L-selectin was undetectable in triple-mutant mice on fluorescence-activated cell-sorter analysis of peripheral neutrophils. Pathological studies revealed moderate cervical lymphadenopathy and lymphoplasmacytic infiltrate, but these were less extensive than in E/P double-mutant mice. Neutrophil emigration during thioglycolate-induced peritonitis was significantly reduced at 4, 8, and 24 hours (35%, 65%, and 46% of wild-type values, respectively). Intravital microscopy of the cremaster muscle revealed almost no rolling at times up to 6 hours after exteriorization, with or without addition of tumor necrosis factor α. The small amount of residual rolling was dependent on α4-integrin. The occurrence of skin and pulmonary disease in E/P double-mutant mice but not E/L/P triple-mutant mice suggests that deficiency of L-selectin alters the inflammatory response in E/P mutants.
Defective expression of the interleukin-2/interleukin-15 receptor β subunit leads to a natural killer cell–deficient form of severe combined immunodeficiency
Authors: Gilmour Kimberly C. , Fujii Hodaka , Cranston Treena , Davies E. Graham , Kinnon Christine , Gaspar Hubert B.
Published: July 2002

Development of T and natural killer (NK) cells is critically dependent on cytokine signaling, and defects in cytokine receptor complex subunits have been shown to result in severe combined immunodeficiency (SCID) syndromes in humans and in murine models. An infant boy had typical clinical features of SCID and was found to lack NK cells in his peripheral circulation. Molecular analysis did not reveal abnormalities in his γc or JAK-3 genes, and he was investigated for defects in the interleukin-15 (IL-15) receptor complex because functional IL-15 signaling is essential for NK cell development. Expression of the IL-2R/IL-15Rβ chain was significantly reduced in the patient's peripheral blood mononuclear cells (PBMCs) by immunoblot, flow cytometry, and Northern blot analysis. Furthermore, IL-2 stimulation of PBMCs showed only minimal tyrosine phosphorylation of JAK-3. These data demonstrate that defects in IL-2R/1L-15Rβ expression can lead to a unique NK-deficient SCID immunophenotype.
Cell surface tissue transglutaminase is involved in adhesion and migration of monocytic cells on fibronectin
Authors: Akimov Sergey S. , Belkin Alexey M.
Published: July 2002

Expression of tissue transglutaminase (transglutaminase II, tTG) was shown to increase drastically during monocyte differentiation into macrophages; however, its role in monocytic cells remains largely unknown. This study describes a novel function of cell surface tTG as an adhesion and migration receptor for fibronectin (Fn). Two structurally related transglutaminases, tTG and the A subunit of factor XIII (FXIIIA), are expressed on the surface of monocytic cells, whereas only surface tTG is associated with multiple integrins of the β1 and β3 subfamilies. Both surface levels of tTG and the amounts of integrin-bound tTG are sharply up-regulated during the conversion of monocytes into macrophages. In contrast, a reduction in biosynthesis and surface expression of FXIIIA accompanies monocyte differentiation. Cell surface tTG is colocalized with β1- and β3-integrins in podosomelike adhesive structures of macrophages adherent on Fn. Down-regulation of surface tTG by expression of antisense tTG construct or its inhibition by function-blocking antibodies significantly decreases adhesion and spreading of monocytic cells on Fn and, in particular, on the gelatin-binding fragment of Fn consisting of modules I6II1,2I7-9. Likewise, interfering with the adhesive function of surface tTG markedly reduces migration of myeloid cells on Fn and its gelatin-binding fragment. These data demonstrate that cell surface tTG serves as an integrin-associated adhesion receptor that might be involved in extravasation and migration of monocytic cells into tissues containing Fn matrices during inflammation.
Chronic graft-versus-host disease after allogeneic blood stem cell transplantation
Authors: Przepiorka Donna , Anderlini Paolo , Saliba Rima , Cleary Karen , Mehra Rakesh , Khouri Issa , Huh Yang O. , Giralt Sergio , Braunschweig Ira , van Besien Koen , Champlin Richard
Published: July 2002

The incidence, characteristics, risk factors for, and impact of chronic graft-vs-host disease (GVHD) were evaluated in a consecutive series of 116 evaluable HLA-identical blood stem cell transplant recipients. Minimum follow-up was 18 months. Limited chronic GVHD occurred in 6% (95% confidence interval [CI], 0%-13%), and clinical extensive chronic GVHD in 71% (95% CI, 61%-80%). The cumulative incidence was 57% (95% CI, 48%-66%). In univariate analyses, GVHD prophylaxis other than tacrolimus and methotrexate, prior grades 2 to 4 acute GVHD, use of corticosteroids on day 100, and total nucleated cell dose were significant risk factors for clinical extensive chronic GVHD. On multivariate analysis, GVHD prophylaxis with tacrolimus and methotrexate was associated with a reduced risk of chronic GVHD (hazard ratio [HR], 0.35; P = .001), whereas the risk was increased with prior acute GVHD (HR, 1.67;P = .046). When adjusted for disease status at the time of transplantation, high-risk chronic GVHD had an adverse impact on overall mortality (HR, 6.6; P < .001) and treatment failure (HR, 5.2; P < .001) at 18 months. It was concluded that there is a substantial rate of chronic GVHD after HLA-identical allogeneic blood stem cell transplantation, that clinical factors may alter the risk of chronic GVHD, and that high-risk chronic GVHD adversely affects outcome.
FLT3 ligand can substitute for macrophage colony-stimulating factor in support of osteoclast differentiation and function
Authors: Lean Jeny Maree , Fuller Karen , Chambers Timothy John
Published: July 2002

Although bone resorption and osteoclast numbers are reduced in osteopetrotic (op/op) mice, osteoclasts are nevertheless present and functional, despite the absence of macrophage colony-stimulating factor (M-CSF). This suggests that alternative factors can partly compensate for the crucial actions of M-CSF in osteoclast induction. It was found that when nonadherent bone marrow cells were incubated in RANKL with Flt3 ligand (FL) without exogenous M-CSF, tartrate-resistance acid phosphatase (TRAP)–positive cells were formed, and bone resorption occurred. Without FL, only macrophagelike TRAP-negative cells were present. Granulocyte-macrophage CSF, stem cell factor, interleukin-3, and vascular endothelial growth factor could not similarly replace the need for M-CSF. TRAP-positive cell induction in FL was not due to synergy with M-CSF produced by the bone marrow cells themselves because FL also enabled their formation from the hemopoietic cells of op/op mice, which lack any M-CSF. FL appeared to substitute for M-CSF by supporting the differentiation of adherent cells that express mRNA for RANK and responsiveness to RANKL. To determine whether FL can account for the compensation for M-CSF deficiency that occurs in vivo, FL signaling was blockaded in op/op mice by the injection of soluble recombinant Flt3. It was found that the soluble receptor induced a substantial decrease in osteoclast number, strongly suggesting that FL is responsible for the partial compensation for M-CSF deficiency that occurs in these mice.
Complement-mediated cell death induced by rituximab in B-cell lymphoproliferative disorders is mediated in vitro by a caspase-independent mechanism involving the generation of reactive oxygen species
Authors: Bellosillo Beatriz , Villamor Neus , López-Guillermo Armando , Marcé Silvia , Esteve Jordi , Campo Elias , Colomer Dolors , Montserrat Emili
Published: July 2002

Mechanisms involving the in vitro effect of rituximab in cells from 55 patients with B-cell lymphoproliferative disorders were investigated. No cytotoxic effect was observed when cells were incubated with rituximab alone, but in the presence of human AB serum rituximab induced complement-dependent cell death (R-CDC). A cytotoxic effect was observed in cells from 9 of 33 patients with B-cell chronic lymphocytic leukemia, 16 of 16 patients with mantle-cell lymphoma, 4 of 4 patients with follicular lymphoma, and 2 of 2 patients with hairy-cell leukemia. R-CDC was observed in cells from patients expressing more than 50 × 103 CD20 molecules per cell, and directly correlated with the number of CD20 molecules per cell. Preincubation with anti-CD59 increased the cytotoxic effect of rituximab and sensitized cells from nonsensitive cases. Neither cleavage of poly-ADP ribose polymerase (PARP) nor activation of caspase-3 was observed in R-CDC. In addition, no cells with a hypodiploid DNA content were detected and R-CDC was not prevented by a broad-spectrum caspase inhibitor, suggesting a caspase-independent mechanism. Incubation with rituximab in the presence of AB serum induced a rapid and intense production of reactive oxygen species (ROS). R-CDC was blocked by the incubation of cells with N-acetyl-L-cysteine (NAC) or Tiron, 2 ROS scavengers, indicating that the cytotoxic effect was due to the generation of superoxide (O2−) radicals. In conclusion, the results of the present study suggest that CD20, CD59, and complement have a role in the in vitro cytotoxic effect of rituximab, which is mediated by a caspase-independent process that involves ROS generation.
Inhibition of fibrinolysis by recombinant factor VIIa in plasma from patients with severe hemophilia A
Authors: Lisman Ton , Mosnier Laurent O. , Lambert Thierry , Mauser-Bunschoten Evelien P. , Meijers Joost C. M. , Nieuwenhuis H. Karel , de Groot Philip G.
Published: July 2002

Recombinant factor VIIa (rFVIIa) is a novel prohemostatic drug for patients with hemophilia who have developed inhibitory antibodies. The postulation has been made that hemophilia is not only a disorder of coagulation, but that hyperfibrinolysis due to a defective activation of thrombin activatable fibrinolysis inhibitor (TAFI) might also play a role. In this in vitro study, the potential of rFVIIa to down-regulate fibrinolysis via activation of TAFI was investigated. rFVIIa was able to prolong clot lysis time in plasmas from 17 patients with severe hemophilia A. The prolongation of clot lysis time by rFVIIa was completely abolished by addition of an inhibitor of activated TAFI. The concentration of rFVIIa required for half maximal prolongation of clot lysis time (Clys½-VIIa) varied widely between patients (median, 73.0 U/mL; range, 10.8-250 U/mL). The concentration of rFVIIa required for half maximal reduction of clotting time (Cclot½-VIIa) was approximately 10-fold lower than the Clys½-VIIa value (median, 8.4 U/mL; range, 1.7-22.5 U/mL). Inhibition of TFPI with a polyclonal antibody significantly decreased Clys½-VIIa values (median, 2.6 U/mL; range, 0-86.9 U/mL), whereas Cclot½-VIIa values did not change (median, 7.2 U/mL; range, 2.2-22.5 U/mL). On addition of 100 ng/mL recombinant full-length TFPI, a nonsignificant increase of Clys½-VIIa values was observed (median, 119.2 U/mL; range, 12.3-375.0 U/mL), whereas Cclot½-VIIa values did not change (median, 8.8 U/mL; range, 2.6-34.6 U/mL). In conclusion, this study shows that rFVIIa both accelerates clot formation and inhibits fibrinolysis by activation of TAFI in factor VIII-deficient plasma. However, a large variability in antifibrinolytic potential of rFVIIa exists between patients.
Follicular lymphoma with a novel t(14;18) breakpoint involving the immunoglobulin heavy chain switch mu region indicates an origin from germinal center B cells
Authors: Fenton James A. L. , Vaandrager Jan-Willem , Aarts Wilhelmina M. , Bende Richard J. , Heering Karel , van Dijk Martin , Morgan Gareth , van Noesel Carel J. M. , Schuuring Ed , Kluin Philip M.
Published: July 2002

With the use of DNA-fiber fluorescent in situ hybridization, a BCL2 protein positive follicular lymphoma with a novel BCL2 breakpoint involving the immunoglobulin heavy chain (IGH) switch mu (Sμ) region instead of the JH orDH gene segments was identified. Sequence analysis showed that the genomic breakpoint is localized between the Sμ region of the IGH complex and the first intron of BCL2. Reverse-transcriptase polymerase chain reaction showed expression of a unique hybrid IGH-BCL2 transcript involving the transcription initiation site Iμ. Sequence analysis of the VH region of the functional nontranslocatedIGH allele showed multiple shared somatic mutations but also a high intraclonal variation (53 differences in 15 clones), compatible with the lymphoma cells staying in or re-entering the germinal center. This is the first example of a t(14;18) translocation that results from an illegitimate IGH class-switch recombination during the germinal center B-cell stage.
Superior survival associated with transplantation of matched unrelated versus one-antigen–mismatched unrelated or highly human leukocyte antigen– disparate haploidentical family donor marrow grafts for the treatment of hematologic malignancies: establishing a treatment algorithm for recipients of alternative donor grafts
Authors: Drobyski William R. , Klein John , Flomenberg Neal , Pietryga Daniel , Vesole David H. , Margolis David A. , Keever-Taylor Carolyn A.
Published: July 2002

The purpose of this study was to compare transplantation outcomes in patients with hematologic malignancies who received marrow grafts from either phenotypically matched unrelated, one- antigen–mismatched unrelated, or highly human leukocyte antigen (HLA)–disparate family donors. Between 1993 and 2000, 139 patients underwent transplantation from unrelated donors (81 matched and 58 mismatched) and 48 patients received marrow grafts from family donors that were mismatched at 2, 3, or 4 of 8 HLA loci. All patients received a standardized conditioning regimen and a graft-versus-host disease (GVHD) prophylaxis schedule with the exception of recipients of haploidentical marrow grafts, who received antithymocyte globulin after bone marrow transplantation as additional immunosuppression. There was no statistically significant difference in the rate of engraftment, or the cumulative incidences of acute and chronic GVHD between any of the 3 groups. The 2-year cumulative incidence of relapse was lower in matched unrelated patients (25%, P = .01) and mismatched unrelated patients (26%,P = .014) than in haploidentical patients (42%). Transplant-related mortality was significantly higher in recipients of mismatched unrelated grafts (45%, P = .01) and haploidentical grafts (42%, P = .001) compared with recipients of matched unrelated marrow grafts (23%). This resulted in a significantly higher probability of overall survival for matched unrelated patients (58%) versus either mismatched unrelated (34%,P = .01) or haploidentical (21%, P = .002) patients. There was no statistically significant difference in survival between patients who received mismatched unrelated grafts versus those who received haploidentical grafts. This study supports a donor selection algorithm whereby patients who lack a closely matched family donor be offered a phenotypically matched unrelated donor if available. There is no apparent advantage to using a mismatched unrelated versus a highly HLA-disparate family donor.
Treatment of adult acute lymphoblastic leukemia (ALL): long-term follow-up of the GIMEMA ALL 0288 randomized study
Authors: Annino Luciana , Vegna Maria Luce , Camera Andrea , Specchia Giorgina , Visani Giuseppe , Fioritoni Giuseppe , Ferrara Felicetto , Peta Antonio , Ciolli Stefania , Deplano Wilma , Fabbiano Francesco , Sica Simona , Di Raimondo Francesco , Cascavilla Nicola , Tabilio Antonio , Leoni Pietro , Invernizzi Rosangela , Baccarani Michele , Rotoli Bruno , Amadori Sergio , Mandelli Franco
Published: July 2002

The GIMEMA ALL 0288 trial was designed to evaluate the impact of a 7-day prednisone (PDN) pretreatment on complete remission (CR) achievement and length, the influence of the addition of cyclophosphamide (random I) to a conventional 4-drug induction on CR rate and duration, and whether an early post-CR intensification (random II) by an 8-drug consolidation could improve CR duration. Median follow-up of this study was 7.3 years. From January 1988 to April 1994, among 794 adult (> 12 but < 60 years) patients registered, 778 were eligible. Their median age was 27.5 years; 73% had B-lineage acute lymphoblastic leukemia (ALL) and 22% had T-lineage disease; 18% showed associated myeloid markers; 47 of 216 analyzed patients (22%) had Philadelphia chromosome–positive ALL. Response to PDN pretreatment was observed in 65% of cases. CR was achieved in 627 patients (82%). Resistant patients and induction death rates were 11% and 7%, respectively. Random II was applied to 388 patients with CR; 201 had maintenance alone and 187 had consolidation followed by maintenance. The relapse rate was 60%; isolated central nervous system relapses were 8% of all CRs and 13% of all relapses. Median survival (overall survival [OS]), continuous complete remission (CCR), and disease-free survival (DFS) were 2.2, 2.4, and 2 years, respectively. PDN pretreatment response resulted the main independent factor influencing CR achievement, OS, CCR, and DFS; the addition of cyclophosphamide in induction significantly influenced CR achievement in a multivariate analysis. Neither induction intensification nor early consolidation appeared to influence CCR and DFS duration. For the first time PDN pretreatment response proved to be a powerful factor predicting disease outcome in adult ALL patients.
Precise quantification of minimal residual disease at day 29 allows identification of children with acute lymphoblastic leukemia and an excellent outcome
Authors: Nyvold Charlotte , Madsen Hans O. , Ryder Lars P. , Seyfarth Jeanette , Svejgaard Arne , Clausen Niels , Wesenberg Finn , Jonsson Olafur G. , Forestier Erik , Schmiegelow Kjeld
Published: July 2002

The postinduction level of minimal residual disease (MRD) was quantified with a competitive polymerase chain reaction (PCR) technique in 104 children with acute lymphoblastic leukemia (ALL) diagnosed between June 1993 and January 1998 and followed for a median of 4.2 years. A significant correlation was found between the MRD level on day 15 (D15) and day 29 (D29) after the start of induction therapy (rs = 0.70, P < .0001). The 15 patients with T-cell disease had higher D29 MRD than those with B-lineage ALL (P = .01). Age was positively related to D29 MRD (rs = 0.32, P = .001). The 16 patients who had a relapse had higher D15 and D29 MRD levels than the patients who stayed in remission (median levels D15, 1% versus 0.1%,P = .03; D29, 0.4% versus 0.01%,P = .0001). No patients with a MRD level less than 0.01% on D29 have so far had a relapse, whereas the 7-year probability of event-free survival for patients with higher MRD levels was 0.52 (P = .0007). The group of patients with a D29 MRD less than 0.01% included patients with T-cell disease, white blood cell count more than 50 × 109/L at diagnosis, or age 10 years or older, and could not be identified by up-front criteria. The best-fit Cox model to predict the risk of relapse included D29 MRD (P = .004) and age (P = .009). These findings indicate that with the present treatment protocol MRD quantification at an early stage of therapy identifies patients with a very low risk of relapse. Further trials are needed to reveal whether such patients with D29 MRD less than 0.01% can be cured with less intensive chemotherapy, which would reduce the risk of serious late effects as well as the costs of therapy.
Severe deficiency of switched memory B cells (CD27+IgM−IgD−) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease
Authors: Warnatz Klaus , Denz Axel , Dräger Ruth , Braun Moritz , Groth Christoph , Wolff-Vorbeck Guido , Eibel Hermann , Schlesier Michael , Peter Hans Hartmut
Published: July 2002

Hypogammaglobulinemia is the hallmark of common variable immunodeficiency (CVID) syndrome, a heterogeneous disorder predisposing patients to recurrent bacterial infections. In this study, we investigated the peripheral B-cell compartment of 30 well-characterized CVID patients in comparison to 22 healthy controls. Flow cytometric analysis of peripheral blood lymphocytes revealed a reduction of class-switched CD27+IgM−IgD− memory B cells below 0.4% in 77% of our patients (group I), while this B-cell subpopulation exceeded 0.5% in all healthy donors and in 23% of CVID patients (group II). These results correlate well with the capacity of peripheral blood lymphocytes to produce immunoglobulins in vitro upon stimulation with Staphylococcus aureus Cowan I (SAC) plus interleukin-2 because the production of immunoglobulin G in vitro is entirely dependent on the presence of switched memory B cells. The subdivision of group I into patients with an increased proportion of CD21− peripheral B cells (> 20%; group Ia) and patients with normal percentages of CD21− B cells (< 20%; group Ib) revealed a significant clustering of patients with splenomegaly and autoimmune cytopenias in group Ia. Based on these observations, we propose a fast and reliable new classification for CVID patients by flow cytometric quantification of class-switched memory and immature B cells in the peripheral blood of patients. Our results point toward defects at various stages of B-cell differentiation in CVID subgroups and support the value of a B-cell–oriented classification principle. A consensus on this new classification system will hopefully provide a tool for rapidly defining homogeneous subgroups of CVID for functional studies and genetic linkage analysis.
Increased sensitivity of T lymphocytes to tumor necrosis factor receptor 1 (TNFR1)– and TNFR2-mediated apoptosis in HIV infection: relation to expression of Bcl-2 and active caspase-8 and caspase-3
Authors: de Oliveira Pinto Luzia Maria , Garcia Sylvie , Lecoeur Hervé , Rapp Christophe , Gougeon Marie-Lise
Published: July 2002

The destruction of CD4 T cells in human immunodeficiency virus (HIV) infection is associated with activation of apoptotic programs, partly mediated by death receptors. The role of CD95L/CD95 in depletion of patients' CD4 T cells is well documented, but the possible contribution of the tumor necrosis factor/tumor necrosis factor receptor (TNF/TNFR) pathway has not been examined. In this study, we found that both TNFR1 and TNFR2 induced marked apoptosis in peripheral T cells from HIV-infected persons, involving both CD4 and CD8 T cells. Longitudinal follow-up of HIV+ patients suggests an association between the in vivo evolution of CD4 T-cell numbers and variations in susceptibility to TNFR-induced apoptosis. Analysis of molecular mechanisms involved showed that it was not related to altered ex vivo expression of TNFR1-associated death domain, receptor interacting protein, or TNFR-associated factor 2. Susceptibility to TNFR-mediated apoptosis was rather related to Bcl-2 expression, because patients' T cells expressing high levels of Bcl-2 were completely protected from TNFR1- and TNFR2-induced cell death, whereas T cells expressing normal levels of Bcl-2 were not protected in patients in contrast to controls. Early recruitment of caspase-8 and caspase-3 is needed to transduce the apoptotic signals, and expression of both caspases in their active form was detected in blood T cells from HIV+ patients, whereas it was hardly detected in controls. Moreover, ligation of TNFRs induced increased activation of both caspases in patients' T cells. Together these data demonstrate that exacerbated TNFR-mediated cell death of T cells from HIV-infected individuals is associated with both alteration of Bcl-2 expression and activation of caspase-8 and caspase-3 and may contribute to the pathogenesis of acquired immunodeficiency syndrome.
A FACETS analysis of rater bias in measuring Japanese second language writing performance
Authors: Kondo-Brown Kimi
Published: July 2002

The purpose of this study is twofold. First, using FACETS (Linacre, 1996), it investigates how judgements of trained teacher raters are biased towards certain types of candidates and certain criteria in assessing Japanese second language (L2) writing. Previous studies that identified significantly biased rater-candidate interactions did not discuss who the candidates were, but this study examines rater-candidate interactions in much more detail. Secondly, since there is no established rating scale for assessing Japanese L2 writing, this study explores the potential for using a modified version of Jacobs et al.’s (1981) rating scale for norm-referenced decisions about Japanese L2 writing ability. The participants in the study comprised 234 university candidates and three trained teacher raters. The raters produced highly correlated scores and were self-consistent, but significant differences in overall severity surfaced. The raters scored certain candidates and criteria more leniently or harshly, and every rater’s bias pattern was different. The highest percentage of significantly biased rater-candidate interactions was found among the candidates whose ability was extremely high or low. This study suggests that the modified version of Jacobs et al.’s scale can be a reliable tool in assessing Japanese L2 writing in norm-referenced settings, but multiple ratings are still necessary.