Organelle proteomics: looking at less to see more.

The recent finding that the human genome comprises between 21 000 and 39 000 genes, a number much lower than expected, has in no way simplified the complexity associated with the understanding of how cells perform their functions. Elucidation of the molecular mechanisms underlying cell functions will require a global knowledge of the expressed proteins, including splice variant products, their post-translational modifications, their subcellular localizations and their assembly into molecular machines as deduced from protein-protein interactions, at any given time during the life of the cell or under any cellular conditions. Current and expected advances in mass spectrometry and bioinformatics might help the realization of these goals in a shorter time than is currently predicted.

Interclade neutralization and enhancement of human immunodeficiency virus type 1 identified by an assay using HeLa cells expressing both CD4 receptor and CXCR4/CCR5 coreceptors.

We report on the development and evaluation of a human immunodeficiency virus (HIV) neutralization assay that uses P4P cells, which are CD4+CXCR4+CCR5+ HeLa cells that carry the lacZ gene under the control of the HIV-1 long terminal repeat. The results of the present study suggest that the P4P assay can be used for the extensive study of both neutralizing and enhancing activity in serum samples from HIV-1-infected patients and from vaccinated individuals.

Evidence for the rare decay B-->K*l+l- and measurement of the B-->Kl+l- branching fraction.

We present evidence for the flavor-changing neutral current decay B-->K*l+l- and a measurement of the branching fraction for the related process B-->K l+l-, where l+l- is either an epsilon+epsilon- or a mu+mu- pair. These decays are highly suppressed in the standard model, and they are sensitive to contributions from new particles in the intermediate state. The data sample comprises 123 x 10(6) Upsilon(4S)-->B(-)B decays collected with the BABAR detector at the SLAC PEP-II epsilon+epsilon- storage ring.

Observation of a significant excess of pi0pi0 events in B meson decays.

We present a study of the decay B0-->pi(0)pi(0) based on a sample of 124 x 10(6) BB pairs recorded by the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC. We observe 46+/-13+/-3 events, where the first error is statistical and the second is systematic, corresponding to a significance of 4.2 standard deviations including systematic uncertainties.

FLT3 mutations are associated with other molecular lesions in AML.

The basic molecular defects underlying acute myeloid leukemias (AML) seem to be caused by inactivating mutations in transcription factors which control normal myeloid differentiation (Class II mutations) and genetic lesions in tyrosine kinases resulting in constitutive activation (Class I mutations). We sought to determine the frequency of associated mutations (Class I + Class II) in a consecutive series of adult de novo AML (353 patients) in order to stress the validity of this model. Mutations and rearrangements at the FLT3, AML1/ETO, CBFbeta/MYH11, AML1, CEBPalpha and MLL genes were investigated using standard molecular methods.

Organelle proteomics: looking at less to see more.

The recent finding that the human genome comprises between 21000 and 39000 genes, a number much lower than expected, has in no way simplified the complexity associated with the understanding of how cells perform their functions. Elucidation of the molecular mechanisms underlying cell functions will require a global knowledge of the expressed proteins, including splice variant products, their post-translational modifications, their subcellular localizations and their assembly into molecular machines as deduced from protein-protein interactions, at any given time during the life of the cell or under any cellular conditions. Current and expected advances in mass spectrometry and bioinformatics might help the realization of these goals in a shorter time than is currently predicted.

Long-term disease-free survival in patients with angioimmunoblastic T-cell lymphoma after high-dose chemotherapy and autologous stem cell transplantation.

Background And Objectives: Patients with angioimmunoblastic T-cell lymphoma (AIL) have a poor prognosis with conventional treatment.

Design And Methods: We initiated an EBMT-based survey studying the impact of high-dose chemotherapy (HDCT) and autologous hematopoietic stem cell transplantation in patients with AIL. Data on 29 patients, who were transplanted between 1992 and 1998 in 16 transplant centers, were collected on standardized documentation forms.

Peripheral blood is safer than bone marrow as a source of hematopoietic progenitors in patients with myelodysplastic syndromes who receive an allogeneic transplantation. Results from the Spanish registry.

Allogeneic hematopoietic stem cell transplantation (HSCT) is the only curative therapy for patients with myelodysplastic syndromes (MDSs). We have analyzed the outcome of 81 patients who underwent an allogeneic transplant from an HLA-identical sibling donor. The overall survival (OS) was 31% and disease-free survival was 30% at 5.

Are exercise social-cognitive factors and behaviours different for adults with diabetes? A randomized community sample.

Exercise plays a key role in the prevention and delay of the onset of Type 2 diabetes and in the management of this disorder. To determine if there are differences in key social-cognitive determinants of exercise and self-reported physical activity levels between adults with diabetes and those without the condition, a random selected sample of adults was surveyed. A telephone interview assessed physical activity behaviour and key social-cognitive constructs from major health behaviour change theories/models.

Strategies to reduce transplant-related mortality after allogeneic stem cell transplantation in elderly patients: Comparison of reduced-intensity conditioning and unmanipulated peripheral blood stem cells vs a myeloablative regimen and CD34+ cell selection.

Objectives: The aim of this study was to compare two approaches used to reduce transplant-related mortality (TRM) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT) in elderly patients.

Patients And Methods: Data from 50 patients, 45 years of age or older, consecutively treated with an HLA-identical sibling allo-PBSCT at the Hospital de Sant Pau were analyzed. We have compared the outcome of patients treated with conventional myeloablative regimens and CD34(+)-selected cells (CD34(+) group; n=23) with those receiving reduced-intensity conditioning regimens, consisting of fludarabine (150 mg/m(2)) plus an alkylating agent, followed by unmanipulated grafts (RIC group; n=27).

Post-transplant Burkitt's leukemia or lymphoma. Study of five cases treated with specific intensive therapy (PETHEMA ALL-3/97 trial).

Burkitt's lymphoma (BL) and Burkitt-like acute lymphoblastic leukemia (ALL) are uncommon lymphoproliferative disorders after solid organ or stem cell transplantation. Although their prognosis is considered to be poor, there are scarce data on the clinical characteristics and the response to specific therapies. We report the main clinical characteristics and the results of a specific intensive chemotherapy in 5 adult patients with postransplant BL/ALL3 included in the PETHEMA ALL3/97 protocol.

Allogeneic stem cell transplantation with reduced-intensity conditioning is potentially feasible as an outpatient procedure.

Allogeneic stem cell transplantation (allo-SCT) after a reduced-intensity conditioning (RIC) protocol is associated with decreased short-term toxicity. This suggests that the procedure could be performed on an outpatient basis. We analysed the incidence and risk factors of grade >or=2 conditioning-related toxicities (CRTs) as a hallmark for hospital admission, in 41 consecutive patients allografted from an HLA identical sibling after RIC.

Functionality of the spindle checkpoint during the first meiotic division of mammalian oocytes.

The spindle checkpoint ensures accurate chromosome segregation by delaying anaphase until all chromosomes are correctly aligned on the microtubule spindle. Although this mechanism is conserved throughout eukaryotic evolution, it is unclear whether it operates during meiosis in female mammals. The results of the present study show that in mouse oocytes spindle alterations prevent both chromosome segregation and MPF (M phase promoting factor) inactivation during the first meiotic M phase.

Phagosomes are competent organelles for antigen cross-presentation.

The ability to process microbial antigens and present them at the surface of cells is an important aspect of our innate ability to clear infections. It is generally accepted that antigens in the cytoplasm are loaded in the endoplasmic reticulum and presented at the cell surface on major histocompatibility complex (MHC) class I molecules, whereas peptides present in endo/phagocytic compartments are presented on MHC class II molecules. Despite the apparent segregation of the class I and class II pathways, antigens from intracellular pathogens including mycobacteria, Escherichia coli, Salmonella typhimurium, Brucella abortus and Leishmania, have been shown to elicit an MHC class-I-dependent CD8+ T-cell response, a process referred to as cross-presentation.

Factors predicting peripheral blood progenitor cell collection from pediatric donors for allogeneic transplantation.

Background And Objectives: Although several studies have reported on the use of children as donors for peripheral blood progenitor cells (PBPC), no specific characteristics have been identified as predictors of PBPC collection in this population. In this study we analyzed predictive factors for PBPC collection in pediatric donors.

Design And Methods: We retrospectively analyzed factors predicting the yield for a target CD34+ cell dose of > or =4x10(6)/Kg donor or recipient body weight, in 105 aphereses from 76 healthy pediatric donors (36 boys and 40 girls) included in the Spanish National Donor Registry.

Efficacy of a canarypox virus-vectored vaccine against feline leukaemia.

Canarypox virus recombinant vaccines have a unique efficacy and safety profile for the vaccinated host because the canarypox virus is non-replicative in mammalian hosts. After the vaccination of a mammalian species, recombinant canarypox viruses express the inserted genes but cannot multiply in the host. They stimulate a strong immune response in the absence of any virus amplification in the host or any viral spread into the environment.

Inflammatory reaction without endogenous antioxidant response in Caco-2 cells exposed to iron/ascorbate-mediated lipid peroxidation.

To characterize the role of intestinal epithelial cells in mucosal host defense, we have examined endogenous antioxidant reactivity and inflammatory response in Caco-2 cell line. When differentiated Caco-2 cells were incubated with iron/ascorbate for 1-24 h, they exhibited increased malondialdehyde levels and decreased polyunsaturated fatty acid proportion in favor of saturated fatty acids. These modifications were accompanied with alterations in membrane fluidity and permeability.

Positive selection for CD34+ reduces the incidence and severity of veno-occlusive disease of the liver after HLA-identical sibling allogeneic peripheral blood stem cell transplantation.

Objective: T-cell depletion (TCD), primarily developed to prevent graft-vs-host disease (GVHD), might reduce early liver dysfunction after allogeneic hematopoietic stem cell transplantation. However, no comparative studies have been performed to investigate this. We analyzed the influence of selection for CD34(+) cells on the incidence and severity of hepatic veno-occlusive disease (VOD).

International and Italian prognostic indices in follicular lymphoma.

Background And Objectives: The International Prognostic Index (IPI), initially designed for aggressive lymphomas, has been successfully used in patients with follicular lymphoma (FL). The Italian Lymphoma Intergroup (ILI) created a new prognostic index specific for FL. The aim of this study was to compare which of these two indices is more useful when applied to a large group of FL patients.

Lack of influence of human immunodeficiency virus infection status in the response to therapy and survival of adult patients with mature B-cell lymphoma or leukemia. Results of the PETHEMA-LAL3/97 study.

Background And Objectives: Short, intensive multiagent chemotherapy has resulted in significant progress in Burkitt's lymphoma and leukemia. A protocol was designed to treat all adult patients with mature B-cell lymphoma or leukemia with the aims of comparing the response to therapy and survival with regards to their HIV infection status.

Design And Methods: Fifty-three adult patients with advanced stage Burkitt's lymphoma or Burkitt's leukemia were treated.

Low transplant related mortality in older patients with hematologic malignancies undergoing autologous stem cell transplantation.

Background And Objectives: Patients over 60 years are frequently excluded from autologous stem cell transplantation (ASCT) programs due to a traditionally high rate of transplant-related mortality (TRM) in such indications. We evaluated the results of ASCT in a group of 49 patients >= 60 years of age [32 males, median age 63 years (range, 60 to 71)] autografted in our institution from January 1995 to December 2001.

Design And Methods: There were 27 patients with multiple myeloma, 13 with non-Hodgkin's lymphoma, 3 with acute myelogenous leukemia, 3 with chronic myelogenous leukemia and 3 with other hematological malignancies.

Impact of CD34+ cell dose on the outcome of patients undergoing reduced-intensity-conditioning allogeneic peripheral blood stem cell transplantation.

We analyzed the impact of CD34+ cell dose on the outcome of 86 patients undergoing reduced-intensity conditioning (RIC) allogeneic peripheral blood stem cell transplantation. The RIC was based on fludarabine 150 mg/m2 and melphalan 140 mg/m2 or busulphan 10 mg/kg. A median of 5.

Patients with biochemical iron overload: causes and characteristics of a cohort of 150 cases.

Biochemical iron overload (IO) is a frequent metabolic abnormality. It may be caused by several diseases, and data regarding the relative frequency of these are scant. A single diagnostic protocol including clinical, biochemical, and genetic data was used to diagnose the cause of biochemical IO in a group of 150 patients referred by general practitioners.