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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Escherichia coli replication terminator TerB was inserted in its two alternate orientations into a Bacillus subtilis fork-arrest assay plasmid. After transferring these new plasmids into B. subtilis, which could overproduce the E. coli terminator protein Tus, it was shown that the E. coli Tus–TerB complex could cause polar replication fork arrest, albeit at a very low level, in B. subtilis. A new B. subtilis–E. coli shuttle plasmid was designed to allow the insertion of either the TerI (B. subtilis) or TerB (E. coli) terminator at the same site and in the active orientation in relation to the approaching replication fork generated in either organism. Fork-arrest assays for both terminator-containing plasmids replicating in both organisms which also produced saturating levels of either the B. subtilis terminator protein (RTP) or Tus were performed. The efficiency of the Tus–TerB complex in causing fork arrest was much higher in E. coli than in B. subtilis. The efficiency of the B. subtilis RTP–TerI complex was higher in B. subtilis than in E. coli, but the effect was significantly less. Evidently a specificity feature in E. coli operates to enhance appreciably the fork-arrest efficiency of a Tus–Ter complex. The specificity effect is of less significance for an RTP–Ter complex functioning in B. subtilis.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK; Malden, USA : Blackwell Science Ltd
    Scandinavian journal of immunology 60 (2004), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The only available vaccine against Mycobacterium tuberculosis, the bacille Calmette-Guérin (BCG) vaccine, is at present being used as a reference for the efficacy of novel vaccines. Herein, we demonstrate that viable BCG can be detected at late time points after vaccination in C57BL/6J mice. If BCG is cleared by antibiotic treatment, the number of mycobacteria-reactive effector cells in the spleen gradually reverts to low levels and consequently immunity in this organ wanes, while resistance in the lung remains stable. The implications for comparing BCG vaccination with experimental vaccines including non-replicating vaccines are discussed.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We have used a synthetic-peptide approach to map epitope regions of the Mycobacterium tuberculosis ESAT-6 antigen recognized by human T cells in relation to major histocompatibility complex (MHC) restriction. ESAT-6-specific CD4+ T-cell lines were established by stimulating peripheral blood mononuclear cells from 25 HLA-DR-typed tuberculosis patients with complete antigen in vitro. The established T-cell lines were then screened for proliferation and interferon-γ (IFN-γ) secretion in response to eight overlapping 20-mer peptides covering the ESAT-6 sequence. The response of the T-cell lines to ESAT-6 and peptides from a human leucocyte antigen (HLA)-heterogeneous group of donors suggested the presence of multiple epitopes and promiscuous recognition of the antigen. Analysis of antigen and peptide recognition in the presence of anti-HLA class I and class II antibodies suggested that the T-cell lines recognized ESAT-6 in association with HLA-DR and -DQ molecules. Furthermore, testing of selected T-cell lines with ESAT-6 and the peptides in the presence of autologous and allogeneic HLA-DR- and -DQ-typed antigen-presenting cells identified HLA-DR2, -DR52 and -DQ2 amongst the HLA molecules involved in the presentation of ESAT-6 and its peptides to human Th1 cells. In addition, the T-cell lines were cytotoxic for monocytes and macrophages pulsed with ESAT-6 and peptides. In conclusion, the recognition of ESAT-6 by IFN-γ-secreting and cytotoxic CD4+ T cells in association with frequently expressed HLA class II molecules supports the application of this antigen to either specific diagnosis or subunit vaccine design.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Scandinavian journal of immunology 56 (2002), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Past attempts to raise resistance against Mycobacterium tuberculosis using various preparations of killed mycobacteria have questioned the specificity of the generated immune response. In the present study, we have focused on the protective efficacy of experimental vaccines based on killed mycobacteria. We demonstrate that killed mycobacteria can confer high levels of protection, which can be adoptively transferred to recipient T-cell-deficient mice. Moreover, protective antigens can be found in the cell wall, membrane and cytosol of the mycobacterial cell, and hence emphasize the importance of searching for protective antigens in various compartments of the mycobacterial cell.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Regulation of T-cell receptor (TCR) cell surface expression levels is probably an important mechanism by which T-cell responsiveness is controlled. Previously, two distinct pathways for TCR downregulation have been described. One is dependent on protein kinase C (PKC) and the leucine-based receptor-sorting motif (l-based motif) of the CD3γ chain but independent of tyrosine kinases, whereas the other is dependent on the tyrosine kinase activation but independent of the PKC and the CD3γl-based motif. In this study, we describe a new pathway for TCR downregulation distinct from both the PKC/CD3γl-based motif-dependent and the tyrosine kinase-dependent pathways. This pathway is dependent on ceramide-induced activation of caspases and correlate with caspase-mediated cleavage of the ζ chain. Thus, a 10–15% downregulation of the TCR was induced following the treatment of the T cells with ceramide for 4 h. A close correlation between TCR downregulation, caspase activation, and cleavage of the ζ chain was found. Furthermore, the caspase inhibitors abolished the cleavage of the ζ chain and TCR downregulation in parallel with the inhibition of the caspase activity.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of food science 69 (2004), S. 0 
    ISSN: 1750-3841
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Peripheral blood mononuclear cells (PBMC) were obtained from culture-proven tuberculosis (TB) patients before and after 2 and 6 months of chemotherapy with a multi-drug regimen. PBMC were tested for cellular responses in antigen-induced proliferation and interferon-γ (IFN-γ) assays in response to complex mycobacterial antigens (whole cell Mycobacterium bovis BCG and M. tuberculosis, cell walls and short-term culture filtrate [ST-CF] of M. tuberculosis), fractionated ST-CF antigens (fractions F1–F10) and ESAT-6. The responses in TB patients before anti-TB treatment were low (median stimulation index (SI)=1–7, median delta IFN-γ=0–12 U ml−1, and percent responders=13–67%) to all the antigenic preparations. Following the administration of anti-TB chemotherapy for 2 months, there were significant (P〈0.05) improvements in the cellular responses (median SI=9–76, median delta IFN-γ=3–70 U ml−1, and percent responders=33–100%) to most of the antigenic preparations tested. However, concanavalin A-induced proliferation responses of PBMC from the same patients before and after 2 months of chemotherapy were high and comparable (median SI=101 and 114, respectively, P〉0.05, 100% responders). A further increase in IFN-γ responses (median delta IFN-γ=14–250 U ml−1 and percent responders=43–100%) to mycobacterial antigens was observed in patients receiving chemotherapy for 6 months. Among the ST-CF fractions, F1 and F2 containing low molecular mass proteins resulted in the highest responses, whereas ESAT-6 showed responses comparable to these fractions only in a minority of the patients. HLA-DR typing of these patients showed heterogeneity in the expression of molecules encoded by HLA-DRB genes. These results show that effective chemotherapy restores cellular responses of TB patients to a large number of M. tuberculosis antigens, which could be useful in monitoring the efficacy of anti-TB treatment.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Journal of geodesy 74 (2000), S. 531-551 
    ISSN: 1432-1394
    Keywords: Key words: Kalman filter – RTS smoother – SRIF/S – VLBI
    Source: Springer Online Journal Archives 1860-2000
    Topics: Architecture, Civil Engineering, Surveying
    Notes: Abstract.  The method of square root information filtering and smoothing (SRIF/S) is reviewed and has been implemented in the combined square root information filter and smoother (CSRIFS) program. CSRIFS is a part of the GEOSAT space geodesy software developed at Forsvarets forskningsinstitutt (FFI, The Norwegian Defence Research Establishment). The state vectors and complete variance–covariance matrices from the analyses of a number of independent arcs of space geodesy data can be combined using CSRIFS. Four parameter levels are available and any parameter can, at each level, be represented as either a constant or a stochastic parameter (white noise, colored noise, or random walk). The batch length (i.e. the time interval between the addition of noise to the SRIF array) can be made time and parameter dependent. CSRIFS was applied in the combination of 623 very long baseline interferometry (VLBI) observing sessions. The purpose of this test was to validate the computer implementation of the SRIF/S method and to give an example of how this method can be used in the analysis of a large number of space geodetic observations. The results show that the implementation is very satisfactory.
    Type of Medium: Electronic Resource
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