54 research outputs found

    Antibodies from women urogenitally infected with C. trachomatis predominantly recognized the plasmid protein pgp3 in a conformation-dependent manner

    Get PDF
    <p>Abstract</p> <p>Background</p> <p><it>C. trachomatis </it>organisms carry a cryptic plasmid that encodes 8 open reading frames designated as pORF1 to 8. It is not clear whether all 8 pORFs are expressed during <it>C. trachomatis </it>infection in humans and information on the functionality of the plasmid proteins is also very limited.</p> <p>Results</p> <p>When antibodies from women urogenitally infected with <it>C. trachomatis </it>were reacted with the plasmid proteins, all 8 pORFs were positively recognized by one or more human antibody samples with the recognition of pORF5 protein (known as pgp3) by most antibodies and with the highest titers. The antibody recognition of the pORFs was blocked by <it>C. trachomatis</it>-infected HeLa but not normal HeLa cell lysates. The pgp3 fusion protein-purified human IgG detected the endogenous pgp3 in the cytosol of <it>C. trachomatis</it>-infected cells with an intracellular distribution pattern similar to that of CPAF, a chlamydial genome-encoded protease factor. However, the human antibodies no longer recognized pgp3 but maintained recognition of CPAF when both antigens were linearized or heat-denatured. The pgp3 conformation is likely maintained by the C-terminal 75% amino acid sequence since further deletion blocked the binding by the human antibodies and two conformation-dependent mouse monoclonal antibodies.</p> <p>Conclusion</p> <p>The plasmid-encoded 8 proteins are both expressed and immunogenic with pgp3 as the most immunodominant antigen during chlamydial infection in humans. More importantly, the human anti-pgp3 antibodies are highly conformation-dependent. These observations have provided important information for further understanding the function of the plasmid-encoded proteins and exploring the utility of pgp3 in chlamydial diagnosis and vaccination.</p

    The value of diffusion-weighted imaging in assessing the ADC changes of tissues adjacent to breast carcinoma

    Get PDF
    <p>Abstract</p> <p>Background</p> <p>To define a threshold value of apparent diffusion coefficient (ADC) with which malignant breast lesions can be distinguished from benign lesions, and to evaluate the ADC change of peri-tumor tissue in breast carcinoma by echo planar-diffusion weighted imaging (EPI-DWI).</p> <p>Methods</p> <p>57 breast lesions were scanned by routine MRI and EPI-DWI. The ADC values were compared between malignant and benign lesions. The sensitivity and specificity of EPI-DWI and the threshold ADC value were evaluated by Receiver Operating Characteristic curve (ROC). The ADC values of malignant lesion and layered peri-tumor tissues (from innermost layer 1 to outermost layer 4 with 5 mm every layer) in different directions were compared and the ADC values among different layers were compared.</p> <p>Results</p> <p>The ADC value of 35 malignant lesions was statistically lower than that of 22 benign lesions (P < 0.05). In ROC curve, the threshold value was 1.24 +/- 0.25*10E-3 mm<sup>2</sup>/s (b = 500) or 1.20 +/- 0.25*10E-3 mm<sup>2</sup>/s (b = 1000). The ADC value of malignant lesions was statistically lower than that of peri-tumor tissues in different directions (P < 0.05). For peri-tumor tissues, the ADC values increased gradually from layer 1 to layer 4 and there was a significant difference between the ADC values of layer 1 and layer 2 (P < 0.05); while from layer 2 outwards, there was no statistical difference among different layers.</p> <p>Conclusion</p> <p>ADC value was a sensitive and specific parameter that could help to differentiate benign and malignant breast lesions. ADC changes in tissues adjacent to breast carcinoma could be detected by EPI-DWI, which made EPI-DWI a promising method for helping to determine surgical scope of breast carcinoma.</p

    Production of a Proteolytically Active Protein, Chlamydial Protease/Proteasome-Like Activity Factor, by Five Different Chlamydia Species

    No full text
    We have previously identified a chlamydial protein, chlamydial protease/proteasome-like activity factor (CPAF), for degrading host transcription factors in cells infected with the human chlamydial species Chlamydia trachomatis or Chlamydia pneumoniae. We now report that functional CPAF was also produced during infection with the species Chlamydia muridarum, Chlamydia psittaci, and Chlamydia caviae, which primarily infect nonhuman hosts

    Cleavage of Host Keratin 8 by a Chlamydia-Secreted Protease

    No full text
    Chlamydiae have to replicate within a cytoplasmic vacuole in eukaryotic cells. Expansion of the chlamydia-laden vacuole is essential for chlamydial intravacuolar replication, which inevitably causes host cell cytoskeleton rearrangements. A cleavage fragment of keratin 8 corresponding to the central rod region was detected in the soluble fraction of chlamydia-infected cells. Since keratin 8 is a major component of the intermediate filaments in simple epithelial cells, cleavage of keratin 8 may increase the solubility of the host cell cytoskeleton and thus permit vacuole expansion in chlamydia-infected cells. A chlamydia-secreted protease designated CPAF (chlamydial protease/proteasome-like activity factor) was both necessary and sufficient for keratin 8 cleavage in chlamydia-infected cells, suggesting that chlamydiae have evolved specific mechanisms for modifying the host cell cytoskeleton

    Intramolecular Dimerization Is Required for the Chlamydia-Secreted Protease CPAF To Degrade Host Transcriptional Factors

    No full text
    We previously identified a chlamydial protein designated CPAF (chlamydia protease/proteasome-like activity factor) that is secreted into host cell cytosol for degrading host transcription factors required for major histocompatibility complex antigen expression. Here we report that CPAF, synthesized as a 70-kDa proprotein, is processed into two fragments (designated CPAFn and CPAFc) to form intramolecular dimers that are much more stable than the naïve CPAF. Precipitation with antibodies that recognized CPAF dimers removed the proteolytic activity responsible for degrading host transcription factor RFX5 from chlamydia-infected host cell cytosol, while precipitation with antibodies that recognized free CPAF fragments alone did not remove this activity. Separation of CPAFn from CPAFc resulted in a loss of proteolytic activity. Furthermore, neither expressed full-length CPAF that was not processed nor coexpressed CPAFn and CPAFc fragments that failed to form dimers degraded RFX5. These observations demonstrate that intramolecular dimerization is required for CPAF to degrade host transcription factors, a strategy that is utilized by an obligate intracellular bacterial species to evade host defenses

    Chlamydia trachomatis Infection Inhibits Both Bax and Bak Activation Induced by Staurosporine

    No full text
    We have previously shown that Chlamydia trachomatis inhibits host cell apoptosis and blocks mitochondrial cytochrome c release. We now report that activation of both Bax and Bak, two proapoptotic members of the Bcl-2 family that regulate mitochondrial cytochrome c release, was inhibited in chlamydia-infected cells. This observation has provided new information on the mechanisms of chlamydial antiapoptotic activity

    Occlusion-Aware Path Planning to Promote Infrared Positioning Accuracy for Autonomous Driving in a Warehouse

    No full text
    Infrared positioning is a critical module in an indoor autonomous vehicle platform. In an infrared positioning system, the ego vehicle is equipped with an infrared emitter while the infrared receivers are fixed onto the ceiling. The infrared positioning result is accurate only when the number of valid infrared receivers is more than three. An infrared receiver easily becomes invalid if it does not receive light from the infrared emitter due to indoor occlusions. This study proposes an occlusion-aware path planner that enables an autonomous vehicle to navigate toward the occlusion-free part of the drivable area. The planner consists of four layers. In layer one, a homotopic A* path is searched for in the 2D grid map to roughly connect the initial and goal points. In layer two, a curvature-continuous reference line is planned close to the A* path using numerical optimal control. In layer three, a Frenet frame is constructed along the reference line, followed by a search for an occlusion-aware path within that frame via dynamic programming. In layer four, a curvature-continuous path is optimized via quadratic programming within the Frenet frame. A path planned within the Frenet frame may violate the curvature bounds in a real-world Cartesian frame; thus, layer four is implemented through trial and error. Simulation results in CarSim software show that the derived paths reduce the poor positioning risk and are easily tracked by a controller
    corecore