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      1. Author :
        Thurlow, L. R.; Hanke, M. L.; Fritz, T.; Angle, A.; Aldrich, A.; Williams, S. H.; Engebretsen, I. L.; Bayles, K. W.; Horswill, A. R.; Kielian, T.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2011
      5. Publication :
        J Immunol
      6. Products :
      7. Volume :
        186
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xen29, Xen 29, Staphylococcus aureus Xen29, Animals; *Biofilms; Catheter-Related Infections/immunology/metabolism/microbiology; Cytokines/immunology/metabolism; Green Fluorescent Proteins/genetics/metabolism; Host-Pathogen Interactions/immunology; Immune Evasion/immunology; Inflammation/*immunology/metabolism; Macrophages/*immunology/metabolism; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Microscopy, Confocal; Microscopy, Electron, Scanning; Models, Immunological; Phagocytosis/*immunology; Staphylococcal Infections/*immunology/metabolism/microbiology; Staphylococcus aureus/*immunology/physiology/ultrastructure; Toll-Like Receptor 2/genetics/immunology; Toll-Like Receptor 9/genetics/immunology
      12. Abstract :
        Biofilms are complex communities of bacteria encased in a matrix composed primarily of polysaccharides, extracellular DNA, and protein. Staphylococcus aureus can form biofilm infections, which are often debilitating due to their chronicity and recalcitrance to antibiotic therapy. Currently, the immune mechanisms elicited during biofilm growth and their impact on bacterial clearance remain to be defined. We used a mouse model of catheter-associated biofilm infection to assess the functional importance of TLR2 and TLR9 in the host immune response during biofilm formation, because ligands for both receptors are present within the biofilm. Interestingly, neither TLR2 nor TLR9 impacted bacterial density or inflammatory mediator secretion during biofilm growth in vivo, suggesting that S. aureus biofilms circumvent these traditional bacterial recognition pathways. Several potential mechanisms were identified to account for biofilm evasion of innate immunity, including significant reductions in IL-1beta, TNF-alpha, CXCL2, and CCL2 expression during biofilm infection compared with the wound healing response elicited by sterile catheters, limited macrophage invasion into biofilms in vivo, and a skewing of the immune response away from a microbicidal phenotype as evidenced by decreases in inducible NO synthase expression concomitant with robust arginase-1 induction. Coculture studies of macrophages with S. aureus biofilms in vitro revealed that macrophages successful at biofilm invasion displayed limited phagocytosis and gene expression patterns reminiscent of alternatively activated M2 macrophages. Collectively, these findings demonstrate that S. aureus biofilms are capable of attenuating traditional host proinflammatory responses, which may explain why biofilm infections persist in an immunocompetent host.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/21525381
      14. Call Number :
        PKI @ kd.modi @ 19
      15. Serial :
        10457
      1. Author :
        Tsunooka, N.; Hirayama, S.; Medin, J. A.; Liles, W. C.; Keshavjee, S.; Waddell, T. K.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2011
      5. Publication :
        Ann Thorac Surg
      6. Products :
      7. Volume :
        91
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xen29, Xen 29, Staphylococcus aureus Xen29, Animals; Disease Models, Animal; Female; Mice; Mice, Inbred C57BL; Pneumonectomy/*adverse effects; Postoperative Complications/*surgery; Stem Cell Transplantation/*methods; Thoracic Cavity/*surgery; Thoracoplasty/*methods; Tissue Engineering/*methods
      12. Abstract :
        BACKGROUND: Transfer of viable tissue flaps and thoracoplasty are effective against pleural space complications after pneumonectomy but highly disfiguring. The aim of this study was to explore the possibility of engineered tissue to treat space complications after pneumonectomy. METHODS: A left pneumonectomy was performed in mice, and the cavity immediately filled with the cellularized collagen matrices. First, bone marrow derived-mesenchymal stroma cells with luciferase expression were used as donor cells to evaluate cell viability and angiogenesis using bioluminescence imaging. Second, using bone marrow cells from GFP mice, histologic evaluation, immunohistochemistry for von Willebrand Factor, and flow cytometric analysis was performed compared with acellular matrix implants. The effect on bacterial clearance was examined using an empyema model with Staphylococcus aureus expressing luciferase. RESULTS: Embedded cells proliferated within the denatured collagen matrices ex vivo. In vivo, bioluminescent imaging activity could be detected till day 8, and the slope (suggesting rate of perfusion with luciferin) increased with time up to day 6 but decreased after day 7. Although GFP-positive donor cells decreased with time, total cellularity increased. Furthermore, vessels stained by von Willebrand factor were significantly increased. Both cellularized and acellularized matrices showed bacterial clearance in vivo. CONCLUSIONS: Cells within collagen matrices survive in the thoracic cavity at early time points. Cellularized matrices quickly lead to neovascularization and recipient cell infiltration. Both cellularized and acellularized matrices show bacterial clearance in vivo. This study indicates the potential feasibility of a novel tissue engineering approach to problems of the postpneumonectomy space.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/21353020
      14. Call Number :
        PKI @ kd.modi @ 3
      15. Serial :
        10458
      1. Author :
        Joh, E. H.; Hollenbaugh, J. A.; Kim, B.; Kim, D. H.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2012
      5. Publication :
        PLoS One
      6. Products :
      7. Volume :
        7
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        A549-luc-C8, A549-luc, IVIS, Bioware
      12. Abstract :
        While pharmacological inhibition of Akt kinase has been regarded as a promising anti-cancer strategy, most of the Akt inhibitors that have been developed are enzymatic inhibitors that target the kinase active site of Akt. Another key cellular regulatory event for Akt activation is the translocation of Akt kinase to the cell membrane from the cytoplasm, which is accomplished through the pleckstrin homology (PH) domain of Akt. However, compounds specifically interacting with the PH domain of Akt to inhibit Akt activation are currently limited. Here we identified a compound, lancemaside A (LAN-A), which specifically binds to the PH domain of Akt kinase. First, our mass spectra analysis of cellular Akt kinase isolated from cells treated with LAN-A revealed that LAN-A specifically binds to the PH domain of cellular Akt kinase. Second, we observed that LAN-A inhibits the translocation of Akt kinase to the membrane and thus Akt activation, as examined by the phosphorylation of various downstream targets of Akt such as GSK3beta, mTOR and BAD. Third, in a co-cultured cell model containing human lung epithelial cancer cells (A549) and normal human primary lung fibroblasts, LAN-A specifically restricts the growth of the A549 cells. LAN-A also displayed anti-proliferative effects on various human cancer cell lines. Finally, in the A549-luciferase mouse transplant model, LAN-A effectively inhibited A549 cell growth with little evident cytotoxicity. Indeed, the therapeutic index of LAN-A in this mouse model was >250, supporting that LAN-A is a potential lead compound for PH domain targeting as a safe anti-cancer Akt inhibitor.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/23189201
      14. Call Number :
        PKI @ kd.modi @ 5
      15. Serial :
        10524
      1. Author :
        Engelsman, Anton F; van Dam, Gooitzen M; van der Mei, Henny C; Busscher, Henk J; Ploeg, Rutger J
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        Annals of surgery
      6. Products :
      7. Volume :
        251
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Abdominal Wall; Animals; Bioware; Female; Luminescent Measurements; Mice; Mice, Inbred BALB C; Polypropylenes; Polytetrafluoroethylene; pXen-5; Staphylococcal Infections; Staphylococcus aureus; Surgical Mesh; Xen29
      12. Abstract :
        OBJECTIVE To study the influence of morphology of surgical meshes on the course of bacterial infection under the influence of the host immune system in an in vivo chronic bacterial infection model. BACKGROUND The use of prosthetic meshes has increased dramatically the last decades in abdominal wall reconstructive surgery. Whereas infection is becoming a more frequent complication, attention is increasingly drawn to the influence of the surgeon's mesh choice on the course of this complication. METHODS Samples of 6 often applied surgical meshes were contaminated with a bioluminescent strain of Staphylococcus aureus and implanted subcutaneously in an immunocompetent BALB/c mouse. The intensity and the spreading of bioluminescence (ie, p/s/cm/sr) were analyzed non-invasively in vivo during a 10-day follow-up period. RESULTS Over the course of infection, multifilament polypropylene and hydrophobic materials showed a significantly higher persistence of bacteria as well as spreading of infection compared to all other meshes. In contrast, infection resolved in almost all animals with a low-weight polyester mesh. CONCLUSION The results of this study are in accordance with circumstantial evidence from limited clinical reports on infection involving surgical meshes and suggest that multifilament and hydrophobic meshes significantly increase bacterial persistence or spreading in the infected area in contrast to monofilament polypropylene and lightweight meshes. Therefore, the surgeon should consider this outcome when choosing a mesh graft for limiting infection in abdominal wall repair.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/19864938
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9017
      1. Author :
        N/A
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2007
      5. Publication :
        PLoS pathogens
      6. Products :
      7. Volume :
        3
      8. Issue :
        6
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Anthrax; Bacillus anthracis; Bioware; Disease Models, Animal; Gastrointestinal Diseases; Inhalation Exposure; Luciferases; Luminescence; Luminescent Measurements; Lymph Nodes; Mice; Mice, Inbred BALB C; Nasal Cavity; Organisms, Genetically Modified; Peyer's Patches; Pharynx; pXen-5; Skin; Spores, Bacterial
      12. Abstract :
        Bacillus anthracis causes three forms of anthrax: inhalational, gastrointestinal, and cutaneous. Anthrax is characterized by both toxemia, which is caused by secretion of immunomodulating toxins (lethal toxin and edema toxin), and septicemia, which is associated with bacterial encapsulation. Here we report that, contrary to the current view of B. anthracis pathogenesis, B. anthracis spores germinate and establish infections at the initial site of inoculation in both inhalational and cutaneous infections without needing to be transported to draining lymph nodes, and that inhaled spores establish initial infection in nasal-associated lymphoid tissues. Furthermore, we found that Peyer's patches in the mouse intestine are the primary site of bacterial growth after intragastric inoculation, thus establishing an animal model of gastrointestinal anthrax. All routes of infection progressed to the draining lymph nodes, spleen, lungs, and ultimately the blood. These discoveries were made possible through the development of a novel dynamic mouse model of B. anthracis infection using bioluminescent non-toxinogenic capsulated bacteria that can be visualized within the mouse in real-time, and demonstrate the value of in vivo imaging in the analysis of B. anthracis infection. Our data imply that previously unrecognized portals of bacterial entry demand more intensive investigation, and will significantly transform the current perception of inhalational, gastrointestinal, and cutaneous B. anthracis pathogenesis.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/17542645
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9022
      1. Author :
        Hardy, Jonathan; Chu, Pauline; Contag, Christopher H
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Disease models & mechanisms
      6. Products :
      7. Volume :
        2
      8. Issue :
        1-2
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Bone Marrow; Bone Marrow Cells; Disease Models, Animal; Female; Host-Pathogen Interactions; Humans; Knee Joint; Listeria monocytogenes; Listeriosis; Mice; Mice, Inbred BALB C; Mutation; pXen-5; Tibia
      12. Abstract :
        Murine listeriosis is one of the most comprehensive and well-studied models of infection, and Listeria monocytogenes has provided seminal information regarding bacterial pathogenesis. However, many aspects of the mouse model remain poorly understood, including carrier states and chronic colonization which represent important features of the spectrum of host-pathogen interaction. Bone marrow has recently been shown to harbor L. monocytogenes, which spreads from this location to the central nervous system. Bone could, therefore, be an important chronic reservoir, but this infection is difficult to study because it involves only a few bacteria and the extent of infection cannot be assessed until after the animal is sacrificed. We employed in vivo bioluminescence imaging to localize L. monocytogenes bone infections over time in live mice, revealing that the bacteria grow in discrete foci. These lesions can persist in many locations in the legs of mice and are not accompanied by a histological indication such as granuloma or a neutrophil infiltratate. We demonstrate that highly attenuated hly mutants, which have defective intracellular replication, are capable of prolonged focal infection of the bone marrow for periods of up to several weeks. These results support the recently proposed hypothesis that the bone marrow is a unique niche for L. monocytogenes.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/19132117
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9018
      1. Author :
        Hardy, Jonathan; Margolis, Jeffrey J; Contag, Christopher H
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2006
      5. Publication :
        Infection and immunity
      6. Products :
      7. Volume :
        74
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bacterial Toxins; Biliary Tract; Bioware; Feces; Food Contamination; Intestines; Listeria monocytogenes; Listeriosis; Mice; Mice, Inbred BALB C; pXen-5
      12. Abstract :
        Listeria monocytogenes is a ubiquitous gram-positive bacterium that can cause systemic and often life-threatening disease in immunocompromised hosts. This organism is largely an intracellular pathogen; however, we have determined that it can also grow extracellularly in animals, in the lumen of the gallbladder. The significance of growth in the gallbladder with respect to the pathogenesis and spread of listeriosis depends on the ability of the bacterium to leave this organ and be disseminated to other tissues and into the environment. Should this process be highly inefficient, growth in the gallbladder would have no impact on pathogenesis or spread, but if it occurs efficiently, bacterial growth in this organ may contribute to listeriosis and dissemination of this organism. Here, we use whole-body imaging to determine the efficacy and kinetics of food- and hormone-induced biliary excretion of L. monocytogenes from the murine gallbladder, demonstrating that transit through the bile duct into the intestine can occur within 5 min of induction of gallbladder contraction by food or cholecystokinin and that movement of bacteria through the intestinal lumen can occur very rapidly in the absence of fecal material. These studies demonstrate that L. monocytogenes bacteria replicating in the gallbladder can be expelled from the organ efficiently and that the released bacteria move into the intestinal tract, where they pass into the environment and may possibly reinfect the animal.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/16495556
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9024
      1. Author :
        Harms, Jerome S; Durward, Marina A; Magnani, Diogo M; Splitter, Gary A
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Journal of immune based therapies and vaccines
      6. Products :
      7. Volume :
        7
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Bioware; pXen-13
      12. Abstract :
        BACKGROUND There is no safe, effective human vaccine against brucellosis. Live attenuated Brucella strains are widely used to vaccinate animals. However these live Brucella vaccines can cause disease and are unsafe for humans. Killed Brucella or subunit vaccines are not effective in eliciting long term protection. In this study, we evaluate an approach using a live, non-pathogenic bacteria (E. coli) genetically engineered to mimic the brucellae pathway of infection and present antigens for an appropriate cytolitic T cell response. METHODS E. coli was modified to express invasin of Yersinia and listerialysin O (LLO) of Listeria to impart the necessary infectivity and antigen releasing traits of the intracellular pathogen, Brucella. This modified E. coli was considered our vaccine delivery system and was engineered to express Green Fluorescent Protein (GFP) or Brucella antigens for in vitro and in vivo immunological studies including cytokine profiling and cytotoxicity assays. RESULTS The E. coli vaccine vector was able to infect all cells tested and efficiently deliver therapeutics to the host cell. Using GFP as antigen, we demonstrate that the E. coli vaccine vector elicits a Th1 cytokine profile in both primary and secondary immune responses. Additionally, using this vector to deliver a Brucella antigen, we demonstrate the ability of the E. coli vaccine vector to induce specific Cytotoxic T Lymphocytes (CTLs). CONCLUSION Protection against most intracellular bacterial pathogens can be obtained mostly through cell mediated immunity. Data presented here suggest modified E. coli can be used as a vaccine vector for delivery of antigens and therapeutics mimicking the infection of the pathogen and inducing cell mediated immunity to that pathogen.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/19126207
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9029
      1. Author :
        Park, Hae-Sun; Francis, Kevin P; Yu, Jun; Cleary, P Patrick
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2003
      5. Publication :
        Journal of immunology (Baltimore, Md.: 1950)
      6. Products :
      7. Volume :
        171
      8. Issue :
        5
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Administration, Intranasal; Animals; Bioware; Disease Models, Animal; Female; Humans; Immunohistochemistry; Intracellular Fluid; Lymphoid Tissue; Mice; Mice, Inbred BALB C; Nasal Mucosa; Nasopharynx; Palatine Tonsil; pXen-5; Streptococcal Infections; Streptococcus pyogenes
      12. Abstract :
        Human tonsils are suspected to be an antibiotic-impervious human reservoir for group A streptococcus. An intranasal infection model in mice and a bioluminescent-tagged strain were used to investigate this possibility. Viable streptococci were predominantly found both intra- and extracellularly in nasal-associated lymphoid tissue (NALT), a human tonsil homologue. Ulex europaeus-1, a membranous (M) cell-specific lectin, identified cells harboring streptococci at the epithelial surface of NALT and blocked bacterial colonization of this tissue. These results suggest that M cells in NALT transport this Gram-positive pathogen across the epithelial layers in a manner similar to those in Peyer's patches, which permit enteric pathogens to invade deeper tissues from the gastrointestinal tract.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/12928403
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9025
      1. Author :
        Rambow-Larsen, Amy A; Rajashekara, Gireesh; Petersen, Erik; Splitter, Gary
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Journal of bacteriology
      6. Products :
      7. Volume :
        190
      8. Issue :
        9
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Amino Acid Sequence; Animals; Bioware; Brucella melitensis; Brucellosis; Disease Models, Animal; Flagella; Gene Deletion; Gene Expression Regulation, Bacterial; Interferon Regulatory Factor-1; Macrophages; Mice; Mice, Mutant Strains; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; pXen-13; Quorum Sensing; Repressor Proteins; Trans-Activators; Virulence Factors
      12. Abstract :
        Brucella melitensis is an intracellular pathogen that establishes a replicative niche within macrophages. While the intracellular lifestyle of Brucella is poorly understood and few virulence factors have been identified, components of a quorum-sensing pathway in Brucella have recently been identified. The LuxR-type regulatory protein, VjbR, and an N-acylhomoserine lactone signaling molecule are both involved in regulating expression of the virB-encoded type IV secretion system. We have identified a second LuxR-type regulatory protein (BlxR) in Brucella. Microarray analysis of a blxR mutant suggests that BlxR regulates the expression of a number of genes, including those encoding the type IV secretion system and flagella. Confirming these results, deletion of blxR in B. melitensis reduced the transcriptional activities of promoters for the virB operon, flagellar genes, and another putative virulence factor gene, bopA. Furthermore, our data suggested that both BlxR and VjbR are positively autoregulated and cross-regulate the expression of each other. The blxR deletion strain exhibited reduced growth in macrophages, similar to that observed for a vjbR deletion strain. However, unlike the vjbR deletion, the blxR deletion did not fully attenuate virulence in mice. More strikingly, bioluminescent imaging revealed that dissemination of the blxR mutant was similar to that of wild-type B. melitensis, while the vjbR mutant was defective for systemic spread in IRF-1(-/-) mice, suggesting that these regulators are not functionally redundant but that they converge in a common pathway regulating bacterial processes.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18310341
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9030
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