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      1. Author :
        Kadurugamuwa, J. L.; Francis, K. P.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Methods in Molecular Biology
      6. Products :
      7. Volume :
        431
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Bioware, Xen29, Animals, Bacteria/chemistry/ genetics, Bacterial Infections/diagnosis/ microbiology, Biofilms/ growth & development, Diagnostic Imaging/methods, Luminescent Measurements/ methods IVIS, Xenogen, Xen5, Xen44
      12. Abstract :
        Whole body biophotonic imaging (BPI) is a technique that has contributed significantly to the way researchers study bacterial pathogens and develop pre-clinical treatments to combat their ensuing infections in vivo. Not only does this approach allow disease profiles and drug efficacy studies to be conducted non-destructively in live animals over the entire course of the disease, but in many cases, it enables investigators to observe disease profiles that could otherwise easily be missed using conventional methodologies. The principles of this technique are that bacterial pathogens engineered to express bioluminescence (visible light) can be readily monitored from outside of the living animal using specialized low-light imaging equipment, enabling their movement, expansion and treatment to be seen completely non-invasively. Moreover, because the same group of animals can be imaged at each time-point throughout the study, the overall number of animals used is dramatically reduced, saving lives, time, and money. Also, as each animal acts as its own control over time, the issues associated with animal-to-animal variation are circumvented, thus improving the quality of the biostatistical data generated. The ability to monitor infections in vivo in a longitudinal fashion is especially appealing to assess chronic infections such as those involving implanted devices. Typically, bacteria grow as biofilms on these foreign bodies and are reputably difficult to monitor with conventional methods. Because of the non-destructive and non-invasive nature of BPI, the procedure can be performed repeatedly in the same animal, allowing the biofilm to be studied in situ without detachment or disturbance. This ability not only allows unique patterns of disease relapse to be seen following termination of antibiotic therapy but also in vivo resistance development during prolonged treatment, both of which are common occurrences with device-related infections. This chapter describes the bioluminescent engineering of both Gram-positive and Gram-negative bacteria and overviews their use in device-associated infections in several anatomical sites in a variety of animal models.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18287760
      14. Call Number :
        139321
      15. Serial :
        5568
      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
      1. Author :
        Chung, HM; Cartwright, MM; Bortz, DM; Jackson, TL; Younger, JG
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Shock
      6. Products :
      7. Volume :
        30
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen, Xen39
      12. Abstract :
        Unlike many localized infections, the development and resolution of bacteremia involves physical and immunological interactions between many anatomic sites. In an effort to better understand these interactions, we developed a computational model of bacteremia as a dynamical system fashioned after multicompartmental pharmacodynamic models, incorporating bacterial proliferation and clearance in the blood, liver, spleen, and lungs, and the transport of pathogens between these sites. A system of four first-order homogeneous ODEs was developed. Blood and organ bacterial burdens were measured at various time points from 3 to 48 h postinoculation using an LD25 murine model of Staphylococcus epidermidis bacteremia. Using these empiric data, solutions to the mathematical model were recovered. A bootstrap resampling method was used to generate 95% confidence intervals around the solved parameters. The validity of the model was examined in parallel experiments using animals acutely immunocompromised with cyclophosphamide; the model captured abnormalities in bacterial partitioning previously described with this antineoplastic agent. Lastly, the approach was used to explore possible benefits to clinically observed hyperdynamic blood flow during sepsis: in simulation, normal mice, but not those treated with cyclophosphamide, enjoyed significantly more rapid bacterial clearance from the bloodstream under hyperdynamic conditions.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18317411
      14. Call Number :
        136975
      15. Serial :
        5976
      1. Author :
        Wunder A and Klohs J.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Basic Research in Cardiology
      6. Products :
      7. Volume :
        103
      8. Issue :
        2
      9. Page Numbers :
        N/A
      10. Research Area :
        Cardiovascular Research
      11. Keywords :
        In vivo imaging; atherosclerosis; bioluminescence imaging; fluorescence imaging; myocardial infarction; stroke; ProSense
      12. Abstract :
        Pathophysiological processes in the vascular system are the major cause of mortality and disease. Atherosclerosis, an inflammatory process in arterial walls, can lead to formation of plaques, whose rupture can lead to thrombus formation, obstruction of vessels (thrombosis), reduction of the blood flow (ischemia), cell death in the tissue fed by the occluded vessel, and depending on the affected vessel, to myocardial infarction or stroke. Imaging techniques enabling visualization of the biological processes involved in this scenario are therefore highly desirable. In recent years, a number of reporter agents and reporter systems have been developed to visualize these processes using different imaging modalities including nuclear imaging techniques, such as positron emission tomography or single photon emission computed tomography, magnetic resonance imaging, and ultrasound. This article comprises a brief overview of optical imaging techniques, such as fluorescence imaging and bioluminescence imaging for the visualization of vascular pathophysiology.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18324374
      14. Call Number :
        PKI @ sarah.piper @
      15. Serial :
        4649
      1. Author :
        N/A
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Microbes and infection / Institut Pasteur
      6. Products :
      7. Volume :
        10
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bacterial Proteins; Biofilms; Bioware; Catheterization, Central Venous; Male; Mice; Point Mutation; Sigma Factor; Staphylococcal Infections; Staphylococcus aureus; Virulence; Xen29
      12. Abstract :
        The impact of the alternative sigma factor sigma B (SigB) on pathogenesis of Staphylococcus aureus is not conclusively clarified. In this study, a central venous catheter (CVC) related model of multiorgan infection was used to investigate the role of SigB for the pathogenesis of S. aureus infections and biofilm formation in vivo. Analysis of two SigB-positive wild-type strains and their isogenic mutants revealed uniformly that the wild-type was significantly more virulent than the SigB-deficient mutant. The observed difference in virulence was apparently not linked to the capability of the strains to form biofilms in vivo since wild-type and mutant strains were able to produce biofilm layers inside of the catheter. The data strongly indicate that the alternative sigma factor SigB plays a role in CVC-associated infections caused by S. aureus.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18328762
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9046
      1. Author :
        Hokaiwado, Naomi; Takeshita, Fumitaka; Naiki-Ito, Aya; Asamoto, Makoto; Ochiya, Takahiro; Shirai, Tomoyuki
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Carcinogenesis
      6. Products :
      7. Volume :
        29
      8. Issue :
        6
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Androgens; Animals; Animals, Genetically Modified; Apoptosis; Bioware; Blotting, Western; Cell Line, Tumor; Cell Proliferation; Glutathione S-Transferase pi; Humans; In Situ Nick-End Labeling; Male; Neoplasm Transplantation; Oligonucleotide Array Sequence Analysis; PC-3M-luc; Prostatic Neoplasms; Rats; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering
      12. Abstract :
        Prostate cancers generally acquire an androgen-independent growth capacity with progression, resulting in resistance to antiandrogen therapy. Therefore, identification of the genes regulated through this process may be important for understanding the mechanisms of prostate carcinogenesis. We here utilized androgen-dependent/independent transplantable tumors, newly established with the 'transgenic rat adenocarcinoma in prostate' (TRAP) model, to analyze their gene expression using microarrays. Among the overexpressed genes in androgen-independent prostate cancers compared with the androgen-dependent tumors, glutathione S-transferase pi (GST-pi) was included. In line with this, human prostate cancer cell lines PC3 and DU145 (androgen independent) had higher expression of GST-pi compared with LNCaP (androgen dependent) as determined by semiquantitative reverse transcription-polymerase chain reaction analysis. To investigate the roles of GST-pi expression in androgen-independent human prostate cancers, GST-pi was knocked down by a small interfering RNA (siRNA), resulting in significant decrease of the proliferation rate in the androgen-independent PC3 cell line. In vivo, administration of GST-pi siRNA-atelocollagen complex decreased GST-pi protein expression, resulting in enhanced numbers of TdT mediated dUTP-biotin nick-end labering (TUNEL)-positive apoptotic cells. These findings suggest that GST-pi might play important roles in proliferation of androgen-independent human prostate cancer cells.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18413363
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8967
      1. Author :
        N/A
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Blood
      6. Products :
      7. Volume :
        112
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Antigen Presentation; Antigens, Neoplasm; B16-F10-luc-G5 cells; Bioware; B-Lymphocytes; Cell Adhesion; Cell Line, Tumor; Cell Movement; dendritic cells; Female; Lectins, C-Type; Lymphatic Metastasis; Lymphatic System; Macrophages; Male; Mannose-Binding Lectins; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mice, Knockout; Receptors, Cell Surface; T-Lymphocytes
      12. Abstract :
        Macrophage mannose receptor (MR) participates in pathogen recognition, clearance of endogenous serum glycoproteins, and antigen presentation. MR is also present on lymphatic vessels, where its function is unknown. Here we show that migration of lymphocytes from the skin into the draining lymph nodes through the afferent lymphatics is reduced in MR-deficient mice, while the structure of lymphatic vasculature remains normal in these animals. Moreover, in a tumor model the primary tumors grow significantly bigger in MR(-/-) mice than in the wild-type (WT) controls, whereas the regional lymph node metastases are markedly smaller. Adhesion of both normal lymphocytes and tumor cells to lymphatic vessels is significantly decreased in MR-deficient mice. The ability of macrophages to present tumor antigens is indistinguishable between the 2 genotypes. Thus, MR on lymphatic endothelial cells is involved in leukocyte trafficking and contributes to the metastatic behavior of cancer cells. Blocking of MR may provide a new approach to controlling inflammation and cancer metastasis by targeting the lymphatic vasculature.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18434610
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9000
      1. Author :
        Inglefield, Jon R; Dumitru, Calin Dan; Alkan, Sefik S; Gibson, Sheila J; Lipson, Kenneth E; Tomai, Mark A; Larson, Chris J; Vasilakos, John P
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Journal of interferon & cytokine research: the official journal of the International Society for Interferon and Cytokine Research
      6. Products :
      7. Volume :
        28
      8. Issue :
        4
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Aminoquinolines; Animals; B16-F10-luc-G5 cells; Bioware; Cell Line, Tumor; Cell Proliferation; Cell Survival; Culture Media, Conditioned; dendritic cells; Humans; Interferon Type I; Leukocytes, Mononuclear; Lung; Melanoma; Mice; Neoplasms; Oligodeoxyribonucleotides; Quinolines; Subcellular Fractions; Sulfonamides; Toll-Like Receptor 7
      12. Abstract :
        Antitumor effects of the toll-like receptor 7 (TLR7) agonist, 852A, were evaluated. Supernatants from human peripheral blood mononuclear cells (PBMC) stimulated with 852A inhibited the proliferation of tumor cell lines Hs294T and 769-P but had no effect on others (786-O and Caki-1). Because addition of 852A directly to the Hs294T cells did not inhibit their proliferation, the mechanism(s) of inhibition of tumor cell proliferation was investigated. Low nanomolar concentrations of 852A stimulated the production of interferon-alpha (IFN-alpha), IFN-inducible protein-10 (IP-10), interleukin-1 receptor antagonist (IL-1Ra), monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) from human PBMCs. Cytokines stimulated by submicromolar concentrations of 852A were sufficient to inhibit Hs294T proliferation. At higher concentrations (3-30 microM), 852A induced the production of IL-12p70, IL-18, and IFN-gamma. PBMC cultures depleted of plasmacytoid dendritic cells (pDC) did not produce IFN-alpha, and their conditioned medium did not inhibit Hs294T proliferation. Anti-IFN-alpha/beta receptor (IFNAR) and anti-IFN-alpha antibodies partially abrogated Hs294T proliferation inhibition by 852A-stimulated PBMC supernatants, whereas separate neutralization of TRAIL, tumor necrosis factor-alpha (TNF-alpha, IFN-gamma, IFN-beta, or IFN-omega had no effect. In vivo, six doses of 852A administration significantly delayed the onset of lung colonies in a B16 melanoma model. Thus, the results demonstrate that the TLR7 agonist 852A inhibits in vitro proliferation of some tumor cells in a pDC-dependent and IFN-alpha-dependent manner and can delay tumor growth in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18439103
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9001
      1. Author :
        Burkatovskaya, Marina; Castano, Ana P; Demidova-Rice, Tatiana N; Tegos, George P; Hamblin, Michael R
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        Wound repair and regeneration: official publication of the Wound Healing Society [and] the European Tissue Repair Society
      6. Products :
      7. Volume :
        16
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Anti-Infective Agents; Bandages; Biocompatible Materials; Bioware; Chitosan; Cyclophosphamide; Male; Mice; Mice, Inbred BALB C; Staphylococcal Skin Infections; Staphylococcus aureus; Wound Healing; Wound Infection; Xen8.1
      12. Abstract :
        HemCon bandage is an engineered chitosan acetate preparation designed as a hemostatic dressing, and is under investigation as a topical antimicrobial dressing. We studied its effects on healing of excisional wounds that were or were not infected with Staphylococcus aureus, in normal mice or mice previously pretreated with cyclophosphamide (CY). CY significantly suppressed wound healing in both the early and later stages, while S. aureus alone significantly stimulated wound healing in the early stages by preventing the initial wound expansion. CY plus S. aureus showed an advantage in early stages by preventing expansion, but a significant slowing of wound healing in later stages. In order to study the conflicting clamping and stimulating effects of chitosan acetate bandage on normal wounds, we removed the bandage from wounds at times after application ranging from 1 hour to 9 days. Three days application gave the earliest wound closure, and all application times gave a faster healing slope after removal compared with control wounds. Chitosan acetate bandage reduced the number of inflammatory cells in the wound at days 2 and 4, and had an overall beneficial effect on wound healing especially during the early period where its antimicrobial effect is most important.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18471261
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9986
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