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      1. Author :
        Kadurugamuwa, J. L.; Sin, L. V.; Yu, J.; Francis, K. P.; Kimura, R.; Purchio, T.; Contag, P. R.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2003
      5. Publication :
        Antimicrobial Agents and Chemotherapy
      6. Products :
      7. Volume :
        47
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals, Anti-Bacterial Agents/ pharmacology, Bacterial Infections/drug therapy/microbiology, Biofilms/ drug effects/growth & development, Bioware; Catheterization/adverse effects, Chemiluminescent Measurements, Ciprofloxacin/pharmacology, Colony Count, Microbial, Disease Models, Animal, Dose-Response Relationship, Drug, Drug Monitoring/methods, Mice, Rifampin/pharmacology, Staphylococcus aureus/drug effects/genetics/growth & development, Tobramycin/pharmacology IVIS, Xenogen; Xen29
      12. Abstract :
        We have developed a rapid, continuous method for monitoring the effectiveness of several antibacterial agents in real time, noninvasively, by using a recently described mouse model of chronic biofilm infection (J. L. Kadurugamuwa et al., Infect. Immun. 71:882-890, 2003), which relies on biophotonic imaging of bioluminescent bacteria. To facilitate real-time monitoring of infection, we used a Staphylococcus aureus isolate that was made bioluminescent by inserting a modified lux operon into the bacterial chromosome. This bioluminescent reporter bacterium was used to study the antimicrobial effects of several antibiotics belonging to different molecular families. Treatment with rifampin, tobramycin, and ciprofloxacin was started 7 days after subcutaneous implantation of catheters precolonized with 10(4) CFU of S. aureus. Three different doses of antibiotics were administered twice a day for 4 consecutive days. The number of metabolically active bacteria in untreated mice and the tobramycin- and ciprofloxacin-treated groups remained relatively unchanged over the 4-week observation period, indicating poor efficacies for tobramycin and ciprofloxacin. A rapid dose-dependent decline in metabolic activity in rifampin-treated groups was observed, with almost a 90% reduction after two doses and nearly undetectable levels after three doses. The disappearance of light emission correlated with colony counts. After the final treatment, cell numbers rebounded as a function of concentration in a time-dependent manner. The staphylococci isolated from the catheters of mice treated with rifampin were uniformly resistant to rifampin but retained their in vitro susceptibilities to tobramycin and ciprofloxacin. Since the metabolic activities of viable cells and a postantibiotic effect could be detected directly on the support matrix nondestructively and noninvasively, the methodology is specifically appealing for investigating the effects of antibiotics on biofilms in vivo. Moreover, our study points to the possible use of biophotonic imaging for the detection of the development of resistance to therapeutic agents during treatment of chronic infections in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14506020
      14. Call Number :
        139345
      15. Serial :
        7448
      1. Author :
        Orihuela, Carlos J; Gao, Geli; McGee, Mackenzie; Yu, Jun; Francis, Kevin P; Tuomanen, Elaine
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2003
      5. Publication :
        Scandinavian journal of infectious diseases
      6. Products :
      7. Volume :
        35
      8. Issue :
        9
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Disease Models, Animal; Female; Lung; Mice; Mice, Inbred BALB C; Pneumococcal Infections; pXen-5; Serotyping; Streptococcus pneumoniae, Xen10, Xen7, Xen35
      12. Abstract :
        The variability of the course of infection by Streptococcus pneumoniae is well known but poorly understood. Most animal models of pneumonia, sepsis or meningitis have been forced to use site-specific bacterial inoculation to mimic localized human infection. This study examined the differences in the progression of disease-causing strains D39 (serotype 2), A66.1 (serotype 3) and TIGR4 (serotype 4) using isolates transformed with the Gram-positive lux transposon cassette, Tn4001 luxABCDE Km(r). Expression of the lux operon results in bioluminescence, permitting the detection of the bacteria within a living animal while using a CCD camera. Mice infected intranasally with A66.1 developed only pneumonia, those challenged with D39 experienced high-grade sepsis, while TIGR4 infection resulted in low-grade pneumonia and bacteremia ultimately progressing to meningitis. Quantitative analysis of bacterial titers confirmed these patterns, which were consistent across different lineages of mice. Mice anesthetized with ketamine and xylazine developed more severe forms of the disease compared with isoflurane. These studies unambiguously characterize 3 distinct models of the natural course of pneumococcal infection. Mapping these models provides a framework for detailed molecular modeling of pneumococcal virulence determinants at specific stages of disease.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14620149
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9026
      1. Author :
        Jenkins, Darlene E; Oei, Yoko; Hornig, Yvette S; Yu, Shang-Fan; Dusich, Joan; Purchio, Tony; Contag, Pamela R
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2003
      5. Publication :
        Clinical & experimental metastasis
      6. Products :
      7. Volume :
        20
      8. Issue :
        8
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        A549-luc-C8; Animals; Bioware; Cell Line, Tumor; Colonic Neoplasms; Fluorouracil; HT-29-luc-D6 cells; Humans; Image Interpretation, Computer-Assisted; Longitudinal Studies; Luciferases; Luminescent Measurements; Lung Neoplasms; Lymphatic Metastasis; Male; Mice; Mice, SCID; Mitomycin; Models, Biological; Neoplasm Transplantation; PC-3M-luc; Prostatic Neoplasms
      12. Abstract :
        Bioluminescent imaging (BLI) permits sensitive in vivo detection and quantification of cells specifically engineered to emit visible light. Three stable human tumor cell lines engineered to express luciferase were assessed for their tumorigenicity in subcutaneous, intravenous and spontaneous metastasis models. Bioluminescent PC-3M-luc-C6 human prostate cancer cells were implanted subcutaneously into SCID-beige mice and were monitored for tumor growth and response to 5-FU and mitomycin C treatments. Progressive tumor development and inhibition/regression following drug treatment were observed and quantified in vivo using BLI. Imaging data correlated to standard external caliper measurements of tumor volume, but bioluminescent data permitted earlier detection of tumor growth. In a lung colonization model, bioluminescent A549-luc-C8 human lung cancer cells were injected intravenously and lung metastases were monitored in vivo by whole animal imaging. Anesthetized mice were imaged weekly allowing a temporal assessment of in vivo lung tumor growth. This longitudinal study design permitted an accurate, real-time evaluation of tumor burden in the same animals over time. End-point bioluminescence measured in vivo correlated to total lung weight at necropsy. For a spontaneous metastatic tumor model, bioluminescent HT-29-luc-D6 human colon cancer cells implanted subcutaneously produced metastases to lung and lymph nodes in SCID-beige mice. Both primary tumors and micrometastases were detected by BLI in vivo. Ex vivo imaging of excised lung lobes and lymph nodes confirmed the in vivo signals and indicated a slightly higher frequency of metastasis in some mice. Levels of bioluminescence from in vivo and ex vivo images corresponded to the frequency and size of metastatic lesions in lungs and lymph nodes as subsequently confirmed by histology. In summary, BLI provided rapid, non-invasive monitoring of tumor growth and regression in animals. Its application to traditional oncology animal models offers quantitative and sensitive analysis of tumor growth and metastasis. The ability to temporally assess tumor development and responses to drug therapies in vivo also improves upon current standard animal models that are based on single end point data.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14713107
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8980
      1. Author :
        Jenkins, Darlene E; Yu, Shang-Fan; Hornig, Yvette S; Purchio, Tony; Contag, Pamela R
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2003
      5. Publication :
        Clinical & experimental metastasis
      6. Products :
      7. Volume :
        20
      8. Issue :
        8
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Antineoplastic Agents; Bioware; Cell Line, Tumor; Disease Models, Animal; Heart Neoplasms; Humans; Injections, Subcutaneous; Luminescent Measurements; Lung Neoplasms; Lymphatic Metastasis; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Recurrence, Local; PC-3M-luc; Prostatic Neoplasms
      12. Abstract :
        We used the bioluminescent human prostate carcinoma cell line PC-3M-luc-C6 to non-invasively monitor in vivo growth and response of tumors and metastasis before, during and after treatments. Our goal was to determine the utility of a luciferase-based prostate cancer animal model to specifically assess tumor and metastatic recurrence in vivo following chemotherapy. Bioluminescent PC-3M-luc-C6 cells, constitutively expressing luciferase, were implanted into the prostate or under the skin of mice for primary tumor assessment. Cells were also injected into the left ventricle of the heart as an experimental metastasis model. Weekly serial in vivo images were taken of anesthetized mice that were untreated or treated with 5-fluorouracil or mitomycin C. Ex vivo imaging and/or histology was used to confirm and localize metastatic lesions in various tissues initially detected by images in vivo. Our in vivo data detected and quantified early inhibition of subcutaneous and orthotopic prostate tumors in mice as well as significant tumor regrowth post-treatment. Local and distal metastasis was observed within seven days following intracardiac injection of PC-3M-luc-C6 cells. Differential drug responses and metastatic tumor relapse patterns were distinguished over time by in vivo imaging depending on the metastatic site. The longitudinal evaluation of bioluminescent tumor and metastatic development within the same cohorts of animals permitted sensitive and quantitative assessment of both primary and metastatic prostate tumor response and recurrence in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14713108
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8981
      1. Author :
        Sadikot, Ruxana T; Zeng, Heng; Yull, Fiona E; Li, Bo; Cheng, Dong-sheng; Kernodle, Douglas S; Jansen, E Duco; Contag, Christopher H; Segal, Brahm H; Holland, Steven M; Blackwell, Timothy S; Christman, John W
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        Journal of immunology (Baltimore, Md.: 1950)
      6. Products :
      7. Volume :
        172
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Cells, Cultured; Dose-Response Relationship, Immunologic; Immunity, Innate; Lung; Macrophages; Membrane Glycoproteins; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred DBA; Mice, Transgenic; NADPH Oxidase; Neutrophil Infiltration; NF-kappa B; Phosphoproteins; Pneumonia, Bacterial; Pseudomonas Infections; Receptors, Cell Surface; Signal Transduction; Toll-Like Receptors; Xen5
      12. Abstract :
        We examined the role of redox signaling generated by NADPH oxidase in activation of NF-kappaB and host defense against Pseudomonas aeruginosa pneumonia. Using mice with an NF-kappaB-driven luciferase reporter construct (HIV-LTR/luciferase (HLL)), we found that intratracheal administration of P. aeruginosa resulted in a dose-dependent neutrophilic influx and activation of NF-kappaB. To determine the effects of reactive oxygen species generated by the NADPH oxidase system on activation of NF-kappaB, we crossbred mice deficient in p47(phox) with NF-kappaB reporter mice (p47(phox-/-)HLL). These p47(phox-/-)HLL mice were unable to activate NF-kappaB to the same degree as HLL mice with intact NADPH oxidase following P. aeruginosa infection. In addition, lung TNF-alpha levels were significantly lower in p47(phox-/-)HLL mice compared with HLL mice. Bacterial clearance was impaired in p47(phox-/-)HLL mice. In vitro studies using bone marrow-derived macrophages showed that Toll-like receptor 4 was necessary for NF-kappaB activation following treatment with P. aeruginosa. Additional studies with macrophages from p47(phox-/-) mice confirmed that redox signaling was necessary for maximal Toll-like receptor 4-dependent NF-kappaB activation in this model. These data indicate that the NADPH oxidase-dependent respiratory burst stimulated by Pseudomonas infection contributes to host defense by modulating redox-dependent signaling through the NF-kappaB pathway.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14734763
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9999
      1. Author :
        Hardy, J.; Francis, K. P.; DeBoer, M.; Chu, P.; Gibbs, K.; Contag, C. H.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        Science
      6. Products :
      7. Volume :
        303
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        animal cell, animal model, article, bacterial colonization, bacterial growth, bacterial virulence, bioluminescence, cell culture, controlled study, extracellular space, gallbladder, in vivo study, Listeria monocytogenes, mouse, nonhuman, priority journal IVIS, Xenogen, Xen32
      12. Abstract :
        The bacterium Listeria monocytogenes can cause a life-threatening systemic illness in humans. Despite decades of progress in animal models of listeriosis, much remains unknown about the processes of infection and colonization. Here, we report that L. monocytogenes can replicate in the murine gall bladder and provide evidence that its replication there is extracellular and intraluminal. In vivo bioluminescence imaging was employed to determine the location of the infection over time in live animals, revealing strong signals from the gall bladder over a period of several days, in diseased as well as asymptomatic animals. The data suggest that L. monocytogenes may be carried in the human gall bladder.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/14764883
      14. Call Number :
        138442
      15. Serial :
        6154
      1. Author :
        Scatena, Caroline D; Hepner, Mischa A; Oei, Yoko A; Dusich, Joan M; Yu, Shang-Fan; Purchio, Tony; Contag, Pamela R; Jenkins, Darlene E
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        The Prostate
      6. Products :
      7. Volume :
        59
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Disease Models, Animal; Humans; LnCaP-luc cells; Luciferases; Luminescent Measurements; Male; Mice; Mice, SCID; Neoplasm Metastasis; Phenotype; Plasmids; Prostatic Neoplasms; Transfection; Transplantation, Heterologous; Tumor Cells, Cultured
      12. Abstract :
        BACKGROUND Animal experiments examining hormone-sensitive metastatic prostate cancer using the human LNCaP cell line have been limited to endpoint analyses. To permit longitudinal studies, we generated a luciferase-expressing cell line and used bioluminescent imaging (BLI) to non-invasively monitor the in vivo growth of primary LNCaP tumors and metastasis. METHODS LNCaP.FGC cells were transfected to constitutively express firefly luciferase. LNCaP-luc-M6 cells were tested for bioluminescent signal intensity and hormone responsiveness in vitro. The cells were implanted in subcutaneous and orthotopic sites in SCID-bg mice and imaged over time. RESULTS The LNCaP-luc-M6 cells formed subcutaneous and orthotopic tumors in SCID-bg mice, and nearly all tumor-bearing animals developed pulmonary metastases. Early detection and temporal growth of primary tumors and metastatic lesions was successfully monitored by BLI. CONCLUSIONS The LNCaP-luc-M6 cell line is a bioluminescent, hormone-sensitive prostate cancer cell line applicable for BLI studies to non-invasively monitor subcutaneous and orthotopic prostate tumor growth and metastasis in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15042605
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9015
      1. Author :
        Shi, Lei; Takahashi, Kazue; Dundee, Joseph; Shahroor-Karni, Sarit; Thiel, Steffen; Jensenius, Jens Christian; Gad, Faten; Hamblin, Michael R; Sastry, Kedarnath N; Ezekowitz, R Alan B
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        The Journal of experimental medicine
      6. Products :
      7. Volume :
        199
      8. Issue :
        10
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Disease Susceptibility; DNA, Bacterial; Lung; Mannose-Binding Lectin; Mice; Mice, Knockout; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; Spleen; Staphylococcal Infections; Xen8.1
      12. Abstract :
        Gram-positive organisms like Staphylococcus aureus are a major cause of morbidity and mortality worldwide. Humoral response molecules together with phagocytes play a role in host responses to S. aureus. The mannose-binding lectin (MBL, also known as mannose-binding protein) is an oligomeric serum molecule that recognizes carbohydrates decorating a broad range of infectious agents including S. aureus. Circumstantial evidence in vitro and in vivo suggests that MBL plays a key role in first line host defense. We tested this contention directly in vivo by generating mice that were devoid of all MBL activity. We found that 100% of MBL-null mice died 48 h after exposure to an intravenous inoculation of S. aureus compared with 45% mortality in wild-type mice. Furthermore, we demonstrated that neutrophils and MBL are required to limit intraperitoneal infection with S. aureus. Our study provides direct evidence that MBL plays a key role in restricting the complications associated with S. aureus infection in mice and raises the idea that the MBL gene may act as a disease susceptibility gene against staphylococci infections in humans.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15148336
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9994
      1. Author :
        Kadurugamuwa, Jagath L; Sin, Lin V; Yu, Jun; Francis, Kevin P; Purchio, Tony F; Contag, Pamela R
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        Antimicrobial agents and chemotherapy
      6. Products :
      7. Volume :
        48
      8. Issue :
        6
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Antibiotics, Antitubercular; Biofilms; Bioware; Colony Count, Microbial; Diagnostic Imaging; DNA-Directed RNA Polymerases; Luminescent Measurements; Mice; Rifampin; Staphylococcal Infections; Staphylococcus aureus; Xen29
      12. Abstract :
        Eradication of Staphylococcus aureus biofilms after rifampin treatment was tested in a mouse model of device-related infection by using biophotonic imaging. Following treatment, the bioluminescent signals decreased to undetectable levels, irrespective of the age of the biofilm. After the final treatment, the signals rebounded in a time-dependent manner and reached those for the untreated mice. Readministration of rifampin was unsuccessful in eradicating reestablished infections, with the rifampin MICs for such bacteria being increased and with the bacteria having point mutations in the rpoB gene.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15155235
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9056
      1. Author :
        Minakuchi, Yoshiko; Takeshita, Fumitaka; Kosaka, Nobuyoshi; Sasaki, Hideo; Yamamoto, Yusuke; Kouno, Makiko; Honma, Kimi; Nagahara, Shunji; Hanai, Koji; Sano, Akihiko; Kato, Takashi; Terada, Masaaki; Ochiya, Takahiro
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        Nucleic acids research
      6. Products :
      7. Volume :
        32
      8. Issue :
        13
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; B16-F10-luc-G5 cells; Bioware; Cell Division; Cell Line, Tumor; Collagen; Humans; Injections; Male; Mice; Mice, Nude; RNA Interference; RNA Stability; RNA, Small Interfering; Testicular Neoplasms; Transduction, Genetic; Xenograft Model Antitumor Assays
      12. Abstract :
        Silencing gene expression by siRNAs is rapidly becoming a powerful tool for the genetic analysis of mammalian cells. However, the rapid degradation of siRNA and the limited duration of its action call for an efficient delivery technology. Accordingly, we describe here that Atelocollagen complexed with siRNA is resistant to nucleases and is efficiently transduced into cells, thereby allowing long-term gene silencing. Site-specific in vivo administration of an anti-luciferase siRNA/Atelocollagen complex reduced luciferase expression in a xenografted tumor. Furthermore, Atelocollagen-mediated transfer of siRNA in vivo showed efficient inhibition of tumor growth in an orthotopic xenograft model of a human non-seminomatous germ cell tumor. Thus, for clinical applications of siRNA, an Atelocollagen-based non-viral delivery method could be a reliable approach to achieve maximal function of siRNA in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15272050
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9003