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      1. Author :
        Kalchenko, V.; Shivtiel, S.; Malina, V.; Lapid, K.; Haramati, S.; Lapidot, T.; Brill, A.; Harmelin, A.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2006
      5. Publication :
        J Biomed Opt
      6. Products :
      7. Volume :
        11
      8. Issue :
        N/A
      9. Page Numbers :
        50507
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen
      12. Abstract :
        We develop an optical whole-body imaging technique for monitoring normal and leukemic hematopoietic cell homing in vivo. A recently developed near-infrared (NIR) lipophilic carbocyanine dye 1,1(')-dioctadecyl-3,3,3('),3(')-tetramethylindotricarbocyanine iodide (DiR) is used to safely and directly label the membranes of human leukemic Pre-B ALL G2 cell lines as well as primary murine lymphocytes and erythrocytes. DiR has absorption and fluorescence maxima at 750 and 782 nm, respectively, which corresponds to low light absorption and autofluorescence in living tissues. This allows us to obtain a significant signal with very low background level. A charge-coupled device (CCD)-based imager is used for noninvasive whole-body imaging of DiR-labeled cell homing in intact animals. This powerful technique can potentially visualize any cell type without use of specific antibodies conjugated with NIR fluorescent tag or loading cells with transporter-delivered NIR fluorophores. Thus, in vivo imaging based on NIR lipophilic carbocyanine dyes in combination with advanced optical techniques may serve as a powerful alternative or complementation to other small animal imaging methods.
      13. URL :
        http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17092148
      14. Call Number :
        139363
      15. Serial :
        8075
      1. Author :
        Qin, S; Seo, JW; Zhang, H; Qi, J; Curry, FRE; Ferrara, KW
      2. Title :
        An imaging-driven model for liposomal stability and circulation
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Molecular Pharmaceutics
      6. Products :
      7. Volume :
        7
      8. Issue :
        N/A
      9. Page Numbers :
        41264
      10. Research Area :
        N/A
      11. Keywords :
        Pharmacokinetic model, imaging-driven model, liposomal stability IVIS, Xenogen
      12. Abstract :
        Simultaneous labeling of the drug compartment and shell of delivery vehicles with optical and positron emission tomography (PET) probes is developed and employed to inform a hybrid physiologically based pharmacokinetic model. Based on time-dependent estimates of the concentration of these tracers within the blood pool, reticuloendothelial system (RES) and tumor interstitium, we compare the stability and circulation of long-circulating and temperature-sensitive liposomes. We find that rates of transport to the RES for long-circulating and temperature-sensitive particles are 0.046 and 0.19 h?1, respectively. Without the application of exogenous heat, the rates of release from the long-circulating and temperature-sensitive particles circulating within the blood pool are 0.003 and 0.2 h?1, respectively. Prolonged lifetime in circulation and slow drug release from liposomes result in a significantly greater drug area under the curve for the long-circulating particles. Future studies will couple these intrinsic parameters with exogenous heat-based release. Finally, we develop a transport constant for the transport of liposomes from the blood pool to the tumor interstitium, which is on the order of 0.01 h?1 for the Met-1 tumor system.
      13. URL :
        N/A
      14. Call Number :
        142306
      15. Serial :
        5387
      1. Author :
        N/A
      2. Title :
        Systemic application of CpG rich DNA suppresses adaptive T cell immunity via induction of IDO
      3. Type :
        Journal Article
      4. Year :
        2006
      5. Publication :
        European Journal of Immunology
      6. Products :
      7. Volume :
        36
      8. Issue :
        N/A
      9. Page Numbers :
        41263
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen
      12. Abstract :
        CpG-rich oligonucleotides (CpG-ODN) bind to Toll-like receptor 9 (TLR9) and are used as powerful adjuvants for vaccination. Here we report that CpG-ODN not only act as immune stimulatory agents but can also induce strong immune suppression depending on the anatomical location of application. In agreement with the adjuvant effect, subcutaneous application of antigen plus CpG-ODN resulted in antigen-specific T cell activation in local lymph nodes. In contrast, systemic application of CpG-ODN resulted in suppression of T cell expansion and CTL activity in the spleen. The suppressive effect was mediated by indoleamine 2,3-dioxygenase (IDO) as indicated by the observation that CpG-ODN induced IDO in the spleen and that T cell suppression could be abrogated by 1-methyl-tryptophan (1-MT), an inhibitor of IDO. No expression of IDO was observed in lymph nodes after injection of CpG-ODN, explaining why suppression was restricted to the spleen. Studies with a set of knockout mice demonstrated that the CpG-ODN-induced immune suppression is dependent on TLR9 stimulation and independent of type I and type II interferons. The present study shows that for the use of CpG-ODN as an adjuvant in vaccines, the route of application is crucial and needs to be considered. In addition, the results indicate that down-modulation of immune responses by CpG-ODN may be possible in certain pathological conditions.
      13. URL :
        N/A
      14. Call Number :
        144382
      15. Serial :
        7721
      1. Author :
        Raikwar, S. P.; Zavazava, N.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2011
      5. Publication :
        Transplantation
      6. Products :
      7. Volume :
        91
      8. Issue :
        N/A
      9. Page Numbers :
        41233
      10. Research Area :
        N/A
      11. Keywords :
        N/A
      12. Abstract :
        BACKGROUND: Whole pancreas and islet transplantation are currently used for the treatment of type 1 diabetes. However, the major limitations of this potentially curative approach are an inadequate supply of cadaveric pancreata, lifelong immunosuppression, and chronic graft rejection. Therefore, there is an urgent need to develop new sources of insulin-producing cells (IPCs). Here, we investigated whether embryonic stem (ES) cells can be exploited for the derivation of IPCs, and whether their transplantation can correct hyperglycemia in diabetic mice. METHODS: ES cells engineered to express pancreatic and duodenal homeobox 1 (Pdx1), a critical pancreatic transcription factor, were differentiated into pancreatic endoderm-like cells (PELCs) and evaluated for their potential to correct hyperglycemia after transplantation in diabetic mice. RESULTS: After systemic injection, PELCs localized to the pancreas, liver, and kidney. They then spontaneously differentiated into IPCs that corrected hyperglycemia in diabetic mice. When transplanted under the kidney capsule, PELC-derived IPCs were equally efficient at correcting hyperglycemia. Real-time noninvasive in vivo bioluminescence imaging (BLI) of rat insulin promoter (RIP)-driven luciferase was used to monitor the fate of the transplanted PELCs. To confirm that the transplanted cells were responsible for the correction of hyperglycemia, kidneys containing the transplanted cells were nephrectomized, causing rapid hyperglycemia. Interestingly, none of the animals transplanted with PELCs developed tumors, a potential consequence of the differentiation and purification procedures. CONCLUSIONS: Our data suggest that Pdx1-expressing PELCs are capable of spontaneously undergoing differentiation in vivo into IPCs and leading to a sustained correction of hyperglycemia in diabetic mice.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/21452407
      14. Call Number :
        142357
      15. Serial :
        7660
      1. Author :
        Li, J. Y.; Wang, H.; May, S.; Song, X.; Fueyo, J.; Fuller, G. N.; Wang, H.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2008
      5. Publication :
        J Neurooncol
      6. Products :
      7. Volume :
        88
      8. Issue :
        N/A
      9. Page Numbers :
        41220
      10. Research Area :
        N/A
      11. Keywords :
        Astrocytoma/drug therapy/pathology, Blotting, Western, Brain Neoplasms/genetics/*metabolism/*pathology, Cells, Cultured, Enzyme Activation/physiology, Glioblastoma/drug therapy/pathology, Glioma/genetics/*metabolism/*pathology, Gliosarcoma/drug therapy/pathology, Humans, Immunohistochemistry, JNK Mitogen-Activated Protein Kinases/*metabolism, Oligodendroglioma/drug therapy/pathology, Receptor, Epidermal Growth Factor/*biosynthesis/genetics IVIS, Xenogen
      12. Abstract :
        The c-Jun NH2-terminal kinase (JNK), which belongs to mitogen-activated protein kinase family, plays a major role in apoptosis in various cell types. JNK activation, however, also contributes to proliferation, survival, and tumorigenesis in some tumors, including gliomas. In this study, we used an immunohistochemical approach to examine the activation status of JNK of 226 gliomas in a high-density tissue microarray comprising all WHO codified WHO diffuse glioma subtypes and grades. The results were correlated with grade and EGFR expression status. Constitutively activated JNK (pJNK) was detected in 90.5%, 62.9% and 17.5% of WHO grade IV, III and II gliomas, respectively (p < 0.001). pJNK expression was not detected in the astrocytes or oligodendrocytes of any of 10 normal cerebral and cerebellar brain tissue samples. Among the 76 diffuse gliomas that exhibited EGFR expression, 63 (82.9%) were positive for pJNK. In contrast, only 50% (36/72) of the gliomas that were negative for EGFR were positive for pJNK (p < 0.0001). Overexpression of EGFR vIII in U87 cells or EGF treatment of U87-EGFR stable cells led to marked increase in JNK activation compared to parental U87 cells. Our data thus provide strong support for the hypothesis that JNK activation plays a role in the tumorigenesis and/or progression of diffuse gliomas, and suggests that EGFR is involved in constitutive JNK activation in diffuse gliomas. The ability to inhibit JNK activation might confer increased sensitivity to therapeutic modalities targeting this pathway.
      13. URL :
        http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18246408
      14. Call Number :
        140311
      15. Serial :
        5780
      1. Author :
        Zhao, H; Doyle, TC; Coquoz, O; Kalish, F; Rice, BW; Contag, CH
      2. Title :
        Emission spectra of bioluminescent reporters and interaction with mammalian tissue determine the sensitivity of detection in vivo
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Journal of biomedical optics
      6. Products :
      7. Volume :
        10
      8. Issue :
        N/A
      9. Page Numbers :
        41210
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen
      12. Abstract :
        In vivo bioluminescence imaging depends on light emitted by luciferases in the body overcoming the effect of tissue attenuation. Understanding this relationship is essential for detection and quantification of signal. We have studied four codon optimized luciferases with different emission spectra, including enzymes from firefly (FLuc), click beetle (CBGr68, CBRed) and Renilla reniformins (hRLuc). At 25 degrees C, the in vitro lambda(max) of these reporters are 578, 543, 615, and 480 nm, respectively; at body temperature, 37 degrees C, the brightness increases and the firefly enzyme demonstrates a 34-nm spectral red shift. Spectral shifts and attenuation due to tissue effects were evaluated using a series of 20-nm bandpass filters and a cooled charge-coupled device (CCD) camera. Attenuation increased and the spectra of emitted light was red shifted for signals originating from deeper within the body relative to superficial origins. The tissue attenuation of signals from CBGr68 and hRLuc was greater than from those of Fluc and CBRed. To further probe tissue effects, broad spectral emitters were created through gene fusions between CBGr68 and CBRed. These resulted in enzymes with broader emission spectra, featuring two peaks whose intensities are differentially affected by temperature and tissue depth. These spectral measurement data allow for improved understanding of how these reporters can be used in vivo and what they can reveal about biological processes in living subjects.
      13. URL :
        N/A
      14. Call Number :
        145102
      15. Serial :
        6052
      1. Author :
        Mansfield, J. R.; Gossage, K. W.; Hoyt, C. C.; Levenson, R. M.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        J Biomed Opt
      6. Products :
      7. Volume :
        10
      8. Issue :
        N/A
      9. Page Numbers :
        41207
      10. Research Area :
        N/A
      11. Keywords :
        Algorithms, Animals, *Artifacts, Image Enhancement/*methods, Image Interpretation, Computer-Assisted/*methods, Luminescent Proteins/metabolism, Male, Mice, Microscopy, Fluorescence/instrumentation/*methods, Neoplasm Proteins/*metabolism, Phantoms, Imaging, Prostatic Neoplasms/*metabolism/*pathology, *Quantum Dots, Reproducibility of Results, Sensitivity and Specificity
      12. Abstract :
        The ability to image and quantitate fluorescently labeled markers in vivo has generally been limited by autofluorescence of the tissue. Skin, in particular, has a strong autofluorescence signal, particularly when excited in the blue or green wavelengths. Fluorescence labels with emission wavelengths in the near-infrared are more amenable to deep-tissue imaging, because both scattering and autofluorescence are reduced as wavelengths are increased, but even in these spectral regions, autofluorescence can still limit sensitivity. Multispectral imaging (MSI), however, can remove the signal degradation caused by autofluorescence while adding enhanced multiplexing capabilities. While the availability of spectral “libraries” makes multispectral analysis routine for well-characterized samples, new software tools have been developed that greatly simplify the application of MSI to novel specimens.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/16178631
      14. Call Number :
        140896
      15. Serial :
        8271
      1. Author :
        Murakami, T; Kobayashi, E
      2. Title :
        Color-engineered rats and luminescent LacZ imaging: a new platform to visualize biological processes
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Journal of biomedical optics
      6. Products :
      7. Volume :
        10
      8. Issue :
        N/A
      9. Page Numbers :
        41204
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen
      12. Abstract :
        The rat represents an excellent mammalian model for broadening medical knowledge, and a wealth of information on its physiology has been obtained from its use as an experimental organism. Furthermore, its ample body size allows various surgical manipulations that cannot be performed on a mouse. Many rat models mimic human diseases and have therefore been used in a variety of biomedical studies, including physiology, pharmacology, and transplantation. In an effort to create specifically designed rats for new biomedical research and the field of regenerative medicine, we develop an engineered rat system on the basis of transgenic technology and succeed in establishing unique rats that possess genetically encoded color probes: green fluorescent protein (GFP), DsRed2 (red liver), Cre/LoxP (red to green), and LacZ (blue and luminescence). In this work, we highlight their characteristics and describe recent applications for tissue engineering and regeneration. Coupled with recent progress in modern imaging systems, these transgenic rats are providing powerful tools for the elucidation of many cellular processes in biomedical science, and may lead to innovative medical treatments.
      13. URL :
        N/A
      14. Call Number :
        141457
      15. Serial :
        5731
      1. Author :
        Troy, T.; Jekic-McMullen, D.; Sambucetti, L.; Rice, B.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2004
      5. Publication :
        Molecular imaging : official journal of the Society for Molecular Imaging
      6. Products :
      7. Volume :
        3
      8. Issue :
        N/A
      9. Page Numbers :
        41175
      10. Research Area :
        N/A
      11. Keywords :
        Adenocarcinoma/metabolism/pathology, Animals, Cell Line, Tumor, Diagnostic Imaging/instrumentation/methods, Female, Fluorescence, Fluorescent Dyes/diagnostic use/metabolism, Hela Cells, Humans, *Luminescent Measurements, Luminescent Proteins/analysis/*diagnostic use/metabolism, Male, Mice, Mice, Inbred Strains, Mice, Nude, Models, Animal, Optics and Photonics, Organic Chemicals/diagnostic use/metabolism, Prostatic Neoplasms/metabolism/pathology, Sensitivity and Specificity IVIS, Xenogen
      12. Abstract :
        Bioluminescent and fluorescent reporters are finding increased use in optical molecular imaging in small animals. In the work presented here, issues related to the sensitivity of in vivo detection are examined for standard reporters. A high-sensitivity imaging system that can detect steady-state emission from both bioluminescent and fluorescent reporters is described. The instrument is absolutely calibrated so that animal images can be analyzed in physical units of radiance allowing more quantitative comparisons to be performed. Background emission from mouse tissue, called autoluminescence and autofluorescence, is measured and found to be an important limitation to detection sensitivity of reporters. Measurements of dual-labeled (bioluminescent/fluorescent) reporter systems, including PC-3M-luc/DsRed2-1 and HeLa-luc/PKH26, are shown. The results indicate that although fluorescent signals are generally brighter than bioluminescent signals, the very low autoluminescent levels usually results in superior signal to background ratios for bioluminescent imaging, particularly compared with fluorescent imaging in the green to red part of the spectrum. Fluorescence detection sensitivity improves in the far-red to near-infrared, provided the animals are fed a low-chlorophyll diet to reduce autofluorescence in the intestinal region. The use of blue-shifted excitation filters is explored as a method to subtract out tissue autofluorescence and improve the sensitivity of fluorescent imaging.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15142408
      14. Call Number :
        143731
      15. Serial :
        7423
      1. Author :
        Wilmink, G. J.; Opalenik, S. R.; Beckham, J. T.; Davidson, J. M.; Jansen, E. D.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2006
      5. Publication :
        J Biomed Opt
      6. Products :
      7. Volume :
        11
      8. Issue :
        N/A
      9. Page Numbers :
        41114
      10. Research Area :
        N/A
      11. Keywords :
        Animals, Biomimetic Materials/*radiation effects, Cells, Cultured, Dose-Response Relationship, Radiation, HSP70 Heat-Shock Proteins/*metabolism, Humans, Lasers/*adverse effects, Luminescent Measurements/*methods, Mice, Microscopy, Fluorescence/methods, Radiation Dosage, Risk Assessment/methods, Risk Factors, Skin/injuries/*metabolism/*radiation effects, *Skin, Artificial IVIS, Xenogen
      12. Abstract :
        Effective medical laser procedures are achieved by selecting laser parameters that minimize undesirable tissue damage. Traditionally, human subjects, animal models, and monolayer cell cultures have been used to study wound healing, tissue damage, and cellular effects of laser radiation. Each of these models has significant limitations, and consequently, a novel skin model is needed. To this end, a highly reproducible human skin model that enables noninvasive and longitudinal studies of gene expression was sought. In this study, we present an organotypic raft model (engineered skin) used in combination with bioluminescent imaging (BLI) techniques. The efficacy of the raft model was validated and characterized by investigating the role of heat shock protein 70 (hsp70) as a sensitive marker of thermal damage. The raft model consists of human cells incorporated into an extracellular matrix. The raft cultures were transfected with an adenovirus containing a murine hsp70 promoter driving transcription of luciferase. The model enables quantitative analysis of spatiotemporal expression of proteins using BLI. Thermal stress was induced on the raft cultures by means of a constant temperature water bath or with a carbon dioxide (CO2) laser (lambda=10.6 microm, 0.679 to 2.262 Wcm2, cw, unfocused Gaussian beam, omegaL=4.5 mm, 1 min exposure). The bioluminescence was monitored noninvasively with an IVIS 100 Bioluminescent Imaging System. BLI indicated that peak hsp70 expression occurs 4 to 12 h after exposure to thermal stress. A minimum irradiance of 0.679 Wcm2 activated the hsp70 response, and a higher irradiance of 2.262 Wcm2 was associated with a severe reduction in hsp70 response due to tissue ablation. Reverse transcription polymerase chain reaction demonstrated that hsp70 mRNA levels increased with prolonged heating exposures. Enzyme-linked immunosorbent protein assays confirmed that luciferase was an accurate surrogate for hsp70 intracellular protein levels. Hematoxylin and eosin stains verified the presence of the thermally denatured tissue regions. Immunohistochemical analyses confirmed that maximal hsp70 expression occurred at a depth of 150 microm. Bioluminescent microscopy was employed to corroborate these findings. These results indicate that quantitative BLI in engineered tissue equivalents provides a powerful model that enables sequential gene expression studies. Such a model can be used as a high throughput screening platform for laser-tissue interaction studies.
      13. URL :
        http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16965142
      14. Call Number :
        144367
      15. Serial :
        5460
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