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  • 1
    Electronic Resource
    Electronic Resource
    Enhanced Maturation and Functional Capacity of Dendritic Cells Induced by Mannosylated L2 Domain of ErbB2 Receptor (2005)
    Oxford, UK; Malden, USA : Blackwell Science Ltd
    Scandinavian journal of immunology 62 (2005), S. 0 
    Details
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The nature of antigens and functional state of dendritic cells (DC) are important in antigen presentation. The ability of DC for the induction of T-cell responses is promoted by maturation. It has been confirmed that mannose receptors mediate highly efficient endocytosis and presentation of mannosylated proteins. In the present study, L2 domain of ErbB2 ectodomain was expressed in Escherichia coli, purified and mannosylated. The maturation and functional capacity of DC induced by mannosylated L2 (mL2) protein were investigated. The results showed that L2 protein could induce DC maturation, which was accompanied by elevated expression of MHC and co-stimulatory molecules. The effect of mL2 protein on DC maturation was more remarkable than that of non-mL2 proteins. Uptake of mL2 antigens by DC was more efficient. Furthermore, the T cells can be stimulated to proliferate in vitro and secrete Th1 and Th2 cytokines. Higher levels of both IFN-γ and IL-10 were detected from the T cells stimulated by mL2-pulsed DC, suggesting a concurrent activation of CD4+ and CD8+ T cells. The results demonstrated that L2 domain of ErbB2 receptor is an immunodominant molecule. The mL2 domain of ErbB2 can induce an enhanced maturation and functional capacity of DC. It may become an effective strategy to induce anti-ErbB2 response.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3083.2005.01642.x
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  • 2
    Electronic Resource
    Electronic Resource
    Thermochemistry of the copper fluorides (1977)
    s.l. : American Chemical Society
    The @journal of physical chemistry 〈Washington, DC〉 81 (1977), S. 2069-2073 
    Details
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/j100537a005
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  • 3
    Electronic Resource
    Electronic Resource
    Carbon monoxide-hydrogen-water: reduction of anthracene, dihydroanthracene, and quinoline (1978)
    s.l. : American Chemical Society
    The @journal of organic chemistry 43 (1978), S. 2991-2994 
    Details
    ISSN: 1520-6904
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/jo00409a012
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  • 4
    Electronic Resource
    Electronic Resource
    .alpha.-(2-Pyridine)benzyl aryl ketones as potential hypocholesteremic agents (1978)
    s.l. : American Chemical Society
    Journal of medicinal chemistry 21 (1978), S. 1339-1340 
    Details
    ISSN: 1520-4804
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/jm00210a038
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  • 5
    Electronic Resource
    Electronic Resource
    063
Electroporation-Facilitated Gene Therapy for Wound Healing (2005)
    Oxford, UK; Malden, USA : Blackwell Publishing Ltd/Inc.
    Wound repair and regeneration 13 (2005), S. 0 
    Details
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Introduction:  Using peptide growth factors to improve upon natural wound healing provides promise, but topical application of growth factors has found limited clinical success. Gene therapy has been limited by low transfection efficiency. We are investigating the use of electroporation (EP)in vivo to enhance transfection efficiency and improve wound healing with DNA expression vectors for growth factors.Methods:  To assess plasmid transfection and wound healing, gWIZ luciferase vector and PCDNA3.1/KGF expression vector were used, respectively. Cutaneous wounds were produced via 8 mm-punch biopsy in Sprague Dawley rats. Healing was impaired by cecal ligation induced sepsis. We used NIH image analysis software and histologic assessment to assess wound closure.Results:  Plasmid Transfection: EP effectively increased expression of gWIZ luciferase vector 53-fold compared to vector without EP (p 〈 0.001). We demonstrated that transfection was localized to skin when transfected skin was reflected to reveal underlying muscle. We found that EP-assisted transfection lasted for 30 days, which is appropriate for treatment of wound healing.Wound Healing:  Using PCDNA3.1/KGF expression vector and EP wounds were 60% smaller on day 12 versus vector without EP (p 〈 0.009). We assessed quality of healing with a histologic scoring system grading quality of epithelial coverage, organization of scar and resolution of inflammation. KGF vector + EP score of 3.0 +/− 0.3 was better than that of 1.8 +/− 0.3 for treatment with vector alone (p 〈 0.05).Conclusions:  These results demonstrate the capacity of electroporation-facilitated transfection with DNA plasmid expression vector for growth factor to improve wound healing.Acknowledgments:  Funding from USAMRMC PRMRP DAMDI7-03-1-0029 and Maryland State Firefighters Fund
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1067-1927.2005.130215bk.x
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  • 6
    Electronic Resource
    Electronic Resource
    067
Electroporation-Enhanced Gene Therapy with KGF-1 Corrects Healing Impairment in Older Diabetic Mice (2005)
    Oxford, UK; Malden, USA : Blackwell Publishing Ltd/Inc.
    Wound repair and regeneration 13 (2005), S. 0 
    Details
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Introduction:  Diabetes is known to impair wound healing. We assessed the effect of aging on this impairment and further explored the possibility of repairing age-dependant impairment by replenishing levels of the growth factor KGF-1 using our system of electroporation enhanced gene therapy.Methods:  Female BKS.Cg-m+/+Leprdb/J mice (diabetic) were obtained from the Jackson Laboratory (Bar Harbor, ME). Mice at time of study were 7, 27, and 84 weeks, respectively. Two wounds were created using a 5 mm punch biopsy. Plasmid DNA encoding KGF-1 was injected at 40 μg / wound. Electroporation was carried out with six square wave pulses, at 1800 volts for 100 μs; with aninterval of 125 ms. Wounds were assessed planometrically over a 14-day period.Results:  We found that wound healing is severely impaired in older, but not young diabetic mice (7-week-old mice displayed a threefold smaller wound area than 84-week-old mice on day 9). Glucose levels were similar in old and young groups ruling out differences in intensity of the metabolic derangement to explain this disparity (old: 334 +/− 41 mg/dl vs. young: 290 +/− 33 mg/dl). Treatment with KGF-1 gene therapy increased wound closure in aged mice 5.9-fold as compared to untreated aged mice 5 days after wounding (treated: 3724 +/− 631 vs. untreated: 632 +/− 517, P = 0.002). Aged KGF-1 gene therapy-treated mice closed wounds 10-fold faster than treated young mice 5 days after wounding (old: 3724: +/− 631 vs. young 358 +/− 871, P = 0.004).Conclusions:  We conclude that progressive damage to the tissues capability to heal occurs in individuals with diabetes. Treatment with KGF-1 gene therapy is more effective in aged mice than young mice. With an increasing geriatric diabetic population, gene therapy to replenish growth factors may be worth exploring.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1067-1927.2005.130215bo.x
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  • 7
    Electronic Resource
    Electronic Resource
    128
A New Method to Evaluate the Biomechanical Properties of Skin in a Porcine Model (2005)
    Oxford, UK; Malden, USA : Blackwell Publishing Ltd/Inc.
    Wound repair and regeneration 13 (2005), S. 0 
    Details
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Aims:  A method to test the axial and transverse tensile properties of skin was developed in order to improve our understanding of the mechanical behavior of skin, and how it changes with the scarring response. This experimental technique was used to investigate the bidirectional mechanics of skin in the juvenile Yorkshire pig model.Methods:  Skin samples were taken upon sacrifice, and all subcutaneous tissue was carefully removed via dissection. Dumbbell-shaped specimens were obtained from the skin using a custom designed stainless steel punch, to provide consistent geometry, and to avoid stress concentrations that can result from specimen gripping. Samples were taken in the axial (cranial-caudal) and transverse (dorsal-ventral) directions. Specimens were secured in an INSTRON universal test machine with serrated soft tissue grips, elongated to a desired preload, subjected to a 10% elongation and held for a 4-min isometric period. They were then returned to the original length and failed in tension at constant velocity.Results:  This method of sample preparation indicated high repeatability in specimen geometry, showing standard deviations of 2% and 3% in gauge length and width, respectively (N = 12). Furthermore, no slippage was observed at the skin/grip interface, and all failures occurred within the specimen gauge length.Conclusions:  This technique shows great promise for evaluating the viscoelastic and failure properties of skin, scar tissue, and pathologic scarring. Combining this biomechanical method with our parallel studies in molecular biology and biochemistry can likely produce correlations between constituent material organization and the mechanical properties of the tissue, as well as provide valuable information for the design of bioengineered tissue constructs.Acknowledgments:  Alberta Ingenuity Fund, CIHR, and NSERC
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1067-1927.2005.130216af.x
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  • 8
    Electronic Resource
    Electronic Resource
    Changing the reaction paths of a metathesis catalyst (1975)
    s.l. : American Chemical Society
    The @journal of organic chemistry 40 (1975), S. 2983-2985 
    Details
    ISSN: 1520-6904
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/jo00908a040
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  • 9
    Electronic Resource
    Electronic Resource
    A domain at the C-terminus of PS1 is required for presenilinase and γ-secretase activities (2005)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 92 (2005), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The structural requirements for presenilin (PS) to produce active presenilinase and γ-secretase enzymes are poorly understood. Here we investigate the role the cytoplasmic C-terminal region of PS1 plays in PS1 activity. Deletion or addition of residues at the PS C-terminus has been reported to inhibit presenilinase endoproteolysis of PS and alter γ-secretase activity. In this study, we use a sensitive assay in PS1/2KO MEFs to define a domain at the extreme C-terminus of PS1 that is essential for both presenilinase and γ-secretase activities. Progressive deletion of the C-terminus demonstrated that removal of nine residues produces a PS1 molecule (458ST) that lacks both presenilinase processing and γ-secretase cleavage of Notch and APP substrates. In contrast, removal of four or five residues had no effect (462ST, 463ST), while intermediate truncations partially inhibited PS1 activity. The 458ST mutant was unable to replace endogenous wtPS1 in HEK293 cells. Although 458ST was able to form a γ-secretase complex, this complex was not matured, illustrated by mutant PS1 instability, lack of endoproteolysis, and little production of mature Nicastrin. These data indicate that the C-terminal end of PS1 is essential for Nicastrin trafficking and modification as well as the replacement of endogenous PS1 by PS1 transgenes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.2004.02945.x
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  • 10
    Electronic Resource
    Electronic Resource
    Two domains within the first putative transmembrane domain of presenilin 1 differentially influence presenilinase and γ-secretase activity (2005)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 94 (2005), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Presenilins (PS) are thought to contain the active site for presenilinase endoproteolysis of PS and γ-secretase cleavage of substrates. The structural requirements for PS incorporation into the γ-secretase enzyme complex, complex stability and maturation, and appropriate presenilinase and γ-secretase activity are poorly understood. We used rescue assays to identify sequences in transmembrane domain one (TM1) of PS1 required to support presenilinase and γ-secretase activities. Swap mutations identified an N-terminal TM1 domain that is important for γ-secretase activity only and a C-terminal TM1 domain that is essential for both presenilinase and γ-secretase activities. Exchange of residues 95–98 of PS1 (sw95–98) completely abolishes both activities while the familial Alzheimer's disease mutation V96F significantly inhibits both activities. Reversion of residue 96 back to valine in the sw95–98 mutant rescues PS function, identifying V96 as the critical residue in this region. The TM1 mutants do not bind to an aspartyl protease transition state analog γ-secretase inhibitor, indicating a conformational change induced by the mutations that abrogates catalytic activity. TM1 mutant PS1 molecules retain the ability to interact with γ-secretase substrates and γ-secretase complex members, although Nicastrin stability is decreased by the presence of these mutants. γ-Secretase complexes that contain V96F mutant PS1 molecules display a partial loss of function for γ-secretase that alters the ratio of amyloid-β peptide species produced, leading to the amyloid-β peptide aggregation that causes familial Alzheimer's disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.2005.03278.x
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