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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
    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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  • 3
    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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  • 4
    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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  • 5
    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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  • 6
    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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  • 7
    Electronic Resource
    Electronic Resource
    Increased oxidative damage in nuclear and mitochondrial DNA in Alzheimer's disease (2005)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 93 (2005), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Increasing evidence suggests that oxidative stress is associated with normal aging and several neurodegenerative diseases, including Alzheimer's disease (AD). Here we quantified multiple oxidized bases in nuclear and mitochondrial DNA of frontal, parietal, and temporal lobes and cerebellum from short postmortem interval AD brain and age-matched control subjects using gas chromatography/mass spectrometry with selective ion monitoring (GC/MS-SIM) and stable labeled internal standards. Nuclear and mitochondrial DNA were extracted from eight AD and eight age-matched control subjects. We found that levels of multiple oxidized bases in AD brain specimens were significantly (p 〈 0.05) higher in frontal, parietal, and temporal lobes compared to control subjects and that mitochondrial DNA had approximately 10-fold higher levels of oxidized bases than nuclear DNA. These data are consistent with higher levels of oxidative stress in mitochondria. Eight-hydroxyguanine, a widely studied biomarker of DNA damage, was approximately 10-fold higher than other oxidized base adducts in both AD and control subjects. DNA from temporal lobe showed the most oxidative damage, whereas cerebellum was only slightly affected in AD brains. These results suggest that oxidative damage to mitochondrial DNA may contribute to the neurodegeneration of AD.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.2005.03053.x
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  • 8
    Electronic Resource
    Electronic Resource
    Surfactants, polymers and their nanoparticles for personal care applications (2005)
    Oxford, UK : Blackwell Science Ltd
    International journal of cosmetic science 27 (2005), S. 0 
    Details
    ISSN: 1468-2494
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A ‘touch me not’ plant folding up rapidly upon being attacked or microbes depositing on teeth or ocean vessels even under hostile conditions are examples in nature that provide inspiration for developing new classes of personal care release or deposition systems. In this paper, development of such systems based on polymer/surfactant colloid chemistry is explored for achieving transport and release of cosmetic and pharmaceutical molecules at desired rates at desired sites. The successful development of products depends upon understanding and utilizing key interactions among surfactants, polymers and hybrid polymers that are relevant to personal care products. Thus, the absorbed layers or tethers on the particulates can be manipulated for desired dispersion of actives or depositions on substrate under any and all conditions. New hybrid polymers and nanogels have been synthesized for tuning up nanodomains that can extract and deliver at will cosmetics/drugs/toxins by perturbing pH, temperature or ionic strength of the system. Particularly, hydrophobically modified polymers have features of both polymers and surfactants and due to the associative nature of the hydrophobic groups, such polymers can form intramolecular nanodomains for performing carrier functions. Nanogels developed recently include that of polyacrylamide, poly(acrylic acid) and starch nanogels modified for extraction and subsequent slow release of fragrances and overdosed toxic drugs. Binding and release processes were investigated using surface plasmon resonance and fluorescence spectroscopies, powerful techniques for monitoring short term and long term changes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1467-2494.2005.00257_2.x
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  • 9
    Electronic Resource
    Electronic Resource
    Cytokine Responses to Fungal Pathogens in Kupffer Cells are Toll-like Receptor 4 Independent and Mediated by Tyrosine Kinases (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: Disseminated fungal infections are increasing. However, the interactions between the body's largest population of tissue macrophages, the Kupffer cells and the fungal pathogens are scarcely understood. The aim of this study was to examine the involvement of Toll-like receptor 4 (TLR4) signalling in cytokine production, using primary cultures of rat and murine Kupffer cells exposed to Aspergillus fumigatus and Candida albicans hyphae and conidia. All fungal components induced the release of tumour necrosis factor-α (TNF-α), but with delayed kinetics compared with lipopolysaccharide (LPS). Candida albicans was the most potent inducer of TNF-α protein and mRNA and the only inducer of interleukin-10 (IL-10) in rat Kupffer cells. All fungal components induced enhanced mRNA levels of macrophage inhibitory protein-2 (MIP-2) in the cells, similar to LPS. Inhibitors of Src tyrosine kinases added to cells prior to stimulation led to attenuation in the release of both TNF-α (60%, P 〈 0.05) and IL-10 (70%, P 〈 0.05) induced by C. albicans conidia but did not influence the LPS-mediated cytokine release. Murine Kupffer cells (C57BL/10J) also released TNF-α as well as the chemokines keratinocyte-derived chemokine (KC) and MIP-2 in response to fungal component. Surprisingly, Kupffer cells from TLR4-deficient C57BL/ScCr mice exhibited significantly enhanced production of KC and MIP-2 upon stimulation by fungal components compared with control littermates (P 〈 0.05). Our study demonstrates that Aspergillus and Candida components induce cytokine production in rat Kupffer cells and that the response to C. albicans conidia involves Src tyrosine kinases. The experiments with TLR4-deficient Kupffer cells suggest that the cytokine response in these cells to fungal component is not mediated by TLR4.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3083.2005.01653.x
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  • 10
    Electronic Resource
    Electronic Resource
    Genetic diversity of the annual weed Monochoria vaginalis in southern China detected by random amplified polymorphic DNA and inter-simple sequence repeat analyses (2005)
    Oxford, UK : Blackwell Science Ltd
    Weed research 45 (2005), S. 0 
    Details
    ISSN: 1365-3180
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Two molecular genetic screening techniques, RAPD (random amplified polymorphic DNAs) and ISSR (inter-simple sequence repeats), were applied to detect the level and pattern of genetic diversity of Monochoria vaginalis, a common weed of rice fields, in seven populations from southern China. Among these populations, 116 bands were amplified by 18 RAPD primers, of which 34 bands (29.31%) were polymorphic, and 14 ISSR primers produced 111 bands with 87 polymorphic bands (78.38%). Within each population, a relatively low level of genetic diversity was detected by both RAPD and ISSR analyses, with a mean genetic diversity (H) of 0.0348 and 0.0551 respectively. Analysis of molecular variance of the data from the RAPD and ISSR markers detected that the majority of total genetic variation existed among populations (73.50% and 76.70% respectively) and only minor genetic variation within populations (26.50% and 23.30% respectively). Cluster analysis divided the seven populations into two groups, indicating that the genetic relationships among populations have relatively low correlation with their geographical distribution (Mantel test; r = 0.45 and 0.48 respectively). Our results indicated that both RAPD and ISSR markers were effective and reliable for accurately assessing the degree of genetic variation of M. vaginalis. Comparing the two techniques, ISSR markers were more efficient than the RAPD assay. The Mantel test gave r = 0.16, suggesting no correlation between these two molecular markers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3180.2005.00475.x
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