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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science
    Wound repair and regeneration 5 (1997), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Products of the eicosanoid pathways, namely prostaglandins and leukotrienes, are known to play a key role in inflammatory and immune responses, as well as other wound cellular activities of wounded tissue. The objective of this study was to determine whether surgically induced intraperitoneal and incisional wounds in the rat express 5-lipoxygenase and contain binding sites for leukotrienes and whether their levels change during the course of healing. With the use of a specific monoclonal antibody generated against 5-lipoxygenase, the enzyme was immunohistochemically localized in various wound cells during postsurgical days 2 to 35. The inflammatory cells within the wound were the major cell types containing 5-lipoxygenase immunoreactive protein, followed by fibroblasts in the incisional and peritoneal fibrous adhesions, striated muscle, and the vasculature. The greatest level of immunostaining was observed during the first 2 weeks after surgery, which decreased to near unwounded levels by day 35. Light microscope autoradiographic binding studies using 3H-leukotrienes indicated that the peritoneal/incisional wounds and unwounded tissues contain specific 3H-leukotriene C4 and 3H-leukotriene D4 but not 3H-leukotriene B4 binding sites. Quantitative grain analysis (net grain density/100 µm2), representing specific 3H-leukotriene C4 and 3H-leukotriene D4 binding sites calculated for different cell types in the wound and unwounded regions showed that 3H-leukotriene C4 binding was highest over the striated muscle proximal to the injury and incisional and peritoneal granulation tissue fibroblasts. The net grain density over these cells increased by 3-, 2.5-, and 2-fold by day 14, respectively, and declined to the control values by day 21 after injury (p 〈 0.05). The pattern of 3H-leukotriene D4 binding was similar to that observed for 3H-leukotriene C4, but with a lower density. The grain density for 3H-leukotriene C4 and 3H-leukotriene D4 in arteriolar endothelial and smooth muscle cells remained unchanged. These data suggest that the products of the lipoxygenase pathway through the presence of their specific receptors may play an important role in peritoneal wound repair and adhesion formation.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The presence and cellular distribution of epidermal growth factor (EGF), TGF-α, and EGF-R were determined in the rat fascial and peritoneal tissue during healing of an incisional injury by means of immunohistochemistry and autoradiographic techniques. The immunostaining intensity for EGF in the regenerating wound area was substantially higher during the first 14 days, then decreased to near prewound levels during 14 to 35 days after surgery. Within the wound area, the most intense immunostaining occurred with inflammatory cells, followed by fascial striated muscle and arterioles, whereas fibroblasts in the regenerating area contained very low immunostaining intensity. The immunostaining pattern for TGF-α with the use of three separate polyclonal antibodies that were directed against the amino and carboxy termini of TGF-α precursor and a fragment of the mature 50-amino-acid form of TGF-α was similar to that seen with EGF and persisted until 28 days after injury. However, fibroblasts in the regenerating area immunostained intensely for TGF-α but not for EGF. Quantitative autoradiography of iodine 125—labeled EGF binding and immunohistochemical studies of the EGF-R with monoclonal antibodies that were directed against the extracellular binding domain of EGF-R demonstrated the presence of specific EGF-R in regenerating fascial and peritoneal tissue. Net grain density (100 µm2), representing specific binding of 125I-EGF, was calculated for different cell types in the wound. The grain density over fascial striated muscle, migratory fibroblasts and peritoneal fibroblasts increased by two and one half, three, and four times, respectively, at 7 days and decreased to the values in adjacent unwounded tissue by 21 days after injury (p 〈 0.05). Immunostaining for the EGF-R generated similar patterns, which persisted for 14 days after injury. The grain density and immunostaining for EGF-R over the arterioles in the wound did not change during the course of healing and was similar to that of the uninjured regions. In summary, these observations indicate that the local levels of EGF, TGF-α, and EGF-R increase during the early phases of healing in fascial and peritoneal injury, which suggests a role for these growth factors in the normal mechanism of fascial/peritoneal wound healing and fibrous adhesion formation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 141 (1989), S. 142-147 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A differentiation-defective mouse myoblast subclone (DD-1), cells of which do not fuse into myotubes nor synthesize muslce-specific proteins, was employed to help define the role of eicosanoids in mouse myoblast differentiation. We observed by hplc, tIc, and radioimmunoassay that the DD-1 cells release strikingly higher levels of cyclooxygenase pathway products prostaglandin E2 and F2α into the culture medium than the parental non-differentiation-defective cells (DZ). In contrast, the levels of 15-hydroxyeicosatetraenoic acid (15-HETE), a lipoxygenase product, and a putatively identified second lipoxygenase product (LLP) did not differ greatly in the two cell types. The DD-1 cells also have strikingly higher levels of cyclooxygenase activity than the parental cells as determined by intact and broken cell assays. Additional fusion-defective clones were isolated on the basis of their flattened appearance and ability to grow in “mitogen-poor” medium and these cells also released strikingly higher levels of prostaglandins E2 and F2α into the growth medium. The “turn on” of the cyclooxygenase pathway in the DD-1 cells and other fusion-defective cells is consistent with the hypothesis that the products of this pathway contribute to the inability of myoblasts to fuse with one another. This hypothesis is supported by the observation that there is a dose-dependent decrease in fusion of DZ cells when PGE2 is added to commitment medium.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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