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  • Immunocytochemistry  (2)
  • Bone marrow transplantation  (1)
  • Holliday structure  (1)
  • 1
    ISSN: 1432-0878
    Keywords: Cathepsin E ; Aspartic proteinase ; Osteoclasts ; Immunocytochemistry ; Rat (WKA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunocytochemical localization of cathepsin E, a non-lysosomal aspartic proteinase, was investigated in rat osteoclasts using the monospecific antibody to this protein. At the light-microscopic level, the preferential immunoreactivity for cathepsin E was found at high levels in active osteoclasts in the physiological bone modeling process. Neighboring osteoblastic cells were devoid of its immunoreactivity. At the electron-microscopic level, cathepsin E was exclusively confined to the apical plasma membrane at the ruffled border of active osteoclasts and the eroded bone surface. Cathepsin E was also concentrated in some endocytotic vacuoles of various sizes in the vicinity of the ruffled border membrane, some of which appeared to be secondary lysosomes containing the phagocytosed materials. These results strongly suggest that this enzyme is involved both in the extracellular degradation of the bone organic matrix and in the intracellular breakdown of the ingested substances in osteoclasts.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Key words: Cathepsin E ; Aspartic proteinase ; Osteoclasts ; Immunocytochemistry ; Rat (WKA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The immunocytochemical localization of cathepsin E, a non-lysosomal aspartic proteinase, was investigated in rat osteoclasts using the monospecific antibody to this protein. At the light-microscopic level, the preferential immunoreactivity for cathepsin E was found at high levels in active osteoclasts in the physiological bone modeling process. Neighboring osteoblastic cells were devoid of its immunoreactivity. At the electron-microscopic level, cathepsin E was exclusively confined to the apical plasma membrane at the ruffled border of active osteoclasts and the eroded bone surface. Cathepsin E was also concentrated in some endocytotic vacuoles of various sizes in the vicinity of the ruffled border membrane, some of which appeared to be secondary lysosomes containing the phagocytosed materials. These results strongly suggest that this enzyme is involved both in the extracellular degradation of the bone organic matrix and in the intracellular breakdown of the ingested substances in osteoclasts.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1619-0904
    Keywords: In vivo tissue engineering ; Bone marrow transplantation ; bFGF ; Release system of angiogenic growth factors ; Revascularization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract For successful in vivo tissue engineering, a growth factor release system will be useful. We adopted autologous bone marrow transplantation as an angiogenic growth factor release system. Bone marrow transplanted into a synthetic vascular prosthesis produced continuous synthesis of angiogenic growth factors, resulting in rapid neointima formation on the prosthesis after implantation. We expected a similar angiogenic phenomenon to occur if bone marrow was transplanted into ischemic myocardium. Bone marrow was transplanted into ischemic myocardium created in dogs. Marrow cells continued synthesis of angiogenic growth factors, which were effective in protecting the capillary network from ischemia, but not myocytes. Autologous bone marrow was injected intramuscularly into ischemic myocardium created in the left ventricular wall of dogs. Control operations were performed without bone marrow. On days 3 and 7, marrow cells survived, and their adjacent cells and the surrounding extracellular matrix were immunohistochemically bFGF reactive. At 3 weeks, no marrow cells were identified. Myocytes disappeared, but the capillary blood vessel networks remained. With some exceptions, these capillaries did not contain blood cell components. In the controls, scar tissue with a very small number of capillaries was formed. In conclusion, marrow cells survived for a short period of time after transplantation, and continued synthesis of angiogenic growth factors, which were effective in protecting endothelial cells from ischemia, but not myocytes. Therefore, the results also suggest that there are limitations in the treatment of ischemic myocardium using angiogenic growth factors alone.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Homologous recombination ; recA ; recF ; Holliday structure ; Mismatch repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gene conversion, the non-reciprocal transfer of sequence information between homologous DNA sequences, has been reported in lower eukaryotes, mammals and in Escherichia coli. In an E. coli rec + strain, we established a plasmid carrying two different deleted neo genes (neoDL and neoDR) in an inverted orientation and then selected for homologous recombination events that had reconstructed an intact neo + gene. We found some plasmids that had apparently experienced intramolecular gene conversion. Further evidence, however, suggests that they are products of multiple rounds of reciprocal crossing-over,apparently involving two plasmid molecules. First, most of the Neo+ clones contained multiple types of Neo+ plasmids, although the frequency of producing the neo + clones was low. Second, all the neo + clones also contained, as a minority, one particular form of dimer, which can be formed by reciprocal crossing-over between neoDL of one plasmid molecule and neoDR of another plasmid molecule. Third, in reconstruction experiments, we cloned and purified this dimer and transferred it back into the rec + cells. The dimer gave rise to clones containing multiple types of neo + recombinant monomers, including those apparent gene conversion types, and containing only few molecules of this dimer plasmid.
    Type of Medium: Electronic Resource
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