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  • 1
    ISSN: 1432-2307
    Keywords: Mast cell ; Compound 48/80 ; Ultrastructure ; Quick-freezing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ultrastructure of mast cells stimulated with compound 48/80 was examined by quick-freezing and deep-etching (QF-DE) or freeze-substitution (QF-FS) methods. Peritoneal cells including mast cells of adult male rats were stimulated in vitro with compound 48/80 at 17° C for 0, 10, 30, 60 or 180 s. The QF-DE replicas revealed that the mast cells stimulated with compound 48/80 for 30 s decreased filamentous actin around secretory granules. In the QF-FS specimens, perigranular membranes in mast cells stimulated for 60 s formed pentalaminar structures between adjacent granules in their cytoplasm prior to degranulation. These findings suggest that preparatory states for degranulation occur in the whole cytoplasm of stimulated mast cells at early stages. Moreover, both QF-FS specimens and QF-DE replicas revealed a compact morphological appearance of discharged granules in the extra-cellular space, indicating the existence of considerable content within the granules. Skeletal structures in the granules were also demonstrated on QF-DE replicas prepared after extracting soluble elements from the cytoplasm. It is suggested that the granular contents associated with the skeletal structures are gradually detached from the discharged granules to ensure local concentration in the tissues.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1335
    Keywords: Boron neutron-capture therapy (BNCT) ; Anti-AFP monoclonal antibody ; Boron-10 ; Prompt-γ-ray spectrometry ; Thermal neutron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We described previously that10B atoms delivered by monoclonal antibody (mAb) exerted a cytotoxic effect on AH66 cells in a dose-dependent manner upon thermal neutron irradiationin vitro. In the present study, the delivering capacity of boronated anti-(α-fetoprotein) (AFP) mAb to carry10B atoms to AFP-producing tumor xenografts in nude mice was determined. Boronated mAb was prepared by conjugating 50 mM 10B compound to an anti-AFP mAb (2 mg/ml) usingN-succinimidyl-3-) (2-pyridyldithio) propionate. The number of10B atoms conjugated directly to the mAb was estimated to be 459/antibody by prompt γ-ray spectrometry. Boron concentrations in tumor tissue obtained 12, 24, 72, and 120 h after injection of 3.0 mg10B-conjugated anti-AFP mAb were 11.10±3.12 (SD,n=6), 29.30±5.11, 33.02±11.8, and 12.91±5.62 ppm respectively. For control10B-conjugated anti-dinitrophenol (DNP) mAb, the values were 9.59±0.99, 10.37±2.86, 10.00±2.95, and 8.83±4.71 ppm respectively. The concentrations in blood were less than 0.40±0.10 ppm with anti-AFP mAb and less than 0.51±0.15 ppm with anti-DNP mAb at each sampling time (12, 24, 72, and 120 h). The number of10B atoms delivered to the tumor cells was calculated to be 0.62×109, 1.63×109, 1.84×109 and 0.72×109 at each sampling time after injection of10B-anti-AFP mAb. The amount of10B atoms necessary for effective boron neutron-capture therapy was estimated to be 109/tumor cell. We were able to carry 1.84×109 10B atoms to AH66 tumor cells by using10B-anti-AFP mAb. The accumulation reached its peak 72 h after injection. These data indicated that the10B-conjugated antitumor mAb could deliver a sufficient amount of10B atoms to the tumor cells to induce cytotoxic effects 72 h after injection upon thermal neutron irradiation.
    Type of Medium: Electronic Resource
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