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IG Visus ME Abad MP Garcia Hernández B Agulleiro 《Canadian Metallurgical Quarterly》1996,102(1):16-27
Somatostatin (SST)- and insulin (INS)-immunoreactive (ir) cells were identified in the gut of sea bass (Dicentrarchus labrax) by immunofluorescence double staining and peroxidase-antiperoxidase (PAP) techniques for light microscopy and by immunogold method for electron microscopy using antisera to mammalian and fish peptides. SST-14 and SST-25 immunoreactivities coexisted in cells mainly located among the mucous neck cells of the gastric glands. Preabsorption controls showed that some SST-25- and, possibly, some SST-14-like peptides appeared in these cells. Immunoreactivity to fish INS, but not to mammalian INS (mINS) or insulin-like growth factor I (mIGF-I), was observed in all the SST-ir cells. The preabsorption controls suggest a cross-reaction of the fish INS antisera with SST-containing or type I cells. These cells displayed ovoid or round secretory granules with fibrous, medium electron-dense or homogeneous osmiophilic materials. Some gastric cells (type II) with round secretory granules of variable electron density, which were gold immunolabeled with bonito INS but not with mINS, mIGF-I, or SST antisera, were also found. INS-related peptide in type II cells of the sea bass stomach is suggested. 相似文献
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Rab3A is a small GTPase implicated in the docking of secretory vesicles in neuroendocrine cells. A putative downstream target for Rab3A, rabphilin-3A, is located exclusively on secretory vesicle membranes. It contains near its C terminus two C2 domains that bind Ca2+ in a phospholipid-dependent manner and an N-terminal, Rab3A-binding domain that includes a Cys-rich region. We have determined that the Cys-rich domain binds two Zn2+ ions and is necessary but not sufficient for efficient binding of rabphilin to Rab3A. A minimal Rab3A-binding domain consists of residues 45 to 170 of rabphilin. HA1-tagged Rab3A and a green fluorescent protein (GFP)-rabphilin fusion were used to examine the roles of Rab3A and of rabphilin domains in the subcellular localization of these proteins. A Rab3A mutant (T54A) that does not bind rabphifin in vitro colocalized with the GFP-rabphilin fusion, indicating that Rab3A targeting is independent of its interaction with rabphilin. Deletion of the C2 domains of rabphilin reduced membrane association of GFP-rabphilin but did not cause mistargeting of the membrane-associated fraction. However, disruption of the zinc fingers, which drastically reduced Rab3A binding, did not reduce membrane association. These results suggest that the C2 domains are required for efficient membrane attachment of rabphilin in PC12 cells and that Rab3A binding may act to target the protein to the correct membrane. 相似文献
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Human ceruloplasmin exhibited different antioxidant effects according to the electron donors in a metal-catalyzed oxidation system. Purified ceruloplasmin did not play a significant role in the protection of DNA strand breaks in the ascorbate/Fe3+/O2 system. However, when ascorbates were replaced with a thiol-reducing equivalent such as dithiothreitol, DNA strand breaks were significantly prevented by the same amount of ceruloplasmin. Ceruloplasmin did not catalyze the decomposition of H2O2 in the absence of reduced glutathione. On the contrary, ceruloplasmin showed a potent peroxidase ability to destroy H2O2 in the presence of reduced glutathione. In conclusion, the removal of H2O2 by human ceruloplasmin is not simply stoichiometric but thiol-dependent. 相似文献
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S. E. Clark (1997) offered a modified signal-detection explanation of the confidence-accuracy inversions observed in E. Tulving's (1981) experiments. In addition to replicating E. Tulving (1981), we had participants make "remember-familiar" judgments. Confidence and accuracy dissociated across subjective reports. Response confidence differed only for judgments based on familiarity, whereas accuracy differed only for "remember" responses. S. E. Clark's model does not predict this, nor can it mimic "remember" performance across all conditions. We propose that although "knowing" can be accommodated within an equal variance signal-detection account, "remembering" is governed by contextual constraints that influence the distinctiveness of information upon which participants rely during reports. The current paradigm is a pictorial analogue to H. L. Roediger and K. B. McDermott's paradigm (1995) in that participants claim to explicitly remember thematically related items that were not actually seen during study. 相似文献
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5'-degenerate 3'-dideoxy-terminated competitors of PCR primers increase specificity of amplification
Amplification of a product in PCR with specific primers may be viewed as an artificial Darwinian-type "selection of the fittest". In other selective systems, such as general evolution, immune system and probably brain cortex, the stringency of selection is not absolute but rather degenerate, with selection of many highly fit units, not limited, however, to only the fittest. In PCR also, annealing of the primers is not absolutely specific. The subsequent amplification frequently leads to amplification of not only the desired product but also to less-specific sequences. Using theoretical analysis of the degenerate mode of selection, we predict theoretically and prove experimentally that 5'-degenerate, 3'-dideoxy-terminated competitors of PCR primers can be used to dramatically improve the specificity of PCR amplification without affecting the quantitation of the final specific product. 相似文献
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