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In this study we examined the developmental potential of reconstructed embryos and the fate of donor mitochondria during preimplantation development after nuclear transfer in cattle. Isolated cumulus cells were used as donor cells in nuclear transfer. Cumulus cells labelled with MitoTracker Green FM fluorochrome were injected into enucleated bovine MII oocytes and cultured in vitro. MitoTracker labelling on donor cells did not have a detrimental effect on blastocyst formation following nuclear transfer. Cleavage rate was about 69% (56/81) and blastocyst formation rate was 6.2% (5/81) at 7 days after nuclear transfer. The labelled mitochondria dispersed to the cytoplasm and became distributed between blastomeres and could be identified up to the 8- to 15-cell stage. Small patches of mitochondria were detected in some 8- to 15-cell stage embryos (5/20). However, donor mitochondria were not detected in embryos at the 16-cell stage and subsequent developmental stages. In the control group, mitochondria could be identified in arrested 1-cell embryos up to 7 days after nuclear transfer. These results suggest that disappearance of the labelled donor mitochondria in nuclear transfer bovine embryos is not due to fading of the fluorochrome marker, but is rather an as yet undefined cytoplasmic event.
A new approach is proposed to obtain fast crystallizing materials based on a conventional GeSbTe alloy for rewritable phase change optical data storage. By means of co-sputtering, Ge1Sb2Te4alloy was mixed with Sn1Bi2Te4alloy so as to form pseudo-binary alloys (Ge1Sb2Te4)1-x(Sn1Bi2Te4)x (x is a mole fraction). From structural and optical analyses of the co- sputtered and annealed alloy films, the formation of stable crystalline single phases was observed along with a Vegard's law behavior, suggesting a homogeneous mixing of the two alloys. By use of a 4 layered disk with (Ge1Sb2Te4)0.85(Sn1Bi2Te4)0.15 recording layer, a preliminary test of writing and erasing was carried out and the results were compared with the case of the disk with Ge1Sb2Te4recording layer. The (Ge1Sb2Te4)0.85(Sn1Bi2Te4)0.15 recording layer was found to yield markedly higher erasibility, especially with increasing disk linear velocity.
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