DNA methylation at CpG sites, which is a epigenetic modification, is associated with gene expression without change of DNA sequences. During early mouse embryogenesis, dynamic changes of DNA methylation occur. In this study, DNA methylation patterns of porcine embryos produced in vivo and in vitro were examined at various developmental stages by the immunocytochemical staining method. Interestingly, active demethylation was not observed on the paternal pronucleus of porcine zygotes. However, differences were detected in the passive demethylation process between in vivo and in vitro embryos. There was no change in the DNA methylation state until the blastocyst stage of in vivo embryos, whereas partial demethylation was observed in several blastomeres from a 4 cell stage to a morula stage of in vitro embryos. The whole genome of inner cell mass (ICM) and trophectoderm (TE) cells in porcine blastocysts were evenly methylated without de novo methylation. Our findings demonstrate that genome-wide demethylation does not occur in pig embryos during preimplantation development unlike murine and bovine embryos. It indicates that the machinery regulating epigenetic reprogramming may be different between species.

Raman spectroscopy has been used to investigate the structure of coal tar pitch heat-treated up to 3000℃ by using 514.5 run Ar ion laser line. Four critical temperature ranges were found on pyrolyzing coal tar pitch, which correspond to four distinct processes from disordered carbons to the well-ordered graphite structure. The range of heat treat temperature (HTT) below 1000℃ corresponds to gas evolution during the pyrolysis of coal tar pitch. Above the HTT are correlated to rearrangements of enlarged molecules, growth of the molecules along the direction of plane, finally stacking in the normal direction of the plane, in the respective HTT ranges of 1000-2000, above 2000 and 2500-3000℃.