The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from 2~6mm follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, 0.5㎍/ml FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at 39℃ in a humidified atmosphere of 5% CO2 in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with 1% orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm (1 x10 5 cells/ml) in mTBM with 0.3% BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with 0.4% BSA. At 6hr of culture, the embryos were fixed in 3.7% formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and ≥3 PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group (67%), but it did not differ among the all added groups (86%, 85%, 79% and 81%, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups (25%, 30%, 33%, 29% and 29%, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased (66% vs 58%, 54%, 52% and 55%, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

We present a progress report on HCN(1-0) line observations toward starless cores to probe inward motions. We have made a single pointing survey toward the central regions of 85 starless cores and performed mapping observations of 6 infall candidate starless cores. The distributions of the velocity difference between HCN(1-0) hyperfine lines and the optically thin tracer N2H+(1-0) are significantly skewed to the blue, meaning that HCN(1-0) frequently detects inward motions. Their skewness to the blue is even greater than that of CS(2-1) Lee et al., possibly implying more infall occurrence than CS(1-0). We identify 19 infall candidates by using several characteristics illustrating spectral infall asymmetry seen in HCN(1-0) hyperfine lines, CS(3-2), CS(2-1), DCO+(2-1) and N2H+ observations. The HCN(1-0) F(O-l) with the least optical depth usually shows a similar intensity distribution to that of N2H+ which closely traces the density distribution of the cores, indicating that HCN(1-0) is less chemically affected and so believed to reflect kinematics occurring in rather inner regions of the cores. Detailed radiative transfer model fits of the spectra are underway to analyze central infall kinematics in starless cores.

Using a developed high-frequency induction heated combustion method. the simultaneous synthesis and densification of WC-xvol.%Co() hard materials was accomplished using elemental powders of W, C and Co. A complete synthesis and densification of the materials was achieved in one step within a duration of 1min. The final relative densities of the composite were over 98.5% for all cases, under the applied pressure of 60 MPa and the induced current. The hardness of the composites decreases and the fracture toughness increases with increasing cobalt content. As the carbon to tungsten ration increases, the hardness increase, but the fracture toughness decreases. The maximum values for the fracture toughness and hardness are 15.1 (at 20vol.%Co, W:C=1:1), and 1928 (at 5vol.%Co, W:C=1:1.3), respectively. Therefore we concluded that the HFIHCS method. which can produce WC-xvol.%Co within 1 minute in one step is superior to conventional ones.