• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.1
• /
• pp.1-11
• /
• 1997

It is well known that the zona pellucidae of mouse oocytes become "hardened" when they are allowed to mature in vitro in the absence of serum components. To see if oocytes already undergone meiotic resumption in vivo exhibit similar zona hardening, hardening of ZP of cumulus-enclosed oocytes(CEOs) was examined after culture in vitro since their release from follicles various hours after hCG injection. When CEOs matured in vivo for 3h or longer were subjected to culture in vitro for 14h with BSA alone, zona hardening was significantly reduced compared to those cultured in vitro from the begining of maturation. However, when CEOs matured in vivo for 5h were freed from cumulus cells and then cultured in vitro with BSA alone, little reduction of zona hardening was observed. Preincubation of CEOs for 5h with fetuin, one of the well known inhibitor of in vitro zone hardening, did not prevent zona hardening during its subsequent culture of CEOs for 14h without fetuin. However, when CEOs precultured with both fetuin and PMSG for 5h and then further cultured with BSA alone for 14h, zona hardening was dramatically reduced. Under these conditions, the expansion of cumulus cell was observed. In addition, CEOs cultured with both BSA and dbcAMP to prevent their meiotic resumption showed a significant increase of zona hardening. Whether the observed zona hardening was correlated with the conversion of ZP2 to $ZP2_{f}$ was examined. Zona pellucida, isolated from CEOs matured for 5h in vivo and then further cultured with BSA alone was subjected to SDS-PAGE. Most of ZP2 molecules from these CEOs did not undergo conversion from ZP2 to $ZP2_{f}$. From these results, it is concluded that CEOs undergone meiotic resumption in vivo do not exhibit zona hardening when they were subsequently cultured in vitro without serum components. It appears that cumulus cells play an important role in this phenomenon.

• Applied Biological Chemistry
• /
• v.19 no.1
• /
• pp.31-35
• /
• 1976

Fifteen naked barley cultivars including radiation breeding lines from three places were analized for crude protein, carbohydrates, P, K, Ca, Mg and tested protein by dye binding method and biuret method. Their content and simple correlation analyses among them were as follows. 1. Protein content was 7.67 for average (max. 10.3 in Baegdong, min. 6.0 in Bangju) that was lower than in milled barley and had significant (at p=0.01) correlation with dye binding capacity (r=0.769) and biuret absorbance (r=0.616). 2. Protein content also had significant correlation with $P_2O_5$(r=0.607, p=0.01) and with MgO(r=0.498, p=0.05). 3. There was great difference in protein content among radiation breeding lines(max. 8.40, min. 6.75%). 4. Naked barley appeared to be lower in carbohydrate content but higher in crude ash to compare with milled barley. 5. There was significant correlation(r=0.560, p=0.01) between Ca and K, indicating competition in uptake or translocation to grain. 6. Carbohydrate content showed the highest negative correlation with protein content but it was not significant. 7. The low protein variety (Bangju) showed higher yield than the high protein one (Baegdong) both with (16%) and without (48%) fertilizers.

• Proceedings of the KWS Conference
• /
• 2009.11a
• /
• pp.72-72
• /
• 2009

전자부품의 내부접속 및 파워반도체의 다이본딩과 같은 1차실장에는 고온환경에서의 사용과 2차실장에서의 재용융방지를 위해 높은 액상선온도 및 고상선온도를 필요로 하여, Pb-5wt%Sn, Pb-2.5wt%Ag로 대표되는 납성분 85%이상의 고온솔더가 널리 사용되고 있다. 생태계와 인체에 대한 납의 유해성이 보고된 이래, 무연솔더에 대한 연구가 활발히 진행되어 왔으나, Sn-Ag-Cu계로 대표되는 Sn계 합금으로 대체 중인 중온용 솔더와는 달리, 고온용 솔더에 대해서는 대체합금에 대한 연구가 미흡한 실정이다. 대체재의 부재로 인해 기존의 납을 다량함유한 솔더로 1차실장이 지속됨으로서, 2차실장의 무연화에도 불구하고 전자부품 및 기기의 재활용에 큰 어려움을 겪고 있다. 지금까지 고온용 무연솔더로서는 융점에 근거해 Au-(Sn, Ge, Si)계, Bi-Ag계, Zn-(Al, Sn)계의 극히 제한된 합금계만이 보고되어 왔다. Au계 솔더는 현재 플럭스를 사용하지 않는 광학, 디스플레이 분야 등 고부가가치 공정에 사용되고 있으나, 합금가격이 매우 비싸며 가공성이 나빠 대체재료로서는 적합하지 않다. Bi-Ag계 솔더 또한 취성합금으로 와이어 및 박판으로 가공하는데 어려움이 크며, 솔더로서 중요한 특성중 하나인 전기전도도 및 열전도도가 나쁜 편이다. 이에 비해, Zn계 합금은 비교적 낮은 합금가격, 적절한 가공성과 뛰어난 인장강도, 우수한 전기전도도 및 열전도도를 지녀, 고온용솔더 대체재료의 유력한 후보로 생각된다.이전 연구에서, 필자의 연구그룹은 Zn-Sn계 합금을 고온용 무연솔더로서 제안한 바 있다. Zn-Sn계 합금은 충분히 높은 융점과 함께, 금속간화합물이 없는 미세조직, 우수한 기계적 특성, 높은 전기전도도 및 열전도도 등의 장점을 나타내었다. 본 연구에서는 기초합금특성상 고온솔더로서 다양한 장점을 지닌 Zn-30wt%Sn합금을 고온용 솔더의 대표적인 적용의 하나인 다이본딩에 적용하여, 접합부의 강도 및 미세조직, 열피로 신뢰성에 대해 분석을 함으로서 실제 공정에의 적용가능성에 대해 검토하였다. Zn-30wt%Sn을 이용해 Au/TiN(Titanium nitride) 코팅한 Si다이를 AlN-DBC(aluminum nitride-direct bonded copper)기판에 접합한 결과, 양측에 완전히 젖은 기공이 없는 양호한 다이접합부를 얻었으며, 솔더내부에는 금속간화합물을 형성하지 않았다. Si다이와의 계면에는 TiN만이 존재하였으며, Cu와의 계면에는 Cu로부터 $Cu_5Zn_8,\;CuZn_5$의 반응층을 형성하였다. 온도사이클시험을 통한 열피로특성평가에서, Zn-30wt%Sn를 이용한 다이접합부는 1500사이클 지점에서 Cu와 Cu-Zn금속간화합물의 사이에서 피로균열이 형성되며, 접합강도가 크게 감소하였다. 열피로특성 향상을 위해 Cu표면에 TiN코팅을 하여 Zn-30wt%Sn 솔더로 다이접합한 결과, Si다이와 기판 양측에 TiN만으로 구성된 계면을 형성하였으며, TEM관찰을 통해 Zn-30wt%Sn과 극히 미세한 접합계면이 형성하고 있음을 확인하였다. Zn-wt%30Sn솔더와 TiN층의 병용으로 2000사이클까지 미세조직의 변화 및 강도저하가 없는 극히 안정된 고신뢰성의 다이접합부를 얻을 수가 있었다.

• Reproductive and Developmental Biology
• /
• v.35 no.3
• /
• pp.307-311
• /
• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG ($33.1{\pm}9.6$ vs $21.7{\pm}8.3%$). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP-treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The mRNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

• Journal of Embryo Transfer
• /
• v.19 no.2
• /
• pp.173-183
• /
• 2004

This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.