• Reproductive and Developmental Biology
• /
• v.35 no.3
• /
• pp.385-390
• /
• 2011

The purpose of this study was undertaken to evaluate of cryopreservation efficiency in ${\alpha}$ 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.

• Reproductive and Developmental Biology
• /
• v.35 no.3
• /
• pp.197-201
• /
• 2011

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

• Reproductive and Developmental Biology
• /
• v.36 no.3
• /
• pp.231-235
• /
• 2012

The objective of this study was to investigate the comparison of transferable embryos and pregnancy rate between Hanwoo and Chickso. The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on Hanwoo 78%, and Chickso 85%, respectively. The mean number of total embryos are each $14.76{\pm}2.16$ and $6.23{\pm}1.07$. So the mean number of transferable embryos are each $10.94{\pm}1.91$ and $4.58{\pm}1.05$. In addition, the mean number of total Hanwoo embryo from <10 and $10{\leq}$ of corpora luteum was $0.50{\pm}0.50$, $11.56{\pm}1.92$, respectively. In case of Chickso, The mean number of transferable embryo from <10 and $10{\leq}$ of CL was $2.75{\pm}1.39$, $6.00{\pm}1.00$, respectively. The pregnancy rates were Hanwoo 40%, and Chickso 37% following transfer of fresh embryos produced in vivo. Also, the pregnancy rates of Chickso 60% were significantly greater (p<0.05) than the Hanwoo 42.48% following transfer of following transfer of frozen embryos, respectively. In conclusion, these results suggest that Chickso may be effectively used for transferable embryos production in Hanwoo. Although the transferable embryos number was not enough, it seems the Chickso greatly affect pregnancy rate. The results indicated that the possibility of transferable embryos from Chickso for embryo transfer could be confirmed in this study. Based on the present findings, it was suggested that it is very important to evaluate in vivo embryo production and pregnancy rate of embryo transfer following superovulation for effective Hanwoo and Chickso production.

• Journal of the Korean Graphic Arts Communication Society
• /
• v.31 no.1
• /
• pp.39-49
• /
• 2013

Nowadays the printing market has shift from 2D(two dimension) industry to 3D(three dimension) industry, along with IT. With rapidly changing market, it's true that the effort to develop the printing materials and upgrade the printing qualities is produced actively in the country. Particularly, the differences in printing materials can often determine the quality of printings in the printability. In the case of the printability of offset, 9 items(paper, ink, order of printing, ink supply, blowing-up halftone, grey balance, key color(base color), overlap efficiency, quality at the halftone dot reproduction etc.) are usually considered as the important factors and used to estimate the printability at abroad. However, in domestic, ink-jet paper is not used for the printing/publishing by ink-jet printing for digital, but done mainly for offset printing. Because the paper of gloss coated type, which is the ink-jet paper, has high price, it's too hard to manufacture massively printing/publishing. Also, in the case of the paper of uncoated type, in spite of its reasonable price rather than the former, the type of uncoated one also has demerit that the ink-jet paper has a clear difference between the front and the back. Therefore, it's tough reality that Ink Jet paper is hardly taken advantage of as the printing/publishing. Thus, what is done in the first to print and/or publish on ink-jet paper is that the front and the back must be printed evenly, and appropriate to the printability of ink-jet. Although there're some kinds of ink-jet paper in the country, it has not been produced in true domestic production yet. In addition, thanks to the brightness of fluorescence brightening agent, the foreign product can offer the good-qualified one-time printing. But when it comes to the printing/publishing, there're difficulties that it may cause the Ocular Fatigue because of long-term exposure of the eyes to text. This research tries to find out the optimum ink-jet paper in domestic for the printing/publishing by studying on the printability of ink-jet on digital web-printing. And it also tries to look for the paper's brightness which can be reduce the possibility to cause the consumers' Ocular Fatigue.

• Reproductive and Developmental Biology
• /
• v.30 no.2
• /
• pp.125-133
• /
• 2006

This study was conducted to detect the apoptosis incidence in blastocysts and to compare the abundance of Bax, Bcl2L1, VEGF and FGFR2 in in vitro fertilized (IVF), parthenogenetic (PAT) and nuclear transfer (NT) embryos. Oocytes matured for 40 hr were enucleated and reconstructed with confluenced fetal fibroblasts (FFs) derived from a ${\sim}45$ day fetus. Reconstructed eggs were then fused with 2 DC pulses (2.0 kV/cm, $30{\mu}sec$) and cultured with $7.5{\mu}g/ml$ cytochalasin B for 3 hr. Parthenotes (PAT) were produced with the same electric strength and culture for NT eggs. The embryos were cultured in NCSU-23 medium at $39^{\circ}C,\;5%\;CO_2,\;5%\l;O_2$ in air. In 3 runs, set of 10 embryos at the 4-cell to blastocyst stages were used to extract total RNA for analyzing the gene expression patterns of pro-apoptotic (Bax), anti-apoptotic (Bcl2L1), vasculogenesis (VEGF), implantation (FGFR2III) using real-time quantitative PCR. Cleavage and blastocyst rates were significantly higher (P<0.05) in IVF and PAT ($79.3{\pm}8.5\;and\;25.5{\pm}6.1,\;and\;85.0{\pm}6.4\;and\;38.6{\pm}5.5$, respectively)than NT counterparts ($65.1{\pm}5.2\;and\;15.6{\pm}3.0$, respectively). Significantly higher (P<0.05) total cells were observed in IVF controls and PAT ($34.7{\pm}5.8\;and\;38.1{\pm}4.1$) than NT embryos ($24.8{\pm}3.2$). Apoptosis index was significantly lower (P<0.05) in IVF than NT embryos. The Relative abundances (RA) of Bax and VEGF were significantly higher (P<0.05) at blastocyst stage in NT than IVF control. The RA of Bcl2L1 and FGFR2III were significantly higher (P<0.05) at blastocyst stage in IVF than NT. The present study observed the abnormal gene expressions in NT embryos at various developmental stages, suggesting certain clues to find out the cause of the low efficiency of NT to term.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.9-9
• /
• 2001

내분비계 장애물질 중 관찰물질로 지정된 bisphenol A(BPA)의 투여가 생쥐의 번식 효율에 미치는 영향을 규명하기 위하여 웅성 ICR 생쥐에 무처리구, corn oil, 0.05, 0.5 및 5.0mg BPA/kg b.w의 BPA을 3일 간격으로 5회 투여했으며, 자성 ICR 생쥐도 웅성생쥐와 동일하게 3일 간격으로 5회 투여한 후, 웅성은 투여후 2일, 자성은 7일후에 모든검사를 실시하였다. BPA의 투여가 웅성 생쥐의 번식기관무게에 미치는 영향을 조사한 결과는 체중, 정소상체, 정낭선 및 응고선의 무게는 투여구간에 커다란 차이가 없었으며(P>0.05), 정소의 무게는 5.0mg 투여구가 여타구보다 다소 낮은 무게를 나타냈다. BPA의 투여가 웅성 생쥐의 정액성상에 미치는 영향을 조사한 결과, 생존율과 유효 정자수는 각 투여구간에 커다란 차이가 없었으나 총정자수는 각각 35.50, 35.73, 36.33, 33.13 및 29.28$\times$$10^{7}$ 정자/$m\ell$로서 5.0mg 투여구가 여타구보다 통계적으로 유의하게 낮은 결과을 나타냈다(P<0.05). 기형율은 5.0mg 투여구가 23.15%로서 무처리구(18.13%), 대조구(18.58%) 및 0.05mg 투여구(19.68%)보다 통계적으로 유의하게 높은 기형율을 나타냈다(P<0.05). 웅성 생쥐에 BPA의 투여가 혈액의 일반성분에 미치는 영향을 조사한 결과, RBC, Hb, HT, MCV, MCH 및 MCHC는 각 투여구간에 차이가 없었으며, WBC는 BPA 투여구가 무처리구 및 대조구보다 다소 낮은 수치를 나타냈으며, PLT는 BPA 투여구가 무처리구 및 대조구보다 높은 수치를 나타냈다. BPA의 투여가 자성 생쥐의 번식기관무게에 미치는 영향을 조사한 결과, 체중은 투여구간에 커다란 차이가 없었으며 자궁의 무게는 BPA 투여구가 대조구에 비해 다소 논은 무게를 나타냈다. 오른쪽 난소의 무게는 0.5mg 투여구와 5.0mg 투여구가 각각 0.0059 및 0.0061g으로 대조구(0.0081g)와 0.05mg 투여구(0.0071g)보다 낮은 무게를 나타냈으며, 왼쪽 난소의 무게는 각각 0.0076, 0.0076, 0.0058 및 0.0061g으로 0.5mg 투여구가 대조구와 0.05mg 투여구보다 낮은 무게를 나타냈다(P<0.05). BPA의 투여가 자성 생쥐의 혈구화학치에 미치는 영향을 조사한 결과, RBC, Hb, HT, MCV, MCH 및 MCHC는 각 투여구간에 차이가 없었으며, WBC는 BPA 투여구가 대조구보다 다소 높은 수치를 나타냈다. PLT는 0.5mg 투여구와 5.0mg 투여구가 대조구보다 다소 높은 수치를 나타냈다. 정소, 난소 및 자궁의 조직, 병리학적인 검사에서는 모든 투여구간에 커다란 차이가 없었다.

• Reproductive and Developmental Biology
• /
• v.33 no.1
• /
• pp.35-40
• /
• 2009

The objectives of this study was to evaluate the efficiency of the bacteria eliminated sperm by percoll gradient method on sperm quality and embryo cleavage in vitro in pig. The semen of miniature pig collected by gloved-hand method pre-warmed ($37^{\circ}C$) in thermos bottle, and separated by 65% percoll. Analysis of sperm ability was estimated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) and the abnormality. Also, fertility of sperm was monitored with cleavage rate of embryo after IVF using separated and un-separated sperm by percoll. The result, viability of separated sperm was significantly(p<0.05) higher($83.6{\pm}$2.0 vs $59.0{\pm}4.4%$) than un-separated sperm. The results of CTC analysis showed the percentage of F- and B-patterned separated sperm was higher in separated that than un-separated sperm. On the contrary, the percentage of AR-patterned form unseparaed sperm was significantly(p<0.05) higher($13.6{\pm}0.8$ vs $8.1{\pm}0.6%$) than separated sperm. Also, abnormality of un-separated sperm was significantly(p<0.05) higher($2.2{\pm}0.4$ vs $16.8{\pm}2.8%$) than separated sperm. However, the cleavage rates of embryo using separated sperm by percoll and un-separated sperm had not significantly difference on 2 cell stage(9.25 vs 11.88%), 4 cell stage(26.76 vs 24.51%) and >4 cell stage(63.99 vs 63.61%) at 48h of IVF. Therefore, the sperm separated by percoll method showed improvement in sperm quality than un-separated sperm in miniature pig.

• Reproductive and Developmental Biology
• /
• v.33 no.2
• /
• pp.107-111
• /
• 2009

The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.18 no.2
• /
• pp.465-471
• /
• 2017

This study examined the effects of neoliberal educational policy on the appointment system of teachers to assess the impacts of this neoliberal education policy. Neoliberal education policy has led to increased private education cost, and strengthened the function of social stratum reproduction in the education system despite increasing the efficiency and flexibility. Since contracted employment was applied to the educational field, the number of regular teachers began to decrease, while that of contracted teachers increased. For example, the number of contracted teachers was 300 in the 1990, which increased to 42,042 in 2015. Neoliberal education policy contributed to teacher's quantitative demand; however, the increase in the number of contracted teachers is ultimately degrading the education quality. In an education market, the issues of balancing between supply and demand often occur. Therefore, it is necessary to strongly control the number of contracted teachers by limiting their contract period. Furthermore, an institutional device that can convert a contracted teacher to a regular one must be established.

• Korean Journal of Ecology and Environment
• /
• v.53 no.3
• /
• pp.221-228
• /
• 2020

We studied the development of the endangered gold-spotted pond frog, Pelophylax chosenicus eggs and analyzed growth and survival rate of the tadpoles while feeding different feed to obtain the basic data on the its biological traits and reproduction. We observed the fertilized eggs of gold-spotted pond frog that naturally spawned in a breeding tank using a microscope. Fertilized eggs were sphere shape surrounded by colloid layers, clearly distinct with animal and plant poles. The fertilized eggs hatched 31 hours after fertilization and operculum complete at 175 hours after fertilization at water temperature of 27.0±1.0℃. As a result of growth and survival rate by different feed from 35 days after mouth open stage, vegetable feed (VF) group and tetra-min (T) group were significantly higher average weight gain rate (WG)(5843.9% and 5736.3%, respectively) and average specific growth rate (SGR) (11.67% and 11.62, respectively) compared with other groups (WG: 641.8~4625.2%, SGR: 5.72~11.01%). The vegetable (V) group showed the lowest growth rate (P<0.05). The average feed efficiency was 177.83% in the VF group, about 17 times higher than the V group. In addition, the average survival rate of the VF and T groups were the high at 97.5% and 100%, respectively, and the V group was the lowest at 32.5%. Therefore, the more efficient feeds for successful breeding is singly with VF or T.