The transcription factor POU5F1, also known as OCT4 plays critical roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is important to establish an OCT4 promoter region-based reporter system to study pluripotency. However, there is still a lack of information about the porcine OCT4 upstream region. To improve our understanding of the porcine OCT4 regulatory region, we identified conserved regions in the porcine OCT4 promoter upstream region by sequence-based comparative analysis using various mammalian genome sequences. The similarity of nucleotide sequences in the 5' upstream region was low among mammalian species. However, the OCT4 promoter and four regulatory regions, including distal and proximal enhancer elements, had high similarity. The putative transcription factor binding sites in the Oct4 5' upstream region nucleotide sequences from mice and pigs also differed. Some of these genes are related to pluripotency, and further research will allow us to better understand the differences in species-specific pluripotency. Next, a functional analysis of the porcine OCT4 promoter region was conducted. Luciferase reporter assay results indicated that the porcine OCT4 distal enhancer and proximal enhancer were highly activated in mouse embryonic stem cells and embryonic carcinoma cells, respectively. Similar to OCT4 upstream-based reporter systems derived from other species, the porcine OCT4 upstream region-based reporter constructs showed exclusive expression patterns depending on the state of pluripotency. This work provides basic information about the porcine OCT4 upstream region and various porcine OCT4 fluorescence reporter constructs, which can be applied to study species-specific pluripotency in early embryo development and the establishment of embryonic stem cells in pigs. This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (NRF-2017R1D1A1B03032256).

Sonic hedgehog (Shh) signaling pathway plays a key role in the development of various vertebrate embryos and remains important in adults. Although Shh signaling pathway has widely been studied in post-implantation stage embryos, only few studies are reported about pre-implantation stage embryos. To investigate the effect of Shh on pre-implantation stage embryos, cyclopamine and purmorphamine were treated to embryos in culture. Cyclopamine acts as an antagonist of the hedgehog signaling because it has a high affinity to Smoothened, a key part of the hedgehog signaling pathway. On the other hand, purmorphamine activate Smoothened and acts as a Shh signaling agonist. The oocytes were collected after superovulation and parthenogenetically activated in Chatot, Ziomek, and Bavister medium (CZB) including 10 mM strontium for 5 hr. The activated oocytes were cultured in potassium simplex optimized medium (KSOM), KSOM with 5 uM of cyclopamine, KSOM with 1 uM of purmorphamine, or KSOM with both 5 uM of cyclopamine and 1 uM of purmorphamine. After 5.5 days in culture, there was no significant difference in blastocyst development among the four experimental groups. However, the hatching rate was increased in the groups containing purmorphamine, and the blastocysts of the purmorphamine-containing groups had higher total cell number than those of other two groups when the cells were counted after Hoechst33342 staining. Quantitative real-time PCR (qRT-PCR) shows the difference of gene expression level which are related to epithelial-mesenchymal transition (EMT). Taken together, this study suggests that the increase of Shh has an effect on the increases of EMT-related genes and hatching rate of pre-implantation stage embryos, and this may improve implantation subsequently.

Caspases are a family of cysteine protease enzymes composed of more than 10 members that play essential roles in apoptosis and inflammation. It has been reported that caspases play a critical role in regulating apoptosis at the maternal-conceptus interface in many species. However, the expression and regulation of caspases have not been determined in the endometrium in pigs. Therefore, we analyzed the expression, localization, and regulation of caspases in the endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy. The expression of CASP6, CASP7, and CASP8 during the estrous cycle and CASP3, CASP6, CASP7, and CASP10 changed during pregnancy. Levels of CASP3 mRNA in the endometrium were higher on Day 12 of pregnancy than the estrous cycle and levels of CASP7 mRNAs were highest on Day 15 of estrous cycle and pregnancy. Immunohistochemistry analysis showed that CASP3 protein was localized to endometrial epithelial cells on Days 12 and 15 the estrous cycle and pregnancy, but cleaved CASP3 was localized only to luminal epithelial (LE) cells on Days 12 and 15 of pregnancy in the endometrium. CASP7 protein was localized to endometrial LE cells only on Day 15 of pregnancy. CASP3, CASP6, CASP7, CASP8, and CASP10 mRNAs were detectable in conceptus on D12 and D15 of pregnancy, and chorioallantoic tissues expressed CASP3, CASP6, CASP7, CASP8, CASP8, and CASP10 with increasing levels toward term pregnancy, except CASP3 mRNA. The effect of steroid hormones and interleukin-1βß (IL1B) on CASP3 expression and the effect of interferon-γ(IFNG) on CASP7 expression was determined by endometrial explant cultures and we found that CASP3 expression was increased by IL1B and CASP7 expression was increased by IFNG in a dose-dependent manner. These results showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy in a stage- and/or pregnancy-specific dependent manner and some caspases were regulated by IL1B or IFNG in the endometrial tissues, suggesting that caspase may play an important role in regulating apoptosis for the establishment and maintenance of pregnancy at the maternal-conceptus interface in pigs.

Successful cryopreservation of bovine oocytes is a very important technology for research and commercial applications. However, the survival and development rate of vitrified-thawed oocytes is lower than non-vitrified oocytes. Hydroxypropyl Cellulose supplementation (HPCs) has extremely high viscosity, which permits transitions to a glassy state at low temperatures. This characteristics of HPCs have been reported to help the survival of human oocytes. In this study, we investigated the survival rate, fertilization rate and ROS levels to confirm the effect of cryoprotectant solutions with HPC for oocyte vitrification in bovine. For vitrification, bovine MII oocytes were pretreated with EG10 (added 0, 10, 50 and 100 ㎍/㎖ HPC) for 5 min, exposed to EG30 (added 0, 10, 50 and 100 ㎍/㎖ HPC) for 30 sec, and then directly plunged into LN2. Thawing was taken by 4-step procedures [1 M sucrose and 10% FBS added D-PBS (SFD) -> 0.5 M SFD -> 0.25 M SFD -> 0.125 M SFD] for 1 min, respectively. After thawing, oocytes were washed with TL-HEPES, incubated in a droplet of previous cultured IVM medium for 1 h to recover. IVF drop (44 ㎕) contained 10 vitrified-thawed oocytes with sperm concentration of 1 × 106 cells ㎖, and then 2 ㎕ heparin and 2 ㎕ PHE were added. At 2 days after IVF, cleaved embryos were cultured in CR1aa + 3 mg/mL FAF-BSA for 48 h and cultured in CR1aa + 10% FBS for 4 days. In the results, in vitro survival rate of bovine vitrified-thawed MII oocyte was significantly higher in 50 (85.5%) and 100 ㎍/㎖ (80.2%) HPC groups than 0 (71.2%) and 10 ㎍/㎖ (71.3%) groups (p<0.05). The ROS level was lower in 50 ㎍/㎖ HPC group than in control group. After in vitro fertilization, cleavage rate and blastocyst development rate were not significantly different among treatment groups. Therefore, these results indicated that HPC treatment has a positive effect on the survival of vitrified-thawed bovine oocytes.

To improve survival rates of vitrified pig oocytes, the treatment of cytoskeletal stabilizer on an appropriate time is one of the possible approaches. However, the exact treatment timing and effect of cytoskeletal stabilizer such as cytochalasin B (CB) is not well known during oocyte vitrification procedures. Thus, the present study was conducted to determine optimal treatment timing of CB during vitrification and warming procedures. In experiment 1, the survival rates of the post-warming pig oocytes were analyzed by fluorescein diacetate (FDA) assay with 4 classifications. In results, post-warming oocytes showed significantly (p<0.05) decreased number of alive oocytes (31.8% vs. 86.4%) compared to fresh control. In detail, the significant difference (p<0.05) was found only in strong fluorescence (18.2% vs. 70.5%) not in intermediate fluorescence groups (13.6% vs. 15.9%). In experiment 2, CB was treated before (CB-Vitri) and after (Vitri-CB) vitrification. In results, group of Vitri-CB showed significantly (p<0.05) higher (91.6%) survival rates compared to group of CB-Vitri (83.7%), significantly (p<0.05) and comparable with group of Vitri Control (88.7%) by morphological inspection. In FDA assay results, group of Vitri-CB showed significantly (p<0.05) higher (44.2%) survival rates compared to groups of CB-Vitri (36.7%) and Vitri Control (35.1%). In conclusion, the increased survival rates of post-warming pig oocyte treated with Vitri-CB method are firstly described here. The main finding of present study is that the CB treatment during recovery could be helpful to refresh the post-warming pig oocyte resulting its improved survival rates.

Polo-like kinase 1 (Plk1) has multiple roles in somatic cell and mammalian oocyte division. In mice, Plk1 distributes to the centromeres from prophase to anaphase and compose spindle apparatus in mitosis stages. Somatic cell nuclear transfer (SCNT) has diverse advantages. However, low cloning efficiency of SCNT procedure causes difficulty to application. The causes of this low efficiency are still unclear. However, they are attributed to the cumulative results of several biological and technical factors. In this study, Plk1, a biological factor, was investigated. B6D2F1 mice (7 weeks old) were superovulated with 10 IU of pregnant mare’s serum gonadotropin and 9 U of human chorionic gonadotropin (HCG) 48 hr later. The oocytes were collected 14 hr after HCG injection and cultured on potassium simplex optimized medium. The BI2536, Plk1-specific inhibitor, was used to understand the influence of Plk1. Also, the embryos were assessed by immunofluorescence. All BI2536-treated embryos failed to the first mitotic division. It showed Plk1 has a critical role in the first mitotic division of the mouse embryo. Moreover, there were significant differences between the control and SCNT embryos in the patterns of Plk1. All SCNT embryos which failed 2-cell development presented incorrect positioning and low expression of Plk1. On the other hand, the control embryos which failed to 2-cell division showed only low expression of Plk1. Taken together, this results demonstrate that Plk1 is critical for successful mitotic division of mouse embryos. Also, correct localization of Plk1 has crucial effect in the development of murine SCNT embryos.

본 연구에서는 신선편이 양배추 내 Bacillus cereus의 최 적 증균 온도를 선정하고 증균배양액에 소수성필터를 적용하여 multiplex PCR의 검출률을 확인하였다. B. cereus 증균온도는 B. cereus 균주 5 개를 혼합하여 1 Log CFU/ mL이 되도록 증균배지에 접종하고 30oC, 37oC, 42oC에서 증균한 뒤 3시간 간격으로 MYP agar에 도말한 후 계수하여 선정하였다. 소수성필터 미적용 그룹은 B. cereus 균 주 5 개를 혼합하여 신선편이 양배추에 접종한 뒤 최적 증균온도에서 증균하였으며, 증균배양액을 가열하여 DNA 를 추출한 뒤 multiplex PCR을 진행하였다. 소수성필터 적용 그룹은 증균배양액을 소수성 필터에 적용하고 필터에 있는 균을 멸균증류수로 현탁한 뒤 가열하여 추출된 DNA 로 multiplex PCR을 진행하였다. 증균온도 확인 결과, 6시간 증균 시 42oC에서 증균된 샘플(5.4 ± 0.3 Log CFU/mL) 과 30oC에서 증균된 샘플(4.6 ± 0.6 Log CFU/mL) 간 유의 차가 확인되었다(p < 0.05). 소수성필터 적용 유무에 따른 multiplex PCR 결과, 1 Log CFU/g 접종된 샘플의 검출률이 소수성 필터 적용 전 60%(3/5)에서 100%(5/5)로 향상 되었다. 2 Log CFU/g 접종 샘플은 소수성필터 적용 전 80%(4/5)에서 소수성 필터 적용 후 100%(5/5)로 검출률이 증가하였으나, 3 Log CFU/g 접종 샘플은 소수성 필터 적용 전후 모두 100%(5/5)로 확인되었다. 이상의 결과를 통해 신선편이 양배추 내 B. cereus 검출 시 증균배양액에 소수성필터를 적용하고 multiplex PCR을 적용했을 때 신속하고 효율적인 검출이 가능할 것으로 판단된다.

We present the first results of the invariant point (IVP) coordinates of the KVN Ulsan and Tamna radio telescopes. To determine the IVP coordinates in the geocentric frame (ITRF2014), a coordinate transformation method from the local frame, in which it is possible to survey using the optical instrument, to the geocentric frame was adopted. The least-square circles are fitted in three dimensions using the Gauss-Newton method to determine the azimuth and elevation axes in the local frame. The IVP in the local frame is defined as the mean value of the intersection points of the azimuth axis and the orthogonal vector between the azimuth and elevation axes. The geocentric coordinates of the IVP are determined by obtaining the seven transformation parameters between the local frame and the east-north-up (ENU) geodetic frame. The axis-offset between the azimuth and elevation axes is also estimated. To validate the results, the variation of coordinates of the GNSS station installed at KVN Ulsan was compared to the movement of the IVP coordinates over 9 months, showing good agreement in both magnitude and direction. This result will provide an important basis for geodetic and astrometric applications.

In the current study, a total of 102 common Todarodes pacificus squid caught in the East Sea were investigated for parasitological research. The results revealed that 33 (32.35%) out of 102 squid were infected by Nybelinia surmenicola, the mean intensity was 5.58 parasites per squid, and the maximum abundance was 11. Morphological analysis using a field emission scanning electron microscope showed the characteristic features of N. surmenicola. Molecular identification based on the 28S rRNA gene confirmed the isolated parasite as N. surmenicola, while phylogenetic analysis revealed that N. surmenicola isolated in this study was clustered with N. surmenicola isolated from Japan. This is the first report of phylogenetic characterization of N. surmenicola isolated from Korea.

Epizootic HPAIV, H5N6, and H5N8 infections produced severe loss in poultry and wild birds in the Republic of Korea from 2016 to 2017. But pathological lesions and antigen distribution of the novel HPAIV H5N6 clade 2.3.4.4 in natural cases have been rarely reported. Herein, we describe the pathological lesions and antigen localization in chickens (layer and Korean native), ducks, and Japanese quail naturally infected by HPAIV H5N6. Grossly, severe reddening, swelling, and some necrotic foci, which were similar to septicemia or viremia, were observed in skin and many visceral organs including trachea, lung, liver, spleen, and pancreas. Histopathologically, pulmonary congestion and edema, as well as necrotizing hepatitis, splenitis, pancreatitis, myocarditis, and encephalitis were observed. Immunohistochemically, numerous HPAIV antigens were detected in necrotic parenchymal cells and in blood vessels of the respiratory, lymphoid, digestive, urinary, nervous, and cardiovascular systems. The results indicate that HPAIV H5N6 spread to the entire body via blood and caused severe damage throughout the entire body. The HPAI H5N6 clade 2.3.4.4 virus was isolated from samples of all four cases.

This study was conducted to improve growth performance of Hanwoo heifer and to produce high quality of meat with dietary means during growing and early fattening period. Particularly, additional energy diet to relieve estrus stress was main purpose in this study. The results of in vitro rumen fermentation indicated that there was no negative effect by additional energy diet as treatments. In the feeding trial, twenty Hanwoo heifers(average 10 months age) were allocated and distributed into two treatments in randomized block design based on body weight. There were three growth stages such as growing, early fattening and late fattening periods in this feeding program, respectively. In growing stage, there were two treatments consisting of only total mixed ration(TMR) as a control and TMR with additional energy treatment. The experimental diets were fed twice a day, and water and mineral were freely accessed. In additional energy treatment, 500 g of concentrate diet was fed daily to relieve estrus stress due to obese with high energy intake. Not outstandingly differences were found across the treatments during entiretrial period. While, unexpectedly greater feed conversion ratio in treatment compared to the control was found during late fattening period. It seems that the blood cortisol decreased with addition energy supplementation compared with the control during trail period. Carcass characteristics including carcass weight, back fat thickness, marbling score, meat color, fat color, maturity and texture were not significantly different each other. Rib-eye area, however, was greater in the control compared to the treatment(p<0.05). In addition, it appears that yield index was tended to be greater in the control. In conclusion, it is suggested that additional energy supplementation to Hanwoo heifer could get a potential in improving meat quality and relieving estrus stress.

First Mover Advantage is already well known. It is when a company gains a position in a certain market or industry, or when it establishes a strong entry barriers through a distribution channel or a monopoly of resources. It is a concept that has been attracting attention for a long time in marketing and strategy. However, although it is possible for the starter to enjoy these various benefits, it is also true that there is a corresponding price. Therefore, the risks and costs that the starter may bear, and thus the relative benefits enjoyed by the latter, can be significant. Late Mover Advantage and so on. The fact that latecomers can enjoy a variety of benefits as well as the profits of the starters is an important consideration that must be taken into account by many companies considering entry into the market. In general, there is a very high risk of overinvestment in technology and market uncertainty. For example, China has skipped wired networks and went wireless, and many African countries have skipped wired communications and built infrastructure for wireless communications. In other words, companies that hastened to invest in fixed-line facilities in order to preoccupy the African telecom market are in a state of failure rather than expecting the interests of the starters. Another thing is that the starter has to bear more risks and costs than the latter, such as the uncertainty of demand, the risk of changing consumer preferences, and the cost of training new consumers. Also, because imitation is generally less costly than development, a latecomer entering through imitation may be in a better position if patents or other technical defenses are not available. Especially, if latecomers have excellent management ability and financial power such as excellent marketing ability, it is relatively easy to catch up with the first candidate.

PURPOSES: Using recycled asphalt materials (called Reclaimed Asphalt Pavement: RAP) from existing asphalt pavement layers in newly constructed asphalt pavement is an essential option not only for lowering the construction budget but also for mitigating environmental pollution for society. For this reason, many pavement agencies in South Korea, the USA, and Canada have observed the effect of RAP on conventional asphalt pavement to evaluate and set proper material specifications and addable amounts. In this paper, effect of recyclable material on low-temperature performance of asphalt materials was investigated with two different mechanical tests. Among the recyclable material sources, RAP and Taconite Aggregate (TA), which is mainly produced in northern Minnesota (USA), were considered. METHODS : To evaluate the low-temperature mechanical performance of a RAP mixture, two different experimental tests (In-Direct Tensile (IDT) low temperature creep test and Semi-Circular Bending (SCB) test) were considered. The mechanical parameters creep-stiffness, relaxation modulus, fracture energy, and fracture toughness were computed then compared. RESULTS: More brittle characteristics were observed with RAP-added asphalt mixtures compared to the conventional asphalt mixtures, as expected. However, the differences of computed mechanical performances were not significantly distinct for RAP mixtures compared to conventional mixtures when the RAP proportion was around 20%, and with the addition of TA up to 20%. CONCLUSIONS : It can be concluded that up to 20% of RAP addition (along with TA up to 20%) in a virgin asphalt mixture does not provide significant performance reduction. This addable proportion can be viewed as a successful minimum level when considering the addition of RAP to hot-mix asphalt (HMA). Moreover, applying TA with RAP could offer a successful alternative for asphalt recycling and the materials industry.