• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.347-356
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• 2020

Gangliosides are glycolipids in which oligosaccharide is combined with sialic acids. Our previous studies have suggested an interplay between ganglioside GD1a/GT1b and meiotic maturation capacity in porcine oocyte maturation. Furthermore, ganglioside GD1a and GT1b are known for its antioxidant activity, but it is still unclear whether possible antioxidant role of GD1a and GT1b is involved in porcine embryos development competence during in vitro culture (IVC). Here, the effects of ganglioside GD1a and GT1b on the embryonic developmental competence during in vitro culture of porcine were investigated. The effects of ganglioside GD1a and GT1b on the expression of ST3GAL2 were confirmed during embryos development (2-cell, 4-cell, 8-cell and blastocyst) using immunofluorescent staining (IF). As a result, the fluorescent expression of ST3GAl2 was higher in embryos at 4-8 cells stage than blastocysts. Blastocyst development rate significantly increased in only 0.1 μM GD1a and GT1b treated groups compared with control group. To investigate the cellular apoptosis, we analyzed TUNEL assay. In case of only 0.1 μM GD1a and GT1b treated groups, the total number of cells in blastocyst compared with control group, but there was no significant difference in the rate of apoptotic cells. We identified the intracellular ROS levels using DCF-DA staining. According to the result, ROS production significantly decreased in blastocysts derived from the 0.1 μM GD1a and GT1b treated groups. These results suggest that ganglioside GD1a and GT1b improve the developmental competence of porcine embryos via reduction of intracellular ROS during preimplantation stage.

• Journal of dental hygiene science
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• v.21 no.4
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• pp.251-259
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• 2021

Background: The purpose of this study was to investigate the effects of commercially available calamansi soju and other alcoholic beverages on the microhardness and erosion of resin restorations. Methods: In this study, we evaluated the effects of Calamansi soju, Chamisul fresh, Cass fresh, and Gancia Moscato D'asti on resin restorations. Jeju Samdasoo and Coca-Cola were used as negative and positive controls, respectively. Specimens to be immersed in the beverages were manufactured using composite resin according to the product instructions. In each group, the surface microhardness was measured using a surface microhardness instrument before and after immersion for 5, 15, 30, and 60 minutes. The pattern of change in the surface of the composite resin was observed under a scanning electron microscope (SEM). Paired t-tests, one-way ANOVA, and repeated measures ANOVA were performed to compare the surface microhardness of the specimens, and the Tukey test was used as a post hoc test. Results: The pH of all beverages except Jeju Samdasoo was <5.5, which is the critical pH that can induce erosion. The difference in surface microhardness of the composite resin before and after immersion for 60 minutes was significant in all groups. In particular, the largest change in surface microhardness was observed in the calamansi soju group. In the SEM analysis, loss of composite resin was observed in all groups except the Jeju Samdasoo group, and rough surfaces with pores of various sizes were observed. Conclusion: In this study, all beverages except Jeju Samdasoo decreased the microhardness of the composite resin surface, and it was confirmed that calamansi soju had the greatest change.

• Korean Journal of Veterinary Research
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• v.63 no.1
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• pp.3.1-3.9
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• 2023

The efficiency of somatic cell nuclear transfer (NT) in pigs is low and requires enhancement. We identified the most efficient method for zona pellucida (ZP) removal and blastomere aggregation in pigs and investigated whether the aggregation of NT and parthenogenetic activation (PA) of blastomeres could reduce embryonic apoptosis and improve the quality of NT-derived embryos by investigating. Embryonic developmental competence after ZP removal using acid Tyrode's solution or protease (pronase E). The embryonic developmental potential of NT-derived blastomeres was also investigated using well-of-the-well or phytohemagglutinin-L. We analyzed apoptosis in aggregate-derived blastocysts. The aggregation rate of protease-treated embryos was lower than that of Tyrode's solution-treated embryos (69.2% vs. 88.3%). No significant difference was observed between phytohemagglutinin-L and well-of-the-well (35.7%-38.5%). However, 2P1N showed a higher number of blastocysts compared to 3N (73.8% vs. 24.3%) and an increased blastocyst diameter compared to the control and 1P2N (274 ㎛ vs. 230-234 ㎛). In blastomeres aggregated using phytohemagglutinin-L, the apoptotic cell ratio was significantly higher in 1P2N and 3N than in 3P (5.91%-6.46% vs. 2.94%, respectively). Our results indicate that aggregation of one NT embryo with two PA embryos improved the rate of blastocysts with increased blastocyst diameter.

• Animal Bioscience
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• v.35 no.9
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• pp.1360-1366
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• 2022

Objective: The present study analyzed the influence of co-transferring embryos with high and low cloning efficiencies produced via somatic cell nuclear transfer (SCNT) on pregnancy outcomes in dogs. Methods: Cloned dogs were produced by SCNT using donor cells derived from a Tibetan Mastiff (TM) and Toy Poodle (TP). The in vivo developmental capacity of cloned embryos was evaluated. The pregnancy and parturition rates were determined following single transfer of 284 fused oocytes into 21 surrogates and co-transfer of 47 fused oocytes into four surrogates. Results: When cloned embryos produced using a single type of donor cell were transferred into surrogates, the pregnancy and live birth rates were significantly higher following transfer of embryos produced using TP donor cells than following transfer of embryos produced using TM donor cells. Next, pregnancy and live birth rates were compared following single and co-transfer of these cloned embryos. The pregnancy and live birth rates were similar upon co-transfer of embryos and single transfer of embryos produced using TP donor cells but were significantly lower upon single transfer of embryos produced using TM donor cells. Furthermore, the parturition rate for TM dogs and the percentage of these dogs that remained alive until weaning was significantly higher upon co-transfer than upon single transfer of embryos. However, there was no difference between the two embryo transfer methods for TP dogs. The mean birth weight of cloned TM dogs was significantly higher upon single transfer than upon co-transfer of embryos. However, the body weight of TM dogs did not significantly differ between the two embryo transfer methods after day 5. Conclusion: For cloned embryos with a lower developmental competence, the parturition rate and percentage of dogs that remain alive until weaning are increased when they are co-transferred with cloned embryos with a greater developmental competence.

• Journal of Animal Science and Technology
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• v.65 no.4
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• pp.767-778
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• 2023

The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.

• Anatomy and Cell Biology
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• v.57 no.2
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• pp.288-293
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• 2024

The temporal fascia is a double lamina sandwiching a thick fat layer above the zygomatic bony arch. To characterize each lamina, their developmental processes were examined in fetuses. We observed histological sections from 22 half-heads of 10 mid-term fetuses at 14-18 weeks (crown-rump length, 95-150 mm) and 12 near-term fetuses at 26-40 weeks (crown-rump length, 215-334 mm). The superficial lamina of the temporal fascia was not evident at mid-term. Instead, a loose subcutaneous tissue was attached to the thin, deep lamina of the temporal fascia covering the temporalis muscle. At near-term, the deep lamina became thick, while the superficial lamina appeared and exhibited several variations: i) a mono-layered thick membrane (5 specimens); ii) a multi-layered membranous structure (6) and; iii) a cluster of independent thick fasciae each of which were separated by fatty tissues (1). In the second and third patterns, fatty tissue between the two laminae was likely to contain longitudinal fibrous bands in parallel with the deep lamina. Varying proportions of the multi-layered superficial lamina were not attached to the zygomatic arch, but extended below the bony arch. Whether or not lobulation or septation of fatty tissues was evident was not dependent on age. The deep lamina seemed to develop from the temporalis muscle depending on the muscle contraction. In contrast, the superficial lamina developed from subcutaneous collagenous bundles continuous to the cheek. Therein, a difference in development was clearly seen between two categories of the fasciae.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.291-298
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• 2008

We separately cultured follicular oocytes collected from individual ovaries of slaughtered Korean native cows and examined both the embryonic development rate and pregnancy rate after embryo transplantation according to the meat yield and quality grades of the source beef carcass. Oocytes from meat yield grade B cows exhibited a higher fertilization rate and embryonic developmental rate to the eight-cell stage than oocytes from grade A or C animals (p<0.05), but there was no significant difference in rate of development to the blastocyst stage among meat yield grades A, Band C. The oocyte cleavage rate and development rate to the eight-cell stage from meat quality grade 3 cattle was higher than grades 1++, 1+, 1 and 2 (p<0.05). Embryos derived from grade animals displayed a development rate to the blastocyst stage of 19.4%, which was also higher than all other meat quality grades (p<0.05). Transplantation of in vitro-cultured oocytes from meat yield grade A ovaries led to a higher pregnancy rate (64.2%) than in vitro-cultured oocytes from meat yield grade B ovaries (56.5%), but there was no significant difference between the two groups in pregnancy or abortion rates. In conclusion, embryonic development rate and pregnancy rate has a close relation to meat quality grades of the source beef carcass, this results is to give information for the Korean native cows improvement of breed.

• Reproductive and Developmental Biology
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• v.28 no.4
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• pp.227-233
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• 2004

This study was conducted to investigate the effect of various addition and exclusion time of glucose (Control: no addition, A: 24～72 h, B: 24～48 h, C: 48～72 h, D: 0～72 h, E: 0～48 h, F: 0～24 h and 48～72 h, G: 0～24 h) on embryonic developmental capacity of 2-cell embryos in mice. Developed blastocysts were assessed for mean cell number by differential staining. The zona-intact blastocyst (ZiB) rates were higher (p<0.05) in group B than control. However, the zona-escape blastocyst (ZeB) rates were not significantly different in all groups. At 72 h, total blastocyst (ZiB + ZeB) formation rates were not significantly different in all groups. The mean cell number was not significantly different among all groups. The inner cell mass (ICM) cell number was higher (p<0.05) in group F than control, group A, B and G. The trophectoderm (TE) cell number was higher (p<0.05) in control than group A and D. The %ICM was higher (p<0.05) in group C, D and F than control. The ICM : TE ratio was not significantly different in all groups. Between control and glucose group, no significant difference was observed in the total blastocysts (ZiB + ZeB) formation rates. Also, no significant difference was observed in the mean cell number, ICM cell number and ICM : TE ratio. However the TE cell number was higher (p<0.05) in control than glucose group and %ICM was higher (p<0.05) in glucose group than control. In conclusion, glucose added in culture medium was not inhibitory on blastocyst formation but glucose added for 48 ～72 h in culture medium increases %ICM of blastocysts in mice.

• Development and Reproduction
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• v.21 no.4
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• pp.457-466
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• 2017

Survival of embryos largely depends on yolk processing during early development. Proteolytic enzymes, cathepsin B & D (ctsb & ctsd) are known to have some important roles in yolk processing of various fish species. Mature female red spotted groupers were injected with human chorionic gonadotropin (HCG) to induce ovulation. The fertilized eggs and embryos were sampled at 0, 4 and 24 HPF (hours post fertilization). Survivals of each groups of embryos were checked at 24 and 48 HPH (hours post hatching). Transcripts of ctsb & ctsd showed the highest level at 0 HPF and relatively high at 4 HPF, but greatly decreased at 24 HPF. In bad egg quality group (BE, embryos survived until 24 HPH), transcript level of ctsb at 4 HPF were significantly lower than the transcript level at the same stage in good egg quality group (GE, embryos survived until 48 HPH) while no significant change of ctsb transcript level was observed at 0 or 24 HPF between BE and GE. Transcript level of ctsd was decreased at 24 HPF, but the difference was not as strong as the case of ctsb transcript. These results suggest that maternal ctsb transcript rather than ctsd transcript is likely to be involved in egg quality resulting in the difference of survival rate of embryos at early developmental period in this species.

• Development and Reproduction
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• v.21 no.4
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• pp.441-448
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• 2017

Bisphenol-A(BPA) is a member of alkylphenol family, and shows adverse effects including reduced fertility, reproductive tract abnormalities, metabolic disorder, cancer induction, neurotoxicity and immunotoxicity. In the present study, we conducted Hershberger assay to evaluate whether the two candidates to replace BPA have androgenic or antiandrogenic activity. The assay was carried out using immature castrated Sprague-Dawley male rats. After 7 days of the surgery, testosterone propionate (TP, 0.4 mg/kg/day) and test materials (low dose, 40 mg/kg/day; high dose, 400 mg/kg/day) were administered for 10 consecutive days by subcutaneous (s.c.) injection and oral gavage, respectively. Test materials were BPA, isosorbide (ISO) and cyclohexanedimethanol (CHDM). The rats were necropsied, and then the weights of five androgen-dependent tissues [ventral prostate, seminal vesicle, levator ani-bulbocavernosus (LABC) muscle, paired Cowper's glands, and glans penis] and three androgen-insensitive tissues (kidney, spleen and liver) were measured. All test materials including BPA did not exhibit any androgenic activity in the assay. On the contrary, antiandrogen-like activities were found in all test groups, and the order of the intensity was CHDM > BPA > ISO in the five androgen-sensitive tissues. There was no statistical difference between low dose treatment and high dose treatment of BPA group as well as ISO group. In CHDM group, high dose treatment exhibited most severe weight reduction in all measured tissues. There was no statistical difference in androgen-insensitive tissue measurements, except BPA groups. Since the effects of ISO treatment on the accessory sex organs were much less or not present at all when compared to those of BPA, ISO could be a strong candidate to replace BPA. CHDM treatment brought most severe weight reduction in all of androgen-sensitive tissues, so this material should be excluded for further screening of BPA substitute selection.