• Journal of Embryo Transfer
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• v.25 no.3
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• pp.149-154
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• 2010

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100\;{\mu}g$ GnRH at the time of Ind insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. 50.0 percentage of the cows (18/36) had normal resumption of ovarian cyclicity (resumption within 40 days after superovulation), and 50.0% (18/36) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II (first ovulation did not occur until $\geq$ 40 days after superovulation, i.e. delayed first ovulation 33.3%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10$\leq$ of corpora lutea (CL) was 7.3 and 13.9, respectively. The number of transferable embryos differed between < 10 and 10$\leq$ CL was 4.2 and 5.1, respectively. 11.1 percentage of the cows (4/36) did not resumption their ovarian cyclicity until 60 days after superovulation treatment.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.320-324
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• 2017

Development of modern agricultural machinery and accompanying agricultural development cause soil compaction and reduce growth by stressing roots. Kalanchoe pinnata was used to investigate the impact of stress on rooting and changes in plant growth and reproduction. K. pinnata forms somatic embryos capable of asexual reproduction at the edge of leaves. Impact of root pressurization of K. pinnata on somatic embryogenesis and organ differentiation according to external stress factors was investigated by using a high concentration of agar and this phenomenon was studied histologically. Agar concentration in culture media ranged from 0.5%-1.5% to induce a compression effect on roots. The stem and leaf of K. pinnata were subjected to a microtechnique process to study changes in tissue. In vivo, K. pinnata produced 2nd and 3rd plantlets at edges of leaves from lack of water and excessive lighting conditions. In in vitro culture studies, the lower the concentration of agar, the higher the population and the higher the biomass, but plantlet did not occur in leaf bends. Conversely, as concentration of agar increased, increase in the number of individuals was low. Plantlet development occurred only in agar 1.5% medium. The difference in agar concentration was a stressor in the root of K. pinnata, and thus the pattern of asexual reproduction changed from the division method in root to a plantlet generation in leaf. This suggests root pressurization may act as stress and change in the plant reproduction pattern.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.131-137
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• 1997

This study was carried out to investigate the in vivo development rates of vitrified-thawed mouse expanding, hatching and hatched blastoc ysts(BL). In vitro fertilization produced blastocysts were vitrified in EFS40(40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). Expanding a and hatching blastocysts were equilibrated in 20% ethylene glyco](EG) for 5 min. before exposure to EFS40 at 25°C for 1 min., they were then vitrified in liquid nitrogen. Hatched blastocysts which cultured in m-CR1 medium supple mented 0.4% bovine serum albumin on day 5. were equilibrated in 10% EG for 5 min. and then vitrified in EFS40 for 30 sec. After thawing, re-expanding blastocysts were transferred to recipients(3 day of pseudopregnant) on one or both uterine horns(6-8 embryos per a horn). Preg¬n nancy rates of recipients and implantation were a assccessed by autopsy on 15 gestation. The res¬u ults obtained in these experiments were summar¬1 ized as follows; 1) The pregnancy and live fetus rates, for vitrified expanding BL(77.8 and 25.0%) and hatching BL(77.8 and 26.4%)n vitro were not significantly difference in those of control BL (66.7 and 42.9%: 83.3 and 40.4%), respectively, 2) in vitro development of vitrified hatched BL was 34.0%. and 3) in vivo developmental rate of vitrified hatched BL was only 33.3%. These results suggested that proposed rapid vitrification p procedures used EFS40 cryoprotectant can be effectively performed in mouse expanding Ihatching blastocysts and that mouse blastocysts a after being hatched from zona pellucida can be successfully cryopreserved.

• Korean Journal of Veterinary Service
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• v.44 no.4
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• pp.227-237
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• 2021

Artificial insemination of Korean native cattle (KNC) is the predominant method for breed improvement. However, industrialization of embryo production and transfer is necessary to utilize the genetic potential of KNC. The aim of this study was to examine associations between KNC donor cows and ovum pick-up (OPU) conditions, in-vivo oocyte recovery, and embryo development. Oocyte recovery and blastocyst development rates were higher at 50 and 60 mmHg OPU vacuum pressure than at 40 mmHg, which was, however, not significant. Regarding follicle growth, injection of 500 ㎍ GnRH 36 hours before OPU significantly increased the number of OPU oocytes from an average of 4.6 to 7.6 (P<0.05); no significant difference in embryo development rates was observed. Significant differences were observed in the numbers of OPU oocytes, embryo development rates, and transplantable blastocysts per individual among nine KNC donors (P<0.05). Furthermore, although there was no difference in OPU oocyte recovery intervals in approximately 2~8 weeks, the number of recovered oocytes significantly decreased at the 12-week interval (P<0.05); there was no difference in embryo development rates. The number of oocytes and embryonic development rates only tended to decrease until the seventh OPU session, but decreased significantly until the eighth session (P<0.05). The average pregnancy rate after transfer of OPU-derived in-vitro embryos into recipient cows was 41.8%. To improve the efficiency of OPU egg recovery and in-vitro embryo production, considering KNC donor characteristics, vacuum pressure of 60 mmHg, GnRH pretreatment to induce follicle growth, and effective OPU egg recovery up to seven times at intervals of 2~4 weeks appears to be most suitable. This study may facilitate the industrialization of KNC embryo production and transfer using high-quality cows.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.35-40
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• 2013

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Holstein Donor Cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_2{\alpha}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II(first ovulation did not occur until ${\geq}40$ days after superovulation, i.e. delayed first ovulation 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and $10{\leq}$ of corpora lutea(CL) was $7.8{\pm}1.8$ and $12.7{\pm}2.7$, respectively. The number of transferable embryos differed between < 10 and $10{\leq}$ CL was $5.4{\pm}1.3$ and $8.1{\pm}3.4$, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. Diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

• Korean Journal of Food Science and Technology
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• v.51 no.2
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• pp.141-146
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• 2019

Sparassis crispa is an edible mushroom that is distributed in Korea, Japan, Europe, and North America. It exerts various biological activities such as immunopotentiation, anti-diabetic, anti-cancer, and anti-inflammatory effects. Recently, we separated the health functional non-aqueous fraction from the chloroform extract of S. crispa (SCF4). In this study, we evaluated the antiangiogenic activity of SCF4 in human umbilical vein endothelial cells (HUVECs). SCF4 effectively inhibited vascular endothelial growth factor (VEGF)-induced cell growth at concentrations ($5-25{\mu}g/mL$) showing no cytotoxic effects. SCF4 inhibited VEGF-induced invasiveness and tube formation ability, which are in vitro angiogenic features of HUVECs, in a dose-dependent manner. In addition, SCF4 markedly suppressed in vivo angiogenesis of chorioallantoic membrane from growing chick embryos without cytotoxicity. Furthermore, SCF4 downregulated the phosphorylation of VEGFR2, AKT, and ERK1/2, which are major angiogenic signal mediators. These results suggest that SCF4 inhibited angiogenesis by suppressing the VEGFR2 signaling pathways without cytotoxicity.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.9
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• pp.1229-1236
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• 2012

Our previous studies showed that kisspeptin-10 (Kp-10) injected in vivo can markedly increase lipid anabolism in liver of quails. In order to investigate the direct effect of Kp-10 on lipid metabolism of hepatocytes in birds, cells were separated from embryos livers and cultured in vitro with 0, 100 and 1,000 nM Kp-10, respectively. The results showed that after 24 h treatment, cells viability was not affected by 100 nM Kp-10, but showed a mild decrease with 1,000 nM Kp-10 compared to the control cells. Based on the results of the cell viability, 100 nM dosage of Kp-10 was selected for the further study and analysis. Compared with control cells, total cholesterol (Tch) contents in 100 nM treated cells were increased by 51.23%, but did not reach statistical significance, while the level of triglyceride (TG), high density of lipoprotein-cholesterol (HDL-C) and low density of lipoprotein-cholesterol (LDL-C) were significantly increased. Real-time PCR results showed that ApoVLDL-II mRNA expression had a tendency to increase, genes including sterol regulatory element-binding protein-1 (SREBP-1), acetyl coenzyme A carboxylase ${\alpha}$ ($ACC{\alpha}$), carnitine palmitoyltransferase 1 (CPT1), 3-hydroxyl-3-methylglutaryl-coenzyme A reductases (HMGCR) and stearyl coenzyme A dehydrogenase-1 (SCD1) mRNA in hepatocytes were significantly down-regulated by 100 nM Kp-10. However, contrary to its gene expression, SREBP-1 protein expression was significantly up-regulated by 100 nM Kp-10. Some of the significant correlations in mRNA expression were found between genes encoding hepatic factors or enzymes involved in lipid metabolism in liver of birds. These results indicate that Kp-10 stimulates lipid synthesis directly in primary cultured hepatocytes of chickens.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.141-149
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• 1997

This study was carried out to investigate the effect of vitrification and slow freezing methods on the post-thaw developmental rate of rabbit zygotes. After exposing rabbit zygotes in EFS solution for 0.5, 1, 2, 3 and S min at room temperature, they were washed with 0.5 M sucrose solution, D-PBS and TCM-199 and then cultured in TCM-199 plus 10% FBS with bovine oviduct epithelial cells(BOEC) to examine whether the cryoprotectant induced injury during the various exposure periods. The embryo development rates to hatched blastocyst after exposing in EFS solution for 3 and 5 min(40.0 and 16.7%) were significantly lower than in 0.5, 1 and 2 min(63.0, 72.0 and 54.5%), respectively. The post-thaw development rates to hatched blastocyst were significantly(P<0.05) higher in in vivo morula with intact mucin coat(85.2%) and mucin seperated morula(77.8%) than those of in vitro morula(58.5%) and zygote(5.9%), hut no difference was shown between in vitro morulae and mucin separated morula. The cryoprotectant dilution procedures showed no effects on the post-thaw development rates to hatched blastocyst under the present culture conditions. The post-thaw development to hatched blastocyst in the rabbit zygotes was not significantly different between the slow freezing(12.8%) and vitrification(5.9%). These results indicated that the rabbit frozen zygotes could he successfully developed in vitro to hatched blastocysts, though their developmental rate was very low, compared with morula stage embryos, in either vitrification or slow freezing procedure under the present conditions.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.234-237
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• 2011

Objective: During stimulated IVF cycles, up to 15% of oocytes are recovered as immature. The purpose of this study was to investigate the trend of oocyte maturity in repeated ovarian stimulation for IVF. Methods: One hundred forty-eight patients were selected who underwent two consecutive IVF cycles using same stimulation protocol during 2008 to 2010. Ovarian stimulation was performed with FSH and human menopausal gonadotropin and flexible GnRH antagonist protocol in both cycles. Oocyte maturity was assessed according to presence of germinal vesicle (GV) and the first polar body. Immature oocyte was defined as GV stage or metaphase I oocyte (GV breakdown with no visible polar body) and cultured up to 48 hours. If matured, they were fertilized with ICSI. Results: Percentages of immature oocytes were 30.8% and 32.9% ($p$=0.466) and IVM rates of immature oocytes were 36.2% and 25.7% ($p$=0.077), respectively. A significant correlation was noted between percentage of immature oocytes in the two cycles (R=0.178, $p$=0.03). Women with >40% immaturity in both cycles (n=21) showed lower fertilization rate of $in$ $vivo$ matured oocytes (56.4% vs. 72.0%, $p$=0.005) and lower pregnancy rate (19.0% vs. 27.1%, $p$=0.454) after the second cycle when compared with women with <40% immaturity (n=70). In both groups, female age, number of total retrieved oocyte and embryos transferred were similar. Conclusion: In repeated ovarian stimulation cycles for IVF, the immature oocyte tended to be retrieved repetitively in consecutive IVF cycles.

• ALGAE
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• v.32 no.3
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• pp.261-273
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• 2017

Fine dust (FD) particles have become a major contributor to air pollution causing detrimental effects on the respiratory system and skin. Although some studies have investigated the effects of FD on the respiratory system, their possible effects on the skin remain under-explored. We investigated the FD mediated inflammatory responses in keratinocytes, present in the outer layers of skin tissues and the transfer of inflammatory potential to macrophages. We further evaluated the anti-inflammatory effects of the polyphenolic derivative, diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae against FD-induced inflammation. Size distribution of FD particles was analyzed by scanning electron microscopy. FD particles induced the production of cyclooxygenase-2, prostaglandin E2 ($PGE_2$), interleukin (IL)-$1{\beta}$, and IL-6 in HaCaT keratinocytes and the expression of nitric oxide (NO), inducible nitric oxide synthases (iNOS), $PGE_2$, tumor necrosis factor-${\alpha}$ expression in RAW 264.7 macrophages. Further, we evaluated the inflammatory potential of the culture medium of inflammation-induced HaCaT cells in RAW 264.7 macrophages and observed a marked increase in the expression of NO, iNOS, $PGE_2$, and proinflammatory cytokines. DPHC treatment markedly attenuated the inflammatory responses, indicating its effectiveness in suppressing a broad range of inflammatory responses. It also showed anti-inflammatory potential in in-vivo experiments using FD-stimulated zebrafish embryos by decreasing NO and reactive oxygen species production, while eventing cell death caused by inflammation.