• Clinical and Experimental Reproductive Medicine
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• v.32 no.1
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• pp.47-54
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• 2005

Objective: Evaluation of embryos using early cleavage to 2-cell stage has been proposed, but a critical time-point for selecting embryos is unclear. The aim of the present study is to provide a guideline including critical time-point in the selection of early cleaving embryo for the reduction of multiple pregnancies as well as the increase of pregnancy rate in human IVF. Methods: This prospective study was performed in 116 cycles from 85 patients who underwent conventional IVF or ICSI at the infertility clinic of Good Moonhwa Hospital from January 2002 to December 2003. Early cleavage (EC) of embryos to 2-cell stage was assessed at 25 h and 27 h postinsemination/microinjection. Embryos that had early cleaved at each time point were designated as EC-1 and EC-2, respectively, while others were designated as non-early cleavage (NEC). Results: At least one early cleavage embryo was observed in 54 (46.6%) for the EC-1 and 84 (72.4%) for the EC-2 of the 116 cycles assessed. Clinical pregnancy rates (PR) were significantly higher in the EC-1 group (66.7%) compared to the EC-2 group (53.6%) or the NEC group (31.2%) (p<0.05). Significant improvement of the pregnancy rate was found when at least two or more embryos were early cleaved at 25 h postinsemination or when the proportion of early cleavage embryo at 25 h postinsemination was higher than 20% (p<0.05). Conclusion: The critical time-point for the selection of early cleavage embryos with high implantation potential is more effective in 25 h postinsemination/microinjection compared to 27 h. The proportion as well as number of early cleavage embryos is also an important factor for the prediction of pregnancy outcome and the chance of multiple pregnancies. These results demonstrated that the evaluation of early cleavage embryos to 2-cell stage is an easy, simple, and objective method for the selection of good quality embryos suitable for embryo transfer.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.123-130
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• 2005

High salt concentrations in the ginseng nursery soil environment of Korea is one of important reducing factors for the stable production of quality ginseng. These studies were accomplished for check the response on germination of ginseng seed, somatic embryogenesis of zygotic embryo, and biosynthesis of ginsenoside from ginseng hairy root against NaCl. Ratio of germination was at the $3\%\;and\;84.5\%$ on the basic media with 0.1M and free of NaCl repectedly, but $0\%$ at the upper of 0.2M NaCl. Somatic embryogenesis from zygotic embryo were the highest when immatured embryo was cultured on free of NaCl concentration, and which was intend to decrease at treatment of NaCl. However, in case of using the matured embryo, treatment of 0.05M NaCl resulted in better embryogenesis than NaCl free media. Red pigment was synthesized from ginseng hairy root cultured on the medium with various NaCl concentration(from 0.04 to 0.08M) and its pigment was analyzed as spectrum of anthocyane by spectrophoto- meter scanning. This cell line biosynthesized lots of crude saponin and total ginsenoside than other cell lines, also had 2 times of panaxadiol than panaxatriol.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.6
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• pp.537-544
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• 1993

Super-giant-embryo mutant line (Hwacheong-ge$^s$) with high nutritional quality was developed from Hwacheongbyeo, a Korean Japonica cultivar. Grain weight of brown rice of Hwacheong-ge$^s$ line was lighter than of Hwacheongbyeo. However, embryo dry weight of the line was 3.22 times heavier than that of Hwacheongbyeo on the single grain base. Hwacheong-ge$^s$ line showed higher protein(8.99%) than the original variety(7.39%), without changes in storage protein. In amino acid composition, lysine content was greatly increased in the mutant, while the contents of methionine, serine and tyrosine were slightly decreased as compared with the original one. Lipid content of Hwacheong-ge$^s$ line was 1.5 times higher compared with that of Hwacheongbyeo. The contents of Vitamin Bl, B2, and E(${\alpha}$-tocopherol) were also greatly increased in Hwacheong-ge$^s$line. The grain yields of the mutant lines ranged from 69.2% to 78.8% compared with that of Hwacheongbyeo, 556kg /10a. Most of the mutant lines of M4 generation were homogeneous for growth characters, and some of them seemed to be readily applicable for farmer's field.

• Journal of the Optical Society of Korea
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• v.13 no.3
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• pp.335-340
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• 2009

Ultrashort pulse laser drilling of polycarbonate track-etched membrane (pTEM) material was used to fabricate a mouse embryo cell trapping device. Holes with a diameter of $2{\mu}m$ to $5{\mu}m$ were fabricated on a $10{\mu}m$ thick membrane using a femtosecond laser with a 150 fs pulse width and 775 nm wavelength and multiple-pulse irradiation. In cell trapping tests, the overall cell occupancy of the machined holes in the fabricated pTEM was found to be more than 80%. The results of a single pulse and multiple pulse irradiation were compared in terms of the surface quality. It was generally found that a single pulse with high energy was less desirable than irradiation with multiple pulses of lower energy.

• Korean Journal of Breeding Science
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• v.41 no.2
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• pp.84-91
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• 2009

Rice consumption per capta, in South Korea, has been decreased dramatically, owing to the changes of living patterns. Because of not only the major energy source of Korean people but also major income source of Korean farmers, diversifying end-use-quality of rice has been demanded. To the context, 'Suweon 472', a high yielding and early mature japonica line and released as 'Namilbyeo' to framers in 2002, was treated with a chemical mutagen, Sodium Azide to find endosperm mutant types. A total of nine endosperm mutat lines, including five waxy, one dull, two floury, and one white core type, were identified from the 3,542 mutatagen treated lines. Amylose contents, iodine reaction, disintegration in alkali solution, gelatinization in urea solution and amylogram properties of those nine endosperm mutant lines were evaluated to address the possibility as new genetic materials for diversifying rice quality of Korean japonica cultivars. All embryo mutants were clearly differentiated from their wild type, 'Suweon 472', in terms of physic-chemical properties evaluated. The endosperm mutant lines would be very useful in expanding untiliztation of rice through opening new rice markets of processed foods from Korean japonica rice.

• Development and Reproduction
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• v.1 no.1
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• pp.67-77
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• 1997

Mammalian ovary consists of various growing stages of follicles. Ovarian follicular growth and differentiation, however, can be distinguished into recruitment, growth, selectiona nd ovulation. while only minute of the selected follicles ovulate their oocytes, all the rest follicles disappear by atresia. this atresia is an important event of which physiological mechanism must be resolved. The present study was carried out to investigate the effects of various doses of pregnant mare's serum gonadotropin (PMSG) on the oocyte quality, ovulation rate, and the early embryonic development in immature mice. Immature mice were administrated with 5, 20, or 40 IU PMSG. At every 12 hour up to 72 hour after treatment, body and ovary weights were measured. Oocytes were flushed from the oviducts under the dissecting microscope and observed under the inverted microscope. Late 2-cell embryos were collected from the mice which were superovulated by the same dosage of PMSG followed by 5 IU hCG 47 hours after PMSG-treatment. The percentage of abnormal oocytes was higher in 20 or 40 IU PMSG-treated animals than 5 IU PMSG-treated ones. Ovulation occured at 12 hours afger PMSG injection in all experimental groups. The percentage of retrieved abnormal oocytes increased in the 20 or 40 IU PMSG-treated goups but not in 5 IU PMSG-treated group. There was no significant difference in the mating rate among the groups [52.6% (10/19), 66.7% (10/15), 44.0% (11/25) : 5, 20, 40 IU group respectively] ; however, ther was a significant (p<0.01) increase of embryo retrieval rates in 5 and 20 IU-treated groups compared with that in 40 IU-treated group [89.2% (239-268), 85.5% (224/262), 40.0% (18/45)]. There was significant (p<0.01) increase of embryo development rates in 5 IU-treated group compared with that in 20 and 40 IU-treated group [231/239(96.7), 179/224(79.9), 77.8(14/18)]. In conclusion, higher doses of PMSG injection increased the occurrence of abnormal oocytes ovulation in immature mice. The most of oocytes collected from 5 or 20 IU-PMSG-treated group has fertilizabioity. But in mice injected iwth higher doses of PMSG, their oocytes exhibit less fertilizability and, even fertilized, all oocytes are not fully capable of development.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.492-499
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• 2009

A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

• Reproductive and Developmental Biology
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• v.37 no.2
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• pp.65-73
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• 2013

The objective of this study was to examine the effects of high concentrations of glucose on porcine parthenotes developing in vitro. Addition of 55 mM glucose to the culture medium of embryos at the four-cell-stage significantly inhibited blastocyst formation, resulting in fewer cells in blastocyst-stage embryos and increased levels of apoptosis and autophagy compared to control. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression of pro-apoptotic genes (Caspase 3, Bax and Bak) and autophagy genes (Atg6 and Atg8/Lc3) were increased significantly by the addition of 55 mM glucose to the culture medium compared to control. MitoTracker Green fluorescence revealed a decrease in the overall mitochondrial mass compared to control. However, the addition of 55 mM glucose had no effect on mRNA expression of the nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1. These results suggest that hyperglycemia reduced the mitochondrial content of porcine embryos developing in vitro and that this may hinder embryonic development to the blastocyst stage and embryo quality by increasing apoptosis and autophagy in these embryos.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.2
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• pp.111-117
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• 2017

Objective: The aim of this study was to evaluate pregnancy outcomes and the live birth rate at 1-year age increments in women aged ${\geq}40years$ undergoing fresh non-donor in vitro fertilization (IVF) and embryo transfer (ET), and to identify predictors of success in these patients. Methods: This retrospective study was performed among women ${\geq}40years$ of age between 2004 and 2011. Of the 2,362 cycles that were conducted, ET was performed in 1,532 (73.1%). Results: The clinical pregnancy rate and live birth rate in women ${\geq}40years$ significantly decreased with each year of increased age (p<0.001). Maternal age (odds ratio [OR], 0.644; 95% confidence interval [CI], 0.540-0.769; p<0.001), basal follicle-stimulating hormone (FSH) levels (OR, 0.950; 95% CI, 0.903-0.999; p=0.047), the number of high-quality embryos (OR, 1.258; 95% CI, 1.005-1.575; p=0.045), and the number of transferred embryos (OR, 1.291; 95% CI, 1.064-1.566; p=0.009) were significant predictors of live birth. A statistically significant increase in live birth rates was seen when ${\geq}3$ embryos were transferred in patients 40 to 41 years of age, whereas poor pregnancy outcomes were seen in patients ${\geq}43years$ of age, regardless of the number of transferred embryos. Moreover, the cumulative live birth rate increased in patients 40 to 42 years of age with repeated IVF cycles, but the follicle-stimulating hormone in those ${\geq}43years$ of age rarely showed an increase. Conclusion: IVF-ET has acceptable outcomes in those < 43 years of age when a patient's own oocytes are used. Maternal age, basal FSH levels, and the number of high-quality embryos and transferred embryos are useful predictors of live birth.