• Journal of Embryo Transfer
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• v.31 no.3
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• pp.191-197
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• 2016

Historically, Korea old cattle had been consisted with various lines of coat color brindle, black and white-brown breeds or more. The two rare lines of black and white coat color are maintained for animal resources and preserved critically. The present study was carried out to evaluate potential usage of cysteamine supplementation during in vitro matration (IVM) and in vitro culture/production of embryo (IVP) by transvaginal ultrasound-guided follicle aspiration (Ovum Pick-Up: OPU) for the establishment of cryo-banking system. Immature slaughterhouse-derived cumulus-oocyte complexes (SL-COCs) were matured in IVM medium supplemented with 0, 0.1, 0.3 or 0.9 mM cysteamine, and then cultured in mSOF-BAS for 8 days after in vitro fertilization. The treatment of 0.1 mM cysteamine on SL-COCs showed higher rate of blastocyst, so OPU-derived COCs from rare breeds were matured in TCM media supplemented with or without 0.1 mM cysteamine, FSH and 5% FBS. The embryos were evaluated their developmental stages on day 8. During IVM, cysteamine treatment significantly increased the embryo production rate of slaughterhouse-derived COCs (19.6% vs. 30.5%). The presence of cysteamine during IVM of OPU-derived COCs from rare Korean cattle breeds (albino white and black line) also increased embryo production rates than those from SL-COCs (27.4% vs. 41.9% and 36.4%). With these results, cysteamine treatment during IVM is one of key factors IVP of blastocysts to establish banking system of endangered rare Koarean cattle with OPU derived transferable blastocysts.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.12
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• pp.1681-1690
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• 2012

The aim of this study was to investigate the impact of a reported p53 inhibitor, pifithrin-${\alpha}$ (PFT-${\alpha}$), on preimplantation porcine in vitro fertilized (IVF) embryo development in culture. Treatment of PFT-${\alpha}$ was administered at both early (0 to 48 hpi), and later stages (48 to 168 hpi) of preimplantation development, and its impact upon the expression of five genes related to apoptosis (p53, bak, bcl-xL, p66Shc and caspase3), was assessed in resulting d 7 blastocysts, using real-time quantitative PCR. Total cell numbers, along with the number of apoptotic nuclei, as detected by the in situ cell death detection assay, were also calculated on d 7 in treated and non-treated control embryos. The results indicate that PFT-${\alpha}$, when administered at both early and later stages of porcine IVF embryo development, increases the incidence of apoptosis in resulting blastocysts. When administered at early cleavage stages, PFT-${\alpha}$ treatment was shown to reduce the developmental competence of porcine IVF embryos, as well as reducing the quality of resulting blastocysts in terms of overall cell numbers. In contrast, at later stages, PFT-${\alpha}$ administration resulted in marginally increased blastocyst development rates amongst treated embryos, but did not affect cell numbers. However, PFT-${\alpha}$ treatment induced apoptosis and apoptotic related gene expression, in all treated embryos, irrespective of the timing of treatment. Our results indicate that PFT-${\alpha}$ may severely compromise the developmental potential of porcine IVF embryos, and is a potent apoptotic agent when placed into porcine embryo culture media. Thus, caution should be exercised when using PFT-${\alpha}$ as a specific inhibitor of p53 mediated apoptosis, in the context of porcine IVF embryo culture systems.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.2
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• pp.91-96
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• 2004

Accumulation of nitrate in edible crops is undesirable due to potential risks to human health. Since nitrate has a role in the osmotic regulation of plants, salt accumulation in soil is expected to stimulate nitrate accumulation in plants. Lettuce (Lactuca sativa L.) was grown in soils of different salinities, 9.69 and $4.49dS\;m^{-1}$, in a greenhouse, and the effect of soil salinity on nitrate accumulation in lettuce was investigated. Content of nitrate in the lettuce increased significantly as soil salinity increased under low light intensity and ample supply of nitrate in root media. Soluble sugar and oxalate contents in lettuce were also significantly higher in the soil of higher salinity. Phosphate, Cl, and $SO_4$ contents in lettuce were not significantly different in soils of different salinities. Among the cations, K content in lettuce was significantly higher in the soil of higher salinity, but Na, Ca, and Mg comtents were not much influenced. Comparing to the lettuce grown in low salinity soil, although the growth of lettuce was decreased by 9% in the soil of higher salinity, nitrate accumulation in the lettuce was increased by 18.6%. These results indicate that higher nitrate content in lettuce of higher salinity soil is a positive accumulation to adapt to the water stress condition. The nitrate accumulation of vegetables grown in plastic film houses is known to be due to the heavy fertilization and low light intensity, but salt accumulation in the soil, which can lower soil water potential, is expected to stimulate the nitrate accumulation further.

• Journal of Bio-Environment Control
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• v.16 no.4
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• pp.365-371
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• 2007

Objective of this research was to investigate the effect of nitrogen concentrations in the fertilizer solution on growth and development of nutrient deficiency in leaf perilla (Perilla frutesens). The nutrient concentrations in above-ground plant tissue, petiole sap and soil solution of root media were also determined. Nitrogen deficiency resulted in dwarfed growth, small leaves, and bright yellow color of older leaves. The leaves of deficient plants became uniform yellowing in color and finally necrosis occurred on the deficient leaves. Elevation of N concentrations in the fertigation solution from 0 to 20 mM increased the crop growth in leaf length and width as well as fresh and dry weights of above ground plant tissue. That also resulted in the increase of chlorophyll contents. However, light toxicity symptoms such as abnormal leaf surface appeared on crops grown in 20 mM N fertilization. The plant growth was commercially acceptable in the treatments of 10 and 15 mM N. The plants with acceptable growth had 0.9 to 1.25% in N contents of above-ground plant tissue, 800 to $3,300mg{\cdot}kg^{-1}$ in the $NO_3-N$ concentrations of petiole sap, and 28.7 to $47.3mg{\cdot}kg^{-1}$ in the $NO_3-N$ concentrations of soil solution (1:2 extract) at 75 days after transplanting.

• Journal of Bio-Environment Control
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• v.16 no.4
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• pp.372-378
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• 2007

This study was carried out to investigate the effect of potassium concentrations in fertigation solution on growth and development of nutrient deficiency symptoms of leaf perilla (Perilla frutesens). The nutrient concentrations in above ground plant tissue, petiole sap and soil solution of root media were also determined. Potassium deficiency symptoms developed in older leaves with marginal necrosis. The brown areas on the lower leaves enlarged rapidly and the margins became scorched. Elevation of K concentrations in the fertigation solution up to 8 mM increased the crop growth in leaf length, stem thickness, and fresh and dry matter production of above ground plant tissue. However, that decreased the chlorophyll contents. The 8.0 mM K treatment which showed the greatest growth had 5.01 g in dry weight and 2.76% in K content of above ground plant tissue, suggesting that maintaining K content higher than 1.7% is necessary for good growth of Perilla frutesens. The K concentrations in petiole sap and soil solution of 8.0 mM treatment were $12,289mg{\cdot}kg^{-1}\;and\;11.65mg{\cdot}L^{-1}$, respectively. These indicated that K fertilization to maintain higher than $8,700mg{\cdot}kg^{-1}$ in petiole sap and $4.5mg{\cdot}L^{-1}$ in soil solution are necessary to ensure good crop growth.

• Horticultural Science & Technology
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• v.18 no.4
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• pp.518-522
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• 2000

The objective of this research was to determine the effect of waste nutrient solution (WNS) on growth and development of poinsettia 'Gutbier V-10 Amy'. To achieve this, WNS collected from rose grown in greenhouse was diluted with various times and 500 mL of each solution was applied every week. Then growth characteristics and nutrient uptake were determined at 180 days after transplanting. The treatment of undiluted WNS had highest plant height and length of branch among treatments tested, but there were no statistical differences in the number of bract and branches. Undiluted WNS had higher leaf number, leaf length, leaf width, fresh weight, and dry weight than any other treatments tested. There was also a trend that increased dilution times of WNS resulted in decreased plant growth. Undiluted WNS had higher chlorophyll content than Hyponex treatment, but diameter of crown did not show significant differences among treatments. In the analysis of root media collected at 6 months after fertilization, the treatment of undiluted WNS had higher electrical conductivity and organic matter content than other treatments, but the Hyponex treatment had higher phosphorus concentration than other treatments tested.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.115-123
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• 2003

The present study was carried out to examine the effect of catalase (CAT) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with CAT on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows - 1 . The rates of nuclear maturation, penetrated oocytes, pronucleus formation rates, polyspermic oocytes and mean numbers of the penetrated sperm were significantly lower in oocytes matured with 100, 500 and 1,000 units/ml CAT than those of central groups (P>0.05). 2. The rates of blastocyst formation and total cell numbers of blastocyst at day 7 after in vitro fertilization were significantly lower in CAT treatment groups than those of the central groups (P>0.05). 3. There were not significant difference in the blastocyst development and total cell numbers of blastocyst on in vitro culture of NCSU-23 media with 0, 100, 500 and 1000 units/ml CAT under the 5% and 20% $O_2$ concentrations. These results suggested that the addition of CAT was not helpful for porcine oocyte maturation and further development, also the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under the 5% and 20% $O_2$ concentrations.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.207-216
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• 1998

Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.3
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• pp.122-125
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• 2013

Objective: The majority of embryo transfers (ETs) to date have been performed on day 3 to reduce the potential risk of developmental arrest of in vitro cultured embryos before ET. Development of sequential media has significantly improved culture conditions and allowed blastocyst transfer on day 5. While day 5 ET provides higher clinical pregnancy outcomes with reduced risks of multiple pregnancies, it still has potential risks of developmental arrest of IVF embryos. The aim of this study was to evaluate the clinical outcomes of day 4 ETs and compare the efficacy of day 4 ET with day 5 ET. Methods: From 2006 to 2009, a total of 747 fresh IVF-ET cycles were retrospectively analyzed (day 4, n=440 or and day 5, n=307). The cycles with any genetic factors were excluded. The rates of matured oocytes, fertilization, good embryos, and clinical pregnancy of the two groups were compared. The chi-square test and t-test were used for statistical analysis. Results: There were no significant differences between the two groups with respect to the mean age of the females and rates of matured oocytes. The pregnancy outcomes of day 4 ET (40.7%) were similar to those of day 5 ET (44.6%). The implantation rate of day 5 ET (24.2%) was significantly higher than that of day 4 ET (18.4%) (p=0.003). Conclusion: Day 4 ET can be chosen to avoid ET cancellation in day 5 ET resulting from suboptimal circumstances in the IVF laboratory, but the decremented quality of embryos for transfer and the decreased pregnancy rate must be taken into consideration.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.237-242
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• 2003

These studies were carried out to establish an effective in vitro embryo transfer methods by analyzing several factors. The base media were TCM-199 solution for in vitro maturation(IVM) of bovine follicular oocytes and Fer-TALP solution for in vitro fertilization(IVF) and CRlaa medium for in vitro culture(IVC). IVC used the fertilized oocytes of 24-hr culture (day 1)after IVF. Embryos were cultured in drop-culture that contained 25 embryos per 10${mu}ell$. Blastocysts cultured for 7 to 9 in vitro were transferred to recipients. The results obtained were as follows: 1. The pregnancy rate according to different region of embryo transfer were 33.8％, 48.1％, 45.0％ and 35.3％ respectively. 2. The pregnancy rate according to the parity of recipient when embryos were transferred to nulliparous (42.9％) was higher than that of 1∼3nd parlous(36.9％), however there were not show significant difference each other. 3. According to the stage of blastocyst, the pregnancy rate when middle blastocysts (MB) (45.5％) were transferred to recipients were higher than that of late blastocysts (LB) (41.0％). 4. When IVF-derived blastocysts cultured for 7 to 9 day were transferred to recipients, the pregnancy rate was higher 7 day of blastocyst than that of 8 day or 9 day of blastocyst. The results of embryo transfer according to the regions, the parity of recipient and the development stage showed that blastocyst formed for 7day transferred to nulliparou were higher pregnancy rate than others.