• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.77-85
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• 2003

This study was undertaken to evaluate the effects of catalase using xanthine (X)-xanthine oxidase (XO) system on in vitro maturation and fertilization in the pig. When follicular oocytes were cultured with X or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obtained in culture with X-XO-catalase system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without that than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, but were not different between the medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with none (P<0.05), XO and X+XO. On the other hand, when sperm were treated with none, X, XO and X+XO, lipid peroxidation were produced with higher rates in medium without that than with catalase, and consequently the changes in sperm penetration and lipid peroxidation showed opposite patterns. Under the above all conditions, however, sperm-SH group were higher detected by catalase. When the activity of sperm binding to zona pellucida was evaluated through binding to salt-stored porcine oocytes, sperm binding to zona pellucida in control group were higher than in medium with X, XO and X+XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO-catalase system may be caused stimulating in vitro maturation and fertilization in the pig.

• Korean Journal of Animal Reproduction
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• v.15 no.2
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• pp.125-132
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• 1991

The studies on the carried out to investigate the effects of capacitation method of spermatozoa on the in vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and FCS for 24~48hrs in an incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18hrs with motile capacitated sperm by preincubation of mKRB, treatment of HIS(high strength ion), Ca-IA(Inophore A), BFF(bovine follicular fluids) and heparin. The results obtained in these experiments were summarized as follows : 1. The in vitro fertilizatin and cleavage rate offollicular oocytes fertilized with capacitated spermatozoas in BO solution by preincubation of mKRB, treatment of HIS, Ca-IA, BFF and heparin method were 53.1%, 33.9%, 50.8%, 48.1%, 58.8% and 28.1%, 17.7%, 26.2%, 22.8%, 32.8%, respectively. And the fertilization and cleavage rate of heparin method was of highest of all. 2. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solutin by both caffeine, BSA and heparin methods were 65.8%, 70.3% and 40.8%, 47.3%, respectively, and those rates were higher treatment of heparin＋BSA, heparin＋caffeine than treatment of heparin. 3. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoa in BO solution with heparin concentrations of 2, 5, 10, 20, 40$\mu\textrm{g}$/ml were 50.0%, 54.7%, 58.1%, 51.7% and 27.9%, 32.8%, 37.1%, 30.0%, respectively. And the fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution with 10$\mu\textrm{g}$/ml of heparin was the highest of all. 4. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with caffeine concentraton of 10, 20, 30, 40$\mu\textrm{g}$/ml were 71.4%, 74.3% and 70.6%, 70.0% and 45.7%, 47.3%, 44.1%, 41.4%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with caffeine and heparin together(70.3~74.3%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%). 5. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with BSA concentration of 5, 10, 20, 30$\mu\textrm{g}$/ml were 63.6%, 62.9%, 66.7%, 60.3% and 44.1%, 43.5%, 48.5%, 42.7%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with BSA and heparin together(60.3~66.73%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.2
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• pp.140-145
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• 2011

In order to establish the application method of slurry composting & biofilteration liquid fertilizer (SCB LF) in rice cultivation, experiments were studied on split application method of it and effects of it on soil chemical properties and rice yields. Land leveling by rotary tillage within 2 days after application of SCB LF, $NH_4$-N concentration in soil was maintained uniformly in all paddy field. Initial concentrations of $NH_4$-N and $NO_3$-N in soil were high at standard fertilization and 100% application of SCB LF as basal fertilization, however, after tillering stage they maintained similar concentrations in all experimental plots. $NO_3$-N content in infiltration water was slightly lower at 70% application of SCB LF as basal fertilization and 30% application of SCB LF as fertilization at panicle initiation stage than at standard fertilization. Yields of rice by split application of SCB LF were lower at 100% application of SCB LF as basal fertilization, however, those of the other application of SCB LF were similar with that of standard fertilization. In case of rice quality, perfect kernel rates were high and protein contents were lower at non-application and 100% application of SCB LF. Rice quality of 70% application of SCB LF as basal fertilization and 30% application of SCB LF as fertilization at panicle initiation stage were similar with that of standard fertilization.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.2
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• pp.209-215
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• 1992

Different from 'Seyokwiryang' (歲易爲良 : fallowing as the best method) which was an agricultural technique of ancient China, fertilization of rice fields was already practiced in the end of Koryo age in Korea. 'Bunjongbeob'(糞種法 : fertilizer applicating method on seed) or 'Bunkwabeob' (糞科法 : fertilizer applicating method on each plant) was practiced before green manure of bean crops and 'Bunjeonbeob'(糞田法 : fertilizer practicing method on fields) were done. In the 15th century 'Dojeonbunjeonbeob'(稻田糞田法 : fertilizer applicating method on paddy) in 'Jikseol' was divided the materials to be used into the soil brought from another place, trees, grasses, and the manure. Also, it discribed the fertilization between first plowing and the second, and proper fertilization for particular soil conditions. In case of transplanting techniques, the fertilization practices were specified into nursery and rice fields, and restoration of organic matter was systemized by plowing for cultivation in the reclaimed areas. In the 17th century, through 'Jikseolbo'($\ulcorner$直說補$\lrcorner$), the habitual practice of Kyungsang province was systemized and 'Bunyangsool'(糞壤術 : technique of fertilization) of 'Jodoangkicheo' (早稻秧基處 : rearing fields of early-ripening rice) was completed. Specific things was the manufacturing and utilizing techniques of 'Bunhoe'(糞灰 : mixture of manure and ash), 'Yohoe'(尿灰 : mixture of urine and ash), and additional fertilizers. In the 18 to 19th century, the materials of fertilization were greatly enlarged to recover the waste lands and to support the reinforcement of soil fertility for increasing the system of two cropping a year. Also, 'Jeobunbeob'(貯糞法 : method of manure storage) and additional fertilization were emphasized, and use of wagons for it was emphasized to improve the theory of fertilization and working efficiency. As mentioned above, limitation of fertilizing materials was conquered by 'Dojeonbunyang'(稻田糞壤 : techniques of practicing fertilizers in paddy) and the system of additional fertilization was established. The fertilization methods were improved with 'Jeobunbeob' due to the theory and recognition necessary for high rates of fertilizers.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.4
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• pp.287-293
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• 2001

Objective : To investigate the efficacy of high infusion frequency of liquid nitrogen on pregnancy in human embryo after freezing and thawing. Materials and Methods: 150 infertile patients underwent 162 consecutive thawing-ET cycles. In the high infusion frequency group (Group A), 47 patients (50 cycles) underwent cryopreservation with high infusion frequency of liquid nitrogen. In the low infusion frequency group (Group B), 103 patients (112 cycles) underwent cryopreservation with low infusion frequency of liquid nitrogen. We analyzed the clinical characteristics, fertilization rates, development of embryo, good quality embryo ratio, implantation rates, and pregnancy rates between these two groups. Results: There was no difference between the groups with regard to clinical characteristics (mean age, infertility duration, infertility factors, hormone profile), mean number of oocyte retrieval, fertilization rates, and mean embryo number of transfers. The survival rates in group A was 64.9% (228 of 350 embryos), and among the 228 embryos 190 embryos (83.3%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 65 (34.2%), 29 (15.3%), 35 (18.4%), and 37 (19.5%) of grades 1, 2, 3, and above 4, respectively. The survival rates in group B was 63.8% (482 of 755 embryos), and among the 482 embryos 465 embryos (96.5%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 106 (22.8%), 94 (20.2%), 89 (19.1%), and 112 (24.1%) of grades 1, 2, 3, and above 4, respectively. There was no difference in embryo quality change after the freezing-thawing procedure between the groups. Implantation rates (31.1% vs. 34.3%) were not significant. However hCG positive rates in group A (40%) were higher than group B, but not statistically significant. Clinical pregnancy rate (26% vs. 25.9%), on going pregnancy rates (>20 weeks) were not significant (26% vs. 25%). Conclusion: We compared embryo quality change, survival rates, and pregnancy rates between high infusion frequency group and low infusion frequency group and the results were similar between the two groups. Therefore, high infusion frequency of liquid nitrogen for cryopreservation is a worthy method to preserve in human embryos.

• Proceedings of the Korean Society of Crop Science Conference
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• 2005.04a
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• pp.106-107
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• 2005

• 대한생식의학회:학술대회논문집
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• 2002.11a
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• pp.93-94
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• 2002

• Korean Journal of Environmental Agriculture
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• v.19 no.4
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• pp.294-299
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• 2000

In order to find out the effects of shading and nitrogen fertilization on the accumulation of $NO_3\;^-$ in leaves of lettuce, lettuce plants were cultivated in the pots under glasshouse condition with different rates of shading(0, 50%) and nitrogen fertilization(100, 180, 200, 300, $400\;kg{\cdot}ha^{-1}$). The pH value was lower in soil after experiment than before experiment, whereas, contents of EC and $NO_3-N$ were higher. As the amounts of nitrogen fertilization were increased, pHs were decreased, but EC and the contents of $NO_3-N$ were increased. At the nitrogen fertilizations of 100, 180, 200, 300 and $400\;kg{\cdot}ha^{-1}$, the germination rates of lettuce were decreased to 84, 78, 76, 72 and 74%, and survival rates were also decreased to 94, 94, 90, 60 and 46%, respectively. However, the fresh weight of lettuce was highest at $45\;g{\cdot}plant^{-1}$ in the recommended fertilizer $plot(180\;kg{\cdot}ha^{-1})$ with non-shading condition. The contents of $NO_3\;^-$ in the leaves of lettuce were increased 2.8-4.1 times under 50% shading conditions than that under non-shading condition. It kept increasing up to seven order of growth phase; however, it started to decrease after eight order phase. Nitrate reductase activity of lettuce in non-shading condition was higher than that in 50% shading condition.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.153-159
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• 2016

(-)-Epicatechin gallate (ECG) is a polyphenol compound of green tea exhibiting biological activities, such as antioxidant and anticancer effects. To examine the effect of ECG on porcine oocytes during in vitro maturation (IVM), oocytes were treated with 0-, 5-, 15-, and $25{\mu}M$ ECG. After maturation, we investigated nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels and subsequent embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF). After 42 hours of IVM, the $5{\mu}M$ group exhibited significantly increased (p< 0.05) nuclear maturation (89.8%) compared with the control group (86.1%). However, the $25{\mu}M$ group observed significantly decreased (p< 0.05) nuclear maturation (83.5%). In intracellular maturation assessment the 5-, 15-, and $25{\mu}M$ groups had significantly increased (p< 0.05) GSH levels and decreased ROS levels compared with the controls. The 5- and $15{\mu}M$ group showed significantly increased (p< 0.05) embryo formation rates and total cell number of blastocysts after PA (18% and 68.9, 15% and 85.1 vs. 12% and 59.5, respectively) compared with controls. Although the $25{\mu}M$ group observed significantly lower blastocyst formation rates after PA (27.6% vs. 23.2%) than control group, the $5{\mu}M$ group showed significantly increased blastocyst formation rates after PA (37.2% vs. 23.2%) compared to the control group. Furthermore, the $5{\mu}M$ group measured significantly increased blastocyst formation rates (20.7% vs. 8.6%) and total cell number after IVF ($88.3{\pm}1.5$ vs. $58.0{\pm}3.6$) compared to the control group. The treatment of $5{\mu}M$ ECG during IVM affectively improved the porcine embryonic developmental competence by regulating intracellular oxidative stress during IVM.

• Journal of Aquaculture
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• v.17 no.1
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• pp.46-50
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• 2004

In the present study, attempts were made to cryopreserve sea urchin, Anthocidaris crassispina sperm in liquid nitrogen, to evaluate the effects of various cryoprotectants and freezing rates on motility, survival rate and fertilization rate of the post-thawing sperm, and the ultrastructural changes of sperm after cryopreservation were observed. The highest values of sperm motility (motility index: 3.3$\pm$0.37) and survival rate (72$\pm$3.5%) were obtained with 15% dimethyl sulfoxide (DMSO), and these values were significantly higher than those of sperm preserved with glycerol. Comparisons of motilities and survival rates between treatments of difference freezing rates showed that there was no difference between procedures (a) 5$0^{\circ}C$/min to -8$0^{\circ}C$ (motility index: 3.3$\pm$0.31 ; survival late 70$\pm$2.7%) and (b) 3$0^{\circ}C$/min to -8$0^{\circ}C$ (motility index: 3.1$\pm$0.29; survival rate 69$\pm$3.7%), while the results of (c) 1$0^{\circ}C$/min to -8$0^{\circ}C$ were significantly lower than the others (motility index: 2.2$\pm$0.33 ; survival rate 42$\pm$4.6%). There was no significant difference in fertilization rate between fresh sperm and sperm preserved with 15% DMSO as cryoprotectant and freezing rate (3$0^{\circ}C$/min to -8$0^{\circ}C$). Some ultrastructural changes of sperm, such as the detachment of plasma membrane, the destruction of mitochondria, and the flagellum rolling up head, were observed after cryopreservation. Morphological normality of the sperm in 15% DMSO frozen at the ratio of 5$0^{\circ}C$/min to -8$0^{\circ}C$ was better than the others.