• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.244-252
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• 2023

Objective: We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Methods: Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. Results: The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Conclusion: Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

• Applied Biological Chemistry
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• v.9
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• pp.119-124
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• 1968

In order to learn the effect of timing and rate of potash fertilization in the rice plant nursery on the root growth and the vegetative growth of later stage a pot experiment was carried out employing 'Jaekun' a rice variety. Potassium Chloride was applied at the rate of $0{\sim}100g\;K_{2}O/3.3m^2$ right before, and 30 days after seeding. Forty-day old seedlings were transplanted three times successively at the one week intervals with the roots cut each time. Each subject was observed in rooting and other useful traits. A part of seedlings after the third root scission were cultured intact thereafter and the yield characters examined. The results were shown as: 1. The rate of sound seedlings was high in the case that a small amount of potassium was applied as the basic placement while the application of the fertilizer over $75g/3.3m^2$ (as $K_{2}O$) yielded a far less crops. 2. The plant height, number of tillers, number of leaves, and grass weight increased as the amount of potassium was elevated. However, the application of $50g\;K_{2}O/3.3m^2$ in the basic placement and that of $75g\;K_{2}O/3.3m^2$ were the peaks over which the above mentioned characters were weakened. 3. As the amount of potassium was increased the increment in number of rooting was notable, especially when the scission of roots was repeated within the limit of $50{\sim}75g\;K_{2}O/3.3m^2$. 4. In the maximum root newly shooted length indicated was about the same tendency although no statistical significance was observed. 5. The plant height, number of tillers, and variation of weight between grass and root indicated a similar tendency as the number of root shoot. 6. The differences in number of ears, grain yield, and weight of straw between the treatments were not so great but showed somewhat similar trend as in the growth of transplanted plants. In the treatment-100g $K_{2}O/3.3m^2$ the yield (ears, grain and straw) decreased as in the non-fertilized.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.2
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• pp.165-176
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• 1994

This study was carried out to investigate the effect of the timing and dose of human chorionic gonadotropin(hCG) on oocyte recovery, in vitro fertilization, and preimplantation development in superovulation of mouse. F1 hybrid($C57BL{\times}CBA$) mice were obtained and superovulation was induced in female mice by sequential intraperitoneal injection of PMSG and hCG. In the first series of experiments, mice received 5 IU of PMSG given intraperitoneally, and 48 hours later were injected 1 IU, 5 IU, or 10 IU of hCG respectively. In the second series of experiments, mice received 5 IU of PMSG given intraperitoneally and were injected 5IU of hCG 36, 48, or 60 hours later respectively. 1. When the mice received 5 IU of PMSG given intraperitoneally and 48 hours later were injected 1 ItT, 5 IU, or 10 IU of hCG respectively, there were no differences in the total number of the oocytes obtained from the three experimental groups. When the cultures were examined 48 hrs after the termination of insemination the proportion of unfragmented oocytes which had developed over two-cell stage was observer to be lowest in 10 IU hCG group. When the cultrues were examined 120 hour after termination of insemination the proportion of embryos which had developed to the blastocyst stage was observed to be significantly higher in 10IU hCG group than 5IU hCG group(p<0.05), but there was no difference between 10 IU hCG group and 1IU hCG group. 2. When the mice received 5 IU of PMSG and were injected 5 IU of hCG 36, 48, or 60 hours later respectively, there were no differences in the total number of oocytes obtained from the three experimental groups. When cultures were examined 48 hour after the termination of insemination the proportion of unfragmented oocytes which had developed over two-cell stage was observed to be significantly lower in 36 hour interval group than 48 hour interval and 60 hour interval group(p<0.05). When the cultures were examined 120 hour after termination of insemination the proportion of embryos which had developed to the blastocyst stage was found to be higher in 60 hour interval group than 36 interval or 48 hour interval group (P<0.05), and the proportion of hatching blastocyst was found to be higher in 60 hour interval group as well. In this study, it was concluded that the administration of adequate dose of hCG, and long (60 hour) PMSG-hCG interval were necessary in superovulation of mice($C57BL{\times}CBA$) in order to get a large number of oocytes which had an early oocytes which had an early embryonic developmental capability when fertilized in vitro, and especially it had better have been avoided to administer a large dose of hCG.

• Korean Journal of Soil Science and Fertilizer
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• v.41 no.5
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• pp.303-309
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• 2008

The physico-chemical properties of the rainfall interception culture (85 sites) and the open field culture soils (85 sites) in the area of Gochang-gun, Jeollabuk-do were surveyed. Soil textural distribution of the rainfall interception culture and the open field culture soils was 74% and 64% for silt loam, 16% and 35% for loam, and 10% and 1% for clay, respectively. The percentage of aggregate rates was higher in the open field culture soils (60.06%) than rainfall interception culture soils (55.84%). Electrical conductivity, exchangeable cations, and anions in the rainfall interception culture soils were higher than those in open field soils. Specially, accumulated amount of anion in rainfall interception culture soils was remarkably higher 2~3 times than open field culture soils. The results from the analyses of rainfall interception culture soils suggested that the most critical problem is the salts accumulation caused by over-fertilization of chemical fertilizer and compost. Therefore, application rates of chemical fertilizer and compost should be controlled in order to conservation of soil and water for sustainable agriculture.

• Korean Journal of Environmental Agriculture
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• v.30 no.2
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• pp.105-110
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• 2011

BACKGROUND: In this study, 5 different treatments such as non-treatment, mixed expeller cake 1.0 N (standard nitrogen fertilizer), rice straw, rice straw+mixed expeller cake 0.5 N, rice straw+mixed expeller cake 1.0 N were performed over 4 cropping seasons over 2 years in order to identify the optimal application rate of mixture of rice straw and mixed expeller cake, organic source in organic cultivation of tomatoes. METHODS AND RESULTS: There was no difference in all treatments in case of 200 mg/kg in the nitrate nitrogen content in soil prior to the first cropping season test under the criteria for nitrogen nutrient based on yield of crops, cultivation without fertilizers seems possible. But in the second cropping season, no treatment and rice straw showed the reduction of yield and in the third cropping season, rice-straw+mixed expeller cake 0.5 N treatment showed the significant difference. The content of nitrate nitrogen in soil prior to cropping seasons was evaluated in 160 mg/kg and standard fertilization such as mixed expeller cake, source of nitrogen, are needed due to the deficiency of nitrogen. In terms of application of organic resources, rice straw showed the effects of improvements on physical properties of soil such as bulk density, cation exchange capacity and humus contents, but the mixed expeller cake did not show any significant differences in improvements on physical properties of soil. CONCLUSION(s): Fertilizer management in organic cultivation of tomatoes is thought to produce the reliable quantity of crops as well as keep the high quality of soils by using the optimal application rate of mixed expeller cake according to the contents of nitrate nitrogen in soil and rice straw which improves the physical properties of soil.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.285-291
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• 2007

Objective: To evaluate the outcomes of in vitro fertilization and embryo transfer (IVF-ET) in women over 40 years of age. Methods: A total of 170 patients (271 cycles) over 40 years of age who underwent IVF-ET at Seoul Women's Hospital (Incheon, Korea) were analyzed in this study retrospectively. The patients were grouped into the women <44 years old group and the women $\geq$44 years old group. Statistical analysis was performed using Student's t-test and Fisher's exact test as appropriate. Results: An overall clinical pregnancy rate per retrieval was 11% (30/271). Of these, clinical miscarriage rate were 33% (10/30) and the overall delivery rate was 7.4% (20/271) per retrieval, respectively. The women $\geq$44 years old group had significantly higher cancellation rate (13% vs. 25%), lower number of retrieved oocytes (6.17$\times$4.62 vs. 4.13$\times$4.07), decreased number of 2PN (4.83$\times$3.61 vs. 3.46$\times$3.12), and reduced embryos for transfer (3.52$\times$1.72 vs. 2.81$\times$1.83) than the women <44 years old group. We found significantly lower clinical pregnancy rate (13.0% vs. 2.1%) and live birth rate (9.0% vs. 0.0%) in the women $\geq$44 years old group than the women <44 years old group. Conclusion: The present study has shown that IVF outcome is seriously impaired in the women $\geq$44 years old.

• Korean Journal of Environment and Ecology
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• v.34 no.1
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• pp.85-92
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• 2020

The purpose of this study is to provide base information that can be utilized in surveying the status of landscape management within apartment complexes and grasping the conditions of the soil for planting plants, as a way to improve the quality of green spaces by repairing poorly vegetated spaces within apartment complexes and remodeling them with diverse species of plants. Considering that trees and shrubs of equal sizes are simultaneously planted upon the completion of apartment complexes, they were divided based on their completion year at the interval of 5 years and differences in the growth of trees and the components of soil over time were analyzed. Out of the species of trees planted in all the 9 apartment complexes surveyed in this study, the growth of 4 species of needleleaf trees and 4 species of broadleaf trees were surveyed. Juniperus chinensis 'Kaizuka' and Pinus densiflora Siebold & Zucc. out of the needleleaf trees and Malus floribunda Siebold ex Van Houtte out of the broadleaf trees showed the highest growth rate when over 5 years passed after planting and their growth rate decreased when over 10 years passed. Platycladus orientalis and Acer palmatum Thunb. in the apartment complexes that were built over 10 years ago showed the highest growth rate, which indicates that the species require a relatively long period of time for growth. The hardness of the soil at the areas where trees were planted but their soil surface was bare was analyzed. When over 5 years passed after the completion of apartment complexes, over 20 mm of the soil was found to be stamped. The physicochemical properties of soil were also surveyed and the pH level was found to have been continuously high ever since the completion. The organic content in the surveyed soil was about 1/3 of the content in fertile soil, which means that additional fertilization is required. These results indicate that the stamped soil and the health of soil can be restored, when replanting plants in bare areas, by adding plans to improve soil, such as designing drain ways around the planted areas, transporting soil for the 50 cm depth of the ground and mixing organic matters such as chaff, and simply by planting groundcover plants in the lower part of tress and shrubs.

• Journal of Korean Society of Forest Science
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• v.66 no.1
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• pp.109-117
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• 1984

Allozyme study for open pollinated seeds of forty nine pitch pine families in an $F_1$-hybrid seed orchard demonstrated that allozyme variants in aspartate aminotransferase(AAT), glutamate dehydrogenase(GDH) and leucine aminopeptidase(LAP) systems are encoded by at Least eight loci; five for AAT, one for GDH and two fur LAP. Allozyme variations showed polymorphisms at seven of the eight loci, except GDH. Average number of alleles examined over six loci were 2.33 and 2.67 for maternal and progeny groups, respectively. Average heterozygosity and genetic diversity computed over six loci were, respectively, 0.235 and 5.409 for maternal tree group, 0.238 and 5.569 for progeny group. The proportion of $F_1$-hybrid seeds estimated by allozyme analysis was 0.77%. The estimated proportion of $F_1$ hybrid seeds by allozyme study is in good agreement with the value 0.73% estimated by morphological study for the proportion of pitch ${\times}$ Loblolly $F_1$ hybrid seedlings at a nursery. Indications for Wahlund effect, high levels of self-fertilization and for non-random matings in the $F_1$ hybrid seed orchard call for cautions in estimating allele frequency changes and mating probabilities for the parental and progeny groups.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.10a
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• pp.37-43
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• 2001

1. About fifty thousand of cattle embryos were transferred and 16000 ET-calves were born in 1999. Eighty percents of embryos were collected from Japanese Black beef donors and transferred to dairy Holstein heifers and cows. Since 1985, we have achieved in bovine in vitro fertilization using immature oocytes collected from ovaries of slaughterhouse. Now over 8000 embryos fertilized by Japanese Black bull, as Kitaguni 7~8 or Mitsufuku, famousbulls as high marbling score of progeny tests were sold to dairy farmers and transferred to their dairy cattle every year. 2. Embryo splitting for identical twins is demonstrated an useful tool to supply a bull for semen collection and a steer for beef performance test. According to the data of Dr. Hashiyada(2001), 296 pairs of split-half embryos were transferred to recipients and 98 gave births of 112 calves (23 pairs of identical twins and 66 singletons). 3. A blastomere-nuclear-transferred cloned calf was born in 1990 by a joint research with Drs. Tsunoda, National Institute of Animal Industry (NIAI) and Ushijima, Chiba Prefectural Farm Animal Center. The fruits of this technology were applied to the production of a calf from a cell of long-term-cultured inner cell mass (1988, Itoh et al, ZEN-NOH Central Research Institute for Feed and Livestock) and a cloned calf from three-successive-cloning (1997, Tsunoda et al.). According to the survey of MAFF of Japan, over 500 calves were born until this year and a glaf of them were already brought to the market for beef. 4. After the report of "Dolly", in February 1997, the first somatic cell clone female calves were born in July 1998 as the fruits of the joint research organized by Dr. Tsunoda in Kinki University (Kato et al, 2000). The male calves were born in August and September 1998 by the collaboration with NIAI and Kagoshima Prefecture. Then 244 calves, four pigs and a kid of goat were now born in 36 institutes of Japan. 5. Somatic cell cloning in farm animal production will bring us as effective reproductive method of elite-dairy- cows, super-cows and excellent bulls. The effect of making copy farm animal is also related to the reservation of genetic resources and re-creation of a male bull from a castrated steer of excellent marbling beef. Cloning of genetically modified animals is most promising to making pig organs transplant to people and providing protein drugs in milk of pig, goat and cattle. 6. Farm animal cloning is one of the most dreamful technologies of 21th century. It is necessary to develop this technology more efficient and stable as realistic technology of the farm animal production. We are making researches related to the best condition of donor cells for high productivity of cloning, genetic analysis of cloned animals, growth and performance abilities of clone cattle and pathological and genetical analysis of high rates of abortion and stillbirth of clone calves (about 30% of periparutum mortality). 7. It is requested in the report of Ministry of Health, labor and Welfare to make clear that carbon-copy cattle(somatic cell clone cattle) are safe and heathy for a commercial market since the somatic cell cloning is a completely new technology. Fattened beef steers (well-proved normal growth) and milking cows(shown a good fertility) are now provided for the assessment of food safety.

• Journal of Embryo Transfer
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• v.30 no.4
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• pp.289-298
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• 2015

Edaravone (Eda) is a potent scavenger of inhibiting free radicals including hydroxyl radicals ($H_2O_2$). Reactive oxygen species (ROS) such as $H_2O_2$ can alter most kinds of cellular molecules such as lipids, proteins and nucleic acids, cellular apoptosis. In addition, oxidative stress from over-production of ROS is involved in the defective embryo development of porcine. Previous study reported that Eda has protective effects against oxidative stress-like cellular damage. However, the effect of Eda on the preimplantation porcine embryos development under oxidative stress is unclear. Therefore, in this study, the effects of Eda on blastocyst development, expression levels of ROS, and apoptotic index were first investigated in preimplantation porcine embryos. After in vitro fertilization, porcine embryos were cultured for 6 days in PZM medium with Eda ($10{\mu}M$), $H_2O_2$ ($200{\mu}M$), and Eda+$H_2O_2$ treated group, respectively. Rate of blastocyst development was significantly increased (P<0.05) in the Eda treated group compared with only $H_2O_2$ treated group. And, we measured intracellular levels of ROS by DCF-DA staining methods and investigated numbers of apoptotic nuclei by TUNEL assay analysis is in porcine blastocyst, respectively. Both intracellular ROS levels and the numbers of apoptotic nucleic were significantly decreased (P<0.05) in porcine blastocysts cultured with Eda ($10{\mu}M$). More over, the total cell number of blastocysts were significantly increased (P<0.05) in the Eda-treated group compared with untreated group and the only $H_2O_2$ treated group. Based on the results, Eda was related to regulate as antioxidant-like function according to the reducing ROS levels during preimplantation periods. Also, Eda is beneficial for developmental competence and preimplantation quality of porcine embryos. Therefore, we concluded that Eda has protective effect to ROS derived apoptotic stress in preimplantation porcine embryos.