• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.245-252
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• 1990

This study was carried out to investigate the factors affecting fertilization in vitro of follicular oocytes with frozen-thawed spermatozoa in Korean Native Cattle. The bovine ovaries were obtained at a slaughter house and the follicular oocytes were recovered by aspirating the follicular fluid from the visible follicles of 3~6mm. The bovine oocytes were matured in vitro for 20~24 hours in TCM-199 containing FCS and hormones. The matured oocytes were fertilized in vitro using Percoll-separated frozen-thawed spermatozoa in BO solution. The effects of dilution and fertilization media, capacitating method, concentration of inseminated sperm and time after insemination of fertilization, were observed. The results obtained are summarized as follows : 1. The fertilization rate of frozen-thawed sperm inseminated in BO solution with caffeine and heparin together(56.4%) was higher than that of sperm inseminated in BO solution with either caffeine(10.5%) or heparin(8.9%) and without both caffeine and heparin(0%)(P<0.05). 2. The fertilization rate(56.3%) of frozen-thawed sperm inseminated in BO solution with both caffeine and heparin without preincubation was higher than that of sperm preincubated(2.9%)(P<0.05). 3. The fertilization with high concentration of frozen-thawed sperm(1.4~1.8$\times$107cells/ml) in BO solution containing caffeine and heparin resulted in higher fertilization rate, 76.7%, than the low concentration of sperm(0.8~1.0$\times$107cells/ml), 32.7%(P<0.01). 4. When the oocytes were inseminated with frozen-thawed sperm in BO solution containing caffeine and heparin without preincubation, fertilization rate increased by time and the rates were 5.9, 46.0 and 59.4% at 8, 16 and 24 hours, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.155-160
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• 2001

Objective: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. Materials and Methods: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. Results: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). Conclusion: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospennic patients.

• Korean Journal of Environmental Agriculture
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• v.31 no.3
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• pp.205-211
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• 2012

BACKGROUND: Generally, nitrogen (N) fertilization higher than the recommended dose is applied during vegetable cultivation to increase productivity. But higher N fertilization also increases the concentrations of nitrate ions and nitrous oxide in soil. In this experiment, the impact of N fertilization was studied on nitrous oxide ($N_2O$) emission to standardize the optimum fertilization level for minimizing $N_2O$ emission as well as increasing crop productivity. Herein, we developed $N_2O$ emission inventory for upland soil region during red pepper and Chinese milk vetch cultivation. METHODS AND RESULTS: Nitrogen fertilizers were applied at different rates to study their effect on $N_2O$ emission during red pepper and Chinese milk vetch cultivation. The gas samples were collected by static closed chamber method and $N_2O$ concentration was measured by gas chromatography. The total $N_2O$ flux was steadily increased due to increasing N fertilization level, though the overall pattern of $N_2O$ emission dynamics was same. Application of N fertilization higher than the recommended dose increased the values of both seasonal $N_2O$ flux (94.5% for Chinese cabbage and 30.7% for red pepper) and $N_2O$ emission per unit crop yield (77.9% for Chinese cabbage and 23.2% for red pepper). Nitrous oxide inventory revealed that the $N_2O$ emission due to unit amount of N application from short-duration vegetable field in fall (autumn) season (6.36 kg/ha) was almost 70% higher than that during summer season. CONCLUSION: Application of excess N-fertilizers increased seasonal $N_2O$ flux especially the $N_2O$ flux per unit yield during both Chinese cabbage and red pepper cultivation. This suggested that the higher N fertilization than the recommended dose actually facilitates $N_2O$ emission than boosting plant productivity. The $N_2O$ inventory for upland farming in temperate region like Korea revealed that $N_2O$ flux due to unit amount of N-fertilizer application for Chinese cabbage in fall (autumn) season was comparatively higher than that of summer vegetables like red pepper. Therefore, the judicious N fertilization following recommended dose is required to suppress $N_2O$ emission with high vegetable productivity in upland soils.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.3
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• pp.293-299
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• 1995

Objective: To investigate the effects of sperm morphology and their co-incubation with oocytes on the outcome of IVF and ICSI. Design: Strict morphology of washed sperm was assessed by Diff-Quick staining method before or after insemination. And the relationships between strict morphology and outcome (fertilization, embryo development and pregnancy) of IVF(with co-incubation) and ICSI (without co-incubation) were determined. Patients: Two-hundreds-and-sixty-three cycles of IVF and ninety-six cycles of ICSI were analyzed in order to clarify the influence of strict sperm morphology of spermatozoa on outcome of IVF and ICSI. These were divided into four groups. according to fertilization method and sperm morphology(Group 1: IVF, ${\geq}$12%, n:227; Group 2: IVF, <12%, n:36; Group 3: ICSI, ${\geq}$ 12%, n=48; Group 4: ICSI, <12%, n=48). Results: The fertilization rates of better morphology groups were higher than those of poor groups: Group 1(68.1%) > Group 2(62.1%), Group 3(78.1%) > Group 4(71.5%). There was no difference in embryo cleavage rates among four groups (>90%), Regarded with the good embryo rates, Group 1(56.8%) was significantly higher than Group 2(42.3%)(P<0.01), but there was no difference between Group 3(64.7%) and Group 4(61.2%). The pregnancy rates were also higher in better morphology groups as well as fertilization rates: Group 1(34.8%)> Group 2(16.7%)(p<0.05), Group 3(40.0%) > Group 4(23.0%)(p=0.08). Conclusion: Co-incubation with poor morphology sperm might adversely affect the quality of embryos. And strict sperm morphology may represent the ability to establish successful pregnancy. In short, the strict sperm morphology can be a good predictor of IVF and ICSI outcome.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.1
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• pp.41-48
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• 2007

Objective: To determine whether the presence of Y-chromosome microdeletion affects the outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. Methods: Fourteen couples with microdeletion in azoospermic factor (AZF)c region who attempted IVF/ICSI or cryopreserved and thawed embryo transfer cycles were enrolled. All of the men showed severe oligoasthenoteratoazoospermia (OATS) or azoospermia. As a control, 12 couples with OATS or azoospermia and having normal Y-chromosome were included. Both groups were divided into two subgroups by sperm source used in ICSI such as those who underwent testicular sperm extraction (TESE) and those used ejaculate sperm. We retrospectively analyzed our database in respect to the IVF outcomes. The outcome measures were mean number of good quality embryos, fertilization rates, implantation rates, $\beta$-hCG positive rates, early pregnancy loss and live birth rates. Results: Mean number of good quality embryos, implantation rates, $\beta$-hCG positive rates, early pregnancy loss rates and live birth rates were not significantly different between Y-chromosome microdeletion and control groups. But, fertilization rates in the Y-chromosome microdeletion group (61.1%) was significantly lower than that of control group (79.8%, p=0.003). Also, the subgroup underwent TESE and having AZFc microdeletion showed significantly lower fertilization rates (52.9%) than the subgroup underwent TESE and having normal Y-chromosome (79.5%, p=0.008). Otherwise, in the subgroups used ejaculate sperm, fertilization rates were showed tendency toward lower in couples having Y-chromosome microdeletion than couples with normal Y-chromosome. (65.5% versus 79.9%, p=0.082). But, there was no significance statistically. Conclusions: In IVF/ICSI cycles using TESE sperm, presence of V-chromosome microdeletion may adversely affect to fertilization ability of injected sperm. But, in cases of ejaculate sperm available for ICSI, IVF outcome was not affected by presence of Y-chromosome AZFc microdeletion. However, more larger scaled prospective study was needed to support our results.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.171-178
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• 1999

The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF. And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF (Table 1). The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF. The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% in control, 8.77% in 10% FF and 20% in 40% FF (Table 2). On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively (Fig. 1, 2, 3). In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after $8{\sim}16$ cell stage.

• Korean Journal of Soil Science and Fertilizer
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• v.15 no.3
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• pp.178-187
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• 1982

This pot experiment was undertaken to find out the optimum fertilization ratios of total anions to total cations, ${\Sigma}A/{\Sigma}C$, and the optimum application rates of total macro-nutrients in various soil conditions. Soil samples were collected from uncultivated mountains and hills where grassland development was under consideration. 1. The optimum application ratios of ${\Sigma}A/{\Sigma}C$ and the optimum application rates of total macro-nutrients for the high yields of mixed grass-clover sward in various grassland soils were computed by the Hom$\acute{e}$s systematic variation techniqu.e. 2. With respect to the optimum application ratios of ${\Sigma}A/{\Sigma}C$ in fertilization in a mixed grass-clover sward, the grass yield and botanical composition were distinctly proportional to ${\Sigma}A$ wheras the regume yield and botanical composition were proportional to ${\Sigma}C$. 3. The optimum fertilization rates of total macro-nutrients for the high legume yields were depended upon ${\Sigma}A/{\Sigma}C$ ratios. These optimum rates were in proportional to ${\Sigma}C$ ratios and were inversely proportional to ${\Sigma}A$ ratios. 4. The efficiencies of ${\Sigma}A$ and ${\Sigma}C$ in relation to the grass and grass plus legume yields were highest with the low ratios of each other and the low fertilization rates of total macronutrients. The ${\Sigma}A$ effieiency in the legume yield tended to be similar to that of ${\Sigma}A$ related to the grass yield noted above except Daegu soil. The ${\Sigma}C$ efficiency, however, was proportional to the ${\Sigma}C$ ratio, although that was varied with the fertilization rates of total macro-nutrients and with the kinds of soils. 5. The yield of mixed forages, yield component, and botanical composition in a mixed sward were greatly influenced by the ${\Sigma}A/{\Sigma}C$ ratios, the fertilization rates of total macronutrients, and the interactions of ratio and rate noted above. In addition, these effects were generally different and opposite according to grass and legume.

• Journal of Korean Society of Forest Science
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• v.109 no.3
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• pp.271-280
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• 2020

This study compared soil CO₂ efflux rates after fertilization, in Pinus densiflora and Quercus variabilis stands. Compound fertilizers were applied to the forest floor in March 2016, following a one-year calibration period (from March 2015 to February 2016). In situ soil CO₂ efflux rates were measured every month during the two-year study periods, using an infrared gas analyzer with a closed chamber system. Mean annual soil CO₂ efflux rates were higher following fertilizer application in the P. densiflora and Q. variabilis stands (P. densiflora: 2.180 μmol m^-2 s^-1; Q. variabilis: 1.977 μmol m^-2 s^-1) as compared with the rates measured during the calibration period (P. densiflora: 1.620 μmol m^-2 s^-1; Q. variabilis: 1.557 μmol m^-2 s^-1). The mean annual soil CO₂ efflux rates in the unfertilized treatments of both stands were not significantly different between the two-year study periods. The Q₁₀ values of fertilized treatments in Q. variabilis stands were higher in the fertilization period (3.41) than in the calibration period (3.14), whereas the Q₁₀ values in P. densiflora stands did not change between the fertilization and calibration periods. The Q₁₀ values of unfertilized treatments in the Q. variabilis stands were lower during the 2016-2017 period (3.69), than in the 2015-2016 period (3.85), whereas the Q₁₀ values in P. densiflora stands were higher during the 2016-2017 period (3.65), than in the 2015-2016 period (3.15). These results indicate that the increase in soil CO₂ efflux rates in P. densiflora stands could be more sensitive to fertilization compared with the rates in Q. variabilis stands.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.127-132
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• 1993

Frozen semen obtained from 5 different Korean Native Cattle were used for in vitro fertilization of oocytes. The proportions(3∼11%) of oocytes penetrated in the basic medium with 13.9 mM glucose were very low in different semen. However, the penetration rates of oocytes in the presence of caffeine and heparin increased to 68∼84%. The proportions(10∼52%) of oocytes penetrated in the presence of caffeine alone were low at various glucose concentrations(0∼27.8 mM). In the medium with heparin, significantly(P<0.05 at least) higher penetration rate was obtained in the absence(94%) than in the presence(28∼45%) of 0∼27.8 mM glucose concentrations. The medium with caffeine and heparin, however, produced similar penetration rates(84∼97%) regardless of the various glucose concentrations.

• Journal of The Korean Society of Grassland and Forage Science
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• v.2 no.1
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• pp.31-34
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• 1980