• Applied Microscopy
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• v.33 no.2
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• pp.105-116
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• 2003

In Vitro fertilization of U. unicinctus eggs observed by immunofluorescence and electron microscopes revealed an overview of the meiotic pattern of the tide animals. The eggs have been fertilized early at germinal vesicle stage, followed by germinal vesicle break down (GVBD), but pre-mitotic aster like structure could not be resolved by the methods employed in this work. The meiotic features, such as rotation-shift movement of spindle fibers, behavior of spermatozoonmonaster in the egg cytoplasm and active spindle fiber of the 1st polar body, have been observed. The antitubulin-FITC fluorescence show the 2nd meiotic apparatus appeared firstly parallel to the tangential line of the oolemma, proceeding the meiosis, its bipolarity is rotated and shifted towards the oolemma. The polar bodysite of the oolemma was not amorphous, but active in a sense of anti-tubulin-FITC reactions during the extrusions of the polar bodies. The immunofluorescence reactions of the spermatozoon centriole appeared at a later stage of the 2nd meiosis. During the time periods, the fertilized spermatozoon resided in the egg cytoplasm. Activating the centrioles, spermatozoon approaches towards the chromosomal materials of the 2nd oocyte. This suggests that spermatozoon centrioles initiate and play a roll to fuse male and female pronuclei.

• Development and Reproduction
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• v.10 no.2
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• pp.115-125
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• 2006

The present experiment was performed to confirm the effects of selenium on maturation and viability of mouse oocyte. Maturation of oocytes was observed by microscope, Germinal vesicle breakdown(GVBD) and polar body formation(PB) were confirmed at 2.5, 13 hours after in vitro culture. Viability of oocytes was observed by microscope. Normal and abnormal oocytes were distinguished by morphological change in vitro culture for 72 hours. Glutathione(GSH) content of collected oocytes from individual stage also was measured by glutathione assay using spectrophotometer. The results obtained were as follows; The low concentration of selenium($0.005\;{\mu}g/mL{\sim}0.5\;{\mu}g/mL$) increased the maturation rate of germinal vesicle(GV) oocytes to GVBD and PB oocytes. The high concentration of selenium($5\;{\mu}g/mL$) decreased the maturation rate. The low concentration of selenium increased the viability rate of PB oocytes. The high concentration of selenium did not affect the viability rate. The low concentration of selenium increased the GSH content in PB oocytes. The high concentration of selenium decreased GSH content. GSH content in PB oocyte was much higher than that in GVBD oocyte. The results indicate that the low concentration of selenium increases the maturation rate by helping quality elevation of oocyte and minimizing damages of oxidative stress generated from metabolism process. The low concentration of selenium also increases the viability rate by increasing GSH content.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.229-237
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• 2004

The main objective of this study was to examine the effects of serum and gonadotropins supplement during in vitro maturation(IVM) of bovine oocytes on nuclear maturation and embryo development, and we also examine the cell number. 1 . The first polar body(PB) extrusion rates of Korean native cow(KNC) oocytes matured in medium with FBS or gonadotropins were similar among treatment groups. The development rate to the blastocyst stage was significantly higher in the group of both supplement FBS and gonadotropins(26.0％) than in the group of non-supplement(9.9％) and gonadotropins (12.0％). The numbers of inner cell mass (ICM) and trophectoderm (TE) cells and total cell numbers of blastocysts were highest in the group of both supplement FBS and gonadotropins, and the number of ICM cells was increased by FBS supplementation (p<0.05). 2. The PB extrusion rates of KNC oocytes matured in medium with FBS in the different duration of IVM was significantly higher in the 0-18hr(63.1％) and in the 9-18hr(63.4％) group than in the 0-9hr.(37.4％) group (p<0.05). The embryo development rates did not differ among treatment groups. The numbers of TE cells and total cell numbers of blastocysts were similar among treatment groups, but the number of ICM cells of the 0-18h. group were significantly higher than the other treatment groups (p<0.05). The results indicate that although TCM199 alone can support bovine oocyte maturation and development to the blastocyst stage, a high quality of blastocysts can be produced from oocytes matured in medium containing serum and gonadotropins.

• Journal of Aquaculture
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• v.10 no.2
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• pp.123-131
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• 1997

Growth of triploid abalone, Haliotis discus hannai induced by cody (3$^{\circ}C$) shock and its feed efficiency were investigated from larva to adult for 51 months. After 51 months from triploidy induction, the triploid abalones have outgrown to diploid abalones in shell length and total weight. Triploid abalones with inhibition of extrusion of first polar body (3n-1pb) were outgrown to diploid abalones, however, triploid abalones with inhibition of extrusion of second polar body (3n-2pb) were not significantly different from diploid controls in shell length and total weight through the whole rearing period (P<0.05), because of their heterozygosity differences. Daily feeding rates and feed conversion rates decreased with the growth of abalones and both rates had no differnce between two experimental groups. After 51 months from inducing triploid, conditin index of triploid abalone (64.1%) was higher than that of diploid control (59.4%) (P<0.05). Survival rate was 63.0% in triploid group (3n-1pb 62.0%, 3n-2pb 64.0%) and 62.0% in diploid group during the experimental period.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.29-36
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• 2004

The objective of this study was to investigate the effects of amino acid supplementation of oocyte maturation medium on 1st polar body(PB) extrusion, embryo development and blastocsyt cell number. In experiment 1, Cumulus oocyte complexes(COCs) were matured in in vitro maturation(IVM) medium supplemented with 1, 2, or 4-fold of 10 $\mu$l/ml MEM non-essential amino acid(NEAA) and 20 Park, $\mu$ l/ml BME essential amino acid(EAA). The PB extrusion rate of oocytes matured in 1-fold amino acid group was significantly higher than that matured in medium without amino acid (p＜0.05), but it was decreased by the increase of the dosage of amino acid. There were no difference in the percentage of embryos reaching 2-cell, 8-cell and blastocyst in all treatments. The number of trophectoderm(TE) cells and total cell number of blastocysts were highest in 2-fold amino acid group, and the number of inner cell mass(ICM) cells was increased by the increase of the dosage of amino acid. In experiment 2, COCs were matured in IVM medium with 1, 5, or 10 mg/ml lactalbumin hydrolysate(LAH). The PB extrusion rate of oocytes matured in medium with 5 mg LAH was significantly higher than that matured in medium with 1 mg LAH (p＜0.05). The development rate to the blastocyst stage was significantly higher in non-supplement and 1 mg LAH group than in 5 mg and 10 mg LAH group (p＜0.05). The number of TE cells and total cell number did not differ among treatment groups, but the number of ICM cells was increased by the increase of LAH supplement. These results suggested that the supplement of certain group of amino acid in IVM medium effective on the quality of blastocyst, and further studies will be accompany with the search of new sources of amino acid used for the use of in vitro embryo production.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.8
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• pp.1112-1121
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• 2019

Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.223-231
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• 2001

The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

• Journal of the Korean Applied Science and Technology
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• v.38 no.4
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• pp.951-962
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• 2021

The purpose of this study was to analyze the effects of smart machine circulation exercise on body composition, lung function, blood lipid and insulin resistance in obesity middle-aged women among 40-60 years by dividing them into a smart machine circulation exercise group(n=8), and control group(n=6). The smart machine circulation exercise program included 55-minutes sessions thrice each week at the following intensities: The strength of aerobic exercise is applied to smart machines by linking the smart machine with the POLAR T31; the 1-4 week is 40-50%HRR, 5-8 week is 50-60%HRR, and 9-10 week is 60-70%HRR. The strength of the resistance exercise was tested using a smart machine based on the constant velocity motion, and then, using the 1-RM data value, applied 40% 1-RM for 1-4 weeks, 60% 1-RM for 5-8 weeks, and 80% 1-RM for 9-10 week. As a results, body composition indicated that weight, BMI, %BF, WHR had a significant interaction effect. Lung function indicated that FVC levels significantly changes in the exercise group and the between groups difference in changes at 10week was significant. Also, FVC and FEV₁ significantly showed interaction effect. TC, TG and HDL-C levels significantly changes in smart machine circulation exercise group and the between-group difference in changes after 10 weeks was significant. TC, TG and HDL-C significantly showed interaction effect. Insulin resistance demonstrated that Insulin, Glucose and HOMA-IR levels significantly showed difference over 10 weeks between group. Therefore, the 10 weeks smart machine circulation exercise positively effects on the body composition, lung function, blood lipids, and insulin resistance in obesity middle-aged women and this smart machine circulation exercise can improve their obesity and prevent obesity.

• Journal of Aquaculture
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• v.6 no.2
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• pp.89-105
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• 1993

In order to investigate the effects of dietary n-3 highly unsaturated fatty acids (n-3HUF A) levels on growth and body composition of the Korean rockfish, six experimental diets with various levels of n-3HUFA, which were adjusted by adding squid liver oil and/or soybean oil at $8\%$ dietary lipid level, were fed to the Korean rockfish (6.2g in mean body weight) for 10 weeks. Daily weight gain, feed efficiency and nutrient retention efficiency were the lowest in the fish fed a diet containing 0% n-3HUFA. These parameters were effectively improved by supplementation with n-3HUFA, and showed linear increase up to $1.2\%$ dietary n-3HUFA level (P<0.01). There was no additional response above this level. A higher concentration of nonpolar lipids in the liver was observed for the fish fed insufficient levels of n-3HUFA in the diets. However the liver glycogen content and hepatosomatic index were slightly deareased. The lipid contents of the whole body and vicera showed significantly higher in the fish fed sufficient levels of n-3HUFA in the diets (P<0.05). The fatty acid compositions of polar lipids in the whole body and liver were affected by dietary fatty acid compositions. The contents of n-3HUFA and 18:1 in the liver increased with increasing the n-3HUFA level in the diets, while the 18:2n-6 and 18: 3n-3 decreased. These results suggest that n-3HUFA plays an essential role for normal growth of the Korean rockfish, and the requirement of n-3HUF A is around $1.2\%$ of the diet.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.5
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• pp.934-944
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• 2011

This study was designed to investigate the effects of Kyungohkgo to improve aerobic capacity and eliminate exercise-induced fatigue in high school soccer players. Twenty four subjects were participated and randomly assigned into two groups [KG, Kyungohkgo group (n=12); PG, Placebo control group (n=12)]. Two groups were completed treadmill exercise protocol using graded exercise test at before and after experimental treatment of 4 weeks. The $VO_2$max and endurance time were measured by gas analysis and heart rate (HR) was measured by polar system at pre, post 0, post 5, post 15, post 30 and post 60 minutes. Blood samples were collected to analyze blood components. 1. The $VO_2$max was significant increased in the group of after intake Kyungohkgo compared to the group of after intake placebo (p<.05). 2. The HR was significant decreased in the group of after intake Kyungohkgo compared to the group of after intake placebo during recovery time at post 5 mins(p<.05), 30 mins(p<.01), 60 mins(p<.01). 3. Weight, body mass index, percent body fat, anaerobic threshold, endurance time, blood lactate concentrate, lactate dyhydrogenase, creatine kinase, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, other energy sources(total-cholesterol, triglyceride, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, creatinine) and electrolyte (Na, K, Cl) were shown no significant differences between groups. These results suggested that Kyungohkgo can be used as ergogenic aids to improve aerobic capacity and eliminate exercise-induced fatigue.