• Korean Journal of Animal Reproduction
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• v.22 no.1
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• pp.1-9
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• 1998

This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

• Journal of the Korean Dietetic Association
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• v.19 no.2
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• pp.112-123
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• 2013

HbA1c was recently adopted as a reliable indicator for screening diabetes. This study investigated the ability of nutrition consultation to prevent diabetes in overweight women (BMI 23 $kg/m^2$ or more) using HbA1c as an indicator. Twenty overweight and obese women (with $HbA1c{\geq}5.7$%) completed the 12-week nutritional study, with individual and personalized nutrition counseling performed every 2 weeks. The main study guidelines involved the following: 1) reducing the intake of high fat foods and alcohol, 2) consuming a large amount of vegetables, 3) reducing the intake of simple sugars and empty-calorie foods, and 4) increasing physical activity to ${\geq}30$ min/day. Anthropometric (height, weight, BMI, body muscle (kg), body fat (%), waist and hip circumference, blood pressure) and biochemical parameters (fasting blood sugar (FBS), HbA1c, lipid profiles, hs-CRP) were measured before and after the nutrition consultation. After 12 weeks, the HbA1c<5.7% group had significant decreases in BMI, WC, HC, WHR, HbA1c, hs-CRP and also dietary intake of energy (P<0.01), carbohydrates, lipids (P<0.01), proteins (P<0.01) and cholesterol was significantly decreased (P<0.05). In the HbA1c ${\geq}5.7$% group, HbA1c, TC, LDL, NON-HDL, hs-CRP and dietary intake of energy, carbohydrate, lipid, protein, and cholesterol significantly decreased (P<0.05). These results suggest that nutrition consultation effectively helps to prevent diabetes in overweight and obese women after applying HbA1c standards. Overall, the improvement in all markers measured suggest that HbA1c is a good indicator for blood glucose regulation, helping to prevent diabetes.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.3
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• pp.647-656
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• 2017

Purpose: This study investigated the relationship between vitamin D level and dyslipidemia in Korean adults aged 19 years and older. The data for analysis were obtained from the sixth Korea National Health Nutrition Examination Survey 2013 and 2014. Result: This study showed that 74.4% of the specified adult population had vitamin D deficiency. Moreover, the prevalence of dyslipidemia was found to be 39.6% in the same population. Dyslipidemia differed significantly according to gender (p<0.001), age (p<0.001), smoking status (p<0.001), subjective health status (p<0.001), BMI (p<0.001), blood pressure (p<0.001), FBS (p<0.001), physical activity (p<0.001), and vitamin D level (p<0.001). When vitamin D deficiency was less than 20.0 ng/mL, there was a significant risk of dyslipidemia with an odds ratio of 1.29 (95% Cl, 1.11-1.51). The relationship between vitamin D level and a diagnosis index of dyslipidemia with controlled status of physical activity was statistically significant; in Korean adults, the odds ratio for total cholesterol was 1.41 (95% CI, 1.04-1.81) and for triglyceride was 1.36 (95% CI, 1.11-1.66). Conclusion: Our results showed the relationship between vitamin D level and dyslipidemia was significant in Korean adults, and total cholesterol and triglycerides, which are indicators of dyslipidemia, was significant. Future studies, such as a more systematic cohort study investigating the relationship between vitamin D level and dyslipidemia may be helpful in confirming the causal relationship between vitamin D and dyslipidemia.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.349-357
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• 1998

This study was to investigate whether the viability of Hanwoo IVM/IVF/IVC blastocysts was maintained after vitrification and thawing. In vitro produced Hanwoo blastocysts were vitrified by two-step method: equilibrated in EG20 for 3 min, and then exposed in EFS40 ［40% ethylene glycol (EG), 18% ficoll and 10.26% sucrose in mDPBS containing 10% FBS ］and vitrified in L$N_2$for 30 - 45 sec. After thawing, in vitro survival was assessed as the re-expanded and hatched rates at 24 hand 48 h, respectively. The results obtained in these experiments were summarized as follows: From the 12 replicates, 52.5% of Hanwoo blastocysts were produced in vitro at day 7 after IVF. When the effects of freezing solution to the embryo survival were examined, there is no significant toxicity in exposure (100.0, 73.8%) compared to that af control group (100.0, 87.0%). However, when embryos were vitrified, high survival (86.2, 55.4%) was obtained although it was significantly lower than those of exposure and control group (p＜0.05). When the in vitro survival of vitrified embryos according to developmental stage and culture day were examined, it showed that more advanced embryo stage exhibited a significantly higher survival rate irrespective of culture day (p＜0.05). Also, even in the same development stage, the in vitro survival of day 7 embryos (re-expanded: 75.0~87.5%, hatched: 21.4~66.7%) was higher than those of day 8 embryos(re-expanded: 58.6~78.3%, hatched: 10.3~52.2%). Therefore, these results suggested that in vitro produced Hanwoo blastocysts can be successfully cryopreserved by simple two-step vitrification method using EFS40 freezing solution, particularly at the expanded and early hatching blastocyst stage regardless of embryo culture duration (day 7 or day 8 after IVF).

• Development and Reproduction
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• v.12 no.3
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• pp.221-229
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• 2008

For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.

• Journal of Nutrition and Health
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• v.48 no.1
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• pp.46-57
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• 2015

Purpose: The aim of this study was to determine an appropriate energy level of breakfast with less risk of chronic disease for Korean adults. Methods: Using data from the 2007~2009 Korean National Health & Nutrition Examination Survey, from a total of 12,238 adults aged 19~64, the final 7,769 subjects were analyzed except subjects who were undergoing treatment for cancer or metabolic disorder. According to the percent of breakfast energy intake versus their estimated energy requirement (EER), the subjects were divided into four groups: < 10% (very low, VL), 10~20% (low, L), 20~30% (moderate, M), ${\geq}30%$ (sufficient, S). All data were analyzed on the metabolic risk and nutritional state after application of weighted value and adjustment of sex, age, residential area, income, education, job or jobless, and energy intake using a general linear model or logistic regression. Results: The subjects of group S were 16.9% of total subjects, group M 39.2%, group L 37.6%, and group VL 6.3%. The VL group included more male subjects, younger-aged (19 to 40 years), urban residents, higher income, higher education, and fewer breakfasts eaters together with family members. Among the 4 groups, the VL group showed the highest waist circumference, while the S group showed the lowest waist circumference, body mass index, and serum total cholesterol. The groups of VL and L with lower intake of breakfast energy showed high percent of energy from protein and fat, and low percent of energy from carbohydrate. With the increase of breakfast energy level, intake of energy, most nutrients and food groups increased, and the percentage of subjects consuming nutrients below EAR decreased. The VL group showed relatively higher intake of snacks, sugar, meat and eggs, oil, and seasonings, and the lowest intake of vegetable. Risk of obesity by waist circumference was highest in the VL group by 1.90 times of the S group and the same trend was shown in obesity by BMI. Risk of dyslipidemia by serum total cholesterol was 1.84 times higher in the VL group compared to the S group. Risk of diabetes by Glu-FBS (fasting blood sugar) was 1.57 times higher in the VL group compared to the S group. Conclusion: The results indicate that higher breakfast energy level is positively related to lower metabolic risk and more desirable nutritional state in Korean adults. Therefore, breakfast energy intake more than 30% of their own EER would be highly recommended for Korean adults.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.105-112
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• 2005

The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.528-533
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• 2009

In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.