• Journal of Embryo Transfer
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• v.21 no.1
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• pp.21-27
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• 2006

This study was conducted to determine the distribution of cat follicles among varying ages and produce oocytes from preantral follicles cultured in vitro. We used ovaries from 41 cats ranging in age from 0.3 to 5 years. Ovaries were obtained from cats undergoing routine ovariectomy at local veterinary clinics. As a prelude to in vitro culture of preantral follicles, the length and the width and the weight of ovaries among cats of varying ages were measured. Ovaries were fixed in 10% formalin, embedded in paraffin, cut into $3{\mu}m$-sections, mounted on slides and stained with hematoxylin and eosin. Follicles were evaluated at 200X and 400X magnification. Distribution of follicles among cats of varying ages were evaluated according to follicle classification: primordial, primary, transitional, preantral and antral follicles. Preantral follicles were isolated by the simple mechanical procedure. Each follicle was cultured in a well containing $100{\mu}l$ of medium 199 supplemented with 10% fetal bovine serum (FBS) or polyvinylalcohol (PVA) for 16 days. Follicle diameters were measured under inverted microscope every 4 days. The length, the width and the weight of ovaries were increased gradually according to ages but there was not significant difference among cats of varying ages. Majority of follicles were primordial follicles (84%) regardless of cat ages (p<0.05). Follicle diameter increased until 4 days of culture. However, period longer than 4 days of culture in vitro had a deleterious effect on follicle survival regardless of supplement (FBS or PVA). A few oocytes were collected from preantral follicles cultured in vitro. These basic reproductive techniques in domestic cats can be a useful tool to save endangered feline species.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.5
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• pp.373-384
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• 2001

Cellular proliferation is an intricately regulated process mediated by the coordinated interactions of critical growth control genes. Two of these factors in mammalian cells are the p53 and mdm-2 genes. A protein product of the mem-2 oncogene has been recently shown to associate with the protein encoded by the tumor suppressor gene p53. The p53 tumor suppressor protein is stabilized in response to DNA damage and other stress signals and causes the cell to undergo growth arrest or apoptosis, thus preventing the establishment of mutations in future cellular generations. Mutation or loss of p53 is a very common event in tumor progression. It occurs in about 50% of all tumors analysed including of colon, lung, breast and liver. The cellular mdm-2 gene, which has potential transforming activity that can be activated by overexpression, is amplified in a significant percentage of human sarcoma and in other mammalian tumors. Proteins encoded by the mdm-2 gene are able to bind to the p53 protein and, when overexpressed, can inhibit p53's transcriptional activation function, thus mdm-2 can act as a negative regulator of p53 function. Experimental study was performed to observe the relationship between p53 gene mutation and mdm-2 protein expression and apply the results to the clinical activity. 36 golden syrian hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek(control side) was treated with mineral oil as the same manner to the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were examined for histology and immunohistochemistry observation, and were completely dissected by microdissection and DNA from both tissue were isolated by proteins K/phenol/chloroform extraction. Segments of the hamster p53 exons 5, 6, 7 and 8 were amplified by PCR using the oligonucleotide primers, and then confirmational change was observed by SSCP respectively. The results were as follows : 1. Dysplasia at 6 weeks, carcinoma in situ at 8 weeks and invasive carcinoma from 10 weeks could be observed in experimental groups. 2. p53 mutations were detected in 10 of the 36(28%) and the exons 6(6 of the 10 : 60%) was the most hot spot area among the highy conserved region(exons 5, 6, 7 & 8). 3. Immunohistochemical study confirmed 22 of the 36(61%) of p53 expression involving 10 of p53 mutations. 4. mdm-2 expression of was showed in 3 of the 36(8%) involving 1 of the 22 of p53 expression and 2 of the 14 of p53 non-expression. From the above results, mutation of p53 gene or expression of p53 protein may have the influence of the DMBA induced carcinoma of hamster buccal pouch but the expression of mdm-2 protein may not have relationship with tumorigenesis.

• Journal of the Mineralogical Society of Korea
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• v.31 no.3
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• pp.161-172
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• 2018

Multi-anvil press (MAP) is one of the high pressure apparatuses and often generates the pressure-conditions ranging from 5 to 25 GPa and temperature-conditions up to $2,300^{\circ}C$. The MAP is, therefore, suitable to explore the pressure-induced structural changes in diverse earth materials from Earth's mantle and the bottom of the mantle transition zone (~660 km). In this study, we present the experimental results for pressure-load calibration of the 1,100-ton multi-anvil press equipped in the authors' laboratory. The pressure-load calibration experiments were performed for the 14/8 step, 14/8 G2, 14/8 HT, and 18/12 assembly sets. The high pressure experiments using ${\alpha}$-quartz, wollastonitestructure of $CaGeO_3$, and forsterite as starting materials were analyzed by powder X-ray diffraction spectroscopy. The phase transition of each mineral indicates the specific pressure that is loaded to a sample at $1,200^{\circ}C$: a transition of ${\alpha}$-quartz to coesite at 3.1 GPa, that of garnet-structure of $CaGeO_3$ to perovskite-structure at 5.9 GPa, that of coesite to stishovite at 9.2 GPa, and that of forsterite to wadsleyite at 13.6 GPa. While the estimated pressure-load calibration curve is generally consistent with those obtained in other laboratories, the deviation up to 50 tons is observed at high pressure above 10 GPa. This is partly because of the loss of oil pressure at high pressure resulting from the differences in a sample chamber, and the frictional force between pressure medium and second anvil. We also report the ${\sim}200^{\circ}C/mm$ of thermal gradient in the vertical direction of the sample chamber of 14/8 HT assembly. The pressure-load calibration curve and the observed thermal gradient within the sample chamber can be applied to explain the structural changes and the relevant macroscopic properties of diverse crystalline and amorphous earth materials in the mantle.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.2
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• pp.197-206
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• 2003

Physiological functions of raw grain diet composed of brown rice and Job's Tear (1 : 1) were evaluated in rats raised with nutritionally unbalanced diet including 1% cholesterol, high proportion of animal lipids (lard: soybean oil : 8 : 2), sub-optimal levels of vitamin and mineral mixture along with 0.5% ethanol in drinking water for 4 weeks. Control rats were fed the AIN-93G diet for 9 weeks, and nutritionally unbalanced rats were divided into 3 groups, and fed one of the following diets with 0.5% ethanol in drinking water for another 5 weeks: unbalanced control diet (UCD), raw grain diet (RGD) (UCD +20% brown rice and Job's Tear mixture, and cooked grain diet (CGD)(autoclaved RGD at 121$^{\circ}C$ for 3 hours). Feeding UCD for 5 weeks significantly lowered the food efficiency ratio (FER) of rats than the value for control animals, and dietary supplementation of brown rice and Job's Tear mixture to UCD significantly restored the FER. Serum total cholesterol concentration was significantly lowered in rats fed RGD (24% decrease) or CGD (16% decrease) compared to the value for rats fed UCD. Feeding RGD for 5 weeks significaly lowered the serum LDL+VLDL-cholesterol concentration (26% decrease), as well as the hepatic cholesterol level (16% decrease) than the values for UCD rats. Animals fed CGD (38% decrease) or RGU (59% decrease) showed significantly lower level of hepatic thiobarbituric acid reactive substances (TBARS) compared to the value for rats fed UCD (p<0.05), although hepatic activities of antioxidative enzymes were not influenced by dietary supplementation. Feeding RGD for 5 weeks significantly increased CD4$^{＋}$ T-cell population along with CD4$^{＋}$/CD8$^{＋}$ ratio of mesenteric lymph nodes compared to those for UCD rats (p<0.05). In conclusion, dietary supplementation of brown rice and Job's Tear mixture as raw grains exhibited superior activity lowering blood and hepatic levels of cholesterol, and improving mesenteric lymph nodes immune function of rats to the cooked grain mixture of identical ingredients.

• Journal of the Korean Institute of Traditional Landscape Architecture
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• v.28 no.2
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• pp.127-136
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• 2010

This study is aimed at reviewed and analyzed in order to suggest the improved plans related to natural monuments. The summary of this study is as followings; First, Replacing the current term 'cultural properties', which denotes the meaning of 'goods', we need to devise an new categorization that separates such properties into cultural heritage and natural heritage under the national heritage framework. Second, the designation criteria for natural monuments should be divided into the individual realm for animals and plants respectively, since they are not divided in the current Act. Third, the guidelines for naming of natural monuments should be established with the following new categories in accordance with the clear standards. Fourth, such imbalances require us to give priority to the relatively neglected types and areas. Fifth, as the big and old trees account for more than a half of the designated plants, it is necessary to search out new resources(wet plant communities, seashores, sand dune plant communities, etc.) such as geological resources, mineral springs, hot springs, and fossils that are in danger of completely being exploited and exhausted. While most of the designated animals are protected nationally, the existing designation system is required to protect habitats and breeding places for the systematic and efficient conservation. Sixth, as long as we need to preserve those historical and cultural resources for the future generations from national and global perspectives, we should enhance their values by designating them as natural monuments even though they are protected by other regulations such as the natural environment area. Seventh, as a result of the survey, we found that more budgets and experts in the local governments, more empowered organizations, more active public participation should be provided for the better Natural Monument management in Korea. Eighth, the Lap of Natural Heritage in the National Research Institute of Cultural Heritage needs to be developed to the Natural Heritage Institute to conduct the diverse activities such as researches, restoration, exhibition and education programs in a systematic and efficient way. Ninth and the last, major damages to natural monuments can be generally categorized into the artificial one and natural one, respectively. The artificial damages include toxics, soil covering, excessive humidity, fire, construction and management works, unlawful damages, fishing, oil spillage, etc, and the natural ones include lightning, storms(typhoons), heavy snowfalls, damage by insects and diseases, lack of prey, etc. This study will become meaningful in that it proposes specific measures for the improvement of the institutions, designation, and management of natural monuments on the basis of the comprehensive analysis on natural monuments. We wish to leave the other subjects related with this study to the future researches.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.365-373
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• 2000

The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P＜0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P＜0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

• Journal of Nutrition and Health
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• v.24 no.3
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• pp.230-242
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• 1991

This research was conducted on the 10 take-out boxed-lunches commercially prepared in the department stores. chain stores. and the public railroad trains in Korea. Sampling was conducted from February 1990 to March 1990. Nutritive values and sodium contents of the 10 boxed-lunch samples are summarized as follows : 1) The average weight(percentage) of the cooked rice and the side dishes were 304.6g(49.4) and 312.4(506%), respectively. The weight of these samples were significantly heavier than that of Japanese style boxed-lunches. 2) The average number of the side dishes was 12. The average numbers of food items classified by the five food groups were 6.1 in protein food group, 0.3 in calcium food group. 6.0 in vitamin and mineral food group. 1.5 in carbohydrate food group, and 1.5 in oil and fat food group. 3) They contained on the average 840.7kcal of energy, 38.9g of protein, 22.7g of fat, 120.4g of carbohydrate. 300.8mg of calcium. 410.8mg of phosphours, 6.61 mg of iron. 219.8 R.E. of vitamin A, 0.46mg of thiamin, 0.67mg of riboflavin, 10.5mg of niacin, 27.5mg of ascorbic acid. Thus. except vitamin t the content of all the nutrients were higher than the value of 1/3 of the RDA for adults. 4) The high priced group(group 2) had more protein, calcuim. iron and niacin contents than the cheaper group(group 1). Probably, it's because the group 2 had more animal foods than the group 1. 5) The average energy content per unit price(100 won) was 37.3kcal and the average protein content per unit price(100 won) was 1.64g. Korena style boxed-lunches had higher energy and protein contents per unit price than Japanese style, and the group 1 higher than the group 2. 6) The average energy Proportions of Protein, carbohydrate. and fat were 18.3%, 57.4%, and 24.3%, respectively. These proportions are good enough. 7) Frequency of cooking methods for the side dishes were found in the decreasing order : pan-frying, frying, braising, seasoning, kimchi, grilling, pickling, stir-frying, steaming and fermenting. Generally simple cooking methods were used, thus the menus were lack or varieties. 8) Frequency of colors for the side dishes were found in the decreasing order : red, brown. yellow, green, black, white. Too much red pepper was used. 9) The average capacity of the containers for the staples and the side dishes were 468.1ml and 590.6ml, respectively. And the containers could not keep the food items well seperated. 10) The average contensts of sodium and salt were 2.287mg and 5.76g, in the range of 1, 398mg to 3, 489mg and 3.53g to 8.80g, respectively. These are much higher values than the recommended amount of salt.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.181-185
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• 2004

This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.