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In Vitro/In Vivo Development after Thawing of Vitrified Mouse Blastocysts by Culture Condition and Embryo Transfer Method (초자화 동결된 생쥐 배반포기배의 융해 후 배양조건과 수정란 이식방법에 따른 체외/체내발달)

  • Kim, M.K.;Kim, E.Y.;Yi, B.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.347-353
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    • 1997
  • This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

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Effect of Semisulcospira libertina Extracts from Different Extraction Processes on Liver Cell Toxicity and Ethanol Metabolism (간세포 독성과 에탄올 대사에서 추출 조건에 따른 다슬기 추출물의 효과)

  • Cho, Kyoung Hwan;Choo, Ho Jin;Seo, Min Gyun;Kim, Jong Cheol;Shin, Yu Jin;Ryu, Gi Hyung;Cho, Hee Young;Jeong, Chi-Young;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.158-166
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    • 2017
  • Although Semisulcospira libertina is generally regarded as a supplement for the alleviation of alcohol hangover, little is known about its effects on cell metabolism. Therefore, this study was conducted to analyze the constituents of the extracts prepared using different extraction methods and to compare their biochemical properties. The amino acid contents were found to be much higher in acidic and enzymatic hydrolysates than hot water extracts from S. libertina. DPPH radical scavenging activities in acidic and enzymatic hydrolysates were higher than those of hot water extracts. Three types of S. libertina hydrolysate was added to HepG2 cells damaged by acetaminophen (AAP), after which the survival rate of HepG2 cell were measured. In addition, lactate dehydrogenase (LDH) activities in the culture media were evaluated. The survival rates of HepG2 cells were $77.0{\pm}4.3%$ and $81.5{\pm}1.3%$ at 3 h and 5h enzymatic hydrolysates, respectively. These cell survival rates were higher compared to those of the negative control group ($67.8{\pm}4.3%$) treated only with acetaminophen. Cellular toxicities induced by treatment with AAP were also significantly alleviated in response to treatment with the extracts of S. libertina. In addition, the activities of 2 key enzymes that metabolize ethanol, alcohol dehydrogenase and aldehyde dehydrogenase, were upregulated by 4.7- and 2.7-fold respectively in response to treatment with a 3 h enzymatic hydrolysate of S. libertina. Taken together, these results provide biochemical evidence of the method by which S. libertina exerts its biological functions, including the alleviation of alcohol hangover and the protection of liver cells against toxic insults.

Effects of Shelf Temperature on the Quality of Red Pepper (Capsicum annuum L.) after Low Temperature Storage (홍고추의 저온저장 후 모의 유통온도가 품질에 미치는 영향)

  • Park, Sung Min;Kang, Won Hee;Lee, Yun Soo;Kim, Il Seop;Jeong, Cheon Soon
    • Horticultural Science & Technology
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    • v.19 no.4
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    • pp.526-529
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    • 2001
  • The study was conducted to investigate quality changes of red pepper by different shelf temperatures after storage at $7^{\circ}C$ for 30 days. Red pepper fruits produced high volume of $C_2H_4$ and $CO_2$ when alternating temperatures exceeded $10^{\circ}C$. Total sugar contents were not different among all the treatments. In contrast, sucrose content decreased about half at higher shelf temperature conditions than at $7^{\circ}C$. Capsaicinoid content was not influenced by alternating temperature, but ascorbic acid content tended to decrease by high shelf temperature after storage at $7^{\circ}C$. Decay did not occur under $13^{\circ}C$, while it occurred in 22.2% of hits at $15^{\circ}C$ and 68.8% at room temperature, respectively. From this results, it is suggested that the optimum shelf temperature of red pepper after low temperature storage should be under $13^{\circ}C$ and that fruits of red pepper stored at $7^{\circ}C$ for 30 days could be sold within 10 days on the shelf.

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Follicular Stimulation and Laparoscopic Ovum Pick-up (LOPU) in Repeatedly Superovulated Korean Black Goats (반복적인 과배란 처치 경험이 있는 한국흑염소에서 난포 자극 및 복강경을 이용한 난포란 채취(LOPU))

  • Lee, Yong-Boum;Lee, Doo-Soo;Cho, Sang-Cheol;Shin, Sang Tae
    • Journal of Embryo Transfer
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    • v.29 no.1
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    • pp.35-41
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    • 2014
  • Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot ($20mg{\times}6/300IU$) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ${\pm}$ SEM) observed which developed to larger than 2 mm in diameter were $1.6{\pm}2.5$, $4.3{\pm}3.1$, $5.5{\pm}4.2$, $6.6{\pm}2.1$ and $8.8{\pm}7.8$, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + $10{\mu}g/ml$ FSH + $1{\mu}g/ml$ $17{\beta}$-estradiol for 27 h at $39^{\circ}C$ in 5% $CO_2$ in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6-DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.

A Study on the Spatial Information and Location Environment of Dead Coniferous Tree in Subalpine Zone in Jirisan National Park -Focus on Korean Fir(Abies koreana) in Banyabong, Yeongsinbong, Cheonwangbong- (지리산국립공원 아고산대 침엽수 고사개체 공간정보 구축 및 입지환경 분석 - 반야봉, 영신봉, 천왕봉 일원 구상나무를 중심으로-)

  • Park, Hong Chul;Moon, Geon Soo;Lee, Ho;Lee, Na Yeon
    • Korean Journal of Environment and Ecology
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    • v.34 no.1
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    • pp.42-54
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    • 2020
  • This study aimed to analyze the rate of increase and spread patterns of dead trees of the conifer (Abies koreana and others) in subalpine zones by using the high-resolution aerial images in Jirisan National Park around 10 years ago. Furthermore, factors affecting the death of conifer were identified by analyzing the altitude, topographical information, solar radiation, and moisture environment of the site where the dead trees are located. The number of dead trees per unit area increased by two to five times in the Banyabong peak, Yeongsinbong peak, and Cheonwangbong peak in Jirisan National Park over the past decade. The increase was about 2 times in the Banyabong peak, about 3.9 times in the Yeongsinbong peak, and about 5.2 times in the Cheonwangbong peak, indicating the most notable increase in the Cheonwangbong peak. It is estimated that dead trees commonly occurred in the environments where the soil moisture content was low due to the high slope, amount of evaporation was high due to strong solar radiation as the location faced south, and the soil was dry due to strong solar radiation and short rain retention time. In other words, dead conifer trees in subalpine zones were concentrated in dry location environments, and the tendency was the same more than ten years ago.

Study on the Manufacturing of Chitosan Using Krill(Euphausia superba Dana) and Quality Characteristics (크릴을 이용한 키토산 제조 및 품질 특성)

  • Do, Jeong-Ryong;Park, In-Sung;Rhee, Seong-Kap;Kim, Dong-Soo
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.309-313
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    • 2000
  • For the use of Antartic krill(Euphausia superba Dana) as food resource, general composition, extracting condition of chitin and quality characteristics of chitosan were investigated. General composition of frozen krill(Euphausia superba Dana) was consisted of moisture 79.0%, protein 13.1%, lipid 4.0%, VBN 7.7mg%, ash 2.7%, others 1.2% and that of dried krill powder was moisture 5.6%, protein 56.1%, lipid 18.8%, ash 11.4%, others 8.1%. The condition of chitin extraction from krill powder was treated with 1N NaOH at $40^{\circ}C$ for removing protein, 1N HCl for excepting mineral substances and methanol for decoloring. The yield of chitin by new procedure developed was 3.7%. The composition of extracted chitin contents was moisture 7.1%, ash 0.4%, protein 3.5%, lipid 3.1%. The results of degree of deacetylation in chitosan at 50% NaOH, $121^{\circ}C$, for 2 hrs was showed 82%. At the same alkali concentration and reaction concentration, a longer reaction time gave a decreased degree of deacetylation. The apparant viscosity was 0.09241 Pa in 1% chitosan from krill and 0.13826 Pa in standard chitosan.

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A Study on the Anaerobic Treatment of the Phenol-bearing Wastewater with two Sludge Blanket-Packed Bed Reactors in Series (2단의 슬러지-고정상 반응기에서 페놀 함유 폐수의 혐시성 처리에 관한 연구)

  • 정종식;안재동;박동일;신승훈;장인용
    • Journal of Environmental Health Sciences
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    • v.21 no.4
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    • pp.1-9
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    • 1995
  • This study was carried to investigate the biodegradability of phenol in the wastewater with the two sludge blanket-packed bed reactor in series. Each reactor had a dimension of 0.09 m i.d. and 1.5 m height and consisted of two regions. The lower region was a sludge blanket of 0.5 m height and the upper region was a packed-bed of 1 m height. The packed bed region was charged with ceramic raschig rings of 10 mm i.d., 15 mm o.d. and 20 mm length. The reactors were operated at 35$\circ$C and the hydraulic retention time(HRT) was maintained 24 hours. The synthetic wastewater composed of glucose and phenol as major components was fed into the reactor in a continuous mode with incereasing phenol concentration. In addition, the nutrient trace metals($Na^+, Mg^{2+}, Ca^{2+}, PO_4^{3-}, NH_4^+, Co^{2+}, Fe^{2+}$ etc.) were added for growing anaerobes. The phenol concentration of the effluent, the overall gas production, the composition of product gas, the efficiency of COD reduction and the duration of acclimation period were measured to determine the performance of the anaerobic wastewater treatment system as the phenol concentration of the influent was increased from 600 to 2400 mg//l. Successfully stable biodegradation of phenol could be achieved with the anaerobic treatment system from 600 to 1, 800 mg/l of the influent phenol concentration. The upper level of influent phenol loading was high enough to meet most of the practical requirement. The duration of acclimation increased with the phenol loading. At steady state of the influent phenol concentration of 1800 mg/l, the treatment performance indicated the phenol reduction efficiency of 99%, the COD reduction efficiency of 99% and the gas production rate of 37 l/day. At the influent phenol concentration of 2400 mg/l, however, the operation of the treatment system was noted unstable. While the concentration of methane in biogas decreased with increasing the influent phenol loading, the carbon dioxide was increased. However, the concentration of hydrogen was varied negligibly. The concentration of methane was high enough to be used as a fuel. As a result, it is suggested that anaerobic phenol wastewater treament was economical in the sense of energy recovery and wastewater treatment.

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Effect of seaweed extracts(GA14), a plant growth regulator, on growth and yield of two rice cultivars (식물생장조절제 Seaweed extracts(GA14)의 수도 품종간 생육 및 수량에 미치는 효과)

  • Song, Jae-Young;Kim, Yong-Il;Park, Bo-Young;Jung, Jae-Young;Choi, Hyun-Gu;Jung, Jong-Tae;Lee, Hee-Bong
    • Korean Journal of Agricultural Science
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    • v.31 no.1
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    • pp.1-8
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    • 2004
  • This study was carried out to identify the effects of seaweed extracts(GA14) on growth of two rice cultivars, Junambyo and Donganbyo. Seedling qualities of two cultivars were better in all items including heading dates at early stage treated at seedling plus 2-3 leaf stages than at single treatment of seedling. Ripening ratio of Junambyo in paddy field was increased 0.3% by seaweed extracts(GA14) treatment, but that of Donganbyo decreased 0.5%. 1,000 grain weight of Junambyo and Donanbyo by seaweed extracts(GA14) treatment was two to four grams higher and the yield of two cultivars was also higher by three to four percent. Appearance characters of two rice cultivars was high in head, while damaged, chalky and crack rate were low at seaweed extracts(GA14) treatment. Protein, moisture and amylose characteristics related to table quality of Junambyo were not different by seaweed extracts treatment, but table values was high in only treatment. Donganbyo was also similar to Junambyo, but table quality was slightly high at non-treatment.

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Comparison of Semen Characteristics, Frozen-Thawed Sperm Viability, Testosterone Concentration and Embryo Development between Yorkshire Boar A and B

  • Yi, Y.J.;Lee, S.H.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.612-616
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    • 2004
  • This study was carried out to compare the semen characteristics, frozen-thawed sperm viability and testosterone concentration and in vitro fertilization (IVF) and development of in vitro matured pig oocytes between two Yorkshire boars. Semen and blood samples were collected once per week from October to November 2002 from two adult Yorkshire boars at 18 months of age with 170 kg body weight. Sperm were deep frozen in 5 ml maxi-straws with lactose-egg yolk and N-acetyl-D-glucosamine (LEN) diluent and stored in liquid nitrogen. Blood samples were obtained at 10 a.m. by inserting a 21 gauge, hypodermic needle attached to 10 ml syringe into surface veins in the ear. The concentration of testosterone was determined by Competitive Enzyme Immunoassay. Ovaries were collected from prepubertal gilts at a local slaughter house. Cumulus oocyte complexes were aspirated from antral follicles (3 to 6 mm in diameter). The medium used for oocyte maturation was modified TCM 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at $38.5^{\circ}C$, 5% $CO_2$ in air. For IVF, one frozen 5 ml straw was thawed at $52^{\circ}C$in 40 sec and was diluted with 20 ml Beltsville thawing solution at room temperature. Sperm were washed 2 times in mTLP-PVA and inseminated without preincubation after thawing. Oocytes were inseminated with $2{\times}10^7$/ml sperm concentration. Oocytes were coincubated for 6 h in 500 ${\mu}$l mTBM fertilization medium. At 6 h after IVF, oocytes were transferred into 500 ${\mu}$l NCSU-23 culture medium for further culture of 48 and 144 h. There were no significant differences in the semen volume, motility, normal acrosome morphology and sperm concentration of raw semen between A and B of Yorkshire boar. However, motility and normal acrosome of boar A were higher than those of boar B at 0.5, 2, 3, 4, 5 and 6 h incubations of frozen-thawed sperm. Testosterone concentration (3.75 ng/ml) of boar A was higher than that (2.34 ng/ml) of boar B. The rate of blastocyst formation (15.1%) of boar A was higher than that (10.4%) of boar B. In conclusion, serum testosterone concentration of boar showed very important role for the frozen-thawed sperm viability and the blastocyst formation of pig oocytes matured in vitro.

Effects of Coated Compound Proteases on Apparent Total Tract Digestibility of Nutrients and Apparent Ileal Digestibility of Amino Acids for Pigs

  • Pan, L.;Zhao, P.F.;Yang, Z.Y.;Long, S.F.;Wang, H.L.;Tian, Q.Y.;Xu, Y.T.;Xu, X.;Zhang, Z.H.;Piao, X.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.12
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    • pp.1761-1767
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    • 2016
  • Two experiments were conducted to evaluate effects of coated compound proteases (CC protease) on apparent total tract digestibility (ATTD) of nitrogen (N) and energy, and apparent ileal digestibility (AID) of amino acids (AA) and nutrients in diets for pigs. In Exp. 1, 12 crossbred barrows (initial body weight: $20.79{\pm}1.94kg$) were housed in individual metabolism crates and allotted into 2 treatments with 6 piglets per treatment according to weight in a randomized complete block design. The 2 diets were corn-soybean meal basal diets with (0.2 g/kg) or without CC protease supplementation. The CC protease supplementation increased (p<0.05) the digestible and metabolizable N and energy values and the digestibility and retention rate of N in the diet. The ATTD of energy and nutrients had been improved (p<0.05) in the diet supplemented with CC protease. In Exp. 2, 12 crossbred barrows (initial body weight: $20.79{\pm}1.94kg$), fitted with T-cannulas at the distal ileum, were blocked by body weight into 2 groups with 6 pigs each. The diets were the same as those in Exp. 1. The CC protease increased (p<0.05) the AID of crude protein and some essential AA including arginine, isoleucine and leucine. The AID and ATTD of energy and nutrients had been improved (p<0.05) by supplemental CC protease, but the hindgut digestibility of nutrients was unaffected. Overall, the CC protease improved the ATTD of N and energy and AID of some indispensible AA and nutrients in the corn-soybean meal diet for pigs. Therefore, the CC protease supplement could improve the utilization of protein in the corn-soybean meal diet and thus contribute to lower N excretion to the environment.