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Effect of Taxol Pre-treatment to In Vitro Matured Bovine Oocytes on Spindle Morphology and Embryonic Development Following Vitrification

  • Park, Sang-Hyoun;Yu, Il-Jeoung
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.269-274
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    • 2008
  • The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.

Characterization of canine coronaviruses isolated from the dogs with diarrhea in Korea (설사증 이환견(犬)으로 부터 분리(分離)한 canine coronavirus의 성상(性狀)에 관한 연구(硏究))

  • Lee, Byung-hyung;Jun, Moo-hyung;Park, Jong-hyeon;Hwang, Eui-kyung;Huh, Won
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.517-527
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    • 1994
  • An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

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Red Ginseng Saponin Fraction A Isolated from Korean Red Ginseng by Ultrafiltration on the Porcine Coronary Artery

  • Jung, Young-Hyun;Park, Kwang-Yeol;Jeon, Jin-Hong;Kwak, Yi-Seong;Song, Yong-Bum;Wee, Jae-Joon;Rhee, Man-Hee;Kim, Tae-Wan
    • Journal of Ginseng Research
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    • v.35 no.3
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    • pp.325-330
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    • 2011
  • Red ginseng saponin fraction-A (RGSF-A) contains a high percentage of panaxadiol saponins that were isolated from Korean red ginseng by ultrafiltration. The aim of this study was to elucidate the effects of RGSF-A on the porcine distal left anterior descending (LAD) coronary artery. The relaxant responses to RGSF-A were examined during contractions induced by 100 nM U46619 (9,11-dideoxy-9a,11a-methanoepoxy-prostaglandin F2a), a stable analogue of thromboxane A2. RGSF-A dose-dependently induced biphasic (fast- and slow-) relaxation in the distal LAD coronary artery in the presence of an intact endothelium. The fast-relaxation was quickly achieved in a minute, and then the slow-relaxation was slowly developed and sustained for more than thirty minutes after the administration of RGSF-A. The slow-relaxation had a tendency to be bigger than the fast-relaxation. Fast relaxation induced by RGSF-A was almost blocked by $N_{\omega}$-Nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase synthase inhibitor and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a guanylate cyclase inhibitor. However slow relaxation induced by RGSF-A was only partially inhibited by L-NAME and ODQ. In the endothelium-removed ring, RGSF-A evoked only slowrelaxation to a certain extent. These data suggest that RGSF-A induced both endothelium dependent fast- and slow-relaxation and endothelium independent slow-relaxation in the porcine distal LAD coronary artery. The endothelium dependent fast-relaxation is mediated by the nitric oxide (NO)-cGMP pathway, and the endothelium dependent slow-relaxation is at least partially mediated by the NO-cGMP pathway. However, the endothelium-independent slow-relaxation remains to be elucidated.

Studies on the Cause and Control for Non-Sprouting in the Mulberry Field of Kyung-pook Province (경북지방의 뽕밭에 발생한 발아불량현상의 원인 및 방제에 관한 연구)

  • Yu, Geun-Seop;Kim, Gyu-Rae;Kim, Nak-Sang
    • Journal of Sericultural and Entomological Science
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    • v.28 no.2
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    • pp.1-8
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    • 1986
  • This study was conducted to bring light on inducing factors for non-sprouting occured in the mulberry field of Kyungpook Province in 1983. The results are as follws ; 1. In spring, winter buds were suddenly died during germination, necrosis appeared in pholem and cortex of non-sprouting stem and measles at the bottom of non-sprouting stem. 2. The depth of available of soil was more shallow in the non-sproutiong field than in healthy field. 3. There was no signifioant difference between the healthy field and non-sprouting field in siol PH, the content of organic matter, available phosphorous and exchangeble cations. 4. Available boron content in soil was significantly more in healthy field than in non-sprouting field. 5. Boron content in leaf and bark was significantly lower in the non-sproutiong tree than in the healthy tree. 6. The non-sprouting fields were completely controlled by the application of 6kg/10a borax. 7. Cold tolerance of the mulberry tree was higher in the mulberry tree with boron than in the mulberry tree without boron. 8. Mulberry tree in Kyungpook Province in 1983 may absorb water earlier compared with the average year because of high temperature at the end of March, 1983 and they had been damaged by frost injuary due to the lowest temperature (-6.4$^{\circ}C$) on the grass at the beginning of April 1983. As the above results, non-sprouting bud in the mulberry fields of Kyungpook Province in 1983 had been occured by low temperature and boron deficiency and boron deficiency of mulberry trees accelerared the frost injury.

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Environmental Factors and Variation Characteristics of Zooplankton Communities in Gamak Bay (가막만 동물플랑크톤 군집의 변동특성과 환경요인)

  • Moon, Seong-Yong;Yoon, Ho-Seop;Soh, Ho-Young;Choi, Sang-Duk
    • Ocean and Polar Research
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    • v.28 no.2
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    • pp.79-94
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    • 2006
  • Spatio-temporal patterns in the zooplankton communities, physicochemical environmental factors, and dominant species were examined in Camak Bay from April, July, September, and November, 2003. A total of 40 taxa were sampled with a mean abundance of total zooplankton ranging from $270{\sim}1,803inds/m^3$. The most abundant species was Acartia erythraea, followed by Eurytemora pacifica, Podon polyphemoides, A. omorii, and Paracalanus parvus s. 1 during the study period. Multivariate analysis revealed significant differences in community structure among three regions: the middle part of the sampling area (B), sites 1, 2 (A) and other sites (C). The number of species and diversity (H') varied significantly among the three regions (p<0.05), but no differences in the mean abundances were observed (p>0.05). In order to investigate factors affecting the correlations between the abundance of dominant species, all zooplankton and environmental factors ware calculated using a correlation analysis. Of these, the abundance of dominant taxon showed positive correlation with seawater temperature, salinity and dissolved oxygen (p<0.05). In statistical analysis showed that the environmental factors influencing the variation of the communities ere predominantly temperature, salinity, and dissolved oxygen.

Response of Rice Growth under $CO_2$ Enrichment ($CO_2$ 농도 증가에 따른 벼의 생육 반응)

  • Kim Young-Guk;Shin Jin-Chul;Choi Min-Gyu;Koo Bon-Cheul;Kim Seok-Dong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.3
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    • pp.179-185
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    • 2005
  • The effects of $CO_2$ enrichment on growth of rice (Oryza sativa L.) were examined. The plants were grown in growth chambers with a 12-h photoperiod and a day/night temperature of $28/21^{\circ}C$ of the seedling stage and $30/23^{\circ}C$ after the panicle initiation stage. The plants were exposed to two elevated $CO_2$ of 500, 700 ppm and ambient levels (350 ppm). At early growth stage of three varieties (IIpumbyeo, Chucheongbyeo, Hwaseongbyeo), the elevated $CO_2$ increased plant height, tiller, leaf area and dry weight. The photosynthetic rate was decreased at 24 days after treatment (DAT) compared to 11 DAT. The elevated $CO_2$ increased plant height and dry weight at panicle initiation stage (PIS) and heading stage (HS) of three varieties (IIpumbyeo, Chucheongbyeo, Hwaseongbyeo). The photosynthetic rate, stomatal conductance, evapotranspiration rate were decreased at the long days of treatment than that of short days. At entire stages, the elevated $CO_2$ increased the water use efficiency of rice plant because evapotranspiration rate was lowered at the elevated $CO_2$ than ambient levels.

Cloning and Characterization of Ginsenoside-Hydrolyzing β-Glucosidase from Lactobacillus brevis That Transforms Ginsenosides Rb1 and F2 into Ginsenoside Rd and Compound K

  • Zhong, Fei-Liang;Ma, Rui;Jiang, Mingliang;Dong, Wei-Wei;Jiang, Jun;Wu, Songquan;Li, Donghao;Quan, Lin-Hu
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1661-1667
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    • 2016
  • The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30℃), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.

Study on the Soil Improvement in the Grassland II. Effects of the dolomite particles and the shell powder application on soil characteristics, dry matter yield and nutritive value of forages in sandy loam soil (초지에서 토양 개량에서 관한 연구 II. 사양토에서 도로마이트 입자도와 패각분 시용이 토양 특성과 목초의 수량 및 사료가치에 미치는 영향)

  • Lee, J.K.;Choi, S.S.;Kim, M.J.;Yoon, S.H.;Park, G.J.;Choi, K.C.;Lee, S.C.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.4
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    • pp.373-378
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    • 1999
  • This study was conducted to investigate the effects of application of the dolomite particles and the shell powder on soil characteristics, dry matter yield and nutritive value of forage in sandy loam soil at the Experimental Field of National Livestock Research Institute, Suweon, from 1994 to 1996. This was compared with control, lime, dolomite 0.5mm, 2.0mm, 4.0mm, and shell powder in mixed pasture. Although there was no significant difference, average dry matter yield for 3 years was increased with the application of lime, dolomite 2.0mm, 0.5mm, control, 4.0mm and shell powder in order. Average crude protein yield of forages was increased with the application of dolomite 2.0mm, 0.5mm, control, lime, shell powder and 4.0mm in order. Lime requirement was slightly increased from 2,410 to 4,853kg per ha with the lapse of time. Although soil hardness was optimum level by second year, it was apt to become hard little by little. Solid phase of soil was lowered with dolomite 0.5mm treatment. The results demonstrated that dolomite and shell powder could be settled a dust problem in farms and a coast pollution as lime substitutes for soil improvement. Therefore, it is desirable for dry matter yield and crude protein yield to applicate the dolomite 2.0mm every 3 years and the shell powder more frequently in sandy loam soil.

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Effect of Genotype and Dietary Protein Level on Growth Performance and Carcass Characteristics of Fattening Pigs in Central Vietnam

  • Pham, Khanh Tu;Hoang, Nghia Duyet;Le Duc, Ngoan;Hendriks, W.H.;Van Der Peet-Schwering, C.M.C.;Verstegen, M.W.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.8
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    • pp.1034-1042
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    • 2010
  • This study aimed to determine the optimum dietary crude protein level in a typical diet for fattening pigs fed ad libitum under normal climate conditions in Central Vietnam. One hundred and ninety two gilts of Mong Cai local breed (MC), $F_1$ Large White${\times}$Mong Cai and $F_2$ crossbreds of (Landrace${\times}$Mong Cai)${\times}$Large White were used. At the start of the experiment, Mong Cai pigs weighed 12 kg at 11 weeks of age, $F_1$ pigs 12.1 kg at 8 weeks of age and $F_2$ pigs 12.2 kg at 8 weeks of age. Four diets differing in crude protein (CP) content (10.1, 13.1, 16.1 and 18.9% in DM) were formulated from rice bran, corn meal, cassava meal and fish meal. Calculated digestible energy content of the diets ranged from 13.5 to 13.8 MJ per kg DM. Pigs were housed individually in pens of $2.5\;m^2$ each and had ad libitum access to feed in a trough as well as water in bowls. The final weights after a growing period of 150 days were 66, 86 and 96 kg for MC, $F_1$ and $F_2$, respectively. Feed intake of MC pigs was highest at 13.1% CP while $F_1$ and $F_2$ had the highest feed intake at 16.1% CP. The results showed that for MC the maximum gain was obtained at levels between 13 to 16% CP. For the $F_1$ the maximum gain was at dietary protein levels of 16-17%. For $F_2$ the max gain was obtained at CP levels of 16 to 18%. Feed conversion was highest in MC pigs (~4.0) followed by $F_1$ (~3.3) and $F_2$ (~3.1), and within genotypes was lowest at the optimum CP level (p<0.05). Back fat thickness in MC (33.1 mm), $F_1$ (23.0 mm) and $F_2$ (20.5 mm) pigs was different and within genotypes was the lowest at intermediate CP levels. In conclusion, increasing the dietary crude protein contents in practical diets for pigs in Vietnam can increase production on small holder farms. Optimal performance for MC, $F_1$ and $F_2$ pigs is achieved at different dietary crude protein contents.

Emission Character of Dioxins and Precursors in the Control Devices of the MSWI (II) (도시쓰레기 소각로 방지시설 중 다이옥신류 및 전구물질의 배출특성(II))

  • Shin, S.K.;Chung, Y.H.;Lee, W.S.
    • Analytical Science and Technology
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    • v.12 no.1
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    • pp.68-74
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    • 1999
  • The Concentrations of PCDDs/PCDFs and their precusors(chlorophenols, chlorobenzenes, PCB) were analyzed from the dioxin control device such as EP and SCR to know the emission patterns of these compounds and find the dioxin index compounds. The dioxin concentration increased 7 times in outlet part than inlet part of EP and the concentration of CBs, CPs and PCBs also were increased through this control device. These phenomia may be related to the operating temperature of Electroprecipitator(EP), which the operating temperature is near the $300^{\circ}C$, the method of the decreasing the operating temperature need to consider to prevent the formation of these compounds. In the selected catalytic reactor with wet scrubber(SCR+WS), these compounds were removed after passing the device over 90% for CPs, 30~40% for CBs and 60% for PCBs. But, the systematic study have to perform to reduce the formation of PCDDs/PCDFs and precusors.

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