• Title/Summary/Keyword: 3-D ICM

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Effect of droplet vitrification on mitochondrial membrane potential and developmental competence in two-cell mouse embryos

  • Kim, Bo-Hyun;Kim, Ji-Su;Ryu, Jae-Sung;Lee, So-Hyun;Lee, Ju-Taek;Kang, Jae-Yul;Chang, Kyu-Tae;Choo, Young-Kug
    • Animal cells and systems
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    • v.15 no.4
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    • pp.287-294
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    • 2011
  • The accelerated cooling rate associated with vitrification reduces injuries attributed to cryopreservation and improves the post-freezing developmental competence of vitrified embryos. In this study, embryos were vitrified and warmed and morphologically evaluated for their development to blastocysts. Survival rates between the fresh ($96.7%{\pm}3.8%$) and vitrified embryos ($90.7%{\pm}5.1%$) did not differ significantly (P>0.05). The mitochondrial membrane potential of fresh control cells measured by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanide iodide staining was similar to that of cryoprotected and vitrified embryos. Mitochondrial staining with rhodamine 123 did not differ among the fresh, cryoprotected, and vitrified embryos. Moreover, the distribution of $H_2O_2$, assessed by 2',7'-dichlorodihydrofluorescein diacetate staining, did not differ among the groups. The results showed that the developmental rate did not differ significantly among the fresh ($87.8%{\pm}11.3%$), cryoprotected ($83.2%{\pm}7.6%$), and vitrified 2-cell embryos ($75.8%{\pm}14.2%$). The mean number of the inner cell mass (ICM), trophectoderm (TE), and apoptotic cells was counted and statistically compared, and although the number of ICM and TE was decreased in the cryoprotected and vitrified embryos, there were no significant differences among the groups (P>0.05). During the cultivation period, randomly selected blastocysts from each group were stained using either 4',6-diamidino-2-phenylindole and bisbenzimide or the terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling technique. The incidence of apoptosis appeared to be almost identical in all the groups. Droplet vitrification could subsequently lead to high survival and developmental rates of cryopreserved mouse embryos.

Effect of 6-Dimethylaminopurine and Cycloheximide Treatments on Parthenogenetic Development of Porcine Follicular Oocytes (6-Dimethylaminopurine 및 Cycloheximide의 처리가 돼지난포란의 단위발생에 미치는 영향)

  • Kim, Jong-Hwa;Park, Byung-Kwon;Han, Man-Hye;Lee, Kyu-Seung
    • Korean Journal of Agricultural Science
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    • v.34 no.1
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    • pp.1-11
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    • 2007
  • This study is aimed at finding an optimum density for 6-Dimethylaminopurine (6-DMAP) and cycloheximide which have an effect on the revitalization of porcine oocytes. The results were summarized as follows: 1. When 6-DMAP was treated with 2 mM for 2 hours, It showed a significantly (P<0.05) different high result in activation rate, cleavage rate and blastocyst growth rate of 51.2%, 52.7% and 25.2% respectively. 2. When Cycloheximide was treated with 5 ug/ml for 6 hours, It showed a significantly (P<0.05) different high result in activation rate, cleavage rate and blastocyst growth rate of 47.7%, 46.8%, and 27.3% respectively. 3. When it was cultured in the culture medium, NCSU, for 7 days after inducing activation with 6-DMAP and cycloheximide, it showed no differences in the number of inner cell mass (ICM) and total cell of blastocysts. To conclude, it has been examined for porcine oocytes to be suitable when 6-DMAP was treated with 2 mM for 2 hours, Cycloheximide with 5 ug/ml for 6 hours.

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Simulation of Circulation and Water Qualities on a Partly Opened Estuarine Lake Through Sluice Gate (배수갑문을 통해 부분 개방된 하구호에서의 순환과 수질모의)

  • 서승원;김정훈;유시흥
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.14 no.2
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    • pp.136-150
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    • 2002
  • To improve the water quality of the recently constructed Siwhaho, sluice gates were operated to allow free exchange of water with the sea. This estuarine lake connected to the outer sea through narrow gates is affected mainly by flushing by gate operation and river flows and wind forcing sometimes. As a predicting tool far the water qualities, a three-dimensional finite volume model CE-QUAL-ICM is incorporated into a finite element hydrodynamic model, TIDE3D. In coupling these two different modules, a new error minimization technique is applied by considering conservation of mass. Model tests for one year after calibration and validation using field observation show that eutrophication and other biological changes reach quasi-steady state after initial 60 days of simulation, thus it would be necessary to consider moderate ramp up option to remove initial uncertainties due to cold start option. Sediment-water interaction might not be a concern in the long-term simulation, since its effect is negligible. Simulated results show the newly applied scheme can be applied with satisfaction not only fur lessening of eutrophic processes in an estuarine lake but also looking for some active circulation to improve water quality.

The Effect of Cryopreservation on the Mouse Embryos at Various-pronuclear Stages

  • Park, M.C.;Kim, J.Y.;Kim, S.B.;Park, Y.S.;Park, H.D.;Lee, J.H.;Oh, D.S.;Kim, Jae-myeoung
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.174-180
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    • 2009
  • This study was carried out to establish an appropriate condition for the efficient cryopreservation of the mouse pronuclear embryo. In vitro cryopreservation of pronuclear embryos was carried out by slow freezing or vitrification methods and development rate of 2-cell, blastocyst and hatched blastocyst was measured as well as survival rate of the thawed pronuclear embryo. After slow freezing, vitrification and thawing of mouse pronuclear embryos, the survival rate and blastocyst development rate for the vitrification group was 97.3 and 53.4%, respectively, which was significantly higher as compared to the slow freezing group with 88.6 and 23.9%, respectively (p<0.05). Blastocyst developmental rate in each experimental group was significantly higher for 21 h in the post-hCG group at 40.5-57.0% than the 24 h post-hCG group at 40.5% (p<0.05). ICM (Inner cell mass) cell numbers of blastocyst-stage embryos during the different stages of mouse pronuclear embryos, slow freezing and vitrification period in the control and vitrification groups were 22.1${\pm}$2.7 and 17.0${\pm}$3.1-22.0${\pm}$3.2, respectively; hence, the slow freezing group (10.2${\pm}$2.0) had significantly higher cell numbers than those of the other two groups (p<0.05). Trophoblast (TE) cell number in the control group, 65.8${\pm}$12.6, was significantly higher than in the slow freezing group, 41.6${\pm}$11.1 (p<0.05). The total cell numbers in the control group and 21 h post hCG group were 87.9${\pm}$13.6 and 81.8${\pm}$14.1, respectively, and were significantly higher than for the slow freezing group (51.8${\pm}$12.6; p<0.05).

Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes II. Effects of EGF and the Number of COCs into Maturation Media on In Vitro Maturation, Fertilization and Development (돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 II. 체외성숙배양시 EGF와 COC의 수가 체외성숙, 체외수정 및 체외발달에 미치는 영향)

  • Yeon, S.-H.;Son, D.-S.;Han, M.-H.;Wee, M.-S.;Choi, S.-H.;Lee, K.-S.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.173-183
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    • 2004
  • This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.

Effect of Amino Acids Supplemented to Culture Medium on Development of Porcine Embryos Culturde in Vitro (아미노산의 첨가가 돼지 체외수정란의 후기배의 발달에 미치는 영향)

  • Kim Y. S.;Song S. H.;Cho S. K.;Kwack D. O.;Kim C. W.;Park C. S.;Chung K. H.
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.201-205
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    • 2005
  • The objective of this study was to investigative the effects of amino acids supplementation on maturation, fertilization and embryo development of pig oocytes. Essential amino acids (EA), non-essential amino acids (NA) or both amino acids (EA + NA) were supple-mented to North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (pFF). When the amino acids were supplemented to the maturation medium, the maturation rates were higher (p<0.05) in the NA group than control ($83.3{\pm}0.04\%\;versus\;70.0{\pm}0.05\%$, but the subsequent cleavage rates and development to morula and blstocyst stage between aminoacid supplement groups and control were not different. The developmental rates to morula and blastocysts stage were not significantly different regardless of amino acid supplementation to culture medium. In addition, supplementation of amino acids did not significantly affect the rate of fertilization and polyspermy. When the amino acids were supplement to culture medium, the number of trophectodermal (TE) cells was significantly (p<0.05) higher in amino acid supplement group than that of control ($18.6{\pm}0.5\;versus\;16.1{\pm}0.6$), whereas the numbers of inner cell mass (ICM) cells were not different among the treaonent groups and control ($29.0{\pm}0.9\~31.5{\pm}1.2$). Total cell number was also significantly (p<0.05) higher in EANA group ($50.0{\pm}1.0$) than that of control group ($44.2{\pm}1.1$). These results indicate that the amino acid supplementation to maturation and culture medium may not significantly stimulate early embryo development, but may improve the TE cell number of blastocyst stage in the pig.

Effect of Essential and Nonessential Amino Acids in North Carolina State University (NCSU)-23 Medium on Development of Porcine In vitro Fertilized Embryos

  • Hashem, Md. Abul;Bhandari, Dilip P.;Hossein, Mohammad Shamim;Jeong, Yeon Woo;Kim, Sue;Kim, Ji-Hye;Koo, Ok-Jae;Park, Seon Mi;Lee, Eu Gine;Park, Sun Woo;Kang, Sung Keun;Lee, Byeong Chun
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.693-700
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    • 2007
  • The present study was conducted to examine the effect of different levels of essential and nonessential amino acid in NCSU-23 medium on the in vitro-produced porcine embryo as it develops from the zygote to the blastocyst stage. Four experiments were performed, each with a completely randomized design involving 5 to 8 replications of treatments. In order to know the effect of nonessential amino acids in NCSU-23 medium, 0, 5, 10 and $20{\mu}/ml$ MEM were supplemented there to, (Exp. 1) and the medium was supplemented with same level of essential amino acids (Exp. 2). The combined effect of nonessential (0, 5, 10 and $20{\mu}/ml$ MEM) and essential amino acids (0, 5, 10 and $10{\mu}/ml$ MEM) in NCSU-23 medium (Exp. 3), first 72 h with non-essential amino acids (at 0, 5, 10 and $20{\mu}/ml$ MEM), and last 4 d with essential amino acids with the same level as NEAA (Exp. 4) were examined. The embryo development was monitored and the quality of blastocysts was evaluated by counting the number of total cells and determining the ratio of inner cell mass (ICM) to trophoectoderm (TE) cells. When Eagle's nonessential amino acids (MEM) added to NCSU-23 medium, it significantly increased the likelihood of development to the 2- to 4-cell stage and subsequent blastocyst development. Supplementation of different levels of essential amino acids in the NCSU-23 medium decreased cleavage rate, rate of morula and blastocyst development and the number of ICMs. In the case of the combined effect of essential and nonessential amino acids, better and significant results were found for blastocysts, hatching blastocysts and for ICM numbers which were also dose dependent. With respect to the biphasic effect of nonessential and essential amino acids, nonessential amino acids increased cleavage whereas essential amino acids increased the total cell number. Neither the nonessential nor the essential group of amino acids, on their own, affected blastocyst cell number or the differentiation of cells in the blastocyst. In conclusion, this study determined the role of nonessential and essential amino acids in the culture of the porcine embryo and showed that the embryo requires different levels of amino acids as it develops from the zygote to the blastocyst stage.

Effect of Glycosaminoglycans on In vitro Fertilizing Ability and In vitro Developmental Potential of Bovine Embryos

  • Kim, Eun Young;Noh, Eun Hyung;Noh, Eun Ji;Park, Min Jee;Park, Hyo Young;Lee, Dong Sun;Riu, Key Zung;Park, Se Pill
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.2
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    • pp.178-188
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    • 2013
  • The glycosaminoglycans (GAGs) present in the female reproductive tract promote sperm capacitation. When bovine sperm were exposed to 10 ${\mu}g/ml$ of one of four GAGs (Chondroitin sulfate, CS; Dermatan sulfate, DS; Hyaluronic acid, HA; Heparin, HP) for 5 h, the total motility (TM), straight-line velocity (VSL), and curvilinear velocity (VCL) were higher in the HP- or HA-treated sperm, relative to control and CS- or DS-treated sperm. HP and HA treatments increased the levels of capacitated and acrosome-reacted sperm over time, compared to other treatment groups (p<0.05). In addition, sperm exposed to HP or HA for 1 h before IVF exhibited significantly improved fertilizing ability, as assessed by 2 pronucleus (PN) formation and cleavage rates at d 2. Exposure to these GAGs also enhanced in vitro embryo development rates and embryo quality, and increased the ICM and total blastocyst cell numbers at d 8 after IVF (p<0.05). A real-time PCR analysis showed that the expression levels of pluripotency (Oct 4), cell growth (Glut 5), and anti-apoptosis (Bax inhibitor) genes were significantly higher in embryos derived from HA- or HP-treated sperm than in control or other treatment groups, while pro-apoptotic gene expression (caspase-3) was significantly lower in all GAG treatment groups (p<0.05). These results demonstrated that exposure of bovine sperm to HP or HA positively correlates with in vitro fertilizing ability, in vitro embryo developmental potential, and embryonic gene expression.

Signal Change of Iodinated Contrast Agents in MR Imaging (요오드화 조영제가 MR영상에 미치는 신호 변화)

  • Jeong, HK;Kim, Seongho;Kang, Chunghwan;Lee, Suho;Yi, Yun;Kim, Mingi;Kim, Hochul
    • Journal of the Institute of Electronics and Information Engineers
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    • v.53 no.12
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    • pp.131-138
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    • 2016
  • In this study, we tried to analyze the influence of ICM(Iodinated Contrast Media) in MR imaging compare to GBCA(Gadolinium Based Contrast Agent), and as this result we discussed whether resonable or not the protocol which is MRI scan after enhanced CT scan without proper time interval in clinical field. For this research, we assembled two phantoms. which one was iodine and another one was gadolinium. We did test two phantoms in conventional MRI scan which is T1, T2, T2 FLAIR and 3D angio. After that, quantitative analysis was progressed. The results of study were as follow : SSI(Saline's Signal Intensity) was shown as each sequences 175, 1231, 333, 37 [a.u] at iodine. and 1297, 123, 757, 232 [a.u] was recorded at gadolinium. BDEPS(the Biggest Difference of EPS) was shown as each sequences 1297, 123, 757, 232 [a.u] at iodine and 793, 6, 1495, 365 [a.u] was recorded at gadolinium. At this time, EPS(Enhancement Percentage to Saline) was shown 641.1, -90.0, 127.3, 527% at iodine and 685.1, 99.4, 365.7, 1077.4% was recorded at gadolinium. BP(BDEPS's point) was shown 900, 900, 477, 900 mmol at iodine and 4, 0.2, 0.2, 40 mmol was recorded at gadolinium. CPSS(Change Point of SI to SSI) was shown 63, 423, 63, 29 mmol at iodine and each [50, 30], [4, 0.2], [4, 1], 0.2 mmol was recorded at gadolinium. According to this research, we could not only discover the fact that was iodine could effect on MR signal, but also the pattern is different as various sequences compare to gadolinium. Therefore, we expect useful diagnostic MR image in clinical field with this quantitative data for deciding protocol regarding MRI and CT scan order.

Ground of the revolutionary change in early 20C American Mathematics (20세기 초 미국수학계의 혁명적변화의 바탕)

  • Lee, Sang-Gu;Hwang, Suk-Geun;Cheon, Gi-Sang
    • Journal for History of Mathematics
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    • v.20 no.3
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    • pp.127-146
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    • 2007
  • From 1876 to 1883, British mathematician James Joseph Sylvester worked as the founding head of Mathematics Department at the Johns Hopkins University which has been known as America's first school of mathematical research. Sylvester established the American Journal of Mathematics, the first sustained mathematics research journal in the United States. It is natural that we think this is the most exciting and important period in American mathematics. But we found out that the International Congress of Mathematicians held at the World's Columbian Exposition in Chicago, August 21-26, 1893 was the real turning point in American's dedication to mathematical research. The University of Chicago was founded in 1890 by the American Baptist Education Society and John D. Rockefeller. The founding head of mathematics department Eliakim Hastings Moore was the one who produced many excellent American mathematics Ph.D's in early stage. Many of Moore's students contributed to build up real American mathematics research power in early 20 century The University also has a well-deserved reputation as the "teacher of teachers". Beginning with Sylvester, we analyze what E.H. Moore had done as a teacher and a head of the new department that produced many mathematical talents such as L.E. Dickson(1896), H. Slaught(1898), O. Veblen(1903), R.L. Moore(1905), G.D. Birkhoff(1907), T.H. Hilderbrants(1910), E.W. Chittenden(1912) who made the history of American mathematics. In this article, we study how Moore's vision, new system and new way of teaching influenced American mathematical society at early stage of the top class mathematical research. and the meaning that early University of Chicago case gave.

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