• Title/Summary/Keyword: 착상인자

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초기 배아의 분리할구에서 중기 염색체상 획득 방법에 대한 연구: 염색체 변이로 인한 착상전 유전자 진단에서 보인자와 정상 핵형 구분을 위한 연구

  • 임천규;전진현;민동미;변혜경;김진영;궁미경;강인수
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.105-105
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    • 2002
  • 염색체의 구조적 이상으로 인한 습관성 유산과 기형아의 출산을 예방하기 위해 착상전 배아에서 할구를 분석하여 정상적인 핵형을 가진 배아만을 이식하는 착상전 유전자 진단 (preimplantation genetic diagnosis, PGD)의 성공적인 임상 적용이 보고되고 있으며, 그 적용 범위가 확대되고 있다. 그러나 일반적인 간기의 핵상을 이용한 PGD에서는 형광직접보합법 probe의 제약으로 보인자와 정상적인 핵형을 구분할 수 없는 단점이 있다. 따라서 본 연구에서는 보다 정확한 PGD를 위해 생쥐 배아를 이용하여 분리한 할구에서 중기 염색체상을 획득하기 위해 미세소관 (microtubule) 형성 저해제를 처리하였으며, 이를 통해 확립된 방법을 인간의 PGD에 적용하고자 하였다. 과배란이 유도된 ICR 생쥐에서 4- 또는 8-세포기 배아를 수획하여 colcemid, nocodazole, vinblastine을 각각 0.1, 0.5, 1.0, 5.0$\mu$M을 처리하고, hoechst 33342로 염색하여 핵상을 관찰하여 최적의 농도를 결정하였다. 또한 각 미세소관 형성 저해제를 혼합 처리하여 가장 높은 중기 염색체상을 획득할 수 있는 혼합 처리를 결정하였다. 이렇게 결정된 혼합 처리 방법을 인간의 체외 수정 및 배아 이식술에서 획득된 3PN 배아에 처리하여 중기 염색체를 획득하였다. Colcemid, nocodazole, vinblastine 모두 1 $\mu$M이 최적 농도임을 확인할 수 있었다 (각각 96.3%, 92.0%, 98,4%). 미세소관 형성저해제를 혼합 처리하였을 경우 nocodazole과 vinblastine (각각 1$\mu$M)을 혼합 처리했을 때 중기 염색체 획득률(97.3%)이 가장 높았다. 인간의 3PN 배아에 1$\mu$M의 nocodazole과 vinblastine을 혼합 처리한 후, 113개의 할구를 분석하여 44개(38.9%)의 할구에서 중기 염색체를 확인할 수 있었다. 본 실험 결과를 통해 중기 염색체를 획득하기 위하여 미세소관 형성 저해제를 처리하는 방법은 생쥐의 배아에서는 효과적이지만, 인간의 배아에서는 그 효율이 다소 낮음을 알 수 있었다. 그러나 이 방법을 개선하여 인간의 할구에서 중기 염색체의 획득률을 높이고, 이를 염색체의 구조적 이상에 대한 착상전 유전자 진단에 적용한다면, 보인자와 정상의 핵상을 구분하여 정상의 핵상만을 갖는 배아의 이식을 통하여 더욱 정확한 착상전 유전자 진단을 시행할 수 있으리라 사료된다.

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난관상피세포의 체외배양시 IL-1$\alpha$$\beta$ 첨가에 의한 IGF-I의 생산

  • 최선호;조상래;한만희;김현종;손동수;류일선;김영곤;정영호;박춘근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.234-234
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    • 2004
  • 수정란이식 기술이 활성화되고, 한우의 가격이 상승하여, 젖소에 한우 체외수정란이식이 성행하고 있으나, 수정란이식에 의한 수태율은 수정란이식의 성과를 내고 있다고 할 정도는 아니다. 쥐, 돼지 및 사람에서 착상에 관련되는 인자에 대한 연구가 활발히 이루어지고 있으며, 가장 착상에 관여를 많이 하고 있는 인자는 LIF, IGF와 TGF-β인 것으로 보고되고 있다. 따라서 본 연구는 임신과 관련된 주요기관인 난관의 상피세포를 체외배양하고, 그때 착상을 유도하는 것으로 알려진 IL-1α와 β를 첨가하여 IGF-I의 생산을 조사하였다. (중략)

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Frost Formation on a Cold Cylindrical Surface in Cross Flow (직교류내 원통형 냉각표면에서의 착상)

  • Lee, Kwan-Soo;Lee, Dong-Hoon;Yang, Dong-Keun
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.28 no.8 s.227
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    • pp.989-995
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    • 2004
  • This paper presents a semi-empirical model to predict the frost growth formed on the cold cylinder surface. The model is composed of the correlations for frost properties including the various frosting parameters and local heat transfer coefficient. The effects of varying the correlations for local heat transfer coefficient on the frost growth are examined to establish the model. The numerical results are compared with experimental data obtained by the previous researchers. The results agree well with the experimental data within a maximum error of $13\%$. As the results, the frost thickness decreases with changing angular position from front stagnation to separation point. Also, the effects of air velocity on the frost growth are negligible, as compared to the other frosting parameters.

Frost formation on a cold cylinder surface in cross flow (원관의 냉각면에서의 착상)

  • Lee, Dong-Hoon;Yang, Dong-Keun;Lee, Kwan-Soo
    • Proceedings of the KSME Conference
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    • 2004.04a
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    • pp.1540-1545
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    • 2004
  • This paper presents a semi-empirical model to predict the frost growth formed on the cold cylinder surface. The model is composed of the correlations for frost properties including the various frosting parameters and local heat transfer coefficient. The effects of varying the correlations for local heat transfer coefficient on the frost growth are examined to establish the model. The numerical results are compared with experimental data obtained by the previous researchers. The results agree well with the experimental data within a maximum error of 13%. As the results, the frost thickness decreases with changing angular position from front stagnation to separation point. Also the effects of air velocity on the frost growth are negligible, as compared to the other frosting parameters.

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The Role of the Endometrium and Embryo in Human Implantation (인간 착상 과정에 자궁내막과 배아의 역할)

  • Jee, Byung-Chul
    • Development and Reproduction
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    • v.13 no.1
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    • pp.1-11
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    • 2009
  • Implantation itself is governed by an array of endocrine, paracrine and autocrine modulators, of embryonic and maternal origin. Window of implantation is the unique temporal and spatial expression of factors allows the embryo to implant via signaling, appositioning, attachment, and invasion in a specific time frame of $2{\sim}4$ days. When the embryo has arrived in the uterine cavity, a preprogrammed sequence of events occurs, which involves the production and secretion of a multitude of biochemical factors such as cytokines, growth factors, and adhesion molecules by the endometrium and the embryo, thus leading to the formation of a receptive endometrium. Cytokines such as LIF, CSF-1, and IL-1 have all been shown to play important roles in the cascade of events that leads to implantation. Integrin, L-selectin ligands, glycodelin, mucin-1, HB-EGF and pinopodes are involved in appositioning and attachment. The embryo also produces cytokines and growth factors (ILs, VEGF) and receptors for endometrial signals such as LIF, CSF-1, IGF and HB-EGF. The immune system and angiogenesis play an important role. The usefulness of these factors to assess endometrial receptivity and to estimate the prognosis for pregnancy in natural and artificial cycles remains to be proven. Integrins, pinopodes, glycodelin and LIF (from biopsies) are promising candidates; from uterine flushings, glycodelin and LIF are also candidates. The ideal serum marker is not available, but VEGF, glycodelin and CSF have some clinical implications. Further evaluation that includes larger groups of infertile women and fertile controls are needed to elucidate whether their presence in plasma, flushing fluid, or endometrial samples can be used as some kind of a screening tool to assess endometrial function and prognosis for pregnancy before and after artificial reproductive therapy. A better understanding of their function in human implantation may lead to therapeutic intervention, thereby improving the success rate in reproduction treatment. New molecular techniques are becoming available for measuring both embryonic and endometrial changes prior to and during implantation. The use of predictive sets of markers may prove to be more reliable than a single marker. Ultimately, the aim is to use these tools to increase implantation in artificial cycles and consequently improve live-birth rates.

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The EST Study of the Peri-implanting Porcine Embryos (Peri-implanting 단계의 돼지배아 EST 연구)

  • Kwak, In-Seok
    • Journal of Life Science
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    • v.19 no.5
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    • pp.587-592
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    • 2009
  • A dramatic morphological change of embryos occurs at peri-implantation. Maternal and embryonic cross-talk during this period, initiated by signals from embryo(s), provides signals for maternal recognition of pregnancy and establishing and maintaining the pregnancy. However, the cellular, biochemical and genetic processes that direct embryo remodeling in mammalian species are not well studied or understood. In order to identify potential genes responsible for morphological change and cross-talk between embryo and uterus, an initial EST analysis was performed. A catalog of expressed genes (Transcriptome) from the d12 peri-implanting porcine embryos was constructed. Six clones were chosen from the initial ESTs for elucidation of their expression patterns during embryogenesis in early pregnancy. A number of these genes demonstrated unique expression profiles in a tissue, cell-type, and temporal fashion, indicating dynamic regulation of embryonic and endometrial gene expressions at different stages of pregnancy. Cross-talk between the embryo and endometrium of the pregnant uterus has provided a suitable micro-environment for the embryo's rapid and dramatic morphological changing process at the peri-implantation stage.

Regulation of LIF Gene Expression by Interleukin-1 in the Mouse Peri-implantation Embryos and Uterine Endometrial Cells (생쥐의 착상시기 배아와 자궁내막세포에서 IL-1에 의한 LIF 유전자 발현 조절)

  • Lee, Jung-Bok;Kim, Joung-Woul;Oh, Eun-Jeong;Yang, Hye-Young;Ryu, Hyoung-Eun;Lee, Ji-Youn;Gye, Myung-Chan;Yoon, Hyun-Soo;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.183-190
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    • 2000
  • 연구목적: 포유류의 착상은 배아가 모체의 자궁벽에 매몰되는 현상으로 부착과 침투 과정을 거쳐 진행되며, 이 과정은 스테로이드 호르몬, 성장인자, 세포점착분자, 그리고 cytokine 등의 상호작용으로 이루어진다. 이 시기에 Interleukin-1 (IL-1)과 leukemia inhibitory factor (LIF) 등이 발현되는 것으로 알려져 있다. 본 실험에서는 이들의 발현이 착상과정에 어떠한 역할을 하는지 그 상관관계를 알아보고자 하였다. 재료 및 방법: 착상 전후의 배아와 자궁내막세포에서 LIF 유전자의 발현양상과 $IL-1{\beta}$와 IL-1 receptor antagonist (IL-1ra)를 처리한 LIF 유전자의 발현양상을 역전사중합효소연쇄반응 (RT-PCR)을 통해 비교하였다. 결과: 배아에서의 LIF 유전자 발현은 in vivo와 in vitro 모두에서 상실기와 포배기에 발현되었고, 자궁내막에서는 임신 1일과 4일째에 발현되었는데, 상실기보다는 포배기에, 그리고 임신 1일보다는 착상시기인 4일째의 자궁내막세포에서 발현양이 많은 것으로 나타났다. 자궁내막세포를 배양한 경우 LIF 유전자는 in vivo에서의 발현양상과 동일하게 임신 1일과 4일에 발현되었으며, 배양액에 $IL-1{\beta}$(500pgml)를 처리하였을 경우 LIF 유전자가 초기 임신 (1~5일) 중 발현되는 것으로 나타났다. 2-세포기 배아의 배양시에 $IL-1{\beta}$를 처리한 경우 8-세포기부터 LIF 유전자가 발현되었으며, 또한 IL-1ea(60 ng/ml)를 배양액에 첨가하였을 경우에는 임신1일째 자궁내막에서는 LIF 유전자가 발현되지 않은 반면, 임신 4일째의 자궁내막세포와 상실기, 포배기 배아 모두에서 LIF 유전자 발현이 감소하는 경향을 보였다. 결론: 이러한 결과는 착상 전후 배아와 자궁내막세포에서 IL-1에 의해 LIF 유전자 발현이 조절되며, 그 결과 착상에 영향을 줄 수 있다는 것을 의미한다. 또한 배아와 자궁내막세포에서 IL-1이 LIF 유전자 발현에 영향을 주는 것으로 보아 착상을 위해 IL-1과 LIF의 상호작용이 중요한 요인이라는 것을 확인할 수 있었다.

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Correlations of Frost Properties Considering the Environmental Parameters over a Cold Flat Plate (수평편판에서의 착상인자를 고려한 서리 물성치의 상관식)

  • Lee, Kwan-Soo;Kim, Yong-Chul;Jhee, Sung
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.25 no.8
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    • pp.1046-1052
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    • 2001
  • This study has been performed experimentally to measure the frost properties over a cold flat plate in order to obtain the correlations of the frost properties with various environmental parameters. Correlations of the frost surface temperature, thickness, density, and thermal conductivity are presented along with the experimental results as a function of environmental parameters, distance from the leading edge of test plate, and time. These correlations can be used to predict the frost properties with a maximum error of 8% in the following ranges : air temperature 15∼25$\^{C}$, relative humidity 60∼80%, air velocity 1∼3m/s and cooling plate temperature -15∼-25$\^{C}$.

cAMP Mediation in Estradiol-induced Uterine Prostaglandin Synthesis During the Delayed Implantation Process in Rats (흰쥐의 착상지연과정중 Estradiol에 의한 자궁내 Prostaglandin 생합성에 미치는 cAMP의 영향)

  • Yoon, Mi-Chung;Kim, Chang-Mee;Choe, Rim-Soon;Ryu, Kyung-Za
    • The Korean Journal of Pharmacology
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    • v.27 no.2
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    • pp.183-189
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    • 1991
  • The present study was performed to elucidate the factors which modulate uterine prostaglandin synthesis during the implantation period in rats, by employing delayed implantation model. Administration of estradiol sharply increased uterine cAMP concentration 4 hrs later during the delayed implantation process. Concentrations of uterine PGE and $PGF_2{\alpha}$ were increased at 12 hrs after the estradiol treatment although an increase in $PGF_2{\alpha}$ was not statistically significant. The concomitant treatment of indomethacin with estradiol significantly suppressed estradiol-induced PGE and $PGF_2{\alpha}$ at 12 hrs, while uterine cAMP concentration was not suppressed. The treatment of dbcAMP without estradiol gradually increased uterine PGE and $PGF_2{\alpha}$ showing the maximum 8 hrs later, suggesting that cAMP minics estradiol effect on uterine prostaglandin synthesis during the implantation process. Furthermore, the pretreatment of theophylline, phosphodiesterase inhibitor, induced significantly greater concentrations of uterine PGE and $PGF_2{\alpha}$, compared with estradiol-only treated group. These results suggest that estradiol stimulates uterine prostaglandin synthesis and this process may be mediated by an elevation of cAMP during the delayed implantation process in rats.

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An Experimental Study on the Effects of Design Factors for the Performance of Fin-Tube Heat Exchanger Under Frosting Conditions (착상시 설계인자에 따른 핀-관 열교환기의 성능변화에 관한 실험적 연구)

  • 이관수
    • Transactions of the Korean Society of Mechanical Engineers
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    • v.19 no.10
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    • pp.2657-2666
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    • 1995
  • In this study, the effects of design factors of finned-tube heat exchanger, such as fin spacing and fin array on the frost growth and heat exchanger performance are investigated under a frosting condition. The results show that the amount of frost, frost density and blockage ratio of air flow passage increase with decreasing fin spacing. Heat transfer rate increases momentarily at the initial stage of frosting and then decreases. After that heat transfer rate continues to increase again to reach a maximum value and then decreases dramatically. It is shown that the time required for heat transfer rate to reach a maximum value becomes shorter with decreasing fin spacing, and after a maximum value, heat transfer rate decreases very fast. The maximum allowable blockage ratio is introduced to determine the operation limit of a finned-tube heat exchanger operating under frosting condition and is obtained as a function of fin spacing. It is also shown that heat transfer rate of heat exchanger with staggered fin array increases about 17% and the amount of pressure drop of air increases about 1~2 mmH$_{2}$O, compared with those of in-line type heat exchanger under frosting condition.