• 제목/요약/키워드: cleavage.

검색결과 2,068건 처리시간 0.025초

다양한 유리화 동결 방법이 각 시간대별 생쥐 전핵기 배아의 발달에 미치는 영향 (The Effect of Various Vitrification Methods on Developmental Rate of Mouse Pronuclear Embryos at Different Recovery Times)

  • 김지철;서병부;박성백;김재명
    • 한국수정란이식학회지
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    • 제27권1호
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    • pp.63-69
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    • 2012
  • The objective of this study was to investigate the effectiveness of cryopreservation methods for the effect of various vitrification containers, such as EM-grid, OPS, or cryo-loop on the survival and developmental rate of vitrified mouse pronuclear embryos, and mouse cleavage embryo, at 21, 24, 27 and 30 hr after hCG injection. Post-thaw cleavage was similar among treatments, while the developmental rates of mouse blastocyst and hatched blastocyst were higher ($p$ <0.05) in 27 hr and 30 hr than 21 hr. The developmental rate of hatched blastocyst at vitrified cleavage mouse embryos in cryo-loop was significantly higher than vitrified pronuclear embryos of control group as well as EM-grid and OPS ($p$ <0.05). The developmental rate using cryo-loop was higher than EM-grid, but in case of OPS at vitrified cleavage and mouse pronuclear embryos, no significant difference was noticed. These results of our study show that the developmental rates of mouse embryos were unaffected by various vitrification containers, but in case of mouse embryos and hatched blastocysts at late vitrified pronuclear embryos the developmental rates were higher than early vitrified pronuclear embryos. Moreover, the developmental rate of hatched blastocyst at vitrified cleavage mouse embryos was significantly higher than vitrified pronuclear embryos. For better execution of this study, it will be mandatory to include improvement of vitrification containers, cryopreservation methods and conditions, higher survival rate, safe preservation, contamination and embryo loss.

카스파제-3 효소를 이용한 폴리-단백질의 정량적 프로세싱 분석 (Caspase-3-facilitated Stoichiometric Cleavage of a Large Recombinant Polyprotein)

  • 김문일
    • 생명과학회지
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    • 제25권4호
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    • pp.385-389
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    • 2015
  • 선행연구에서 카스파제-3 효소가 DEVD 기질을 완전히 절단하는 반면 IETD 기질은 DEVD의 약 50%정도만을 부분적으로 분해한다는 사실을 보고하였다. 본 연구에서는 정제된 폴리-단백질이 카스파제-3 단백질 분해효소의 기질에 따른 차별적인 분해활성에 의해 프로세싱 되는 양상을 분석하였다. 모델 단백질로서 GST, MBP, RFP 세 종류의 단백질을 DEVD 및 IETD 펩타이드로 연결시킨 폴리-단백질을 제작하였으며, 폴리-단백질은C-말단에 6개의 히스티딘 태그가 결합되도록 클로닝 되었다. IMAC 크로마토그래피를 이용하여 분리 및 정제된 재조합 단백질은 SDS-PAGE 분석을 통해 분자량이 약 93 kDa으로 나타났으며, 카스파제-3 효소의 처리에 의해 각각 MBP:RFP, MBP 그리고 GST 3종류의 단백질 절편으로 절단 및 분리되었다. 이 연구를 통해 단백질 분해효소와 기질 간의 반응성의 차이를 이용하여 폴리-단백질을 정량적으로 프로세싱 할 수 있다는 예를 보여주었다.

융합단백질 절단반응을 위한 고정화된 enterokinase의 고체상 재접힘 (Solid-phase Refolding of Immobilized Enterokinase for Fusion Protein Cleavage)

  • 서창우;나세진;박신혜;박승국;이은규
    • KSBB Journal
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    • 제18권4호
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    • pp.306-311
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    • 2003
  • 융합단백질의 절단을 위해 EK를 고정화하여 액상 절단반응과 같은 80%의 절단수율을 얻을 수 있었다. 그리고 니켈 친화칼럼을 이용하여 간단한 정제공정을 구축하였다. 공유결합한 EK의 경우 니켈친화 결합한 EK보다 높은 재접힘 수율을 나타내었고 풀림과 재접힘을 이용하여 효소의 초기 활성을 회복함에 따라서 반복사용을 통한 경제적인 절단공정을 구축할 수 있게 되었다. 그러나 고정화 과정에서 효소의 활성이 감소하는 문제점과 고정화 수율을 높이기 위한 연구가 필요하다.

${\zeta}-Carotene$의 산화개열산물 (Oxidative Cleavage Products of ${\zeta}-Carotene$)

  • 김선재
    • 한국식품과학회지
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    • 제32권5호
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    • pp.985-990
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    • 2000
  • ${\zeta}-Carotene$를 dichloromethane에 용해하고 dry ice 상에서 ozonolysis을 행하였다. Ozonolysis에 의해 생성된 화합물은 silica gel chromatography를 행하여 분획하고, photodiode array detector를 이용하여 ODS-HPLC로 분석하였다. Toluene에 ${\zeta}-carotene$을 용해하고 $37^{\circ}C$, 72시간 자동산화시킨 결과, 다수의 carbonyl 및 acidic화합물이 생성되었다. ${\zeta}-Carotene$의 자동산화로부터 생성된 carbonyl 화합물의 대부분은 ozonolysis에 의하여 얻어진 산화개열산물이 나타내는 HPLC상의 거동과 분광학적 특성이 서로 잘 일치하였다. 또한 ${\zeta}-carotene$의 자동산화에 의해 생성된 중앙개열 acidic 화합물은 생물활성을 나타내는 4,5-didehydrogeranyl geranyl acid 표준품과 동일한 분광학적 특성을 나타냈다. 이러한 결과는 in vitro상의 산화적 조건하에서 ${\zeta}-carotene$ 자동산화에 의해 eccentric cleavage가 생성됨을 알 수 있었다.

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Human embryos derived from first polar body nuclear transfer exhibit comparatively abnormal morphokinetics during development

  • Leila Heydari;Mohammad Ali Khalili;Azam Agha Rahimi;Fatemeh Shakeri
    • Clinical and Experimental Reproductive Medicine
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    • 제50권3호
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    • pp.177-184
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    • 2023
  • Objective: Reconstructed oocytes after polar body genome transfer constitute a potential therapeutic option for patients with a history of embryo fragmentation and advanced maternal age. However, the rescue of genetic material from the first polar body (PB1) through introduction into the donor cytoplasm is not yet ready for clinical application. Methods: Eighty-five oocytes were obtained following in vitro maturation (IVM) and divided into two groups: PB1 nuclear transfer (PB1NT; n=54) and control (n=31). Following enucleation and PB1 genomic transfer, PB1 fusion was assessed. Subsequently, all fused oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured in an incubator under a time-lapse monitoring system to evaluate fertilization, embryonic morphokinetic parameters, and cleavage patterns. Results: Following enucleation and fusion, 77.14% of oocytes survived, and 92.59% of polar bodies (PBs) fused. However, the normal fertilization rate was lower in the PB1NT group than in the control group (56.41% vs. 92%, p=0.002). No significant differences were observed in embryo kinetics between the groups, but a significant difference was detected in embryo developmental arrest after the four-cell stage, along with abnormal cleavage division in the PB1NT group. This was followed by significant between-group differences in the implantation potential rate and euploidy status. Most embryos in the PB1NT group had at least one abnormal cleavage division (93.3%, p=0.001). Conclusion: Fresh PB1NT oocytes successfully produced normal zygotes following PB fusion and ICSI in IVM oocytes. However, this was accompanied by low efficiency in developing into cleavage embryos, along with an increase in abnormal cleavage patterns.

Nano-identification for the Cleavage of Disulfide Bond during the Self-Assembly Processes of Unsymmetric Dialkyl Disulfides on Au(111)

  • Noh, Jae-Geun
    • Bulletin of the Korean Chemical Society
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    • 제26권4호
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    • pp.553-557
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    • 2005
  • The formation of striped phases of unsymmetric hexyl octadecyl disulfide ($CH_3(CH_2)_5SS(CH_2)_{17}CH_3$, HOD) and 1-hydroxyundecyl octadecyl disulfide ($CH_3(CH_2)_{17}SS(CH_2)_{11}$OH, HUOD) on Au(111) and graphite has been investigated by scanning tunneling microscopy (STM) to understand the self-assembly processes of dialkyl disulfides. STM imaging clearly shows the formation of striped phases having corrugation periodicities that are nearly consistent with the molecular length of alkanethiolate moieties formed after the S-S bond cleavage of dialkyl disulfide on a gold surface. On the other hand, self-assembled monolayers (SAMs) of dialkyl disulfides on a graphite surface displayed long-range, well-ordered monolayers with one striped pattern that shows periodicity as a function of molecular length via nondissociative adsorption. From a nonoscopic viewpoint, we have clearly demonstrated that dialkyl disulfide SAMs on gold form via S-S bond cleavage of disulfide.

Blocking the 1st Cleavage in Mud Loach, Misgurnus mizolepis

  • Yoon Kwon Nam;Gyeong Cheol Choi;Dong Soo Kim
    • 한국양식학회지
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    • 제12권3호
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    • pp.167-173
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    • 1999
  • Blocking the 1st mitotic cleavage was performed in mud loach (Misgurmus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and ternal shocks Optimum UV range for inactivation of cyprinid loach sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to 4,050 ergs/m$m^2$. Heat shock treatment ($41^{\circ}C$ for 3mins) with various treatment initiation times ranged from 22 to 50 min post insemination resulted wide range of success for induced gynogenesis. Best result was obtained when haploid egges were shocked at 28 min after insemination (corresponding to metaphase division of the 1st cleavage); 26% of total eggs inseminated were viable diploid gynogens. The hatching success and early survival of the both meiotic and mitotic gynogenetic groups were significantly lower than those of control crosses (P<0.05). Maternal origin of induced gynogenetic mud loach was verified by multi-locus DNA fingerprinting.

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General Fragmentations of Alkaloids in Electrospray Ionization Tandem Mass Spectrometry

  • Shim, Hee Jung;Lee, Ji Ye;Kim, Byungjoo;Hong, Jongki
    • Mass Spectrometry Letters
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    • 제4권4호
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    • pp.79-82
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    • 2013
  • Various types of alkaloids observed in several herbal medicines were analyzed by electrospray ionization tandem mass spectrometry in positive ion mode. In the present study, MS/MS spectralpatterns were investigated for eight-types of alkaloids (aporpine, protoberberine, tetrahydroprotoberberine, benzylisoquinoline, protopine, phthalide, morpine, and bisbenzylisoquinoline). For aporpine- and protoberberine-type alkaloids, main fragmentations occurred at substituted groups on rigid ring structures, not showing ring fusion. Interesting fragmentations due to iminolization and retro-Diels-Alder (RDA) reaction were observed in MS/MS spectra of protopine- and tetrahydroprotobereberine-type alkaloids. Also, several types of fragmentations such as inductive cleavage and ${\alpha}$-cleavage, or bond cleavage between two ring structures were observed depending on their structural characteristics. These fragmentation patterns are expected to allow instant classification of the specific alkaloid type in various MS/MS spectra of alkaloids.

In-situ Crack Propagation Observation of a Particle Reinforced Polymer Composite Using the Double Cleavage Drilled Compression Specimens

  • Lee Yeon-Soo;Yoon Young-Ki;Jeong Bo-Young;Yoon Hi-Seak
    • Journal of Mechanical Science and Technology
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    • 제20권3호
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    • pp.310-318
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    • 2006
  • In this study, we investigate the feasibility of in-situ crack propagation by using a double cleavage drilled compression (DCDC) specimen showing a slow crack velocity down to 0.03 mm/s under 0.01 mm/s of displacement control. Finite element analysis predicted that the DCDC specimens would show at least 4.3 fold delayed crack initiation time than conventional tensile fracture specimens under a constant loading speed. Using DCDC specimens, we were able to observe the in-situ crack propagation process in a particle reinforced transparent polymer composite. Our results confirmed that the DCDC specimen would be a good candidate for the in-situ observation of the behavior of particle reinforced composites with slow crack velocity, such as the self-healing process of micro-particle reinforced composites.

Temperature-dependent Index of Mitotic Interval (τ0) for Chromosome Manipulation in Korean Rose Bitterling Rhodeus uyekii

  • Kim, Bong-Seok;Lim, Sang-Gu;Gil, Hyun-Woo;Park, In-Seok
    • Fisheries and Aquatic Sciences
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    • 제14권4호
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    • pp.429-433
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    • 2011
  • Eggs of Korean rose bitterling Rhodeus uyekii were collected and fertilized to observe temperature-related cleavage rates and mitotic intervals (${\tau}_0$). As the water temperature was increased, the slope of first cleavage frequency with elapsed time after fertilization increased, and approximately 30% of fertilized eggs reached first cleavage frequency at every 15 min. At higher temperatures, eggs developed faster and underwent further identical developmental processes. There were strong, negative correlations between ${\tau}_0$ and water temperatures at all temperatures studied (Y = -1.225X + 70.05, $r^2=0.988$, where Y is ${\tau}_0$ and X is temperature).