• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.58-58
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• 2001

This study was carried out to investigate the optimal activation condition for parthenogenetic development. In order to activate oocytes at 24 hrs post onset of maturation, the oocytes were cultured 3 - 13 μM Ca for 5 min., 5-8 ㎍/㎖ cytoclacin(CH) for 6 hrs, 0.5-2.0 mM 6-dimethylaminopurine(DMAP) for 3 hrs alone or combination. The activated oocytes were cultured in TCM-199 media at 5% CO₂, 95% N₂, 38℃. 1. The cleavage rate after 48 hrs culture of oocytes treated with 3-13 μM Ca for 5 min. were 9.6%-20.0% and 3.8-7.3%, respectively. When oocyte were treated with 10 μM Ca, the blastocyst formation rate was significantly higher than other group. 2. The cleavage rate after 48 hrs culture of oocytes treated with 5-8 ㎍/㎖ cytoclacin(CH) for 6 hrs, were 9.4%-21.8% and 0.0-7.3%, respectively. When oocyte were treated with 10㎍/㎖ CH, the blastocyst formation rate was significantly higher than other group. 3. The cleavage rate after 48 hrs culture of oocytes treated with 0.5-2.0 mM 6-dimethylaminopurine(DMAP) for 3 hrs were 9.1%-21.8% and 0.0-7.3%, respectively. When oocyte were treated with 2.0mM DMAP, the blastocyst formation rate was significantly higher than other group. 4. The cleavage rate after 48 hrs culture of oocytes treated with Ca＋CH, Ca＋DMAP, CH＋DMAP were 75.9%-93.5% and 9.7 -13.3%, respectively. When oocytes were treated with Ca followed by DMAP, the blastocyst formation rate was significantly higher than other group(p〈0.05). 5. When necleus transferred embryos co-cultured with BSA, EGF and CS, the developmental rate to blastocyst were higher than control group.

• 한국수정란이식학회지
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• 제24권1호
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• pp.65-69
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• 2009

The aim of the present study was to investigate the cleavage pattern, its developmental ability and apoptosis of porcine embryo in vitro. Morphology data on a total of 919 embryos were analyzed retrospectively. Forty-eight hours after insemination, embryos were classified into five groups based on the cleavage state as follows; 1 cell, 2 cell, 4 cell, 5 to 8 cell and fragmentation. These groups were cultured another 120 hours and then evaluated for blastocyst formation. Blastocyst formation rates were significantly higher in 4 cell (42.5%) and 5 to 8 cell (48.6%) cleaving groups than in other groups (p<0.05). On the other hand, 2 cell and fragmentation groups produced 4.9% and 3,9% blastocysts, respectively. And we could verify that in the event of 2 cell block and fragmentation of embryo. To analyze the apoptotic frequency in preimplantation development of porcine IVF embryos, all cells of each blastocyst were performed by TUNEL assay. There were no significantly differences in the total cell numbers of embryos and apoptotic cell rate in blastocysts among the each classified groups. Data suggest that 4 cell and 5 to 8 cell cleaving embryos at 48 hour after insemination have high developmental competence, and may be an useful parameter to predict the development of preimplantation embryos and to study using preimplanation embryonic research.

• 대한화장품학회지
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• 제43권2호
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• pp.87-92
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• 2017

본 연구는 머위 추출물의 화장품소재로서의 가능성을 확인하기 위하여 자체의 독성과 카드뮴으로부터 유도된 세포손상에 미치는 영향에 대하여 평가하였다. 카드뮴으로부터 손상을 유도한 각질형성세포에 머위 추출물을 처리하여 세포사멸 인자인 Bcl-2와 procaspase-3의 단백질 발현을 측정하였다. 그 결과 머위 추출물 $200{\mu}g/mL$를 제외한 모든 농도에서 98% 이상의 높은 생존율을 나타내었으며 세포사멸인자인 Bcl-2와 procaspase-3 단백질 발현이 증가한 것으로 보아 머위 추출물이 카드뮴 독성 시 일어나는 세포자멸사에 대한 보호기전을 나타낸 것으로 평가되었다. 또한 카드뮴으로 12 h 동안 PARP cleavage를 유도한 각질형성세포에 머위 추출물을 전처리한 결과, 카드뮴을 매개로 하는 PARP cleavage를 억제하는 것으로 나타났다. 이러한 결과를 통해 머위 추출물이 피부세포 보호 효능을 나타내는 천연소재로서의 활용가능성을 제안한다.

• BMB Reports
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• 제35권2호
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• pp.206-212
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• 2002

The NS2B-NS3(pro) polyprotein segment from the dengue virus serotype 2 strain 16681 was purified from overexpressing E. coli by metal chelate affinity chromatography and gel filtration. Enzymatic activity of the refolded NS2B-NS3(pro) protease complex was determined in vitro with dansyl-labeled peptide substrates, based upon native dengue virus type 2 cleavage sites. The 12mer substrate peptides and the cleavage products could be separated by reversed-phase HPLC, and were identified by UV and fluorescence detection. All of the peptide substrates (representing the DEN polyprotein junction sequences at the NS2A/NS2B, NS2B/NS3, NS3/NS4A and NS4B/NS5 sites) were cleaved by the recombinant protease NS2B-NS3(pro). No cleavage was observed with an enzymatically inactive S135A mutant of the NS3 protein, or with a modified substrate peptide of the NS3/NS4A polyprotein site that contained a K2093A substitution. Enzymatic activity was dependent on the salt concentration. A 50% decrease of activity was observed in the presence of 0.1M sodium chloride. Our results show that the NS3 protease activity of the refolded NS2B-NS3(pro) protein can be assayed in vitro with high specificity by using cleavage-junction derived peptide substrates.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.140-146
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• 2020

Objective: To investigate whether the degree of post-warming embryo or blastocyst development is associated with clinical pregnancy in vitrified embryo or blastocyst transfer cycles. Methods: Ninety-six vitrified cleavage-stage embryos and 58 vitrified blastocyst transfer cycles were selected. All transfer cycles were performed from February 2011 to March 2019, and all vitrified embryos or blastocysts were warmed from 4 PM to 6 PM and then transferred the next morning from 9 AM to 10 AM. The scores of the cleavage-stage embryos and blastocysts were assessed at warming and at transfer using the modified Steer method and the Gardner method, respectively. The mean embryo or blastocyst score, score of the single top-quality embryo or blastocyst, and the difference in the score between warming and transfer were compared between nonpregnant and pregnant women. Results: In the cleavage-stage embryo transfer cycles, both the top-quality embryo score at transfer and the difference in the score between warming and transfer were significantly associated with clinical pregnancy. A top-quality embryo score at transfer of ≥ 60.0 (area under the curve [AUC], 0.673; 95% confidence interval [CI], 0.531-0.815) and a difference in the score between warming and transfer of ≥ 23.0 (AUC, 0.675; 95% CI, 0.514-0.835) were significant predictors of clinical pregnancy. In blastocyst transfer cycles, the top-quality blastocyst score at transfer was the only significant factor associated with clinical pregnancy. A top-quality blastocyst score at transfer of ≥ 38.3 was a significant predictor of clinical pregnancy (AUC, 0.666; 95% CI, 0.525-0.807). Conclusion: The top-quality embryo score at transfer and the degree of post-warming embryo development were associated with clinical pregnancy in vitrified cleavage-stage embryo transfer cycles. In vitrified blastocyst transfer cycles, the top-quality blastocyst score at transfer was the only significant factor affecting clinical pregnancy.

• 한국발생생물학회지:발생과생식
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• 제22권1호
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• pp.111-117
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• 2018

The objective of this study was to determine the mitotic intervals (${\tau}_0$) of two consecutive cell divisions and synchronous embryonic cleavage in grass puffer, Takifugu niphobles at different water temperatures (18, 20, 22, and $24^{\circ}C$). The color of the fertilized egg was light yellowish. The egg type was demersal and unadhesive. Egg weight was $0.09{\pm}0.002mg$. The sizes of unfertilized eggs were smaller than fertilized eggs in major axis and minor axis at $20^{\circ}C$ (p<0.05). The size of the fertilized egg of $18^{\circ}C$ water temperature group at the blastodisc stage was the smallest (p<0.05), but no significant differences were observed in the other water temperatures group except $18^{\circ}C$ water temperature group (p>0.05). The first cleavage stages at 18, 20, 22, and $24^{\circ}C$ were at 75, 90, 105, and 120 mins, respectively. As water temperature was increased, embryonic development and formation time of the first cleavage furrow were accelerated. There were negative correlation between ${\tau}_0$ and water temperature for grass puffer (Y=-1.225X+70.05, $R^2=0.988$, n=10, where Y was ${\tau}_0$ and X was temperature). This study confirmed that successful hatching of grass puffer was related to water temperature. Chromosome manipulation will be helpful for this species using cleavage frequency and ${\tau}_0$.

• 한국정보과학회논문지:소프트웨어및응용
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• 제34권7호
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• pp.595-602
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• 2007

본 논문에서는 미지의 아미노산 서열이 신호 펩티다제 I에 의해 절단되는 분비성 단백질인지를 판별하고, 분비성 단백질일 경우에는 절단 위치를 예측하는 방법을 제안한다. 아미노산의 소수성을 이용한 전처리를 수행하여 분비성 단백질의 선도서열인 신호서열의 존재와 절단 위치를 추정한다. 전처리를 통해서 신호서열 아닌 서열을 초기에 제외함으로써 신호서열 예측의 정확도를 높인다. 지지벡터기계를 신호서열의 예측에 효과적으로 적용하기 위해서, 생물학적 정보와 관련된 아미노산 서열간의 거리를 제안한다. 아미노산의 세포내 위치를 예측할 수 있는 소수성 척도와 아미노산의 진화적인 관계를 나타낼 수 있는 치환행렬을 이용하여 아미노산 서열간의 거리를 정의한다. Swiss-Prot release 50 단백질 자료에 대하여 교차타당성 기법을 사용하여 실험한 결과 제안한 방법은 신호서열중에 98.9%를 신호서열로 판별하였고, 88%의 절단위치 예측정확도를 보였다. 기존의 방법과의 비교실험을 통해서 제안한 방법이 신호서열의 예측에 더욱 효과적임을 확인하였다.

• 대한금속재료학회지
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• 제48권8호
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• pp.705-716
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• 2010

Effects of specimen thickness and notch shape on fracture mode appearing in drop weight tear test (DWTT) specimens of API X70 and X80 linepipe steels were investigated. Detailed microstructural analysis of fractured DWTT specimens showed that the fractures were initiated in normal cleavage mode near the specimen notch, and that some separations were observed at the center of the fracture surfaces. The Chevron-notch (CN) DWTT specimens had broader normal cleavage surfaces than the pressed-notch (PN) DWTT specimens. Larger inverse fracture surfaces appeared in the PN DWTT specimens because of the higher fracture initiation energy at the notch and the higher strain hardening in the hammer-impacted region. The number and length of separations were larger in the CN DWTT specimens than in the PN DWTT specimens, and increased with increasing specimen thickness due to the plane strain condition effect. As the test temperature decreased, the tendency to separations increased, but separations were not found when the cleavage fracture prevailed at very low temperatures. The DWTT test results, such as upper shelf energy and energy transition temperature, were discussed in relation with microstructures and fracture modes including cleavage fracture, shear fracture, inverse fracture, and separations.

• Journal of Genetic Medicine
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• 제20권1호
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• pp.25-29
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• 2023

The CYP11A1 gene encodes for the cholesterol side-chain cleavage enzyme (P450scc), which initiates steroid hormone biosynthesis. Defective P450scc activity results in severe glucocorticoid and mineralocorticoid deficiencies. We describe a case of P450scc deficiency due to a novel homozygous CYP11A1 variant inherited from the mother with a possibility of uniparental disomy (UPD). The patient was a female, had no family history of endocrine disease, and showed adrenal insufficiency at 13 days of age. Hormonal analysis with an adrenocorticotropic hormone stimulation test showed both glucocorticoid and mineralocorticoid deficiencies, presumed to be a defect of the early stage of steroidogenesis. Exome sequencing reported a novel homozygous frameshift variant of CYP11A1 (c.284_285del, p.Asn95Serfs*10), which was inherited from the mother. Additionally, homozygosity in 15q22.31q26.2, which included CYP11A1, was identified using a chromosomal microarray. It was suggested that the possibility of maternal UPD was involved as the cause of a P450scc deficiency by unmasking the maternally derived affected allele. To our understanding, P450scc deficiency associated with UPD encompassing CYP11A1 had not been reported in Korea before. Genetic analysis can help diagnose rare causes of primary adrenal insufficiency, including P450scc deficiency.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1606-1614
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• 2023

Biochemical gut metabolism of dietary bioactive compounds is of great significance in elucidating health-related issues at the molecular level. In this study, a human gut bacterium cleaving C-C glycosidic bond was screened from puerarin conversion to daidzein, and a new, gram-positive C-glycoside-deglycosylating strain, Dorea sp. MRG-IFC3, was isolated from human fecal sample under anaerobic conditions. Though MRG-IFC3 biotransformed isoflavone C-glycoside, it could not metabolize other C-glycosides, such as vitexin, bergenin, and aloin. As evident from the production of the corresponding aglycons from various 7-O-glucosides, MRG-IFC3 strain also showed 7-O-glycoside cleavage activity; however, flavone 3-O-glucoside icariside II was not metabolized. In addition, for mechanism study, C-glycosyl bond cleavage of puerarin by MRG-IFC3 strain was performed in D₂O GAM medium. The complete deuterium enrichment on C-8 position of daidzein was confirmed by ¹H NMR spectroscopy, and the result clearly proved for the first time that daidzein is produced from puerarin. Two possible reaction intermediates, the quinoids and 8-dehydrodaidzein anion, were proposed for the production of daidzein-8d. These results will provide the basis for the mechanism study of stable C-glycosidic bond cleavage at the molecular level.