• 제목/요약/키워드: In Vitro Development

검색결과 2,954건 처리시간 0.03초

연노랑색 FA 종간잡종 나리 신품종 '골든센터' 육성 (A FA Intersectional Hybrid Lily 'Golden Center' with Light Yellow Petals)

  • 이혜경;조해룡;김미선;박상근;임진희
    • 한국육종학회지
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    • 제43권6호
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    • pp.509-512
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    • 2011
  • '골든센터' 품종은 국립원예특작과학원에서 2008년에 육성된 절화용 FA 아속간 잡종 나리 이다. 연황색 FA 잡종인 'Migreen' 품종을 모본으로 하고, 적색의 아시아틱나리 'Sanzio'품종을 부본으로 교배하였다. 6주 후에 미숙 꼬투리를 채취하여 기내에서 배주 배양하여 2004년 'FA04-24'계통을 선발하였다. 이 계통은 2005년부터 조직배양에 의한 대량증식, 순화 및 양구를 거쳐 2007년까지 생육 및 특성검정을 수행하였다. '골든센터' 품종의 개화기는 6월 중순이다. 꽃은 상향으로 개화하고, 화색은 연황색이다. 초장은 144 cm로 신장이 우수하고, 꽃의 크기는 15.8 cm이다. 잎의 길이는 12 cm이다. 구근의 무게는 56.3 g이고, 구주는 17.4 cm로 양호하다. 주년재배를 위해서는 $-1.5^{\circ}C$에 구근을 동결 저장하여 정식시기를 달리하여 활용할 수 있다.

Gaseous signal molecule SO2 regulates autophagy through PI3K/AKT pathway inhibits cardiomyocyte apoptosis and improves myocardial fibrosis in rats with type II diabetes

  • Zhao, Junxiong;Wu, Qian;Yang, Ting;Nie, Liangui;Liu, Shengquan;Zhou, Jia;Chen, Jian;Jiang, Zhentao;Xiao, Ting;Yang, Jun;Chu, Chun
    • The Korean Journal of Physiology and Pharmacology
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    • 제26권6호
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    • pp.541-556
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    • 2022
  • Myocardial fibrosis is a key link in the occurrence and development of diabetic cardiomyopathy. Its etiology is complex, and the effect of drugs is not good. Cardiomyocyte apoptosis is an important cause of myocardial fibrosis. The purpose of this study was to investigate the effect of gaseous signal molecule sulfur dioxide (SO2) on diabetic myocardial fibrosis and its internal regulatory mechanism. Masson and TUNEL staining, Western-blot, transmission electron microscopy, RT-qPCR, immunofluorescence staining, and flow cytometry were used in the study, and the interstitial collagen deposition, autophagy, apoptosis, and changes in phosphatidylinositol 3-kinase (PI3K)/AKT pathways were evaluated from in vivo and in vitro experiments. The results showed that diabetic myocardial fibrosis was accompanied by cardiomyocyte apoptosis and down-regulation of endogenous SO2-producing enzyme aspartate aminotransferase (AAT)1/2. However, exogenous SO2 donors could up-regulate AAT1/2, reduce apoptosis of cardiomyocytes induced by diabetic rats or high glucose, inhibit phosphorylation of PI3K/AKT protein, up-regulate autophagy, and reduce interstitial collagen deposition. In conclusion, the results of this study suggest that the gaseous signal molecule SO2 can inhibit the PI3K/AKT pathway to promote cytoprotective autophagy and inhibit cardiomyocyte apoptosis to improve myocardial fibrosis in diabetic rats. The results of this study are expected to provide new targets and intervention strategies for the prevention and treatment of diabetic cardiomyopathy.

Utilizing cell-free DNA to validate targeted disruption of MYO7A in rhesus macaque pre-implantation embryos

  • Junghyun Ryu;Fernanda C. Burch;Emily Mishler;Martha Neuringer;Jon D. Hennebold;Carol Hanna
    • 한국동물생명공학회지
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    • 제37권4호
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    • pp.292-297
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    • 2022
  • Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.

느릅나무 초임계 추출물의 항균, 항산화 및 항염증 활성 (Anti-fungal, anti-oxidant, and anti-inflammatory effects of supercritical fluid extracts from Ulmus davidiana)

  • 서주희;이용조;조영익;고정윤;문명재;박광현;최선은
    • 한국융합학회논문지
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    • 제9권8호
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    • pp.225-233
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    • 2018
  • 천연물 유래 난치성 피부질환 기능성 소재를 탐색하기 위해 느릅나무 가지를 초임계 유체 기술을 활용하여 추출을 수행하였고, 확보된 느릅나무 초임계 추출물을 수종의 피부상재균에 대하여 항균력 실험을 실시하였고, 모든 균종에 대해서 항균력이 확인이 되었으며, ABTS 라디컬 소거능과 NO 소거능 실험을 통해서 느릅나무 초임계 추출물의 항산화 활성과 항염증 활성의 우수한 활성이 확인이 되어서 산화적 스트레스 질환과 염증성 피부질환에 대해서 예방 및 치료 보완 기능성 소재로 개발이 가능성이 높다는 것을 알 수 있었다. 결론적으로 느릅나무 초임계 추출물은 피부상재균에 대한 항균활성, 라디컬 소거능에 의한 항산화 활성, 과 생성된 NO 소거능에 의한 항염증활성이 각각 확인이 되었다. 이 상의 결과로 느릅나무 초임계 추출물은 항균효능, 항산화효능, 항염증 효능을 갖는 기능성 원료로서의 가능성을 확인할 수 있었고, 향후 만성 염증성 피부면역 질환 개선 및 치료 보조제 관련 의약품 또는 기능성 향장품 개발을 위한 기초 연구 자료가 될 것으로 사료된다.

Subcellular Characterization of Porcine Oocytes with Different Glucose-6-phosphate Dehydrogenase Activities

  • Fu, Bo;Ren, Liang;Liu, Di;Ma, Jian-Zhang;An, Tie-Zhu;Yang, Xiu-Qin;Ma, Hong;Zhang, Dong-Jie;Guo, Zhen-Hua;Guo, Yun-Yun;Zhu, Meng;Bai, Jing
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권12호
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    • pp.1703-1712
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    • 2015
  • The in vitro maturation (IVM) efficiency of porcine embryos is still low because of poor oocyte quality. Although brilliant cresyl blue positive (BCB+) oocytes with low glucose-6-phosphate dehydrogenase (G6PDH) activity have shown superior quality than BCB negative (-) oocytes with high G6PDH activity, the use of a BCB staining test before IVM is still controversial. This study aimed to shed more light on the subcellular characteristics of porcine oocytes after selection using BCB staining. We assessed germinal vesicle chromatin configuration, cortical granule (CG) migration, mitochondrial distribution, the levels of acetylated lysine 9 of histone H3 (AcH3K9) and nuclear apoptosis features to investigate the correlation between G6PDH activity and these developmentally related features. A pattern of chromatin surrounding the nucleoli was seen in 53.0% of BCB+ oocytes and 77.6% of BCB+ oocytes showed peripherally distributed CGs. After IVM, 48.7% of BCB+ oocytes had a diffused mitochondrial distribution pattern. However, there were no significant differences in the levels of AcH3K9 in the nuclei of blastocysts derived from BCB+ and BCB- oocytes; at the same time, we observed a similar incidence of apoptosis in the BCB+ and control groups. Although this study indicated that G6PDH activity in porcine oocytes was correlated with several subcellular characteristics such as germinal vesicle chromatin configuration, CG migration and mitochondrial distribution, other features such as AcH3K9 level and nuclear apoptotic features were not associated with G6PDH activity and did not validate the BCB staining test. In using this test for selecting porcine oocytes, subcellular characteristics such as the AcH3K9 level and apoptotic nuclear features should also be considered. Adding histone deacetylase inhibitors or apoptosis inhibitors into the culture medium used might improve the efficiency of IVM of BCB+ oocytes.

웅황과 자황의 소화 반응과 인체내 존재형태에 대한 예측 모델링 (Gastric juice and Realgar and Orpiment Mineral Medicine Reaction; Reaction Path and Speciation Modeling in Human Body)

  • 김선옥;박맹언;신순식;김경철
    • 동의생리병리학회지
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    • 제16권2호
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    • pp.365-372
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    • 2002
  • The mineral medicines mean a sort of mineral or rock for medical treatment and natural material using their chemical components and physical properties. In this study, it was apprehended the mineralogical characteristics of As-bearing group mineral medicines. The extraction test is an vitro test system for predicting the bioavailability of the major and minor elements from mineral medicines and incorporates gastrointestinal tract parameters representative of a human(including stomach and small intestinal pH, stomach mixing time and velocity). The results of the extraction test are used for reaction path modeling in human body. Reaction path modeling in human body can predict digestion with gastric juice as well as bioavailability, speciation. Also, it can predict accumulation of arsenic as pH condition. As the results of the extraction test for digestion, the amounts of Fe extraction was the highest, followed by As, Ca, Ni. In addition, as the results of the reaction path modeling between arsenic compounds and gastric juice using thermodynamic data, when absorbed, major species are followed by H₃As₃S/sub 6/(aq), As₃S/sub 6/ (aq), AsO/sup +/, H₂As₃S/sup 6-/, H₂AsO/sup 3-/, HAs₃S6/sup 2-/, HAsO/sub 3//sup 2-/ and AsO/sub 3//sup 3-/. Specifically the concentration of H₃As₃S/sub 6/(aq) is the highest. As pH increases, the concentration of H₂AsO/sup 3-/, HAsO/sub 3//sup 2-/, HAsO/sub 3//sup 3-/, HAs₃S/sub 6//sup 2-/, H₂As₃S/sup 6-/, and H₃As₃S/sub 6/ increases, whereas the concentration of H₃As₃S/sub 6/ and AsO/sup +/ decreases. On the results of this study, it is able to find out effective and toxic components of poisonous arsenic group of mineral medicines and expected to be widely used for the development of new medicines.

A Tuber Lectin from Arisaema jacquemontii Blume with Anti-insect and Anti-proliferative Properties

  • Kaur, Manpreet;Singh, Kuljinder;Rup, Pushpinder Jai;Kamboj, Sukhdev Singh;Saxena, Ajit Kumar;Sharma, Madhunika;Bhagat, Madhulika;Sood, Sarvesh Kumar;Singh, Jatinder
    • BMB Reports
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    • 제39권4호
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    • pp.432-440
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    • 2006
  • A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

두 개의 다른 마우스 배아줄기세포의 차별적인 조혈세포 분화능 (Different Potential of Hematopoietic Differentiation in Two Distinct Mouse Embryonic Stem Cells)

  • 김진숙;강호범;송지연;오구택;남기환;이영희
    • 한국발생생물학회지:발생과생식
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    • 제9권2호
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    • pp.105-114
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    • 2005
  • 배아 줄기세포는 세포 치료 목적을 위한 재료로써 매우 큰 잠재력을 가지고 있으며, 이러한 잠재력의 실현을 위해서 세포의 운명에 결정적인 역할을 하는 요소들을 확인하고 특정 세포의 대량 생산을 위한 방법을 개발하여야 한다. 조혈과정은 폭넓게 연구되어 왔으며, 배아 줄기세포로부터 조혈세포의 분화는 lineage commitment에 관한 연구에 좋은 모델이 된다. 본 연구에서는, 두 종류의 마우스 배아 줄기세포주 TC-1과 B6-1를 이용하여 그 특성과 조혈세포 분화능을 비교하여 보았다. 두 세포주는 작은 차이는 있으나 줄기세포로서의 특성을 공통적으로 가지고 있었다. 그러나 methylcellulose 배양 system을 사용하여 embryonic body 형성능을 확인한 결과 TC-1이 B6-1에 비해 월등함을 확인하였다. 조혈세포 분화의 추적을 위해 blast colony의 형성, progenitor assay, RT-PCR을 통한 조혈세포 분화 관련 marker의 발현 분석을 수행한 결과, TC-1은 정상적으로 조혈세포를 생성해 내지만, B6-1은 제대로 분화되지 못함을 확인할 수 있었다. 이러한 결과들은 in vitro에서 배아 줄기세포로부터 조혈세포로 분화를 유도할 때, 보다 적합한 세포주의 탐색이 요구됨을 제시하며 이는 향후 인간 배아 줄기세포주에서도 마찬가지로 적용될 수 있음을 암시한다고 사료된다.

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점망둑(Chasmichthys dolichognathus)의 성숙기 난모세포에서의 $C_{21}$-스테로이드 생성 (Production of $C_{21}$-Steroids in Longchin Goby, Chasmichthys dolichognathus during Oocyte Maturation)

  • 백혜자
    • 한국발생생물학회지:발생과생식
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    • 제12권1호
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    • pp.107-112
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    • 2008
  • 점망둑의 난모세포 성숙과정에서 생성되는 주요 성 스테로이드 호르몬, $C_{21}$-스테로이드를 분석하고자 전구물질 $^3H-17{\alpha}hydroxyprogesterone$ ($^3H-17{\alpha}OHP$)를 성숙기 난모세포(난경 $0.74{\sim}0.97\;mm$) 배양초기에 첨가하여 24시간 배양하였다. 스테로이드 대사물질 분석과 동정은 thin layer chromatography와 gaschromatography-mass spectrometry로 이루어졌다. $^3H-17{\alpha}20{\alpha}P$로부터 생성된 주요 성 스테로이드 대사물질은 $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$)와 $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$)로 확인되었다. 이들 대사물질은 난경 0.80 mm 이상에서 관찰되었으며, GVBD (germinal vesicle breakdown) 유도 효능 테스트에서 점망둑의 난모세포는 $17{\alpha}20{\beta}P$에 더 민감하게 반응하였다. 이러한 결과는 점망둑의 난소성숙 과정에 $17{\alpha}20{\alpha}P$$17{\alpha}20{\beta}P$이 모두 관여하나, MIS (maturation inducing steroid)로서의 가능성은 $17{\alpha}20{\beta}P$이 더 큰 것으로 관찰되었다.

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숙근(宿根)안개초의 조직배양(組織培養)에 있어서 Agar 농도(濃度) 및 Auxins의 농도(濃度)가 Vitrification에 미치는 영향(影響) (Effects of the Agar and Auxins Concentrations on Vitrification in Tissue Culture of Gypsophila paniculata L. cv. 'Bristol Fairy')

  • 전재기;최상태;박인환;신학기
    • Current Research on Agriculture and Life Sciences
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    • 제6권
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    • pp.37-41
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    • 1988
  • 숙근(宿根)안개초(草)의 조직배양(組織培養)에 있어 배지(培地)에 첨가하는 agar농도(濃度) 및 auxins의 농도(濃度)가 vitrification에 미치는 영향(影響)을 조사(調査)한 결과(結果)는 다음과 같다. 1. 초장(草長), 생체중(生體重), 분지수(分枝數)는 agar농도(濃度)가 높아질수록 감소(減少)되는 경향(傾向)이었으며 auxins의 효과(效果)는 미약(微弱)하였다. 그러나 초장신장(草長伸長)에 있어서 IAA 1.0ppm+NAA 0.5ppm 및 IAA 2.0ppm+NAA 1.0ppm 첨가구에서는 그 결과(結果)가 매우 양호(良好)하였다. 2. 발근소요일수(發根所要日數)에 있어서는 auxins의 첨가효과(效果)가 적었다. 그러나 근(根)의 생육(生育)에 있어서는 IAA 1.0ppm+NAA 0.5ppm 및 IAA 2.0ppm+NAA 1.0ppm 첨가구에 있어서 분화(分化)된 근(根)의 생장(生長)이 매우 양호(良好)한 경향(傾向)이었다. 3. 건전주율(健全株率)에 있어서는 agar농도(濃度)가 높아질수록 증가(增加)하는 경향(傾向)이었으나 배지(培地)가 너무 경화(硬化)되면(1.5%) 지상부(地上部) 및 지하부(地下部)의 생육(生育)이 극(極)히 저조(低調)하였다. 4. 이상(以上)을 종합(綜合)해 보면 숙근(宿根)안개초의 조직배양시(組織培養時) 외부환경(外部環境)에 대(對)한 적응력(適應力)이 높은 강건한 묘(苗)의 육성(育成)을 위해서는 agar농도(濃度) 1.2%로 한 MS배지(培地)(RM1962)에 IAA 1.0ppm+NAA 0.5ppm 첨가배지(培地)를 사용(使用)하는 것이 바람직하다고 생각된다.

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