• Title/Summary/Keyword: fluorescent lipid

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Synthesis of Nanomaterials such as fluorescent-magnetic Nanoclusters and Frequency Doubling Nanowires for Bioapplications

  • Jeong, Jinhoo;Kim, Seungwook;Kim, Woong
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.132.1-132.1
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    • 2014
  • Fluorescent-magnetic nanoclusters were synthesized for biomedical applications. The nanoclusters consisted of superparamagnetic core-nanoclusters, highly fluorescent shell of nanocrystals, and lipid A. Magnetic cores were used for both magnetic resonance imaging (MRI) and cell separation. Fluorescent shell was used for optical imaging. The lipid-A-loaded nanoclusters were up-taken by dendritic cells via phagocytosis, which successfully activated dendritic cells. The dendritic cells were migrated to lymph nodes and spleen of mice. The results showed that our novel nanoclusters can play a role as an efficient optical and magnetic imaging, a cell separating and a pathogen mimetic agent at the same time. Additionally, synthesis of wavelength conversion nanowires will be discussed, which may be used as an optical nanoprobe in biological studies.

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Green Fluorescent Protein(GFP)의 Fluorescence-Activated Cell Sorter(FACS) 분석을 통한 유전자 이입의 최적화 (Optimization of Gene Transfection Using Fluorescence-Activated Cell Sorter(FACS) Analysis of Green Fluorescent Protein(GFP))

  • 김태경;박민태;이균민
    • KSBB Journal
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    • 제14권3호
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    • pp.377-379
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    • 1999
  • CHO/dhfr- 세포에 대해 LipofectAmine$^{TM}$을 이용한 유전자 이입 효율을 증가시키기 위하여 지질과 DNA의 최적 농도를 구하였다. Reporter 유전자로서 GFP 유전자를 이용하였으며, 여러 농도의 지질 DNA로 유전자 이입된 각 세포군에서 나타나는 green fluorescence intensity를 FACS 분석함으로써 유전자 이입 효율을 정량화 할 수 있었다. 그 결과 24-well plate에서 $2.0{\mu}L$LipofectAmine$^{TM}$$0.4{\mu}g$ DNA를 조합하여 사용했을 때 최적의 유전자 이입 효율이 나타남을 알 수 있었다. 또한, GFP는 유전자 이입 최적화를 수행하는 데에 여러가지 면에서 유용한 수단이 될 수 있음을 확인할 수 있었다.

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형광펩타이드를 이용한 Oligosaccharyltransferase Assay 방법 연구 (Comparison of Oligosaccharyltransferase Assay Methods Using a Fluorescent Peptide)

  • 김성훈
    • 미생물학회지
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    • 제46권1호
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    • pp.96-103
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    • 2010
  • 단백질의 N-글리코실화는 대표적인 번역 후 변형 중의 하나로 진핵생물 뿐 아니라 원핵생물에서도 발견된다. N-글리코실화는 단백질 상의 N-글리코실 서열인 N-x-S/T 위치에 지질과 연결된 올리고당(lipid-linked oligosaccharide, LLO)으로부터 올리고당 전이효소(oligosaccharyltransferase, OTase) 활성에 의해 글리칸(glycan)이 전달되어 당단백질의 합성이 이루어진다. 본 연구에서는 OTase의 세포내 활성을 측정하기 위하여 5/6-carboxyltetramethylrhodamine (TAMRA)이 도입된 형광펩타이드 TAMRA-DA$\underline{N}$Y$\underline{T}$K-$NH_2$를 이용하였다. OTase활성 측정은 단일 서브유닛으로 효소의 활성을 갖는 운동핵 편모충류인 Leishmania major Stt3p와 병원성 미생물인 Campylobacter jejuni PglB를 진핵생물과 원핵생물의 모델 효소로 각각 사용하여 Saccharomyces cerevisiae와 C. jejuni 유래 LLO와 형광 펩타이드를 반응시켜 당-펩타이드를 합성하였다. 합성된 당-펩타이드를 미반응한 형광펩타이드와 분리 및 당-펩타이드의 정량 분석을 위하여 Tricine SDS-PAGE, ConA 렉틴 컬럼 및 fluorospectrophotometer, HPLC를 사용하였으며, 당-펩타이드 분석을 통해 각 방법의 장단점을 비교하였다. 비교 분석 결과 Tricine SDS-PAGE를 이용한 형광 이미지 분석과, 렉틴 컬럼을 통해 분리된 당-펩타이드의 fluorospectrophotometer 정량법에 비해, HPLC를 이용한 방법이 OTase에 의해 생성된 당-펩타이드를 분석하는데 더 정확하고 정량적인 값을 제시하는 것으로 확인되었다.

Synthesis of NBD-Labeled DOTAP Analog to Track Intracellular Delivery of Liposome

  • Doh, Kyung-Oh;Kim, Bieong-Kil;Lee, Tae-Jin;Park, Jong-Won;Seu, Young-Bae
    • Journal of Microbiology and Biotechnology
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    • 제23권1호
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    • pp.131-135
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    • 2013
  • A DOTAP analog labeled by NBD on the head group (DTNBD) was designed and synthesized to label DOTAP liposome. The structure was confirmed by $^1H$ NMR and FAB-MS, and the fluorescence of the newly synthesized DT-NBD was observed by fluorescent microscopy. The transfection efficiency of DOTAP liposome containing DT-NBD was comparable to commonly used NBD PE in COS7 and MCF7 cells. Furthermore, the level of cellular uptake and fluorescent intensity of fluorescent liposome containing DT-NBD was higher than NBD PE. Therefore, the novel NBD-labeled DOTAP analog seems to be effectively used for investigation of the cellular interaction and transfection mechanism of DOTAP liposome.

육가공 제품의 저장 및 조리방법이 지질산패에 미치는 영향 (Effects of Storage Days and Cooking Method on Lipid Oxidation in Processed Meat Products)

  • 이재민;조정순
    • 한국응용과학기술학회지
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    • 제12권1호
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    • pp.69-79
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    • 1995
  • Ham, sausage and bacon were treated with common household processing techniques including refrigerated storage(0, 14, 28 days) and cooking(pan-frying, microwaving, boiling). Lipid oxidation was evaluated by measuring fatty acid composition, malonaldehyde(MA), TBA values and by measuring fluorescent products. Major fatty acid composition were oleic acid and followed respectively palmitic acid, stearic acid, linoleic acid, linolenic acid. There was no significant difference in fatty acid composition by cooking method but there was a tendency of being increased of unsaturated fatty acid during 28days storage. Ma, TBA and fluorescent products showed a tendency of being increased continually according to storage days rather than cooking method.

Improved Fluorescent Determination Method of Cellular Sphingoid Bases in High-performance Liquid Chromatography

  • Yoon, Hong-Tak;Yoo, Hwan-Soo;Shin, Bum-Kyu;Lee, Woo-Jin;Kim, Hwan-Mook;Hong, Seon-Pyo;Moon, Dong-Cheul;Lee, Yong-Moon
    • Archives of Pharmacal Research
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    • 제22권3호
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    • pp.294-299
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    • 1999
  • Precolumn orthophthaldehyde (OPA) labeling method of sphingoid bases, sphingosine and sphinganine, was investigated to obtain high fluorescent detectability. In order to improve the fluorescent yield, we investigated the optimal solubility of sphingoid bases for five pre-incubation solvents by incorporating the heating procedure before OPA derivatization. The pre-incubation in ethanol prominently increased the fluorescent peak height of OPA derivative for each sphingoid bases in high performance liquid chromatography. About tenfold increase of detectability was archived by pre-incubating lipid extracts pellets in ethanol at $60^{\circ}C$ for 30 min. Optimal derivatization was performed in 30 min at ambient temperature and the fluorescent intensity of OPA derivative was stable for two weeks at $4^{\circ}C$. The detection limit of sphingosine was 0.1 pmol as injected amount. This method was applied to the determination of cellular sphingosine and sphinganine in various human lung cancer cells. This OPA procedure was prospective to be useful for quantitating the amount of sphingoid bases in other cancer cells.

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Acidity Enhances the Ability of 5-Aminoimidazole-4-carboxamide Ribonucleotide to Increase Respiration and Lipid Metabolism in Daphnia magna

  • Han, Chloe;Kottapalli, Aarthi;Boyapati, Keerti;Chan, Sarah;Jeong, Yong-Joo
    • 대한화학회지
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    • 제63권4호
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    • pp.253-259
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    • 2019
  • 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR), a structural analog of adenosine monophosphate (AMP), promotes oxidative remodeling in muscle cells. AICAR activates AMP-dependent protein kinase (AMPK), thus increasing lipid metabolism, respiration, and mitochondrial counts. This process is called oxidative remodeling, which enhances the physical endurance of mice. To test this drug on an invertebrate that is genetically similar to humans, we used the small water crustacean Daphnia magna, which is sensitive to changes in water conditions. We tested the effects of pH on the efficacy of AICAR using two methods. One method measured oxygen consumption of Daphnia in oxygen chambers. The other method determined lipid levels of Daphnia through fluorescent tagging of lipids. The results showed that when exposed to AICAR at pH 6.58, D. magna consumed more oxygen and had lower overall levels of lipids, which is consistent with the expected effects of AICAR, such as increased respiration and lipid metabolism.

STUDIES OF CELL COMMUNICATION BY USING GAP JUNCTION CHANNELS RECONSTITUTE IN UNILAMELLAR LIPID VESICLES

  • Joe, Cheol-O
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
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    • pp.6-6
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    • 1996
  • Gap junction channels were reconstituted into unilamellar liposomes using immunoaffinity purified connexin 32 gap junction protein from rat liver. Vesicles containing open channels and close channels were separated by means of iso-osmolar sucros density gradient sedimentation. The open channels formed in lipid vesicles were permeable to a fluorescent dye molecule, lucifer yellow of which the hydrodynamic size is similar to pore size of gap junctions in vivo. (omitted)

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고추 가공 및 저장 조건에 따른 지질성분의 변화 (Changes in Lipid Composition of Korean Red Pepper(Capsicum annuum) during Processing and Storage)

  • 황성연;조대희;조재선
    • 한국식생활문화학회지
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    • 제13권5호
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    • pp.519-528
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    • 1998
  • This study was conducted to investigate the changes in lipid components of red pepper under various processing conditions such as drying, exposing to lights, extraction and storage conditions. The material used in this study were Chungyang cultivars which contained the largest amount of capsaicinoids and Dabok cultivars which is most popular among the various cultivars produced in Korea. Total lipid content of Chungyang was 15.7% and those of Dabok, Udeungsang and Hongilpum were about 17%. The neutral lipid content in free lipid of red pepper was $78.5{\sim}80.3%$ Phospholipid and glycolipid content in bound lipid were $53.8{\sim}56.9%$ and $35.3{\sim}38.7%$ respectively. The major fatty acids of lipid were linoleic, palmitic and oleic acid. Linoleic acid was presented mainly in pericarp, seed and placenta, whereas the most of palmitic acid was presented in stem. Lipid content of cut and whole red pepper were decreased $24.7{\sim}28%$ and $18.1{\sim}21.5%$ by drying for 48hrs at $65^{\circ}C$ and $95^{\circ}C$. And lipid content was also decreased $3.5{\sim}3.6%$ when the red pepper powder was exposed to 15,000 lux of incandescent light for 30 days at $40^{\circ}C$ and $4{\sim}4.9%$ to fluorescent light. The lipid was extracted the highest content of $50.5{\sim}51.7%$ by acid solution(pH2) and the lowest content by neutral solution, and the higher the salt concentration, the greater the lipid was extracted. During storage at $4^{\circ}C,\;25^{\circ}C,\;40^{\circ}C$, for 30 days the lipid content was not much changed but linoleic acid content was decreased during storage at the same conditions.

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콩기름 저장 중 휘발성분에 대한 LED와 형광등 광원 조사의 영향 (Effect of Light Emitting Diode and Fluorescent Light on Volatile Profiles of Soybean Oil during Storage)

  • 박인선;최덕주;윤예리;이윤정;김윤경;김문호;최소례;김기화;동혜민;한현정;노봉수
    • 한국식품과학회지
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    • 제45권6호
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    • pp.763-769
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    • 2013
  • 본 연구는 전자코를 이용하여 시판 콩기름의 휘발성분이 파장에 영향을 받는지 알아보고자 암실과 형광등 및 파장이 다른 LED 광원을 PE 용기에 조사하면서 12주 동안 변화되는 휘발성분의 패턴 변화를 분석하였다. 콩기름의 휘발성분의 변화는 저장기간동안 암실 처리구에서 가장 적고 천천히 일어났으며, 형광등 처리구에서 가장 크고 급격한 변화가 일어나는 것을 보여주었다. 저장 초기인 4주에는 형광등 처리구를 제외한 모든 처리구에서 미세한 변화가 일어났으며, 저장 12주에는 형광등 처리구 다음으로 파란색과 백색 LED 처리구에서 큰 변화를 보여준 반면 초록색 LED 처리구에서는 변화가 적었다. 파란색과 흰색 LED 처리구는 저장 12주째에서 급격한 휘발성분의 변화가 나타났다. 하지만 빨간색과 초록색 LED 조사 시에는 8주째에 급격한 휘발성분이 생성된 것에 비하여, 저장 12주째에는 휘발성분의 변화가 매우 감소하는 것을 보여주었다. 또한 초록색 LED 처리구에서 가장 적은 변화를 보였다. 이러한 결과를 토대로 볼 때 초록색 LED 광원에서 콩기름을 전시 판매하는 것이 바람직하며, 형광등과 파란색, 백색 LED 광원은 바람직하지 못한 것으로 판단된다. 또한, 전시 판매가 아니라면 암실에서 저장하는 것이 가장 바람직할 것으로 예상되는 바이다.