• Title/Summary/Keyword: carboxyfluorescein

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A Permeability Measurement of Small Unilamellar Vesicles by 6-Carboxyfluorescein$^*$

  • Lee, Choong-Hee;Choi, Myung-Un
    • Bulletin of the Korean Chemical Society
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    • v.5 no.4
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    • pp.154-158
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    • 1984
  • In order to characterize the permeability of small unilamellar vesicles (SUV), efflux of 6-carboxyfluorescein (6-CF) from the vesicles was monitored spectrophotofluorometrically. Since the entrapped highly quenched 6-CF (200 mM) became fluorescent upon release from the vesicles, the 6-CF could be used as an efflux probe. SUV containing entrapped 6-CF was prepared from egg phosphatidylcholine and separated by gel filtration on Sepharose 4B. Observed change of relative fluorescent intensity with time was sigmoidal. From this curve, the parameter of permeability was determined either by half-time or a released amount per unit time from the initial slope. Half-time of efflux of prepared SUV having 302 ng phospholipid/ml in 10 mM Tris-HCl buffer pH 7.4 was 21.0 min at $37{\circ}C$. Various factors which could affect the half-time were examined including temperature, pH, salt, and vesicle concentration. In particular the effect of vesicle concentration on the efflux revealed that the permeability can be a function of the concentration.

A study of stabilization effect of .alpha.-tocopherol incorporated into liposomal phospholipid membrane

  • Bae, Song-Ja;Kim, Nam-Hong
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.64-68
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    • 1990
  • The stabilization effect of .alpha.-tocopherol incorporated into liposomal phospholipid membrane was investigated by fluorospectrophotometry and UV-visible spectretarded by the presence of .alpha.-tocopherol in the bilayer of liposomal phospholipid membrane relative to cholesterol-containing liposomes and pure phospholipid liposomes. .alpha.-tocopherol-containing liposomes prolonged the oxidation of liposomes-embedded heme as those of cholesterol-containing liposomes and pure phospholipid liposomes. Thus .alpha.-tocopherol-containing liposomes may be useful for the carrier systems of nutrients and drugs to phospholipid bilayer and stabilized liposomes.

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Coordinatively Polymerized Bilayer Membranes Prepared with an Amphiphilic Derivative of 2-Thiazolyazophenol in Water

  • 서정훈;신승훈;심현보
    • Bulletin of the Korean Chemical Society
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    • v.18 no.2
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    • pp.190-193
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    • 1997
  • In search of new amphiphiles that form coordinatively polymerized bilayer membranes (CPBMs) in water, N-dodecyl-3-[2-hydroxy-3-(4-carboxymethyl-2-thiazolylazo)-5-methylphenyl]propanamide (5) was prepared. The bilayer membranes of 5 prepared in the presence of 2 equivalents of NaOH were further sonicated in the presence of transition metal ions such as Ni(Ⅱ), Cu(Ⅱ), Co(Ⅱ), or Co(Ⅲ) at > 80 ℃ to obtain CPBMs. The CPBMs of 5 were characterized by transmission electron microscopy, gel filtration, resistivity against disruption in aqueous ethanol, visible spectra, and release of entrapped 5(6)-carboxyfluorescein. Remarkable stabilization of the bilayer membranes through coordinative polymerization was evidenced by the resistivity of the Ni(Ⅱ)-CPBM of 5 against disruption in 40%(v/v) ethanol upon incubation for 20 hr.

A Study on the Preparation of 5(6)-Carboxyflurescein-supported Phospholipid Liposomes and Evaluation of Bacterial Sensing Ability (5(6)-Carboxyflurescein을 담지한 인지질 리포좀의 제조 및 박테리아 센싱 능력 평가에 대한 연구)

  • Han, Minho;Jeon, Jaewoo;Lee, Junyoung;Shin, Eunsuk;Kim, Woojin;Kim, Samsoo
    • Textile Coloration and Finishing
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    • v.33 no.4
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    • pp.338-347
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    • 2021
  • In the case of occlusive dressings currently used in dressings for burn treatment, it is impossible to confirm the replacement time, so replacement is delayed, resulting in additional infection. To solve this problem, liposomes capable of bacterial sensing were prepared using 5(6)-Carboxyfluorescein, Phosphatidylcholine, 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, Cholesterol, and 10,12-Tricosadiynoic acid. In this study, evaluation of changes in drug encapsulation rate in liposomes according to changes in three types of phosphatidylcholine phospholipids during liposome production, high-performance phosphatidylcholine phospholipids selected through vesicle size analysis, low and high temperature stability evaluation, bacterial sensitization ability evaluation, animals cell responses were assessed.

Studies on the structure-activity of antimicrobial peptide isolated from horseshoe crab (투구게로부터 단리된 항균성 펩티드의 구조-활성에 관한 연구)

  • Lee, Hyung-Ho;Park, Jang-Su;Park, Nam-Gyu
    • Journal of fish pathology
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    • v.9 no.1
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    • pp.65-77
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    • 1996
  • Tachyplesin I is an antimicrobial peptide isolated from horseshoe crab. To investigate the mechanism of action of tachyplesin I for phospholipid bilayers, tachyplesin I and five analogs have been synthesized by the solution method. The synthesized five analogs are [$Phe^2$]-tachyplesin I, [$Phe^{8,13}$]-tachyplesin I, [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I with no disulfide bonds, 7(Acm) and 10 (Acm) which denote the fragments [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I. Circular dichroism spectra showed that tachyplesin I took an antiparallel $\beta$-structure in buffer solution and a less ordered structure in acidic liposomes. The carboxyfluorescein leakage experiment indicated that tachyplesin I interacted strongly with neutral and acidic phospholipid bilayers. In fluorescence experiment, the hydrophobic part of the peptide was shown to be embedded in lipid bilayers. All the peptides except for 7(Acm) and 10(Acm) were almost equally active in lipopolysaccharide binding. Therefore, the present study suggested that phospholipid bilayers induced a conformational change of tachyplesin I from the stable $\beta$-structure to a less ordered one.

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Stimulatory Effects of Ginsan on the Proliferation and Viability of Mouse Spleen Cells

  • Ko, Eun-Ju;Joo, Hong-Gu
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.133-137
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    • 2010
  • Ginsan is an acidic polysaccharide purified from Panax ginseng, a famous oriental herb. Although a variety of biological activities of ginsan have been studied, the effects of ginsan on spleen cells are not fully elucidated. We investigated the effect of ginsan on the viability and proliferation of spleen cells. Using Cell Counting $Kit-8^{(R)}$ solution and trypan blue solution, we found that ginsan significantly enhanced viability and proliferation. Multiple clusters, indicating proliferation, were observed in ginsan-treated spleen cells and, carboxyfluorescein succinimidyl ester and surface marker staining assay revealed that ginsan promoted proliferation from $CD19^+$ B cells rather than $CD4^+$ or $CD8^+$ T cells. In addition, ginsan decreased the percentage of late apoptotic cells. Ginsan increased the surface expression of CD25 and CD69 as well as production of interleukin-2 from spleen cells, suggesting increased activation. Taken together, these results demonstrate that ginsan increases the viability and proliferation of spleen cells via multiple mechanisms, valuable information for broadening the use of ginsan in clinical and research settings.

Characterization and Release Behavior of Polymersomes of PEG-Poly(fumaric-sebacic acids)-PEG Triblock Copolymer in Aqueous Solution (PEG-Poly(fumaric-sebacic acids)-PEG 삼중 블록 공중합체로 수용액에서 만들어진 폴리머솜의 분석과 방출특성)

  • Pourhosseini, Pouneh S.;Saboury, Ali A.;Najafi, Farhood;Divsalar, Adeleh;Sarbolouki, Mohammad N.
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.294-301
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    • 2013
  • Polymersomes made of biodegradable triblock copolymers based on poly(fumaric acid-co-sebacoyl chloride)/PEG (PEG-co-P(FA/SC)-co-PEG) were prepared and studied in aqueous solutions. TEM confirmed the formation of vesicles in aqueous media. Aggregation behavior of the copolymers was studied by fluorescence spectroscopy of 8-anilino-1-naphthalenesulfonic acid, and the critical aggregation concentration (c.a.c.) of the copolymer was found to be ${\sim}26.2{\mu}M$ indicating desirable stability of the vesicles. Dynamic light scattering revealed that the size of the vesicles was distributed within the range of 170-270 nm. Turbidity measurements confirmed the relative short-term stability of the polymersomes. Carboxyfluorescein, a hydrophilic compound, was simply encapsulated in the vesicles during polymersome preparation. The release of encapsulant from the polymersomes at 25 and $37^{\circ}C$ lasted about 3 weeks, and the rate of release followed a first-order kinetics. The release is speculated to be primarily carried out through diffusion. These results confirm that these polymersomes are promising as controlled-release carriers of various drugs.

Real time-PCR을 이용한 내분비계장애 생체지표의 발현감수성 비교

  • 이철우;구자민;전성환;나진균;박광식;이문순
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.164-164
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    • 2002
  • 에스트로젠을 처리한 송사리의 간으로부터 choriogenin vitellogenin estrogen receptor의 발현량을 전사수준에서 Real-time을 사용하여 정량.비교하였다. 시험어종으로는 부화 후 5개월 이상된 성숙한 수컷 송사리(Oryzias latipes)를(체중 약 250mg/마리)를 사용하여 17$\beta$-estradiol(25ppt, 50ppt, 100ppt)에 24시간 노출시켰다. Fluorescence dye는 choriogenin vitellogenin estrogen receptor의 경우 FAM (6-carboxyfluorescein)을 사용하였으며, $\beta$-actin의 경우는 VIC를 사용하였다. 프로브에 사용하는 quencher dye는 TAMRA(6-carboxy-N',N',N',N'-tetramethyl rhodamine)을 사용하였다. Internal control로 사용된 $\beta$-actin은 17$\beta$-estradiol의 농도에 상관 없이 0~10pM 범위에서 일정하게 발현됨을 보여주었다. vitellogenin choriogenin L 및 choriogenin H는 17$\beta$-estradiol의 농도에 의존하여 발현이 증가되는 용량-반응양상(Dose-dependent)을 나타내었다. 반면, estrogen receptor는 모든 처리군에서 $10^{-2}$pM 정도로 발혐됨에 따라 본 시험농도의 17$\beta$-estradiol에 의해서는 거의 유도발현이 되지 않음을 보여주었다. choriogenin L, choriogenin H, vitellogenin I 및 estrogen receptor 발현감수성을 비교한 결과, 25ppt 및 50ppt의 17$\beta$-esoadiol 농도에서는 ChgL > ChgH > VTG I >ER의 순으로 감수성이 높았으며, 100ppt 노출에서는 ChgL > VTG I > Chg H > ER의 순으로 감수성이 높게 나타났다. 결론적으로 choriogenin이 에스트로젠물질에 의한 가장 민감한 생체지표임을 알 수 있었다.

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Identification of Hanwoo and Holstein meat using MGB probe based real-time PCR associated with single nucleotide polymorphism (SNP) in Melanocortin 1 receptor (MC1R) gene (소 모색관련 MC1R 유전자의 SNP와 관련한 MGB probe에 기초한 real-time PCR을 이용한 한우육과 Holstein육의 판별)

  • Park, Sung-Do;Kim, Tae-Jung;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.25-28
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    • 2005
  • The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo meat, we performed a single nucleotide polymorphism (SNP) analysis in Melanocortin 1 receptor (MC1R) gene using TaqMan$^{(R)}$ MGB probe-based real-time PCR. Two specific probes (one for Hanwoo and the other for Holstein and Black angus) were designed. At the 5' end of 2 TaqMan$^{(R)}$ MGB probes, 6-carboxyfluorescein (FAM) was labeled for Hanwoo, and VIC for Holstein and Black angus. As a result, Hanwoo samples showed FAM-positive signal only, whereas other samples showed VIC-positive. This result suggests that the TaqMan$^{(R)}$ MGB probe based real-time PCR technique would be very accurate, easy and reproducible method to discriminate between Hanwoo meat and Holstein/Black angus meat.

Effect of Cisplatin on the Frequency and Immuno-inhibitory Function of Myeloid-derived Suppressor Cells in A375 Melanoma Model

  • Huang, Xiang;Guan, Dan;Shu, Yong-Qian;Liu, Lian-Ke;Ni, Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4329-4333
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    • 2015
  • Background: To investigate the change of frequency and immuno-inhibitory function of myeloid-derived suppressor cells (MDSCs) after treatment of cisplatin (DDP) in A375 human melanoma model. Materials and Methods: BALB/c nude mice were inoculated with A375 cells to establish the human melanoma model and randomly divided into control group given normal saline (NS) and experimental group treated with DDP (5 mg/kg). The percentages of MDSCs in the tumor tissue and peripheral blood after DDP treatment were detected by flow cytometry. The proliferation and interferon-${\gamma}$ (IFN-${\gamma}$) secretion of T cells co-cultured with MDSCs were analyzed through carboxyfluorescein succinimidyl ester (CFSE) labeling assay and enzyme-linked immunospot (ELISPOT) assay, respectively. Results: In A375 human melanoma model, DDP treatment could significantly decrease the percentage of MDSCs in the tumor tissue, but exerted no effect on the level of MDSCs in peripheral blood. Moreover, DDP treatment could attenuate the immuno-inhibitory function of MDSCs. T cells co-cultured with DDP-treated MDSCs could dramatically elevate the proliferation and production of INF-${\gamma}$. Conclusions: DDP can decrease the frequency and attenuate immuno-inhibitory function of MDSCs in A375 melanoma model, suggesting a potential strategy to augment the efficacy of combined immunotherapy.