• Title/Summary/Keyword: poly-L-lysine

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Liquid Crystal Based Optical Sensor for Imaging Trypsin Activity at Interfaces Between Aqueous Phases and Thermotropic Liquid Crystals

  • Zhang, Minmin;Jang, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.10
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    • pp.2973-2977
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    • 2013
  • In this study, we developed a liquid crystal (LC)-based optical sensor for monitoring enzymatic activity through orientational changes in liquid crystals (LCs) coupled to the properties of a poly-${\small{L}}$-lysine (PLL)-based polymeric membrane. We prepared a PLL-based polymeric membrane at the planar interface between the thermotropic liquid crystal and aqueous phases. The PLL-based polymeric membrane was obtained by contacting the PLL solution with water immiscible LCs, 4-cyano-4'-pentyl-biphenyl (5CB) doped with adipoyl chloride. We then investigated the membrane properties by examining the permeability of the membrane to phospholipids, 1,2-didodecanoyl-rac-glycero-3-phosphocholine (DLPC). The permeability of the membrane to transport phospholipids was monitored through the orientational transition of 5CB in contact with the dispersions of DLPC. Since trypsin can enzymatically catalyze the hydrolysis of PLL, we incubated an aqueous trypsin solution with the membrane for 2 h at room temperature to cause an increase in the permeability of the polymeric membrane to DLPC. As a result, a bright to dark optical shift of LCs was observed, which implied that an enzymatic reaction between trypsin and PLL-based membrane occurred. Two control experiments using chymotrypsin and bovine serum albumin (BSA) revealed no sign of improved permeability based on the orientational transition of LCs.

Optimization of Experimental Conditions for In vitro P-glycoprotein Assay Using LLC-GA5 Cells

  • Ahn, A-Ra;Oh, Ju-Hee;Lee, Joo-Hyun;Lee, Young-Joo
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.363-366
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    • 2010
  • Identification of compounds that function as P-glycoprotein (P-gp) substrates or inhibitors can facilitate the selection and optimization of new drug candidates. The purpose of this study is to optimize the experimental conditions for in vitro P-gp assay using LLC-GA5 cells, which is a well-known transformant cell line derived by transfecting LLC-PK1 with human MDR1. The amount of rhodamine123 transported by the LLC-GA5 and LLC-PK1 cells was evaluated under the following experimental conditions: 3 different types of transport media, colchicine pretreatment or nontreatment of the cells in the culture media, and with and without poly-L-lysine coating of the culture plates. The assay sensitivity was found to considerably differ depending on the diluents used in the transport media. P-gp-mediated transport in LLC-GA5 cells was most clearly characterized in the Hanks' balanced salt solution based transport media. The sensitivity of P-gp-mediated transport was not changed by colchicine pretreatment or poly-L-lysine coating of the culture plates.

Preparation of Enzyme Electrodes for Biofuel Cells Based on the Immobilization of Glucose Oxidase in Polyion Complex (폴리이온복합체를 이용하여 글루코스 산화효소를 고정화한 바이오전지용 효소전극 제조)

  • Nguyen, Linh Thi My;Li, Nan;Yoon, Hyon Hee
    • Applied Chemistry for Engineering
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    • v.24 no.1
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    • pp.99-103
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    • 2013
  • An emzymatic bioanode for a glucose/oxygen biofuel cell was prepared by the sequential coating of carbon nanotube (CNT), charge transfer complex (CTC) based on tetracyanoquinodimethane (TCNQ) and tetrathiafulvalene (TTF), glucose oxidase (GOx), and polyion complex (mixture of poly-L-lysine hydrobromide and poly (sodium 4-styrenesulfonate)) on a glassy carbon electrode. A biocathode was also prepared by the sequential coating of CNT, bilirubin oxidase (BOD), 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), and polyion complex. The effect of CNT and CTC on the electrochemical performance was investigated. The biofuel cell exhibited a promising performance with maximum power densities of 3.6, 10.1, and $46.5{\mu}W/cm^2$ at 5, 20, and 200 mM of glucose concentration, respectively. The result indicates that the biofuel cell architecture prepared in this study can be used in the development of biofuel cells and biosensors.

Formation of Complex Between Polyelectrolytes and pH/Temperature Sensitive Copolymers (고분자전해질과 pH/온도감응성 고분자 사이의 복합체 형성에 관한 연구)

  • Yoo, Mi Kyong;Sung, Yong Kiel
    • Journal of the Korean Chemical Society
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    • v.42 no.1
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    • pp.84-91
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    • 1998
  • Random copolymers of N-isopropyl acrylamide (NIPAAm) and acrylic acid (AAc) which exhibit temperature- and pH-responsive behavior were synthesized by free-radical polymerization. The copolymers were characterized by means of FT-IR spectrometry and titration. The influence of polyelectrolyte on the lower critical solution temperature (LCST) of pH/temperature-sensitive polymers was investigated in the pH range of 2-12. The LCSTs of PNIPAAm/water in poly(NIPAAm-co-AAc) were determined by cloud-point measurements. A polyelectrolyte complex was prepared by mixing poly(NIPAAm-co-AAc) with poly(allylamine) (PAA) or poly(L-lysine) (PLL) solutions as anionic and cationic polyelectrolytes, respectively. Back titration was performed to determine the content of AAc and to study the effect of comonomer ionization on the LCST. The LCSTs of PNIPAAm/water in the copolymers were strongly affected by pH, presence of polyelectrolyte, AAc content, and charge density on the polymer. The polyelectrolyte complexes were formed at neutral condition. The influence of more hydrophobic PLL as polyelectrolyte on the cloud-point of PNIPAAm/water in the copolymer was stronger than that of poly(allylamine) (PAA).

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Poly(L-lysine) Based Semi-interpenetrating Polymer Network as pH-responsive Hydrogel for Controlled Release of a Model Protein Drug Streptokinase

  • Park, Yoon-Jeong;Jin Chang;Chen, Pen-Chung;Victor Chi-Min Yang
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.5
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    • pp.326-331
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    • 2001
  • With the aim of developing of pH-sensitive controlled drug release system, a poly(Llysine) (PLL) based cationic semi-interpenetrating polymer network (semi-IPN) has been synthesized. This cationic hydrogel was designed to swell at lower pH and de-swell at higher pH and therefore be applicable for achieving regulated drug release at a specific pH range. In addition to the pH sensitivity, this hydrogel was anticipated to interact with an ionic drug, providing another means to regulate the release rate of ionic drugs. This semi-IPN hydrogel was prepared using a free-radical polymerization method and by crosslinking of the polyethylene glycol (PEG)-methacrylate polymer through the PLL network. The two polymers were penetrated with each other via interpolymer complexation to yield the semi-IPN structures. The PLL hydrogel thus prepared showed dynamic swelling/de-swelling behavior in response to pH change, and such a behavior was influenced by both the concentrations of PLL and PEG-methacrylate. Drug release from this semi-IPN hydrogel was also investigated using a model protein drug, streptokinase. Streptokinase release was found to be dependent on its ionic interaction with the PLL backbones as well as on the swelling of the semi-IPN hydrogel. These results suggest that a PLL semi-IPN hydrogel could potentially be used as a drug delivery platform to modulate drug release by pH-sensitivity and ionic interaction.

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Interaction of Bone Marrow Stromal Stem Cells with Adhesive Protein and Polypeptide-adsorbed Poly(lactide-co-glycolide) Scaffolds (골수유래 간엽줄기세포와 점착성 단백질 및 폴리펩타이드가 흡착된(락티이드/글리콜라이드) 공중합체 지지체와의 상호작용)

  • Choi, Jin-San;Lee, Sang-Jin;Jang, Ji-Wook;Khang, Gil-Son;Lee, Young-Moo;Lee, Bong;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.27 no.5
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    • pp.397-404
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    • 2003
  • The interaction of cell adhesive protein and polypeptide with bone marrow stromal stem cells (BMSCs) grown in tissue engineered films and scaffolds were examined. Several proteins or polypeptide known as cell-adhesive were coated adsorption on poly(lactide-co-glycolide) (PLGA) films and scaffolds and adhesion and proliferation behavior of BMSC on those surfaces were compared. The protein and polypeptide used include collagen IV, fibrinogen, laminin, gelatin, fibronectin, and poly(L-lysine). The protein and polypeptide were adsorbed on the PLGA film surfaces with almost monolayer coverage except poly(L-lysine). BMSCs were cultured for 1, 2, and 4 days on the protein- or polypeptide-adsorbed PLGA films and scaffolds. The cell adhesion and proliferation behaviors were assessed by sulforho damine B assay. It was observed that the protein- or polypeptide-adsorbed surfaces showed better cell adhesion and proliferation than the control.

Effects of $Ca^{2+}$ and Polyamine on Callose Contents in Carrot Suspension Cultured Cells (당근 현탁배양세포에서 $Ca^{2+}$과 Polyamine이 Callose 함량에 미치는 영향)

  • 강영희
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.215-226
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    • 1989
  • The effects of Ca2+ on polyamines on callose contents of carrot suspension cultured cells were studied. The regeneration process of the cell wall of carrot protoplast observed through the electron microscope. Treatment of the carrot suspension cultured cells with Ca2+ and polyamines resulted in considerable increase on callose contents at 0.1 mM of Ca2+ and polyamines, particulary spermidine. Poly-L-lysine and poly-L-ornithine increased about 30% and 100% of callose contents than that of the control respectively, whereas verapamil and flunarizine markedly decreased the callose contents. These effects of Ca2+ of free ion rather than as Ca2+-calmodulin complex. During the cultivation of the protoplast, the regeneration of the cell wall was somewhat observed on the 4th day, however, it was inhibited by verapamil. These results suggested that the promotive action of Ca2+ and polyamines were manifested in the callose contents and the regeneraton of the cell wall.

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Preparation of Insulin-Immobilized Polyurethane Films and Their Interaction with Fibroblasts (인슐린을 고정화한 폴리우레탄 막의 제조 및 섬유아세포와의 상호작용)

  • 맹만;김우식;이동호;박이순;강인규
    • Polymer(Korea)
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    • v.28 no.6
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    • pp.538-544
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    • 2004
  • Polyurethanes containing L-lysine segments in the main chain (PULL) were synthesized from 4,4'-diphenymethyl diisocynate, poly(tetramethylene glycol), and z-lysine oligomer as a chain extender. Insulin-immobilized polyurethanes (PULL-In) were prepared by a coupling reaction of PULL surface amino groups with insulins. The amount of immobilized insulin was about 0.30 nmol/$\textrm{cm}^2$, as determined by Bradford method. The interactions of NIH/3T3 fibroblasts with surface-modified PULLs were investigated using $^3$H-thymidine incoporation and optical microscopy. The cell growth rate on PULL-In film was higher than those on other substrates. The cell proliferation by the immobilized insulin was almost same as that by the free one.

Consistent and Specific Suppression of Mucin Release from Cultured Hamster Tracheal Surface Epithelial Cells by Poly-L-Lysine

  • Lee, Choong-Jae;Lee, Jae-Heun;Seok, Jeong-Ho;Hur, Gang-Min;Park, Ji-Sun;Bae, So-Hyun;Jang, Hyeon-Seok;Park, Sang-Cheol
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.3
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    • pp.143-147
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    • 2003
  • Poly-L-lysine (PLL) was reported to suppress mucin release from airway goblet cells during 30 min treatment period. In this study, we investigated whether PLL consistently suppresses mucin release from cultured airway goblet cells during 24 h after 30 min treatment and also specifically suppresses the release of mucin without any effects on the other releasable glycoproteins. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3H$-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of PLL to assess the effects on $^3H$-mucin release and on the total elution profile of the treated culture medium. The total mucin content during 24 h after 30 min treatment of PLL was assesed to investigate the consistency of effects. PLL did not affect the release of the other releasable glycoproteins whose molecular weights were less than mucin, and decreased the total mucin content during 24 h after 30 min treatment. We conclude that PLL can specifically suppress mucin release from cultured airway goblet cells and the suppression on mucin release is consistent. This finding suggests that PLL might be used as a specific airway mucin-regulating agent by directly acting on airway mucin-secreting cells.