• 제목/요약/키워드: protein delivery

검색결과 297건 처리시간 0.022초

수분거동 패턴에 따른 차폐막 설정을 통한 모발단백질 소실방지 (Prevention of Protein Loss Using A Shield Coating According to Moisture Behavior in Human Hair)

  • 송상훈;임병택;손성길;강내규
    • 대한화장품학회지
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    • 제46권1호
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    • pp.57-65
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    • 2020
  • 세정 과정에서 물에 의한 단백질 소실을 차폐막을 통해서 방지 하고자 먼저 모발 내부의 물 거동 경로를 파악하였다. 이를 위해서 소수성을 가지는 건강한 모발과, 모발 내부는 정상이지만 표면의 최외곽 지질층에서 18-methyleicosanoic acid (18-MEA)만 소실시켜 친수성이 된 모발과, 다시 표면에 18-MEA를 결합시킨 회복모를 이용해서 모발 수분의 거동을 파악하였다. 모발에 열을 가할 때 일어나는 수분 증발에 따른 질량 변화를 관찰한 결과 표면이 소수성인 모발들은 39 s와 151 s의 두개의 시간 상수로 감소하였다. 이에 반해 표면이 친수성으로 변한 손상 모발에서는 83 s의 하나의 빠른 시간 상수로 감소함을 확인하였는데, 이는 모발 내부에 있는 비결합과 결합 수분들이 외곽으로 빠져나올때 모발 표면에 소수성 막이 없음으로써 바로 용출됨을 반증한다. 따라서 세정과정에서 모발 내부의 단백질 용출을 효율적으로 방지하기 위해서는 모발 표면을 소수성 코팅하여 물분자의 거동을 저지시킬 필요가 있을 것이라는 가정한 결과 친수성 polyethylene glycol (PEG)를 코팅한 경우 179 ㎍/mL의 단백질이 용출한데 반해서 소수성 polydimethylsiloxane (PDMS)를 코팅한 모발은 보다 적은 148 ㎍/mL를 용출하였으며, 표면을 소수성 및 친수성으로 코팅하여 모발 내부 단백질을 검량하여 비교하였다. 마침내 소수성 차폐막이 세정 과정에서 모발 단백질 소실을 방어하는 방법이 됨을 확인하였고, 이 소수성 지표를 lateral force microscopy (LFM) 값으로 환산하여 정리하였다. 마침내 이 연구 결과물은 세정 과정에서 발생하는 단백질 소실을 소수성 코팅막을 부여함으로써 막을 수 있는 모발 세정제품 개발에 기여할 수 있다.

Anti-calcification of Bovine Pericardium for Bioprosthetic Heart Valves after Surface Modification with Hyaluronic Acid Derivatives

  • Hahn Sei Kwang;Ohri Rachit;Giachelli Cecilia M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권3호
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    • pp.218-224
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    • 2005
  • Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.

생분해성 폴리락티드/글리콜리드 미립구를 이용한 재조합 소 성장호르몬(rBST)의 지속성주사제 설계 (Sustained Release Injectable of Recombinant Bovine Somatotropin in Biodegradable Poly(D,L-lactide-co-glyceride) Microspheres)

  • 전홍렬;이봉상;권도우;윤미경;전현주;신택환;최영욱
    • Journal of Pharmaceutical Investigation
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    • 제32권3호
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    • pp.199-207
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    • 2002
  • In order to develop a sustained release formulation of bovine somatotropin (BST), which has been used to increase the body weight of oxen or the milk production of dairy cows, poly(D,L-lactide-co-glyceride)(PLGA) microspheres were made by W/O/W multiple emulsification method and solvent extraction method. Physical properties including particle size, drug entrapment, drug release, protein denaturation, and in vivo body weight increase in rats were characterized. The size of the microspheres was increased as the molecular weight of PLGA increased. When Span 65 and stearic acid during preparation were added, the size was decreased but the amount of surface protein was increased, resulting in a high loading efficiency, with fast release of BST from the microspheres. Aggregation or fragmentation of BST by SDS-PAGE during microsphere preparation and drug release study was not observed. Body weight of Sprague-Dawley's male rats was significantly increased after subcutaneous administrations of BST-loaded PLGA microspheres. There was a good correlation between in vivo weight gain and in vitro release rate of microspheres. PLGA microspheres with a high surface protein ratio could be a good candidate for the sustained delivery of BST.

siRNA Silencing EZH2 Reverses Cisplatin-resistance of Human Non-small Cell Lung and Gastric Cancer Cells

  • Zhou, Wen;Wang, Jian;Man, Wang-Ying;Zhang, Qing-Wei;Xu, Wen-Gui
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권6호
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    • pp.2425-2430
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    • 2015
  • Clinical resistance to chemotherapeutic agents is one of the major hindrances in the treatment of human cancers. EHZ2 is involved in drug resistance and is overexpressed in drug-resistant cancer cell lines. In this study, we investigated the effects of EHZ2 on cisplatin -resistance in A549/DDP and AGS/DDP cells. EHZ2 mRNA and protein were found to be significantly overexpressed in A549/DDP and AGS/DDP cells, compared to parental cells. EHZ2 siRNA successfully silenced EHZ2 mRNA and protein expression. Proliferation was inhibited and drug resistance to cisplatin was improved. Flow cytometry showed that silencing of EHZ2 arrested A549/DDP and AGS/DDP cells in the G0/G1 phase, increasing apoptosis, rh-123 fluorescence intensity and caspase-3/8 activities. Silencing of EHZ2 also significantly reduced the mRNA and protein expression levels of cyclin D1 and MDR1,while up-regulating p15, p21, p27 and miR-218 in A549/DPP cells. Furthermore, silencing of EHZ2 also significantly increased the expression level of tumor suppressor factor miR-218. We also found down-regulating EHZ2 expression increased methylation in A549/DDP and AGS/DDP cells. This study demonstrates that drug resistance can be effectively reversed in human cisplatin-resistant lung and gastric cancer cells through delivery of siRNAs targeting EHZ2.

열변성 단백질이 결합된 음이온성 리포솜의 혈장 내 안정성 및 세포 내 이입 평가 (Stability in Plasma and Intracellular Uptake of Thermally Denatured Protein-coated anionic Liposomes)

  • 이미정;황인영;김성규;정석현;정서영;성하수;조선행;신병철
    • Journal of Pharmaceutical Investigation
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    • 제39권6호
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    • pp.423-429
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    • 2009
  • Liposomes have been used as one of the efficient carriers for drug delivery. In this study, anionic liposomes of which surface was modified by using both electrostaic interaction between anionic liposomes and cationically charged BSA molecules at lower pH than isoelectric point (pI) of BSA and denaturation of the BSA-coated liposomes by thermal treatment. The thermally denatured BSA-coated liposomes (DBAL) had mean particle diameter of 125.2${\pm}$1.7 nm and zeta potential value of -22.4${\pm}$4.5 mV. Loading efficiency of model drug, doxorubicin (DOX), into liposomes was 83.0${\pm}$2.6%. Results of in vitro stability study of DBAL in blood plasma showed that the mean particle diameter of DBAL 400 did not increase in blood plasma and adsorption of plasma protein was much less than plain or anionic liposomes. Intracellular uptake of DBAL 400 evaluated by confocal microscopy observation was higher than that of PEG liposomes.

Roles of ERK and NF-${\kappa}$ B in Interleukin-8 Expression in Response to Heat Shock Protein 22 in Vascular Smooth Muscle Cells

  • Kang, Seung-Hun;Lee, Ji-Hyuk;Choi, Kyung-Ha;Rhim, Byung-Yong;Kim, Koan-Hoi
    • The Korean Journal of Physiology and Pharmacology
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    • 제12권4호
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    • pp.171-176
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    • 2008
  • Heat shock proteins (HSPs) serve as molecular chaperones and play a role in cell protection from damage in response to stress stimuli. The aim of this article is to investigate whether HSP22 affects IL-8 expression in vascular smooth muscle cells (VSMCs), and which cellular factors are involved in the HSP-mediated IL-8 induction in that cell type in terms of mitogen activated protein kinase (MAPK) and transcription element. Exposure of aortic smooth muscle cells (AoSMCs) to HSP22 not only enhanced IL-8 release but also induced IL-8 transcript via promoter activation. HSP22 activated ERK and p38 MAPK in AoSMCs. HSP22-induced IL-8 release was inhibited by U0126, but not by SB202190. A mutation in the IL-8 promoter region at the binding site of NF-${\kappa}$ B, but not AP-1 or C/EBP, impaired promoter activation in response to HSP22. Delivery of I ${\kappa}$ B, but not dominant negative c-Jun, lowered HSP22-induced IL-8 release from AoSMCs. These results suggest that HS P22 induces IL-8 in VSMCs via ERK1/2, and that transcription factor NF-kB may be required for the HSP22-induced IL-8 up-regulation.

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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    • 제6권5호
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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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Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale

  • Lee, Chang-Soo;Lee, Sang-Ho;Kim, Yun-Gon;Choi, Chang-Hyoung;Kim, Yong-Kweon;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권2호
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    • pp.146-153
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    • 2006
  • A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.

Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)

  • Park, Yoon Mi;Kim, Kyung A;Kang, Min Uk;Park, Kwan Ho;Nho, Si Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제28권1호
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    • pp.10-18
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    • 2014
  • Baculoviruses base vectors come to be regarded as methods for in vivo gene delivery and transient expression to the silkworm. In the case of silkworm, B. mori, two types of baculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV (Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently, AcNPV showed promising results with some silkworm strains despite different host-specificities. We searched for a highly-permissive silkworm strain in the B. mori stocks of Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on the measured luciferase activity, the strains could be divided into three groups, high-, middle-, and low-permissive strains, according to their relative recombinant protein expression levels. At 48 hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greater than that of the low-permissive strains. At 72 hours post-injection, a significant elevation in luciferase activity was observed in the hemocytes of all strains. Then, based on the above results, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 were pick up and was carried out Dot blotting, RT-PCR and Real time PCR.

Antidepressant effects of aqueous extract of saffron and its effects on CREB, P-CREB, BDNF, and VGF proteins in rat cerebellum

  • Asrari, Najmeh;Yazdian-Robati, Rezvan;Abnous, Khalil;Razavi, BiBi Marjan;Rashednia, Mrazieh;Hasani, Faezeh Vahdati;Hosseinzadeh, Hossein
    • 대한약침학회지
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    • 제21권1호
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    • pp.35-40
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    • 2018
  • Objective: The role of BDNF (brain-derived neurotrophic factor), CREB (cAMP response element binding) and VGF neuropeptide has been proved in antidepressant activity of long term saffron administration in the rat hippocampus. In this study we evaluated the role of these proteins in antidepressant activity of saffron in long term administration in the rat cerebellum. Methods: Saffron aqueous extract (40 and 80 mg/kg/day) and imipramine (10 mg/kg/day) were administered intraperitoneally for 21 days to rats. At the end of experiment, animals were sacrificed and cerebellums were separated. The protein levels of BDNF, VGF, CREB and P- CREB in the rat cerebellum were evaluated using western blot analysis. Results: Saffron aqueous extract (80mg/kg/day) caused significant increase in protein level of P-CREB in long term treatment in the rat cerebellum. The increases in the protein levels of VGF, CREB and BDNF were not significant. Conclusion: In summary, our results showed that antidepressant effect of saffron in rat cerebellum might be due to the enhanced phosphorylation of CREB.