• BMB Reports
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• 제44권10호
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• pp.674-679
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• 2011

Bacillus sphaericus produces mosquito-larvicidal binary toxin composed of BinA and BinB. While BinB is expected to bind to a specific receptor on the cell membrane, BinA interacts to BinB or BinB receptor complex and translocates into the cytosol to exert its activity via unknown mechanism. To investigate functional roles of aromatic cluster in BinA, amino acids at positions Y213, Y214, Y215, W222 and W226 were substituted by leucine. All mutant proteins were highly produced and their secondary structures were not affected by these substitutions. All mutants are able to insert into lipid monolayers as observed by Langmuir-Blodgett trough and could permeabilize the liposomes in a similar manner as the wild type. However, mosquito-larvicidal activity was abolished for W222L and W226L mutants suggesting that tryptophan residues at both positions play an important role in the toxicity of BinA, possibly involved in the cytopathological process after toxin entry into the cells.

• BMB Reports
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• 제48권3호
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• pp.147-152
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• 2015

Small interfering RNA (siRNA) functions through pairing with specific mRNA sequences and results in the mRNA's degradation. It is a potential therapeutic approach for many diseases caused by altered gene expression. The delivery of siRNA is still a major problem due to its rapid degradation in the circulation. Various strategies have been proposed to help with the cellular uptake of siRNA and short or small hairpin RNA (shRNA). Here, we reviewed recently published data regarding local applications of siRNA. Compared with systemic delivery methods, local delivery of siRNA/shRNA has many advantages, such as targeting the specific tissues or organs, mimicking a gene knockout effect, or developing certain diseases models. The eye, brain, and tumor tissues are 'hot' target tissues/organs for local siRNA delivery. The siRNA can be delivered locally, in naked form, with chemical modifications, or in formulations with viral or non-viral vectors, such as liposomes and nanoparticles. This review provides a comprehensive overview of RNAi local administration and potential future applications in clinical treatment.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2005년도 춘계학술대회 논문집
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• pp.1501-1506
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• 2005

This paper presents the design of a multiple beam optical tweezers instrument used for manipulating micro/nano-sized components. The basic equations used in designing the optical tweezers are derived and the stable and time-sharing multiple beam optical tweezers are constructed with scanning mirrors. The laser beam passes through a series of optical components such as lenses, mirrors, and scanning mirrors, and overfills the entrance aperture of microscope objective, which gives a stable trap. By rotating the laser beam with the scanning mirror, the focal positions are translated in the specimen plane and multiple micro/nano-sized objects can be moved. The constructed optical tweezers is used to manipulate cells and liposomes simultaneously and to trap multiple nano-wires. The experiments prove that the developed optical tweezers can be a very versatile manipulation tool for studying gene therapy and nano device fabrication.

• BMB Reports
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• 제28권2호
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• pp.129-137
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• 1995

The ryanodine-receptor $Ca^{2+}$ release channel protein in the sarcoplasmic reticulum membrane of rabbit skeletal muscle plays an important role in muscle exitation-contraction (E-C) coupling. Various types of detergents were tested, including Chaps, cholate, octylglucoside, Zwittergents, Mega-9, Lubrol PX, and Triton X-100 for solubilization of this protein. Among these, Chaps and Triton X-100 were found to optionally solubilize the channel complex. Optimum conditions for this solubilization were pH 7.4 with a salt concentration of 1 M. The addition of phospholipid in the solubilization step helped in stabilizing the protein. The purification of the receptor was performed using sucrose density gradient centrifugation. Various methods [dilution, freeze-thaw, adsorption (Biobeads), and dialysis] were investigated to incorporate the Chaps-solubilized and purified $Ca^{2+}$ release channel protein into liposomes made from different types of phospholipids. Of these, a combined method consisting of a dialysis, freeze-thaw and sonication steps yielded the best results. Reconstituted vesicles produced by this method with 95% phosphatidylcholine (from soybean extract) had good function.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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• pp.286-286
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• 1996

In order to elucidate the mechanism of action of transdermal absorption-enhancing compounds, i.e., pyrrolidone derivatives (2-pyrrolidone, 1-methyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, 1,5-dimethyl-pyrrolidone and 5-methyl-2-pyrrolidone), multilamellar liposome was prepared from the simulated stratum corneum lipid and employed as a model system for the barrier function of the stratum corneum. The liposomal membrane of the stratum corneum lipid liposome (SCLL) behaves as an osmometer and has an excellent barrier function. In addition, its phase transition temperatures are similar to those of human stratum corneum intercellular lipid region. Therefore, SCLL seems to be a useful skin model. To estimate the barrier function of SCLL, the osmotic behavior of SCLL was measured in the presence of pyrrolidone derivatives and the effect on the phase transition temperature of SCLL was also investigated using differential calorimetry. Above a certain concentration (MLAC), enhances perturb the barrier function of the liposome. The relationship between MLACs and the partition coefficient of the pyrrolidone derivatives was observed; the greater the partition coefficients, the smaller the MLAC. This suggests that the more hydrophobic enhancers penetrate into the lipid layer more easily and reduce the barrier function of membrane more effectively. The results of differential scanning thermograms of the SCLL suggest that the pyrrolidone derivatives had incorporated into the lipid layer in the liposome and increased the fluidity of the lipid layer in the liposome. Such activity might have some correlation with the transdermal absorption-enhancing activity these compounds.

• Journal of Microbiology and Biotechnology
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• 제20권8호
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• pp.1185-1188
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• 2010

Previously, papiliocin was isolated from the swallowtail butterfly Papilio xuthus and its antimicrobial activity was suggested. In this study, the antifungal mechanism of papiliocin against Candida albicans was investigated. Confocal laser scanning microscopy (CLSM) and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence analysis indicated that papiliocin disturbed the fungal plasma membrane. Moreover, the assessment of the release of FITC-dextran (FD) from liposomes further demonstrated that the antifungal mechanism of papiliocin could have originated from the pore-forming action and that the radius of the pores was presumed to be anywhere from 2.3 to 3.3 nm.

• 한국응용과학기술학회지
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• 제29권2호
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• pp.295-301
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• 2012

콜라겐 성분을 최대로 보호하면서 안정하게 체내로 흡수 될 수 있도록 고순도 수첨 포스 파티딜콜린과 용매사출방법을 이용하여 콜라겐 리포좀을 제조하였다. 리포좀 막의 안정성을 높이기 위해 포스파티딜콜린에 콜레스테롤을 혼합하여, 에탄올과 프로필렌글리콜 혼합용매에 용해하였으며, 이온의 안정화를 위하여 PBS Buffer를 사용하였다. 다양한 변수에 의해 제조된 콜라겐 리포좀의 특성은 동적광산란광도계(DLS), 주사현미경(SEM), 편광현미경(POM)로 분석하였다.

• 한국식품영양과학회지
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• 제22권1호
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• pp.91-95
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• 1993

Vibrio vulnificus가 생산하는 용혈독소의 단백화학적, 면역화학적 연구에 이용할 목적으로 이 균의 용혈독소에 대한 항혈청을 간편하게 만드는 방법을 실험한 결과는 다음과 같다. Vibrio vulnificus가 생산하는 조용혈독소를 인위적으로 만든 liposome (cholesterol-phos-phatidyl-liposome)에 혼합하여 반응시킨 결과 분자량 50kD의 단백질인 용혈독소만 선택적으로 liposome에 결합되었다. 그러므로 liposome에 결합시킨 조용혈독소를 면역원으로 하고 이 면역원을 토끼의 등근육에 주사하여 항혈청을 간편하게 만들 수 있었으며 이 항혈청은 용혈독소에 대한 특이성이 높았다. 환자와 환경에서 분리된 Vibrio vulnificus의 용혈독소와 liposome를 이용하여 제조한 항혈청을 gel 내 침강반응으로 확인한 결과 단일 침강선을 형성하였다.

• 대한의생명과학회지
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• 제15권4호
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• pp.265-272
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• 2009

Recently, mesenchymal stem cells (MSCs) began to be utilized as a vehicle for ex vivo gene therapy based on their plasticity. Effective and safe transfection of therapeutic genes is a critical step for genetic modification of MSCs. Therefore, optimization of in vitro gene delivery into MSCs is essential to provide genetically modified stem cells. In this study, various cationic liposomes, O,O'-dimyristyl-N-lysyl aspartate (DMKD), DMKD/cholesterol, O,O'-dimyristyl-N-lysyl glutamate (DMKE), DMKE/cholesterol, and N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammonium propane methyl sulfate (DOTAP)/cholesterol, were mixed with plasmid DNA encoding luciferase (pAAV-CMV-Luc) at varied ratios, and then used for transfection to MSCs under varied conditions. The MSCs were also transfected by electroporation under varied conditions, such as voltage, pulse length, and pulse interval. According to the experimental results, electroporation-mediated transfection was more efficient than cationic liposome-mediated transfection. The best MSC transfection was induced by electroporation 3 times pulses for 2 ms at 200 V with 10 seconds of a pulse interval.

• Archives of Pharmacal Research
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• 제25권3호
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• pp.229-239
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• 2002

Recent progress in understanding the molecular basis of cancer brought out new materials such as oligonucleotides, genes, peptides and proteins as a source of new anticancer agents. Due to their macromolecular properties, however, new strategies of delivery for them are required to achieve their full therapeutic efficacy in clinical setting. Development of improved dosage forms of currently marketed anticancer drugs can also enhance their therapeutic values. Currently developed delivery systems for anticancer agents include colloidal systems (liposomes, emulsions, nanoparticles and micelles), polymer implants and polymer conjugates. These delivery systems have been able to provide enhanced therapeutic activity and reduced toxicity of anticancer agents mainly by altering their pharmacokinetics and biodistribution. Furthermore, the identification of cell-specific receptor/antigens on cancer cells have brought the development of ligand- or antibody-bearing delivery systems which can be targeted to cancer cells by specific binding to receptors or antigens. They have exhibited specific and selective delivery of anticancer agents to cancer. As a consequence of extensive research, clinical development of anticancer agents utilizing various delivery systems is undergoing worldwide. New technologies and multidisciplinary expertise to develop advanced drug delivery systems, applicable to a wide range of anticancer agents, may eventually lead to an effective cancer therapy in the future.