• Title/Summary/Keyword: Cytosolic delivery

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Macromolecular Cytosolic Delivery: Cell Membranes as the Primary Obstacle

  • Larson, Gretchen M.;Lee, Kyung-Dall
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.621-628
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    • 1998
  • The "evolution" of a thing, a custom, an organ is thus by no means its progressus toward a goal, even less a logical progressus by the shortest route and with the least expendit ure of force, but a succession of more or less profound, mutually independent processes of subduing, plus the resistances they encounter, the attempts at transformation for the purpose of defense and reaction, and the results of successful counteractions. The form is fluid, but the "meaning" is even more so (Friedrich W. Nietzsche).

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Delivery of Chicken Egg Ovalbumin to Dendritic Cells by Listeriolysin O-Secreting Vegetative Bacillus subtilis

  • Roeske, Katarzyna;Stachowiak, Radoslaw;Jagielski, Tomasz;Kaminski, Michal;Bielecki, Jacek
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.122-135
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    • 2018
  • Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of $CD8^+$ lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

Hydrolysis , Skin Permeation and In Vivo Whitening Effect of Kojic Acid Monostearate as an Antimelanogenic Agent (멜라닌생성억제제인 코직산 모노스테아레이트의 가수분해와 피부투과성 및 in vivo 미백효과)

  • Ha, Yong-Ho;Yu, Sung-Un;Kim, Dong-Sup;Lim, Se-Jin;Choi, Young-Wook
    • YAKHAK HOEJI
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    • v.42 no.1
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    • pp.39-45
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    • 1998
  • Kojic acid, antimelanogenic agent, has been widely used in cosmetics to lighten the skin color. However, it has skin irritancy and instability against pH, temperature and light. To overcome these problems and optimize the molecular structure of kojic acid (KA), a prodrug, kojic acid monostearate(KMS), has been synthesized to modify the topical drug delivery in the point of sustained release of the parent drug via enzymatic hydrolysis during skin absorption. The prodrug was tested for enzymatic hydrolysis with cytosolic fraction of hairless mouse, skin. From the in vitro skin permeation study through hairless mouse skin, we found that KMS was retained in the skin and generated KA continuously by the skin esterase cleavage. In addition, topical formulations of o/w type creams and polyolprepolymer-containing cream were further tested for whitening effects using in vivo yellow skin guinea pig model.

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Inhibition of Melanin Synthesis by Enhanced Cytosolic Delivery of N-glycosylation Inhibitors Using pH-Sensitive Nano-carrier (pH 감응형 나노입자를 이용한 멜라닌 합성저해 연구)

  • Park, Ju-Young;Park, Hyun-Jung;Shim, Jong-Won;Ahn, Soo-Mi;Kim, Junoh;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.1
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    • pp.29-32
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    • 2004
  • Inhibition of the early N-glycosylation process in the endoplasmic reticulum prevents the activation of tyrosinase, a key enzyme for melanin biosynthesis. This work aims at evaluating the increased activity of N-glycosylation inhibitors in vitro b, employing a nano-sized pH-sensitive liposome as a delivery carrier. Melexsome, a pH-sensitive nano carrier loaded with glycosylation inhibitos, was prepared by the hydration method with phospholipids and cholresterol-based amphiphiles. Inhibitory effects of Melexsome on the N-glycosylation process were evaluated by EndoH & PNGaseF digestion and the western blotting. Melanin synthesis was also monitored after treatment with Melexsome Interestingly, Melexsome effectively increased the efficacy of N-glycosylation inhibitors. Melexsome was also much more efficiently translocated into the cytoplasm as observed in CLSM. These results demonstrated that the amphiphilic lipid-based pH-sensitive nano-carriers could be, used as an efficient delivery system for N-glycosylation inhibitor to enhance the effects of skin whitening cosmetics.

A Study of the Generation of Transgenic Chickens That Express Human SOD-3 Protein (사람의 SOD-3 단백질을 발현하는 형질전환 닭 생산 연구)

  • Byun, S.J.;Park, C.;Kim, J.A.;Woo, J.S.;Lee, H.C.;Kim, T.Y.;Kim, S.H.;Seong, H.H.;Park, J.K.;Jeon, I.S.
    • Korean Journal of Poultry Science
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    • v.35 no.3
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    • pp.241-245
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    • 2008
  • Lentiviral vector system is efficient vehicles for the delivery of exogenous genes, and it is generally used in the generation of transgenic chickens. In this study, we used recombinant lentiviral vectors to generate transgenic chicks that express the human superoxide dismutase-3 gene driven by the chicken ovalbumin promoter. It is well known that superoxide dismutases(SODs) are believed to play a crucial role in protecting cells against oxygen toxicity. There are three forms of SOD proteins: cytosolic Cu-Zn SOD, mitochondrial Mn SOD, and extracellular SOD(SOD-3). The recombinant lentivirus containing the human SOD-3 gene was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. From 341 injected embryos, the 78 chicks hatched after 21 days incubation. The hatched chicks were screened for the human SOD-3 gene by using PCR. Two of 47 male chickens that survived to sexual maturity contained the human SOD-3 gene in their semen. These results showed that our transgenic chicken generation system was completely established.

Betaine Induces Epidermal Differentiation by Enhancement of Autophagy through an mTOR-independent Pathway (Betaine의 mTOR 비의존적 자가포식 작용 촉진에 의한 표피 분화 유도 효과)

  • Choi, Seon-Guk;Kim, Mi-Sun;Kim, Jin-Hyun;Park, Sun Gyoo;Lee, Cheon Koo;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.95-101
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    • 2018
  • The epidermis which is stratified by epithelial tissue renewal based on keratinocyte differentiation protects the organism from various environmental insults by forming a physical barrier. Autophagy is a mechanism which mediates lysosomal delivery and degradation of protein aggregates, damaged organelles and intracellular microorganisms. Recent reports have shown that autophagy has critical roles for proper terminal differentiation to stratum corneum via removing metabolic organelles and nuclei. However, whether increasing autophagy can activate epidermal differentiation is unknown. Here, we screened a library of natural single compounds and discovered that betaine specifically increased the LC3 positive cytosolic punctate vesicles and LC3-I to LC3-II conversion in HaCaT human keratinocyte cell line, indicating increased autophagy flux. mTOR pathway, which negatively regulates autophagy, was not affected by betaine treatment, suggesting betaine-induced autophagy through an mTOR-independent pathway. Betaine-induced autophagy was also observed in primary human keratinocyte and skin equivalent. Furthermore, epidermal thickness was increased in skin equivalent under betaine treatment. Overall, our finding suggests that betaine as a novel regulator of autophagy may induce epidermal turnover and improve the skin barrier abnormality of the aged epidermis.