• 제목/요약/키워드: Nucleic Acids

검색결과 285건 처리시간 0.027초

Extracellular vesicles as novel carriers for therapeutic molecules

  • Yim, Nambin;Choi, Chulhee
    • BMB Reports
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    • 제49권11호
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    • pp.585-586
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    • 2016
  • Extracellular vesicles (EVs) are natural carriers of biomolecules that play central roles in cell-to-cell communications. Based on this, there have been various attempts to use EVs as therapeutic drug carriers. From chemical reagents to nucleic acids, various macromolecules were successfully loaded into EVs; however, loading of proteins with high molecular weight has been huddled with several problems. Purification of recombinant proteins is expensive and time consuming, and easily results in modification of proteins due to physical or chemical forces. Also, the loading efficiency of conventional methods is too low for most proteins. We have recently proposed a new method, the so-called exosomes for protein loading via optically reversible protein-protein interaction (EXPLORs), to overcome the limitations. Since EXPLORs are produced by actively loading of intracellular proteins into EVs using blue light without protein purification steps, we demonstrated that the EXPLOR technique significantly improves the loading and delivery efficiency of therapeutic proteins. In further in vitro and in vivo experiments, we demonstrate the potential of EXPLOR technology as a novel platform for biopharmaceuticals, by successful delivery of several functional proteins such as Cre recombinase, into the target cells.

Selection and Characteristics of Fermented Salted Seafood (jeotgal)-Originated Strains with Excellent S-adenosyl-L-methionine (SAM) Production and Probiotics Efficacy

  • Kim, Min-Jeong;Park, Sunhyun;Lee, Ran-Sook;Lim, Sang-Dong;Kim, Hyo Jin;Lee, Myung-Ki
    • 한국축산식품학회지
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    • 제34권1호
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    • pp.65-72
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    • 2014
  • This study is executed to develop probiotics which produce S-adenosyl-L-methionine (SAM), a methyl group donor of the 5-methyltetrahydrofolate methylation reaction within the animal cell. SAM is an essential substance for the synthesis, activation, and metabolism of hormones, neurotransmitters, nucleic acids, phospholipids, and cell membranes of animals. The SAM is also known as a nutritional supplement to improve brain functions of the human. In this study, the SAM-producing strains are identified in 18 types of salted fish, and then, the strains with excellent SAM productions are being identified, with 1 strain in the Enterococcus genus and 9 strains in the Bacillus genus. Strains with a large amount of SAM production include the lactic acid bacteria such as En. faecium and En. durans, En. sanguinicola, as well as various strains in the Bacillus genus. The SAM-overproducing strains show antibacterial activities with certain harmful microbes in addition to the weak acid resistances and strong bile resistances, indicating characteristics of probiotics. It is possible that the jeotgal-originated beneficial strains with overproducing SAM can be commercially utilized in order to manufacture SAM enriched foods.

A DPL (DNA/peptide/liposomes) Tripartite Complex Effective for Transfection in Serum

  • Kim Young-Cheol;Park Jong-Gu
    • 대한의생명과학회지
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    • 제10권3호
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    • pp.187-194
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    • 2004
  • A short peptide corresponding to the nuclear localization signal (NLS) of human immunodeficiency virus (HIV)-l Tat protein, Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg, was employed to improve the efficiency of cellular uptake of nucleic acids. The peptide was first mixed with a reporter plasmid and then with cationic liposomes to form a tripartite complex of DNA/peptide/liposomes (DPL). Transfection efficiency of the DPL complex was compared with that of the conventional DNA/liposomes (DL) complex. When the DPL complex was formed with various cationic liposomes, DOTAP/DOPE (DP) liposome exhibited superior transfection efficiency to other liposomes tested in vitro. With the inclusion of the peptide, the DPL complex showed much enhanced transfection in various cancer cell lines. Particularly, transfection of the DPL complex in serum increased cellular uptake of a transgene up to 2 fold when compared with that in a serum free condition. Further, when the DPL complex was infused through the ureteric route of a rat, transfection efficiency was shown to be better in reporter gene expression than that obtained with the DL complex. This study shows that the DPL complex that is easy to formulate can be employed for much enhanced cellular uptake of a trans gene.

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Molecular Detection and Analysis of Sweet potato feathery motile vims from Root and Leaf Tissues of Cultivated Sweet Potato Plants

  • Ryu, Ki-Hyun;Park, Sun-Hee
    • The Plant Pathology Journal
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    • 제18권1호
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    • pp.12-17
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    • 2002
  • For the molecular detection of Sweet potaio feathery mottle virus (SPFMV) from diseased sweet potato plants, reverse transcription and polymerase chain reaction (RT-PCR) was performed with the use of a set of virus-specific primers to amplify an 816 bp product. The viral coat protein gene was selected for the design of the primers. No PCR product was amplified when Turnip mosaic virus, Potato vims Y or Cucumber mosaic virus were used as template in RT-PCR with the SPFMV-specific primers. The lowest concentration of template viral RNA required for detection was 10 fg. The vim was rapidly detected from total nucleic acids of leaves and roots from the virus-infected sweet potato plants as well as from the purified viral RNA by the RT-PCR. Twenty-four sweet potato samples were selected and analyzed by RT-PCR and restriction fragment length polymorphism (RFLP). RFLP analysis of the PCR products showed three restriction patterns, which resulted in some point mutations suggesting the existence of quasi-species for the vims in the infected sweet potato plants.

Elimination of Aster Yellows Phytoplasma from Dendranthema grandiflorum by Application of Oxytetracycline as a Foliar Spray

  • Chung, Bong-Nam;Park, Gug-Seoun
    • The Plant Pathology Journal
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    • 제18권2호
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    • pp.93-97
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    • 2002
  • Aster yellows phytoplasma-infected chrysanthemums showing stunt, rosette, and excessive branching were treated with a foliar spray of 400 mg/I oxytetracycline at three-day interval for 1,2,3 and 4 months. Two months after the final treatment, new shoots from the recovered chrysanthemums showed the recurrence of the disease symptoms. However, cuttings from chrysanthemums treated with oxytetracycline did not express any photoplasma infection symptoms for more than 10 months. Also, chrysanthemums dipped in 100 mg/I oxytetracycline solution combined with a foliar spray of 400 mg/I oxytetracycline for 4 weeks showed the same results. Using an electron microscope, ultrathin sections of leaf midribs of chrysanthemum cuttings treated with oxytetracycline for 4 months did not show phytoplasma bodies 10 months after treatment. Nucleic acids from chrysanthemums, which did not express phytoplasma infection symptoms for more than 10 months, did not amplify 16S rRNA gene of phytoplasma by polymerase chain reaction. These results may have implications in the propagation of phytoplasma-free healthy stocks for a wide range of plant species.

식품, 영양과 암의 관계 (Food, Nutrition and Cancer)

  • 류태형
    • 한국식품영양과학회지
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    • 제14권3호
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    • pp.305-313
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    • 1985
  • There is a trend that the total number of cancer cases is steadily increasing as the population grows. It has been estimated that 85% of the cancer rate in the U.S. is attributed to environmental factors. Among the environmental factors, diet and nutrition appear to be related to the largest number of human cancers. Diet and nutrition might be related to cancer by several mechanisms. Food may contain a direct carcinogen or precursors that become carcinogens by spontanous reactions, or by host metabolism, or through the actions of microbial flora. Chemicals that cause cancers generally have reactive electrophilic centers which can combine with electron-rich atoms in nucleic acids and cause cancers by changing the genetic activity of the cells. A variety of factors in foods might be involved in the etiology of carcinogenesis. Chemicals in food that cause cancers include carcinogens of plants and animal origin and also those in drinking water. Other then these, fungal metabolites alcohol, asbestos, heavy metals, pesticides, and food additives might be included as food carcinogenesis. The method of cooking foods also might contribute to carcinogenesis. Some chemicals in foods act as promoters in carcinogenesis. Prevention of cancers by dietary practises have received much interest. Consumption of certain vegetables or cellulose can reduce carcinogenic activity of several compounds. A variety of antioxidants or micronutrients may be effective anticarciongens. Glutathione in the soluble fraction of the cells, is a major defense against oxidative and alkylating carcinogens. Recently anticarcinogenic activity of chlorophyll was demonstrated. Daily consumption of milk appears to effectively reduce stomach cancer.

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Parenteral Formulations Based on Albumin Particulate Technology

  • Lee, Hong-Hwa;Lee, Min-Jung;Heo, Sun-Ju;Sah, Hong-Kee
    • Journal of Pharmaceutical Investigation
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    • 제40권spc호
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    • pp.83-95
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    • 2010
  • Over the years, nanoparticle drug delivery systems have demonstrated versatile potentials in biological, medical and pharmaceutical applications. In the pharmaceutical industry nanotechnology research has mainly focused on providing controlled drug release, targeting their delivery to specific organs, and developing parenteral formulations for poorly water soluble drugs to improve their bioavailability. Achievement in polymer industry has generated numerous polymers applicable to designing nanoparticles. From viewpoints of product development, a nanocarrier material should meet requirements for biodegradability, biocompatibility, availability, and regulatory approval crieteria. Albumin is indeed a material that fulfills such requirements. Also, the commercialization of a first albumin-bound paclitaxel nanoparticle product (Abraxane$^{TM}$) has sparked renewed interests in the application of albumin in the development of nanoparticle formulations. This paper reviews the intrinsic properties of albumin, its suitability as a nanocarrier material, and albumin-based parenteral formulation approaches. Particularly discussed in detail are albumin-based particulate injectables such as Abraxane$^{TM}$. Information on key roles of albumin in the nab$^{TM}$ technology and representative manufacturing processes of albumin particulate products are provided. It is likely that albumin-based particulate technology would extend its applications in delivering drugs, polypeptides, proteins, vaccines, nucleic acids, and genes.

1H, 15N and 13C resonance assignment and secondary structure prediction of ss-DNA binding protein 12RNP2 precursor, HP0827 from Helicobacter pylori

  • Jang, Sun-Bok;Ma, Chao;Chandan, Pathak Chinar;Kim, Do-Hee;Lee, Bong-Jin
    • 한국자기공명학회논문지
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    • 제15권1호
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    • pp.69-79
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    • 2011
  • HP0827 has two RNP motif which is a very common protein domain involved in recognition of a wide range of ssRNA/DNA.We acquired 3D NMR spectra of HP0827 which shows well dispersed and homogeneous signals which allows us to assign 98% of all $^1H_N$, $^{15}N$, $^{13}C_{\alpha}$, $^{13}C_{\beta}$ and $^{13}C$=O resonances and 90% of all sidechain resonances. The sequence-specific backbone resonance assignment of HP0827 can be used to gain deeper insights into the nucleic acids binding specificity of HP0827 in the future study. Here, we report secondary structure prediction of HP0827 derived from NMR data. Additionally, ssRNA/DNA binding assay studies was also conducted. This study might provide a clue for exact function of HP0827 based on structure and sequence.

Leakage of Cellular Materials from Saccharomyces cerevisiae by Ohmic Heating

  • Yoon, Sung-Won;Lee, Chung-Young-J.;Kim, Ki-Myung;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • 제12권2호
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    • pp.183-188
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    • 2002
  • The ohmic heating of foods for sterilization provides a shorter come-up time compared to conventional thermal processes. The electric fields as well as the heat generated by ohmic heating facilitate germicidal effects. In the present study, the effect of ohmic heating on the structure and permeability of the cell membrane of yeast cells, Saccharomyces cerevisae, isolated from Takju (a traditional Korean rice-beer), was investigated. The ohmic heating was found to translocate intracellular protein materials out of the cell wall, and the amount of exuded protein increased significantly as the electric field increased from 10 to 20 V/cm. As higher frequencies were applied, more materials were exuded. Compared to conventional heating, more amounts of proteins and nucleic acids were exuded when these cells were treated with ohmic heating. The molecular weights of the major exuded proteins ranged from 14 kDa to 18 kDa, as analyzed by Tricine-SDS PAGE. A TEM study also confirmed the leakage of cellular materials, thus indicating irreversible damage to the cell wall by ohmic heating. It was, therefore, concluded that the electric fields generated by ohmic heating induced electroporation, causing irreversible damage to the yeast cell wall and promoting the translocation of intracellular materials.

Advanced Glycation End Products and Diabetic Complications

  • Singh, Varun Parkash;Bali, Anjana;Singh, Nirmal;Jaggi, Amteshwar Singh
    • The Korean Journal of Physiology and Pharmacology
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    • 제18권1호
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    • pp.1-14
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    • 2014
  • During long standing hyperglycaemic state in diabetes mellitus, glucose forms covalent adducts with the plasma proteins through a non-enzymatic process known as glycation. Protein glycation and formation of advanced glycation end products (AGEs) play an important role in the pathogenesis of diabetic complications like retinopathy, nephropathy, neuropathy, cardiomyopathy along with some other diseases such as rheumatoid arthritis, osteoporosis and aging. Glycation of proteins interferes with their normal functions by disrupting molecular conformation, altering enzymatic activity, and interfering with receptor functioning. AGEs form intra- and extracellular cross linking not only with proteins, but with some other endogenous key molecules including lipids and nucleic acids to contribute in the development of diabetic complications. Recent studies suggest that AGEs interact with plasma membrane localized receptors for AGEs (RAGE) to alter intracellular signaling, gene expression, release of pro-inflammatory molecules and free radicals. The present review discusses the glycation of plasma proteins such as albumin, fibrinogen, globulins and collagen to form different types of AGEs. Furthermore, the role of AGEs in the pathogenesis of diabetic complications including retinopathy, cataract, neuropathy, nephropathy and cardiomyopathy is also discussed.