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Helper virus-free gutless adenovirus (HF-GLAd): a new platform for gene therapy

  • Liu, Jida;Seol, Dai-Wu
    • BMB Reports
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    • v.53 no.11
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    • pp.565-575
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    • 2020
  • Gene therapy is emerging as a treatment option for inherited genetic diseases. The success of this treatment approach greatly depends upon gene delivery vectors. Researchers have attempted to harness the potential of viral vectors for gene therapy applications over many decades. Among the viral vectors available, gutless adenovirus (GLAd) has been recognized as one of the most promising vectors for in vivo gene delivery. GLAd is constructed by deleting all the viral genes from an adenovirus. Owing to this structural feature, the production of GLAd requires a helper that supplies viral proteins in trans. Conventionally, the helper is an adenovirus. Although the helper adenovirus efficiently provides helper functions, it remains as an unavoidable contaminant and also generates replication-competent adenovirus (RCA) during the production of GLAd. These two undesirable contaminants have raised safety concerns and hindered the clinical applications of GLAd. Recently, we developed helper virus-free gutless adenovirus (HF-GLAd), a new version of GLAd, which is produced by a helper plasmid instead of a helper adenovirus. Utilization of this helper plasmid eliminated the helper adenovirus and RCA contamination in the production of GLAd. HF-GLAd, devoid of helper adenovirus and RCA contaminants, will facilitate its clinical applications. In this review, we discuss the characteristics of adenoviruses, the evolution and production of adenoviral vectors, and the unique features of HF-GLAd as a new platform for gene therapy. Furthermore, we highlight the potential applications of HF-GLAd as a gene delivery vector for the treatment of various inherited genetic diseases.

INCOTERMS 2000 and Maritime Term (INCOTERMS 2000과 해상매매조건(海上賣買條件))

  • Oh, Won-Suk
    • THE INTERNATIONAL COMMERCE & LAW REVIEW
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    • v.13
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    • pp.105-129
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    • 2000
  • INCOTERMS 2000 was not changed in its basic system but much improved in many minor aspects, reflecting the expansion of customs free zone, the spread of electronic communications and the changes of transportation practices. Among many points, following revisions are to be considered important to examine : First, the term 'delivery' is clearly defined in the preamble of each term, which is very significant since the risk is connected with the delivery in international trade. Second, the obligation of carrying out export customs formalities in FAS and import customs formalities in DEQ are imposed to seller and buyer respectively according to current trade practices. Third, since the customs formalities are no more needed among the countries of customs free zone, the phrase 'where applicable' is inserted in A.2 and B.2 of each term. Fourth, the method of delivery(for example, loaded or unloaded) became very concrete and clear in each term. Fifth, in the obligation of buyer's contracting insurance, the 'no obligation' is added even though the buyer has no insurance obligation to the seller. Finally the I.C.C. tried to make unification of the special terminologies be in accordance with those of CISG to achieve international harmony between the uniform law and uniform usages.

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Expansion of coffee shop untact service and research on delivery service - Focusing on coffee delivery keywords that utilize big data - (언택트 서비스 증가와 커피전문점 배달서비스 연구 - 빅 데이터를 활용한 커피배달 키워드 중심으로 -)

  • Lim, Miri;Ryu, Gihwan
    • The Journal of the Convergence on Culture Technology
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    • v.8 no.3
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    • pp.183-189
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    • 2022
  • COVID-19 is also influencing the coffee industry. This will increase untact consumption, a new consumption trend. Consumption utilizing online channels and delivery service applications that represent untact consumption is becoming commonplace. The coffee industry is also increasingly using coffee shops with drive-through and smart ordering systems that can be ordered with minimal contact. While most of the untact services are preempted at franchise stores, many independent coffee shops still offer differentiated services by communicating directly with customers. However, along with the prolonged COVID-19 infection, coffee shops in the present era, which cannot be free from infectious diseases, have no choice but to worry about delivery services. Therefore, this study analyzed the factors that influence coffee delivery services. Research results due to the influence of COVID-19, regular delivery services have increased along with coffee delivery services. Regular delivery services will play a central role in coffee delivery services due to increased use of home cafes by consumers who want to enjoy coffee in various ways.

Development of Hydrophobically Modified Casein Derivative-Based Delivery System for Docosahexaenoic Acids by an Acid-Induced Gelation

  • Ho-Kyung Ha;Dan-Bi Woo;Mee-Ryung Lee;Won-Jae Lee
    • Food Science of Animal Resources
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    • v.43 no.2
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    • pp.220-231
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    • 2023
  • Although omega-3 fatty acids including docosahexaenoic acid (DHA) contain various health-promoting effects, their poor aqueous solubility and stability make them difficult to be induced in dairy foods. The aims of this research were to manufacture casein derivative-based delivery system using acid-induced gelation method with glucono-σ-lactone and to investigate the effects of production variables, such as pH and charged amount of linoleic acid, on the physicochemical properties of delivery systems and oxidative stability of DHA during storage in model milk. Covalent modification with linoleic acid resulted in the production of casein derivatives with varying degrees of modification. As pH was reduced from 5.0 to 4.8 and the charged amount of linoleic acid was increased from 0% to 30%, an increase in particle size of casein derivative-based delivery systems was observed. The encapsulation efficiency of DHA was increased with decreased pH and increased charged amount of linoleic acid. The use of delivery system for DHA resulted in a decrease in the development of primary and secondary oxidation products. An increase in the degree of modification of casein derivatives with linoleic acid resulted in a decrease in the formation of primary and secondary oxidation products than of free DHA indicating that delivery systems could enhance the oxidative stability of DHA during storage in model milk. In conclusions, casein derivatives can be an effective delivery system for DHA and charged amount of linoleic acid played a key role determining the physicochemical characteristics of delivery system and oxidative stability of DHA.

Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis

  • Cha, Eun Bi;Shin, Keun Koo;Seo, Jinho;Oh, Doo-Byoung
    • BMB Reports
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    • v.53 no.8
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    • pp.442-447
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    • 2020
  • The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.

Effects of San-Yin-Jio(SP-6) pressure on duration of delivery time and quantity of blood loss for primipara (삼음교(三陰交) 자극이 초산부(初産婦)의 분만(分娩) 소요시간과 실혈량(失血量)에 미치는 효과(效果))

  • Kim, Woo-hwan;Kim, Won-ill;Lee, Kyung-hee;Youn, Hyoun-min
    • Journal of Acupuncture Research
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    • v.20 no.5
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    • pp.82-92
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    • 2003
  • Objective: To appreciate the effect of San-Yin-Jiao(SP-6) pressure on duratof delivery time and quantity of blood loss in order to verify the possibility of application to clinic of San-Yin-Jiao pressure. Methods: The design of this study is nonequivalent control group non- synchronized design. The subjects are 39 persons who are made up of SP-6 pressure applied(experimental) group 20 persons and to control group 19 persons. Collected data were analyzed as frequency, percentage, t-test, $x^2-test$ using SPSS 10.0 WIN Program. Conclusions: San-Yin-Jiao(SP-6) pressure not only make short duration of delivery time, but also decline quantity of blood loss in effect. 1. It could be necessary that the study of the effects of San-Yin-Jio(SP-6) pressure on duration of delivery time and quantity of blood loss for primipara be done repeatedly. 2. It could be necessary to increase this sort of study through free random experimental design in order to generalize this experimental result.

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Comparison of Inhibitory Effects of 17-AAG Nanoparticles and Free 17-AAG on HSP90 Gene Expression in Breast Cancer

  • Ghalhar, Masoud Gandomkar;Akbarzadeh, Abolfazl;Rahmati, Mohammad;Mellatyar, Hassan;Dariushnejad, Hassan;Zarghami, Nosratallah;Barkhordari, Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7113-7118
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    • 2014
  • Background: HSP90 may be overexpressed in cancer cells which are greatly dependent on Hsp90 function. Geldanamycin derivative 17 allylamino-17-demethoxygeldanamycin (17-AAG) inhibits the function and expression of HSP90. 17-AAG has poor water-solubility which is a potential problem for clinical practice. In this study for improving the stability and solubility of molecules in drug delivery systems we used a ${\beta}$-cyclodextrin-17AAG complex. Materials and Methods: To assess cytotoxic effects of ${\beta}$-cyclodextrin-17AAG complexes and free 17AAG, colorimetric cell viability (MTT) assays were performed. Cells were treated with equal concentrations of ${\beta}$-cyclodextrin- 17AAG complex and free 17AAG and Hsp90 gene expression levels in the two groups was compared by real-time PCR. Results: MTT assay confirmed that ${\beta}$-cyclodextrin- 17AAG complex enhanced 17AAG cytotoxicity and drug delivery in T47D breast cancer cells. The level of Hsp90 gene expression in cells treated with ${\beta}$-cyclodextrin- 17AAG complex was lower than that of cells treated with free 17AAG (P=0.001). Conclusions: The results demonstrated that ${\beta}$-cyclodextrin- 17AAG complexes are more effective than free 17AAG in down-regulating HSP90 expression due to enhanced ${\beta}$-cyclodextrin-17AAG uptake by cells. Therefore, ${\beta}$-cyclodextrin could be superior carrier for this kind of hydrophobic agent.

Experimental Investigation on Key Parameters in Air-powered Needle-free Injection System for Skin Treatment (피부 치료를 위한 공압식 무침 주사 시스템의 주요 인자에 관한 실험적 연구)

  • Kim, Jung Kyung;Mohizin, Abdul;Lee, Seung Ku
    • Journal of the Korean Society of Visualization
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    • v.16 no.1
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    • pp.42-47
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    • 2018
  • A needle-free injector is one of the new non-invasive players in impregnating the biological barriers. It is considered as the next phase in drug delivery and therapeutic applications. One of the major fields of application is in skin remodeling procedures. Although many studies were carried out in understanding the principle in the needle-free injection procedure, fewer studies were done with the aim of therapeutic applications. In the present study, we tried to identify key parameters that affect the jet divergence and peak stagnation pressure on the skin surface in a conventional needle-free injector for skin treatment. A summary of the working principle and effect of the key parameters are presented.

Calculation of Free Nicotine by Determination of pH and Nicotine in Tobacco (담배 중 pH와 Nicotine 함량에 의한 Free nicotine 측정)

  • Lee Jeong-Min;Jang Gi-Chul;Hwang Keon-Joong;Kim Yong-Ha;Rhee Moon-Soo
    • Journal of the Korean Society of Tobacco Science
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    • v.27 no.2
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    • pp.219-225
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    • 2005
  • Characterizing nicotine delivery from tobacco products is important in the understanding of their addictive potential. Most previous studies report total nicotine and have not differentiated between nicotine in its protonated or free-base form. The amount of free nicotine calculated by determining pH and nicotine contents. The pH and nicotine contents in smokeless tobacco product, tobacco products and tobacco leaves were analyzed by Health Canada-Official Method T-310 and CORESTA Recommended Method $N^{\circ}62$. The content of free nicotine was calculated according to the Henderson-Hasselbalch equation and the value of $\alpha_{fb}$(the fraction of nicotine that is in the free base form) by using a pKa value of 8.02 for nicotine. The percentage of free nicotine then was calculated by dividing the free nicotine content by total nicotine content. The pH value and percentage of free nicotine ranged from 5.01 to 5.45 and $0.10\%\;to\;0.27\%$ in cut tobacco and 5.10 to 7.10 and $0.12\%\;to\;10.73\%$ in tobacco leaves, respectively.

Surfactant-free microspheres of poly($\alpha$-caprolactone)/poly(ethylene glycol)/poly($\varepsilon$-caprolactone) triblock copolymers as a novel protein carriers

  • Sun, Sang-Wook;Jeong, Young-Il;Jung, Sun-Woong;Kim, Sung-Ho
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.408.2-409
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    • 2002
  • The aim of this study is to prepare biodegradable microspheres without use of any kind of surfactants or emulsifiers for a novel sustained delivery carriers of protein drugs. Poly(e-caprolactone)/poly(ethylene glycol)/poly(e-caprolactone) (CEC) triblock copolymer was synthesized by ring-opening of e-caprolactone with dihydroxy poly(ethylene glycol) and was used to make surfactant-free microspheres. (omitted)

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