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Enhanced Anti-inflammatory Effects of γ-irradiated Pig Placenta Extracts
김강창,허재혁,윤종광,장유연,김연규,김창규,오유경,김영봉 한국축산식품학회 2015 한국축산식품학회지 Vol.35 No.3
Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti-inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.
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좀개구리밥 ( Lemna gibba G3 ) 의 개화에 미치는 Polyamine 의 영향
김강창(Kang Chang Kim),맹주선(Jue Son Maeng) 한국식물학회 1992 Journal of Plant Biology Vol.35 No.4
The flowering in Lemna gibba G3, a long-day plant, was promoted under continuous light by agmatine, putrescine, spermidine and spermine present in the culture medium. Methylglyoxal-bis (guanylhydrazone) (MGBG) and cyclohexylamine (CHA), inhibitors of polyamine biosynthesis, were found to suppress the flowering in the plants. The vegetative growth rate was kept constant while the flowering was being promoted by the polyamines, and the inhibitors with depressive effect on flowering showed stimulatory effect on vegetative growth. The pattern of vegetative growth during floral promotion or depression was an indication that the promotive action of the polyamines and the suppressive effect of the inhibitors may be outcome of their possible involvement specifically in the flowering process rather than in broad spectrum of growth of L. gibba G3. The degree of promotive action of spermdine and spermine could not be altered (or lessened) by simultaneous application of their inhibitors to the medium. This phenomenon indicates that the flowering process in L. gibba G3 may largely be dependent to the status of endogenous spermidine and spermine. Endogenous level of spermidine in florally induced Lemna, was found to rapidly increase. In 24 h of floral induction, the content reached at the level 2 times higher than that in non-induced plants. The elevated level of spermidine provides an additional, though premature, evidence supporting the postulation that endogenous polyamine status might play an important role in the very early stage of floral induction in L. gibba G3.
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Therapeutic potential of an AcHERV-HPV L1 DNA vaccine
이희정,윤종관,허윤기,조한샘,조연동,권용대,김강창,최지원,이재성,오유경,김영봉 한국미생물학회 2015 The journal of microbiology Vol.53 No.6
Cervical cancer is strongly associated with chronic human papillomavirus infections, among which HPV16 is the most common. Two commercial HPV vaccines, Gardasil and Cervarix are effective for preventing HPV infection, but cannot be used to treat existing HPV infections. Previously, we developed a human endogenous retrovirus (HERV)-enveloped recombinant baculovirus capable of delivering the L1 genes of HPV types 16, 18, and 58 (AcHERV-HP16/18/58L1, AcHERV-HPV). Intramuscular administration of AcHERVHPV vaccines induced a strong cellular immune response as well as a humoral immune response. In this study, to examine the therapeutic effect of AcHERV-HPV in a mouse model, we established an HPV16 L1 expressing tumor cell line. Compared to Cervarix, immunization with AcHERVHPV greatly enhanced HPV16 L1-specific cytotoxic T lymphocytes (CTL) in C57BL/6 mice. Although vaccination could not remove preexisting tumors, strong CTL activity retarded the growth of inoculated tumor cells. These results indicate that AcHERV-HPV could serve as a potential therapeutic DNA vaccine against concurrent infection with HPV 16, 18, and 58.
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최지원,김훈미,윤종관,조연동,이희정,김강창,김창규,김계웅,김영봉 한국미생물학회 2015 The journal of microbiology Vol.53 No.5
Studies of the retroviruses have focused on the specific interaction of the nucleocapsid protein with a packaging signal in the viral RNA as important for this selectivity, but the packaging signal in porcine endogenous retrovirus (PERV) has not been defined. Herein, we identified and analyzed this packaging signal in PERV and found hairpin structures with conserved tetranucleotides in their loops and nucleocapsid recognition sequences; both of which are key elements in the viral packaging signal of MLV. We evaluated packaging efficiency of sequence variants isolated from viral and proviral integrated genomes. All viral packaging sequences (Ψ) were identical, while five distinct packaging sequences were identified from proviral sources. One proviral sequence (Ψ1) was identical to that of the viral Ψ and had the highest packaging efficiency. Three variants (Ψ2, Ψ3, Ψ4) maintained key elements of the viral packaging signal, but had nucleotide replacements and consequently demonstrated reduced packaging efficiency. Despite of the same overall hairpin structure, the proviral variant (Ψ5) had only one GACG sequence in the hairpin loop and showed the lowest packaging efficiency other than ΔΨ, in which the essential packaging sequence was removed. This result, thus, defined the packaging sequences in PERV and emphasized the importance of nucleotide sequence and RNA structure in the determination of packaging efficiency. In addition, we demonstrate efficient infection and gene expression from the PERVbased viral vector, which may serve as a novel alternative to current retroviral expression systems.
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