• 한국식품과학회지
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• 제35권4호
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• pp.556-560
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• 2003

CP4 EPSPS에 대한 특이항체(다클론 및 단클론)을 이용하여 확립한 전보의 sandwich ELISA 분석법을 원료 콩의 GMO 분석에 적용가능한 가를 검토하였다. 원료 콩은 수입산 47점, 국산 20점을 분석하였다. 그 결과, 수입산 콩의 CP4 EPSPS 분석치는 크게 두 군으로 나뉘어 높은 군$(39.1{\pm}13.5{\mu}g/g,\;n=33)$과 낮은 군$(2.6{\pm}1.2{\mu}g/g,\;n=14)$으로 나타났다. 수입산 콩의 GMO 비율은 약 70.2%로 나타났다. 한편, 국산 콩의 CP4 EPSPS의 함량 $(0.9{\pm}0.5{\mu}g/g,\;n=20)$은 아주 낮은 것으로 나타났고 이는 non-GMO인 것으로 판단되었다. 콩나물의 경우에 GM과 non-GM 간 CP4 EPSPS의 차이를 보이고 있었고 GM 콩나물의 자엽부에서 뿌리보다 높은 CP4 EPSPS의 함량을 보여주고 있다. 국산 콩으로 만든 콩나물의 경우 CP4 EPSPS의 함량은 아주 낮은 것으로 나타나 GM과 non-GM 콩간의 구분이 가능하였다. 또한 PCR에 의해 sandwich ELISA가 정확하다는 것을 보여주었다. 이상과 같이 sandwich ELISA 방법은 신속, 간편하게 많은 콩 시료의 GMO 여부를 판단할 수 있음을 보여주었다.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.93-97
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• 2011

Previously, we reported that the osmolarity conditions in the satellite region were affected CpG DNA methylation status while Pre-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. This study was conducted to investigate the DNA methylation status of repeat sequences in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaCl or 0.05 M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. The DNA methylation status of the Pre-1 sequences in blastocysts was characterized using a bisulfite-sequencing method. Intriguingly, in the present study, we found the unique DNA methylation at several non-CpG sequences at the Pre-1 sequences in all groups. The non-CpG methylation was hypermethylated in all three groups, including in vivo group (86.90% of PZM-3; 83.87% of NaCl; 84.82% of sucrose; 90.94% of in vivo embryos). To determine whether certain non-CpG methylated sites were preferentially methylated, we also investigated the methylation degree of CpA, CpT and CpC. Excepting in vivo group, preference of methylation was CpT>CpC>CpA in all three groups investigated. These results indicate that DNA methylation of Pre-1 sequences was hypermethylated in CpG as well as non-CpG site, regardless modification of osmolarity in a culture media.

• Molecular & Cellular Toxicology
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• 제5권3호
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• pp.207-215
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• 2009

Despite wide therapeutic use of CpG ODN against infection, allergy and cancer, the safety and toxicity of CpG ODNs were poorly delineated. Thus, we investigated whether optimal dosing of CpG ODN would affect immunotoxicological parameters in NZB/NZW F1 mice. Comparisons were made among control, non-CpG ODN and mouse CpG ODN ($10{\mu}g$)-treated groups for 4 weeks. To gauge the immunotoxicity of CpG ODNs, we measured nonspecific parameters, degree of lupus nephritis, proteinuria, or autoantibody, and cytokine expression in mRNA level of lymphocytes. We found that there were no significant differences among groups in nonspecific immunotoxicological profiles and in evaluation profiles of glomerulonephritis. However, titer of anti-dsDNA and anti-cardiolipin antibodies in mouse CpG ODN group rose three or eight-fold higher than in control group. Collectively, CpG ODN might be clinically less immunotoxic in terms of clinical profiles in lupus-prone NZB/NZW F1 mice, in spite of high autoantibody titer in CpG ODN treated groups.

• Fisheries and Aquatic Sciences
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• 제10권1호
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• pp.1-7
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• 2007

The induction of cellular and humoral immunity and cytokine gene expression by synthetic CpG oligodexoynucleotides (CpG-ODNs) has not been investigated systematically in olive flounder Paralichthys olivaceus in vivo. We optimized the proper concentration of CpG-ODNs using an in vitro assay for the superoxide anion $(O_2^-)$. CpG-ODNs induced $O_2^-$ and nitric oxide (NO) production, lysozyme activity, and the proinflammatory cytokine gene expression of $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder significantly in vivo, whereas non-CpG-ODNs did not produce these effects or produced them to a lesser extent. This implied that CpG-ODNs could stimulate cellular and humoral immunity and cytokine gene expression in olive flounder. This is the first evidence of NO production and the first study on the mRNA expression of the proinflammatory cytokine genes $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder in response to CpG-ODNs. Comparison of the variation in NO production and lysozyme activity to that of other studies led us to postulate that a group-specific difference exists in the immune responses of olive flounder against CpG-ODNs. Furthermore, the detailed immunostimulatory spectrum of CpG-ODNs in olive flounder could be a useful index with which to analyze the effect of CpG-ODNs against the challenge test prior to field applications.

• BMB Reports
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• 제40권4호
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• pp.486-493
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• 2007

Atopic dermatitis (AD) is a chronic inflammatory skin disease and the pathogenesis of AD is associated with the release of various cytokines/chemokines due to activated $Th_2$ immune responses. Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG dinucleotide in the context of particular base sequence (CpG motifs) are known to have the immunostimulatory activities in mice and to convert from Th2 to Th1 immune responses in AD. We aimed to investigate that CpG ODN, especially phosphodiester form, can stimulate the protective immunity in NC/Nga mice with AD. We isolated BMDCs from NC/Nga mice and then, cultured with GM-CSF and IL-4 for 6 days, and treated for 2 days by either phosphorothioate ODN or phosphodiester ODN. CpG ODN-treated DCs resulted in more production of IL-12. When CpG ODN-treated DCs were intravenously injected into the NC/Nga mice, the NC/Nga mice with CpG ODN-treated DCs showed significant improvement of AD symptoms and decrease of IgE level. Histopathologically, the NC/Nga mice skin with CpG ODN-treated DCs showed the decreased IL-4 and TARC expression comparing with non-injected mice. These results may suggest that phosphodiester CpG ODN-treated DCs might function as a potent adjuvant for AD in a mouse model.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.637-644
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• 2013

This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.

• KIEE International Transaction on Electrical Machinery and Energy Conversion Systems
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• 제5B권2호
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• pp.118-122
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• 2005

Electro-chemo-mechanical devices or artificial muscles based on conducting polymers (CP) are presented as bilayers, CP/adhesive polymer, or as triple layers, CP/adhesive polymer/CP. Those soft and wet materials, working in aqueous solutions of a salt, mimic the composition of most organs from animals. Under electrochemical control, so working as new electrical machines, they produce continuous, reverse and elegant bending movements, mimicking those produce by animal muscles. By means of the current a perfect controls of the movement rate is attained giving soft and continuous movements. Muscles able to sense the chemical and mechanical conditions of work or muscle having tactile sense, as will be presented here, are being developed. All of them are founded on the non-stoichiometric nature of the soft and wet materials.

• BMB Reports
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• 제53권8호
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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.

• 한국식품과학회지
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• 제31권4호
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• pp.1115-1119
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• 1999

키토산을 처리하여 건조 김을 제조하고 건조 김의 관능특성, 미생물 변화를 조사하였으며 2차 가공품인 김밥에 키토산김의 적용 가능성을 검토하였다. 키토산을 처리하여 제조한 건조김과 무처리 건조김을 관능평가에 의해 비교하였을 때 색깔 조직감 및 맛에 있어서는 큰 차이가 없었으나 김에서 나는 갯비린내는 키토산 처리김에 있어 현저하게 감소되었다. 건조김의 총균수에 있어서는 무처리 건조김의 경우 약 37,000 cells/g이 검출되었으나, 30 cp 키토산을 0.1% 농도로 처리하여 건조김을 만들 경우 2,100 cells/g으로 균체수가 1/10 정도로 감소하였으며, 균체수의 감소경향은 처리 키토산 농도가 높을수록 효과적이었다. 이런 경향은 평균 분자량이 더 큰 90 cp 키토산을 처리하였을 경우에도 비슷하게 나왔다. 대장균군에 있어서는 무처리 건조김에서는 240 cells/g 정도가 검출되었으나, 30 cp 키토산을 0.1% 처리하였을 경우에는 대장균군이 검출되지 않았으며, 90 cp 키토산을 0.005%와 0.04%로 처리하여 건조김을 제조하였을 때 각각, 93 cells/g과 3.6 cells/g이 검출되었다. 키토산 처리 건조김으로 김밥을 제조하여 $20^{\circ}C$에서 총균수의 변화를 관찰하였을 때 30 cp, 90 cp 키토산으로 처리된 건조김으로 만든 김밥에 있어 모두 무처리 건조김으로 만든 김밥보다 총균수 증가가 현저하게 지연되었으며, 키토산처리 건조김의 이용은 김밥의 유통기간의 증대에 기여할 것으로 기대 된다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권10호
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• pp.1404-1410
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• 2012

In vitro experiments were conducted to evaluate the suitability of several mixtures of high tanniniferous non legumes with low tanniniferous legumes on in vitro gas production (IVGP), dry matter degradation, Ammonia-N, methane production and microbial population. Eight treatments were examined in a randomized complete block design using four non-legumes and two legumes (Carallia integerrima${\times}$Leucaena leucocephala (LL) (Trt 1), C. integerrima${\times}$Gliricidia sepium (GS) (Trt 2), Aporosa lindeliyana${\times}$LL (Trt 3), A. lindeliyana${\times}$GS (Trt 4), Ceiba perntandra${\times}$LL (Trt 5), C. perntandra${\times}$GS (Trt 6), Artocarpus heterophyllus${\times}$LL (Trt 7), A. heterophyllus${\times}$GS (Trt 8). The condensed tannin (CT) content of non legumes ranged from 6.2% (Carallia integerrima) to 4.9% (Ceiba perntandra) while the CT of legumes were 1.58% (Leucaena leucocephala) and 0.78% (Gliricidia sepium). Forage mixtures contained more than 14% of crude protein (CP) while the CT content ranged from 2.8% to 4.0% respectively. Differences (p<0.05) were observed in in vitro gas production (IGVP) within treatments over a 48 h period dominated by C. perntandra${\times}$G. sepium (Trt 6). The net gas production (p<0.05) was also high with Trt6 followed by A. heterophyllus${\times}$L. leucocephala (Trt 7) and A. heterophyllus${\times}$G. sepium (Trt 8). Highest (p>0.05) NH3-N (ml/200 mg DM) production was observed with the A. heterophyllus${\times}$G. sepium (Trt 8) mixture which may be attributed with it's highest CP content. The correlation between IVGP and CT was 0.675 while IVGP and CP was 0.610. In vitro dry matter degradation (IVDMD) was highest in Trt 8 as well. Methane production ranged from 2.57 to 4.79 (ml/200 mg DM) to be synonimous with IVGP. A higher bacteria population (p<0.05) was found in C. perntandra${\times}$G. sepium (Trt 6) followed by Artocarpus heterophyllus+G. sepium (Trt 8) and the same trend was observed with the protozoa population as well. The results show that supplementing high tannin non leguminous forages by incremental substitution of legume forage increased gas production parameters, NH3-N, IVDMD and microbial population in the fermentation liquid. Methane production was not significantly affected by the presence of CT or different levels of CP in forage mixtures. Among non legumes, Ceiba perntandra and Artocarpus heterophyllus performed better in mixture with L. leucocephala and G. sepium.