• 한국식품영양과학회지
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• 제29권5호
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• pp.875-880
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• 2000

Reactive oxygen species are produced under diabetic conditions and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study was to examine the effects of high glucose on the alloxan-induced beta cell injury. The insulinoma (RINm5F) cells were clutured either with high glucose (22.2 mM) or normoglucose (5.6 mM) in RPMI 1460 media for 3 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The effects of high glucose on the cytotoxicity of alloxan were also investigated in RINm5F cells and the cells viability were determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methods. Results showed that the CuZn-SOD activity was decreased but Mn-SOD activity was increased significantly in RINm5F cells cultured with high glucose (22.2 mM) media. The cytotoxicity of alloxan was increased by high glucose compared with normoglucose in RINm5F cells. Diethyl-dithiocarbarmate (DDC), as inhibitor of CuZn-SOC, also potentiate the alloxan-induced cytotoxocity in RINm5F cells. These results suggest that, in RINm5F cells, short term culture with high glucose media decreases Cu-Zn-SOD activity and the decreased activity of CuZn-SOD many one of the causative factors of beta-cell injury induced by high glucose.

• 식물조직배양학회지
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• 제24권1호
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• pp.57-61
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• 1997

Superoxide dismutase (SOD) 고생산세포주로 선발된 토마토(Lycopersicun esculentum) 배양세포를 사용하여 현탁배양에 따른 SOD 활성과 isoenzyme변화를 조사하고 토마토 식물체의 것과 비교하였다. 현탁배양은 세포생중량 2 g을 1 mg/L 2,4-D, 30 g/L sucrose를 함유한 MS 배지 50 mL과 함께 mL flask에서 $25^{\circ}C$암상태로 배양(100 rpm)하였다. 세포생장은 계대배양후 20일에 최고점에 도달한 후, 급격히 감소하며 배양 후 23일부터 세포가 검게 변하였다. 세포 단위무게당 SOD활성(unit/g dry cell wt)은 배양 후 23일부터 증가하여 28일째에 최고활성(52,400 unit)을 나타낸 후 급격히 감소하였다. 세포 밖으로 분비되는 extracellular SOD활성은 배양 후 25일에 최고치(27,800 unit/so mL medium)를 나타낸 후 감소하였다. Flask 전체의 SOD활성은 배양 후 25일에 최대치(35,700 unit)를 나타내었으며 extracellular SOD 활성이 약 75%을 차지하였다. 토마토 배양세포에는 4개의 SOD isoenzyme이 존재하며, isoenzyme의 패턴변화는 세포생장에 따른 효소활성의 변화와 일치하였다. 토마토 식물체는 배양세포에 없는 CuZnSOD가 존재하며 배양세포와 식물체 조직사이에는 서로 다른 isoenzyme 패턴이 존재함을 알 수 있었다.

• 한국토양비료학회지
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• 제44권5호
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• pp.667-673
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• 2011

우리나라 밭은 대부분 산지를 개간하여 조성한 경사 밭으로 작물이 재배되는 여름철 집중강우에 의해 많은 양의 토양이 유실된다. 특히 경사 밭 전면경운에 의한 토양 교란은 무의 발아, 생육초기 단계로 쏟아지는 강우에 표토가 직접 노출되어 토양유실 위험이 가중되고 있다. 본 연구에서는 고랭지 무 재배의 경우 토양피복방법 개선에 의한 토양유실 저감기술을 개발하고자 수행하였다. 토양피복 방법으로 호밀 예취피복, 호밀 초생피복, 흑색비닐멀칭 등 8 처리를 두었으며, 경사 17%내외 무저라이시메타에서 유거수 및 토양유출 특성과 무 생육특성을 평가하였다. 시험결과 호밀예취피복을 포함한 토양피복처리는 유거수 유출량에 있어 ha당 관행 $2,994m^3$인 반면, $773{\sim}2,325m^3$이 유거되어 관행보다 26~78% 수준으로 유거수 량이 감소하였다. 이와 같은 결과는 토양유실량에도 영향을 미쳐 관행 처리구가 ha당 68.2톤인 반면, 호밀예취피복을 포함한 피복처리구는 0.3~16.1톤으로 관행 대비 76~99%에 해당하는 토양유실 경감 효과가 있는 것으로 평가되었다. 무 수확기 수량 구성요소인 근중은 관행의 경우 ha당 92.5톤 수준을 보였는데, 호밀 초생피복은 40.1톤, 벌개미취 초생피복은 46.1톤, 곰취 초생피복은 78.5톤 수준으로 낮은 경향을 보였다. 반면, 호밀 예취피복은 92.9톤, 들묵새 초생피복은 102.6톤, 흑색비닐멀칭은 109.8톤으로 관행과 비슷하거나 오히려 증수되는 결과를 보였다. 본 연구 결과 호밀 예취피복 및 들묵새 초생피복은 무 수량에 는 영향을 미치지 않으면서 토양유실 경감효과가 매우 높아 경사 밭 토양보전기술로 유용하게 적용 되리라 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3015-3023
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• 2012

The mitochondrial antioxidant protein manganese superoxide dismutase (MnSOD) may represent a new type of tumor suppressor protein. Overexpression of the cDNA of this gene by plasmid or recombinant lentiviral transfection in various types of cancer leads to growth suppression both in vitro and in vivo. We previously determined that changes in MnSOD expression had bidirectional effects on adriamycin (ADR) when combined with nitric oxide (NO). Radiation induces free radicals in a manner similar to ADR, so we speculated that MnSOD combined with NO would also have a bidirectional effect on cellular radiosensitivity. To examine this hypothesis, TE-1 human esophageal squamous carcinoma cells were stably transfected using lipofectamine with a pLenti6-DEST plasmid containing human MnSOD cDNA at moderate to high overexpression levels or with no MnSOD insert. Blastidicin-resistant colonies were isolated, grown, and maintained in culture. We found that moderate overexpression of MnSOD decreased growth rates, plating efficiency, and increased apoptosis. However, high overexpression increased growth rates, plating efficiency, and decreased apoptosis. When combined with NO, moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells, whereas high MnSOD overexpression had the opposite effect. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.

• 한국식품영양학회지
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• 제22권2호
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• pp.313-319
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• 2009

To improve the growth of human mesenchymal stem cells(hMSCs) under general cell culture conditions(20% $O_2$ and 5% $CO_2$), we examined the effect of s-allylcysteine(SAC), which is known as an antioxidant and the main component of aged-garlic extract, on hydrogen peroxide-induced cellular stress in hMSCs. We found that SAC blocked hydrogen peroxideinduced cell death and cellular apoptosis, but that SAC did not improve the growth of hMSCs during short-term culture. To evaluate the protective effect of SAC, we examined the endogenous expression of the antioxidant enzymes catalase (CAT), superoxide dismutase(SOD), and glutathione peroxidase(Gpx) in hMSCs. Hydrogen peroxide was found to downregulate the expression of CAT, SOD, and Gpx at the protein level. However, in the pre-treatment group of SAC, SAC inhibited the hydrogen peroxide-induced down-regulation of CAT, SOD, and Gpx. Unfortunately, treatment with SAC alone did not induce the up-regulation of antioxidant enzymes and the cell proliferation of hMSCs. Surprisingly, SAC improved cell growth in a single cell level culture of hMSCs. These results indicate that SAC may be involved in the preservation of the self-renewal capacity of hMSCs. Taken together, SAC improves the proliferation of hMSCs via inhibition of oxidative-stress-induced cell apoptosis through regulation of antioxidant enzymes. In conclusion, SAC may be an indispensable component in an in vitro culture system of human MSCs for maintaining self-renewal and multipotent characterization of human MSCs.

• 한국식생활문화학회지
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• 제33권2호
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• pp.104-111
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• 2018

This study aims to compare and analyze a willow tree (Salix Koreensis andersson) extract's antioxidant and anti-inflammatory activity by investigating its: total polyphenol, flavonoid content, SOD-like activity, DPPH vitality. the willow tree was induced with LPS to determine its active anti-inflammatory effects. as a result, the willow methanol extract showed a higher total polyphenol and flavonoid content than those of willow distilled water extract, but the willow distilled water extract showed a higher SOD than that of willow methanol extract. in its DPPH scavenging ability, the willow methanol extract's antioxidant activity was higher than that of the willow distilled water extract. the willow extract's measurements such as the production of NO, inflammatory cytokine ($TNF-{\alpha}$, IL-6 measurement) were significantly reduced as its concentration level went down. according to the research outcomes, when induced, he will extract's macrophage produces mediator-like substances such as NO and inflammatory cytokine that can be used to alleviate the inflammatory response. therefore, the willow tree proved to be a useful raw plant material for the products designed to combat inflammatory activities due to its natural antioxidant and anti-inflammatory response substances such as NO and cytokine.

• BMB Reports
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• 제44권7호
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• pp.462-467
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• 2011

Up-regulation of selected matrix metalloproteinases (MMPs) such as MMP-9 contributes to inflammatory processes during the development of various skin diseases, such as atopic dermatitis. In this study, we examined the effect of a cell-permeable superoxide dismutase (Tat-SOD) on TNF-${\alpha}$-induced MMP-9 expression in human keratinocyte cells (HaCaT). When Tat-SOD was added to the culture medium of HaCaT cells, it rapidly entered the cells in dose- and time-dependent manners. Tat-SOD decreased TNF-${\alpha}$-induced reactive oxygen species (ROS) generation. Tat-SOD also inhibited TNF-${\alpha}$-induced NF-${\kappa}B$ DNA binding activity. Treatment of HaCaT cells with Tat-SOD significantly inhibited TNF-${\alpha}$-induced mRNA and protein expression of MMP-9, as measured by RT-PCR and Western blot analysis. In addition, Tat-SOD suppressed TNF-${\alpha}$-induced gelatinolytic activity of MMP-9. Taken together, our results indicate that Tat-SOD can suppress TNF-${\alpha}$-induced MMP-9 expression via ROS-NF-${\kappa}B$-dependent mechanisms in keratinocytes, and therefore can be used as an immunomodulatory agent against inflammatory skin diseases related to oxidative stress.

• 한국가금학회지
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• 제35권3호
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• pp.241-245
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• 2008

형질 전환 닭 생산 방법들 가운데 목적 유전자 운반에 탁월한 능력이 있는 것으로 알려진 렌티바이러스는 배반엽 단계 수정란을 이용한 형질 전환 닭 생산 연구에 활발하게 이용되고 있다. 본 연구는 재조합 렌티바이러스를 이용하여 사람의 SOD-3 단백질이 닭의 ovalbumin 프로모터에 의해서 유도되는 형질 전환 닭을 생산하고자 하였다. 사람의 SOD-3 단백질은 호흡 과정에서 체내에서 생성되는 활성산소를 중화시키는 탁월한 기능이 있는 것으로 알려져 있다. 후보 병아리의 생산은 앞서 언급한 유전자를 가지는 $1{\times}10^6$ cfu/mL 재조합 렌티바이러스를 배반엽 단계 수정란의 미세 주입하고 대리난각 배양법을 이용하여 배양기에서 21일 동안 배양하는 방법으로 생산하였다. 유전자를 미세주입한 341개의 수정란에서 78수의 후보 형질 전환 병아리를 생산하였으며, 생산된 후보 형질 전환 병아리들의 유전 분석은 PCR 방법을 이용하여 검증하였다. 유전 분석 결과는 성 성숙에 이른 47수의 수컷들 가운데 2수의 정액에서 사람의 SOD-3 유전자가 존재함을 보였다. 이상의 연구 결과는 완전한 형태의 형질전환 닭 생산의 가능성을 보여주고 있다.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1881-1890
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• 2016

Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

• 한국식품영양학회지
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• 제23권3호
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• pp.299-305
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• 2010

주박 추출물을 열수와 에탄올로 추출하여 항균활성, 항산화, DPPH 라디컬 소거능 그리고 SOD 유사활성을 실험하였다. 주박의 추출물은 B. subtilis, Staph. aureus, P. aeruginosa 그리고 E. coli 균주에 대해서 모든 항균활성을 가지고 있었다. 대두유 유화물을 이용한 주박의 항산화력은 control보다 추출물이 더 높았다. 그리고 주박 추출물의 농도가 200 ppm일때 효과가 있었다. 주박 추출물의 DPPH의 소거능은 80% 이상의 저해 효과 나타냈다. SOD 유사활성은 에탄올 추출물보다 열수 추출물이 더 높게 나타났다. 주박의 지속적인 연구를 함으로써 화장품 원료로서 가능성이 있고, 또한 주박을 다른 용매 추출법에 의해 식품의 기능성 원료로 기대할 수 있을 것으로 사료된다.