• 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.

• 한국환경보건학회지
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• 제43권5호
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• pp.438-445
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• 2017

Objectives: The purpose of this study was to develop exposure factor data for the Korean child population, with a specific focus on behavior characteristics such as hand-to-mouth and object-to-mouth and an analysis of influential factors. Methods: We divided environments into two places, which were house/indoor and nursery. A total of 400 children (house/indoor) and a total of 162 children (nursery) were recruited from the cities of Seoul, Incheon, Daegu, and Gwangju. The children were divided into two groups. We conducted direct measurement by using one hour of videotaping alongside questionnaire surveys. This was performed to calculate behavior rates, such as how many children perform hand-to-mouth and object-to-mouth behaviors. Results: The respective average frequencies of hand-to-mouth and object-to-mouth were $0.8{\pm}2.23$ and $0.82{\pm}2.64contacts/hr$ for house/indoor. The respective average frequencies of hand-to-mouth and object-to-mouth were $2.87{\pm}4.63$ and $1.47{\pm}3.84contacts/hr$ in the nursery group. For the mouthing participants, the average frequencies of hand-to-mouth and object-to-mouth were 3.31 and 3.20 contacts/hr in house, and 4.80 and 3.26 in nursery. Compared to other countries such as the USA, the frequencies of hand-to-mouth and object-to-mouth behaviors found in this study were relatively lower. Conclusions: Children have the potential for exposure to toxic substances through non-dietary ingestion pathways by mouthing objects or their fingers. In this study, the mouthing frequency was relatively lower than that found in Western countries. This can be explained that mouthing behaviour may be affected by culture and lifestyle characteristics.

• 한국식품영양과학회지
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• 제46권5호
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• pp.552-561
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• 2017

본 연구에서는 가시오갈피 물 추출물(ASW)를 이용하여 아직 시도된 바가 없는 HepG2 세포 내 포도당 유입과정 및 glucokinase(GK) 활성을 통한 포도당 이용대사 실험을 수행하였다. 포도당의 세포 내 유입은 GLUT2의 transcription factor들 중 하나인 $HNF-1{\alpha}$의 활성화로 GLUT2의 유전자 발현이 증가하여 이루어지는 것을 확인하였다. GK 활성 측정 결과 ASW가 GK를 활성화하여 포도당의 인산화에 영향을 주는 것을 확인하였고 AMP-activated protein kinase의 인산화 증가로 glycolysis에 관여하는 효소인 GK의 단백질 발현은 증가하고, gluconeogenesis에 관여하는 phosphoenolpyruvate carboxykinase의 단백질 발현은 감소하는 것을 확인하였다. 그리고 인산화된 포도당이 glycogen으로 전환 저장되는 메커니즘을 pPI3k-pAkt-pGSK-$3{\beta}$의 단계별 단백질 발현을 확인함으로써 검증하였으며, glycogen 함량 측정을 통해 확인하였다. 본 연구를 통해 ASW가 다양한 메커니즘에 작용하여 당뇨의 예방 및 개선에 활용할 수 있는 잠재적 소재임을 확인하였고, 이는 ASW가 천연 기능성 소재로서의 개발가치가 높음을 시사한다.

• 대한지리학회지
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• 제42권2호
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• pp.258-279
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• 2007

도시 경쟁력 제고 수단으로 문화 및 예술에 대한 관심이 고조되고, 창조경제에서 문화 및 예술 산업의 중요성이 주목 받고 있다. 서유럽과 북미를 중심으로 도시 재활성화 및 젠트리피케이션 과정에서 아티스트 및 미술관의 역할에 대한 다양한 연구가 이루어졌으나, 예술 작품과 문화 소비자를 매개하는 상업 화랑들의 입지 패턴과 요인에 대한 경험적 연구는 거의 이루어지지 않았다. 특히 아시아 대도시의 도심 재개발 및 젠트리피케이션 과정과 문화 소비 공간 확산에 대한 경험적 연구는 부족한 상황이다. 본 연구에서는 서울시 상업 화랑을 중심으로 아트 클러스터의 성장과 쇠퇴 과정을 이해하고, 그 분포 특성을 도시의 역사성과 시대적 특성, 예술가 집단의 성격과 연결시켜 설명하고자 하였다. 이를 위해 런던과 뉴욕과 같은 대도시에서 아티스트 및 화랑이 클러스터를 형성하는 배경을 아티스트의 생태, 아트 비즈니스의 특성, 세계화의 영향을 중심으로 고찰하였다. 서울의 화랑 입지는 예술가나 문화적 요인보다는 상업적 논리에 의해 결정되고 문화소비공간으로 쉽게 전환되는 경향이 있으며, 화랑의 분포는 도시 개발 정책과 맞물린 지대의 상승, 문화 소비 취향의 변화, 국내외 미술 시장의 경기 변동에 따라 역동적으로 변화하는 입지 주기 순환 양상을 보인다.

• KSBB Journal
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• 제19권2호
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• pp.118-124
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• 2004

본 연구에서는 효모에서 분리한 melanoston이라고 명명된 멜라닌 생성을 억제하는 물질의 작용 기전을 밝히기 위한 것이다. $\alpha$-MSH를 처리한 B16 melanoma 세포에서 melanoston은 tyrosinase mRNA 발현양을 10% 정도 저해되는데 그쳤으며 western blotting을 이용한 단백질 측정에서도 이와 비슷한 정도의 단백질 생성 억제를 보였다. 그러나 B16 세포 배양액에 melanoston을 첨가할 경우 세포내 tyrosinase 활성이 33% 까지 감소되는 것으로 나타나 metanoston이 tyrosinase inhibitor는 아니지만 세포내 tyrosinase 활성화 (activation) 과정을 억제하는 것으로 추측할 수 있었다. 또한 광학 현미경을 이용한 morphology 관찰에서 $\alpha$-MSH를 처리한 세포에서는 많은 dentrite가 형성되면서 세포분화가 일어나는 반면 melanoston를 처리한 경우에는 dendrite가 감소하면서 세포형태가 대조군과 비슷하게 회복 되는 것을 알 수 있었다. 또 FITC-anti-tyrosinase-Ab를 이용한 형광 염색을 통해서는 $\alpha$-MSH만 처리한 세포에서는 tyrosinase의 분포가 dendrite를 포함한 세포 전체로 퍼져나가는 것을 관찰할 수 있었고 $\alpha$-MSH와 melanoston을 동시에 처리한 세포에서는 대조군과 비슷하게 tyrosinase가 핵 주변에서만 관찰되어 melanoston이 B16 melanoma 세포의 분화과정에서 이를 억제하는 효과를 주고 있음을 알 수 있었다. 이상의 결과들을 종합해 볼 때 melanoston은 $\alpha$-MSH에 진행되는 B16 세포의 분화를 억제하고 이 과정에서 멜라닌 생성의 주된 효소인 tyrosinase의 활성화를 억제하며 결과적으로는 멜라닌 생성을 저해하는 것으로 사료된다.

• 대한본초학회지
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• 제30권6호
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• pp.77-82
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• 2015

Objectives : This study was designed to investigate of the anti-inflammatory effects of Schisandra chinensis seed oil(SSO) on the production of pro-inflammatory substances in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.Methods : SSO was measured the production of pro-inflammatory factor (NO, PGE2, IL-1β iNOS and, COX-2) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. we used the following methods : cell viability assay, Griess reagent assay, enzyme-linked immunosorbent assay, Western blotting analysis.Results : The cell viability of SSO(0∼500 μl/mL) processing group was 96.9% and the processing of SSO didn't have an effect on the cytotoxicity. The inhibitory effect of the nitric oxide (no) production of SSO(500 μg/mL, 50 μg/mL, 10 μg/mL) was each 70.3%, 37.6% and 26.5%. IL-1β production inhibition ability of SSO(500 μg/mL, 100 μg/mL) was each 49.88% and 48.8%. PGE2 production inhibition ability of SSO(500 μg/mL, 100 μg/mL) was each 49.88% and 73.1%, 70.5%. By using SSO, it experimented about iNOS protein expression inhibition ability, that is the NO production enzyme. iNOS protein expression increased in the group processing LPS independently. iNOS protein expression decreased in the group processing SSO together. The expression of the COX-2 protein decreased 89.6%, 81.8% in the group processing SSO. The significance was in the relationship with NO formation inhibition with the relationship with the PGE₂ formation inhibition and iNOS protein, it confirmed in SSO with the COX-2 protein.Conclusions : Stimulation of the RAW 264.7 cells with LPS caused an elevated production of nitric oxide (NO), IL-1β and PGE₂ which was markedly inhibited by the pretreatment with SSO without causing any cytotoxic effects. The reduced expressions of iNOS protein were consistent with the reductions in NO production in the culture media. SSO may be useful for the treatment of various inflammatory diseases.

• Preventive Nutrition and Food Science
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• 제8권1호
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• pp.24-28
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• 2003

There is evidence to suggest that cranberry juice supplements improve the health of the urinary tract by inhibiting the binding of fimbriated uropathogenic E. coli to the bladder mucosa. In patients with neurogenic bladders, urinary tract infections (UTI) are particularly common and often poorly managed by antibiotic treatment. A double-blind, randomized, placebo-controlled trial was undertaken on 29 geriatric and spinal cord injured patients with dysfunctional bladders. They received three times daily at mealtimes a 4 oz bottle of cranberry juice (Ocean Spray Cranberries, USA) or a specially prepared synthetic placebo drink. Two episodes of UTI arose in week one of cranberry intake and none thereafter, compared to four episodes of UTI in 4 placebo patients in weeks four, six and 10. Mean bacterial adhesion counts on bladder cells of the patients rose during the first month of treatment in 71 % of the placebo patients compared to only 31 % of cranberry patients (p < 0.001). The difference persisted to some extent for the second and third months. Bacterial adhesion levels correlated with culture findings (higher adhesion and higher viable counts in urine) (p < 0.001), positive leukocyte nitrite tests (136$\pm$131 bacteria per cell versus 52$\pm$86 in negative tests) (p < 0.001), and higher white blood cell counts (> 10) per high power field (126$\pm$125 versus 48$\pm$85 bacteria per cell) (p<0.001). E. coli was the most frequently isolated organism (40% samples) followed by K. pneumoniae (17%) and a number of other uropathogens. Group B Streptococci, and coagulase negative Staphylococcus were recovered from urine in 4 samples but were not associated with any red blood cell presence. The daily intake of cranberry juice, in amounts which are not detrimental to long term compliance, appeared to have a role in reducing the risk of bladder colonization and infection in a highly susceptible patient population.

• 대한가정학회지
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• 제46권1호
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• pp.47-61
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• 2008

The purpose of this study is to provide information about domestic living of Korean immigrants in Australia and Canada who have the same cultural background for comparative study. For this, usage of domestic space and living style in housing of 52 Korean households in Melbourne of Australia and 32 Korean households in the region of Waterloo of Canada were analyzed. Ethnographic research with questionnaire were used. Results of the research were as follows. 1. Korean immigrants in each countries were living in houses which was built by company of Australia and Canada. 44.2% of Korean immigrants in Australia were using L+D K and 53.1% of Korean immigrants in Canada were using L D K. 2. Laundry was indispensable for Korean immigrants in both countries and they all wanted to use the laundry as a utility room which could dry, ironing and so on. 3. Drain hole on the floor of the bathroom was not indispensable for most Korean immigrants in both countries for hygienic reason. 4. Korean immigrants in both countries were ironing in master bedroom and they all wanted to separate it from there through renovation and extension and so on. 5. Korean immigrants in Canada were more active to use the formal lounge which has been planned as a traditional element of western house. 6. The seating style of Korean immigrants in both countries belong to chair-seating style mostly. But it was clear that they were making Kimchi with floor seating style in both countries. 7. A level of satisfaction about using carpet was not high for Korean immigrants in both countries cause of uneasiness to clean and it was considered to relate to the floor seating style of them. 8. Almost Korean immigrants were took off the shoes inside of the house and they had shoes cabinet beside the entrance or basement usually. 9. The most popular heating system was ducted heating in both countries. The level of satisfaction about this was different for Korean immigrants in Australia and Canada but most desirable heating system was Ondol for them in both countries commonly.

• 대한예방한의학회지
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• 제13권2호
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• pp.51-64
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• 2009

Objective : This research was aimed to investigate the anti-tumor effect, safety, mechanism and metabolizing enzyme of Agrimonia pilosa LEDEB(APL) in female C57B/L mouse. Methods : At first, to evaluate the anti-tumor activity of APL, we divided into four groups, normal, control, APL100(100mg/kg), APL150(150mg/kg). LLC obtained American Type Culture Collection was used. LLC had been inoculated to induce tumor. To measure the anti-tumor effect of APL, we calibrate tumor size and weight. To study for mechanism of anti-tumor in APL, we used western blotting and to know metabolizing enzyme in APL we used to real-time PCR. Results : APL100, APL150 inhibited tumor growth after medicine injected. APL did not only induced caspase-dependent apoptosis in LLC-bearing mouse tumor. In APL100, it were decreased 72% in CYP3A11. In APL150, it were decreased 62%, 75% in CYP3A11 and MRP1a respectively. Conclusion : These results suggests that APL has some anti-tumor effects in female C57B/L mouse tumor. APL should be careful use with other drugs related with CYP3A11 or MRP1a.

• 한국식품과학회지
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• 제30권6호
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• pp.1415-1419
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• 1998

시설 원예산물 중 곰팡이 독소 생성능이 있는 균주의 오염여부를 조사하기 위해 진주를 비롯한 서부 경남 일원과 경북 안동 근교에서 가지, 메론, 배추, 상추, 오이, 수박, 참외, 토마토, 딸기, 및 그 토양 등의 시료를 수집하여 ochratoxin A 생성균의 오염정도를 관찰하였다. 분석방법은 indirect competitive ELISA 법을 사용하였으며, HPLC법과도 비교 분석하였다. 그 결과 경남 일원과 경북 안동 근교에서 총 192점의 시료를 분리하여 균분리를 실시한 결과 Aspergillus속 142균주, Penicillium속 153균주를 분리하였다. 분리된 균은 SLS배지로 $28^{\circ}C$, 15일간 배양한 후 indirect competitive ELISA 법에 의해 ochratoxin A의 생성여부를 검색하였다. 그 결과 Penicillium속 5균주에서 ochratoxin A 생성이 확인되었으며 검출량은 $0.084{\sim}2.128\;{\mu}g/mL$이었고, 수박에서 분리한 균주가 $2.128\;{\mu}g/mL$로 가장 많은 ochratoxin A를 생산하였다. 그러나 Aspergillus속은 모두 음성반응을 나타내었다. ELISA법에서 ochratoxin A를 생성하는 것으로 나타난 균주를 HPLC법에 의해 확인한 결과 ochratoxin A의 생성이 확인되었으며 생성량은 ELISA 결과와 비슷하였다.