• 생명과학회지
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• 제26권2호
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• pp.147-154
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• 2016

본 연구에서는 인간 골육종암세포주인 Saos-2 세포를 이용하여 Eucheuma cottonii 추출물(Extract of Eucheuma cottonii, EE)의 항암 활성 및 분자적 작용기전을 분석하였다. 먼저 EE가 세포증식에 미치는 영향을 WST-1^® assay를 통해 확인한 결과 EE는 인간 위암세포 AGS, 인간 간암세포 SK-Hep 1, 인간 뇌교모세포종 U87MG, 인간 정상 신장세포 HEK-293의 생존율에는 영향을 미치지 않고 Saos-2 세포주의 생존율만을 농도의존적으로 감소시킴을 확인 하였다. 또한 처리 농도가 증가함에 따라 Saos-2 세포의 외형적 변화가 나타남을 도립현미경을 통해 확인할 수 있었다. 그리고 DAPI staining을 통해 apoptosis classical hall marker라고 할 수 있는 DNA fragmentation이 EE 처리 농도 의존적으로 나타남을 관찰할 수 있었다. 이를 토대로 EE가 Saos-2 세포에서 세포 내의 어떠한 기작을 통해 apoptosis를 유도하는지 Western blot analysis를 통해 확인한 결과, Fas-Associated Death Domain(FADD)에 의한 caspase cascade signal pathway 발현이 증가하였고, apoptosis의 key protein 이라고 할 수 있는 cleaved caspase-3와 하위 인자인 cleaved PARP가 증가 함을 확인할 수 있었다. 이와 관련된 분자적 기전 분석을 위한 immunofluorescence staining과 Flow cytometry analysis를 추가로 수행한 결과, EE 처리시 caspase cascade signal pathway의 시발점인 FAS와 cleaved caspase-3의 발현증가가 실제 세포 내에서 일어남을 관찰 할 수 있었으며 apoptosis 유발군인 sub G1기가 증가 함을 알 수 있었다. 이상의 결과를 통해 EE는 인간 골육종암세포에서 FADD에 의한 caspase cascade signal pathway 발현증가가 유도되어 apoptosis를 유발시킨다는 것을 증명하였으며 새로운 골육종암 치료제로서의 개발 가능성과 기전연구를 위한 중요한 기초자료가 될 수 있음을 시사한다.

• Korean Chemical Engineering Research
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• 제56권4호
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• pp.496-523
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• 2018

본 연구의 목표는 톱밥의 건조, 급속열분해, 바이오오일 응축, 바이오오일 수첨개질, 전기생산, 폐수처리를 포함하는 2개의 바이오오일 생산공정들에 대한 경제적 타당성을 평가하는 것이다. 첫번째 공정은 수소생산을 위한 steam-methane reforming (SMR)을 포함하는 바이오오일 생산공정이다(Case 1). 두번째 공정은 SMR을 포함하지 않고 수소를 외부로부터 공급받아 수첨개질을 수행하는 바이오오일 생산공정이다(Case 2). 상용공정모사기인 ASPEN Plus를 이용하여 이두 공정에 대한 공정흐름도를 구축하였고, 물질 및 에너지 수지식을 계산하였다. 원료로서 40%의 수분을 포함하는 톱밥 100 t/d, 30% 자기자본비율, 자기자본에 상응하는 자본지출, 수소 구입가 $1,050/ton, 완전건조된 원료대비 수송용 연료 수율 20% (Case 1) 및 25% (Case 2) 로 가정하고, 4단계 경제성 분석기법인 4-level EP를 사용하여 기술경제성 분석을 수행하였다. 총투자비(TCI), 총생산비(TPC), 연간판매금액(ASR), 그리고 개질 바이오오일의 최소판매가격(MFSP)은 Case 1에 대하여 $22.2 million, $3.98 million/yr, $4.64 million/yr, 그리고 $1.56/l 이고, Case 2에 대하여 $16.1 million, $5.20 million/yr, $5.55 million/yr, $1.18/l로 산출되었다. 수소를 직접 생산하는 Case 1과 수소를 외부로 부터 공급받는 Case 2의 투자회수율(ROI)와 투자회수기간(PBP)은 큰 차이를 보여주지 않았고, Case 1과 Case 2의 원료 공급량을 1,500 t/d로 증가할 경우, ROI는 15% 이상으로 향상될 것으로 예상되었다.

• Biomolecules & Therapeutics
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• 제28권3호
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• pp.259-266
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• 2020

The present research work primarily investigated whether spinosin has the potential of improving the pathogenesis of Alzheimer's disease (AD) driven by β-amyloid (Aβ) overproduction through impacting the procession of amyloid precursor protein (APP). Wild type mouse Neuro-2a cells (N2a/WT) and N2a stably expressing human APP695 (N2a/APP695) cells were treated with spinosin for 24 h. The levels of APP protein and secreted enzymes closely related to APP procession were examined by western blot analysis. Oxidative stress related proteins, such as nuclear factor-erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) were detected by immunofluorescence assay and western blot analysis, respectively. The intracellular reactive oxygen species (ROS) level was analyzed by flow cytometry, the levels of Aβ_1-42 were determined by ELISA kit, and Thioflavin T (ThT) assay was used to detect the effect of spinosin on Aβ_1-42 aggregation. The results showed that ROS induced the expression of ADAM10 and reduced the expression of BACE1, while spinosin inhibited ROS production by activating Nrf2 and up-regulating the expression of HO-1. Additionally, spinosin reduced Aβ_1-42 production by impacting the procession of APP. In addition, spinosin inhibited the aggregation of Aβ_1-42. In conclusion, spinosin reduced Aβ_1-42 production by activating the Nrf2/HO-1 pathway in N2a/WT and N2a/APP695 cells. Therefore, spinosin is expected to be a promising treatment of AD.

• 디자인학연구
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• 제20권1호
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• pp.45-54
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• 2007

최근 각 지역 지자체는 지역진흥을 위해 지자체가 가지고 있는 전통과 문화유산을 개발시켜 지역의 특성으로 나타내어 관광자원화 하려는 연구가 활발히 진행되고 있다. 연구 대상지역인 약령시는 대구시의 도심지역 재활성화를 위한 정책 및 사업 계획 중 가장 핵심적인 장소로, 전통과 기능이 유지되고 있다. 선행연구인 "지역진흥을 위한 대구 약령시 재활성화 기초연구"를 통해 문제점 도출과 개발방향을 제시한바, 이를 바탕으로 약령시의 재활성화 할 수 있는 구체적인 방법을 제안하는 연구이다. 연구 방법은 (1)선행연구를 바탕으로 한 현장조사 및 확인 (2) 약령시의 심층 분석을 통한 개선 과제 및 잠재력을 파악하고 (3)재래시장 활성화를 위한 요소 및 방향 연구 (4) 연구결과를 토대로 지역디자인 센터의 환경디자인 디자인개발프로세스를 활용하여 디자인을 제안하였다. 디자인 제안으로 첫째, 기반시설 정비 및 확충으로 장소의 접근성을 용이하게 하고 둘째, 동선 계획에 의한 무계획한 도로망 정비로 공간 활용성의 극대화 추구 셋째, 장소의 특수성을 감안한 통일성과 정체성을 추구하며 넷째, 체험의 장과 이벤트 공간 확보를 통한 찾아가고 싶은 거리, 즐길 수 있는 거리를 제안하므로 선행연구에서 제시된 발전 과제인 도심 재개발 사업에 의한 재활성화 및 관광 자원의 산업화 개발을 도모하였다. 이렇게 볼 때 지역 진흥을 위해서는 지역마다 갖고 있는 지역 특성의 심층 분석을 통해 재활성화를 위한 연구가 지속되어 지역진흥을 꾀하여야 할 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3651-3657
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• 2014

Hepatocellular carcinoma (HCC) has a relatively higher incidence in many countries of Asia. Globally, HCC has a high fatality rate and short survival. Epirubicin, a doxorubicin analogue, may be administered alone or in combination with other agents to treat primary liver cancer and metastatic diseases. However, the toxic effects of epirubicin to normal tissues and cells have been one of the major obstacles to successful cancer chemotherapy. Here, we investigated the effects of epirubicin in combination with kappa-selenocarrageenan on mice with H22 implanted tumors and HepG-2 cell proliferation, immune organ index, morphology, cell cycle and related protein expressions in vivo and in vitro with sequential drug exposure. The inhibitory rate of tumor growth in vivo was calculated. Drug sensitivity was measured by MTT assay, and the King's principle was used to evaluate the interaction of drug combination. Morphological changes were observed by fluorescent microscopy. Cell cycle changes were analyzed by flow cytometry. Expression of cyclin A, Cdc25A and Cdk2 were detected by Western blotting. In vivo results demonstrated that the inhibitory rate of EPI combined with KSC was higher than that of KSC or EPI alone, and the Q value indicated an additive effect. In addition, KSC could significantly raise the thymus and spleen indices of mice with H22 implanted tumors. In the drug sensitivity assay in vitro, exposure to KSC and EPI simultaneously was more effective than exposure sequentially in HepG-2 cells, while exposure to KSC prior to EPI was more effective than exposure to EPI prior to KSC. Q values showed an additive effect in the simultaneous group and antagonistic effects in the sequential groups. Morphological analysis showed similar results to the drug sensitivity assay. Cell cycle analysis revealed that exposure to KSC or EPI alone arrested the cells in S phase in HepG-2 cells, exposure to KSC and EPI simultaneously caused accumulation in the S phase, an effect caused by either KSC or EPI. Expression of cyclin A, Cdc25A and Cdk2 protein was down-regulated following exposure to KSC and EPI alone or in combination, exposure to KSC and EPI simultaneously resulting in the lowest values. Taken together, our findings suggest that KSC in combination with EPI might have potential as a new therapeutic regimen against HCC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.769-773
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• 2014

Fangchinoline (Fan) inhibits cell proliferation and induces apoptosis in several cancer cell lines. The effects of Fan on cell growth and proliferation in breast cancer cells remain to be elucidated. Here, we show that Fan inhibited cell proliferation in the MDA-MB-231 breast cancer cell line through suppression of the AKT/Gsk-3beta/cyclin D1 signaling pathway. Furthermore, Fan induced apoptosis by increasing the expression of Bax (relative to Bcl-2), active caspase 3 and cytochrome-c. Fan significantly inhibited cell proliferation of MDA-MB-231 cells in a concentration and time dependent manner as determined by MTT assay. Flow cytometry analysis demonstrated that Fan treatment of MDA-MB-231 cells resulted in cell cycle arrest at the G1 phase, which correlated with apparent downregulation of both mRNA and protein levels of both PCNA and cyclin D1. Further analysis demonstrated that Fan decreased the phosphorylation of AKT and GSK-3beta. In addition, Fan up-regulated active caspase3, cytochrome-c protein levels and the ratio of Bax/Bcl-2, accompanied by apoptosis. Taken together, these results suggest that Fan is a potential natural product for the treatment of breast cancer.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.126-135
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• 2001

Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma HeLa cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration- and time-dependent, and preceded activation of caspase-9 and -3. Flow cytometric FACScan analysis using fluorescence intensities of $DiOC_6$/ demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (${\blacktriangle}{\Psi}_m$). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector caspase-3, Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5,8,11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDCA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDCA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.

• 한국수자원학회논문집
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• 제51권7호
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• pp.565-574
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• 2018

아시아 지역의 인구증가와 하천주변 및 삼각주 평야 저지대의 인구밀도 집중으로 인해 우기 시 아시아의 많은 지역에서 홍수로 인한 대규모 인명 및 재산 피해가 발생하고 있다. 본 연구에서는 NOAA에서 제공하는 1992년부터 2003년까지의 나이트라이트 위성영상자료를 수집하여 인명 및 재산피해 정보와의 상호 공간분석을 통해 메콩강 유역의 홍수피해에 대한 분석을 실시하였다. 인명 및 재산피해 자료는 EM-DAT에서 제공하는 지역 공간분포자료를 활용하였으며, 메콩강 주하천 격자로 부터 모든 임의 격자의 떨어진 거리를 계산하여 해당 격자에서의 나이트라이트 강도와 인명 및 재산피해와의 상관분석을 수행하였다. 그 결과, 나이트라이트 강도가 클수록 홍수피해가 큰 것으로 분석되었으며, 특히, 하천으로부터 가까운 거리에서 나이트라이트 강도가 높게 나타났다. 이는 높은 나이트라이트 강도를 갖는 격자, 즉 인구밀집도가 상대적으로 높은 격자가 메콩강 하천주변으로 분포되어 있으며, 홍수피해와 양의 상관관계를 갖고 있음을 의미한다. 이와 같이 나이트라이트 위성영상정보는 에너지소비, 재해 등 다양한 공간분석을 통해 사회경제적 영향성 평가를 위한 대리변수로 사용이 가능할 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권11호
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• pp.1776-1788
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• 2019

Objective: The demands for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). Methods: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells ($M{\Phi}$) were activated by phorbol 12-myristate 13-acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B ($NF-{\kappa}B$) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative polymerase chain reaction was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 $M{\Phi}$ cells was evaluated by the colony forming unit assay. Results: ASF upregulated the cell viability and growth rate of 3D4/31 $M{\Phi}$, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of $NF-{\kappa}B$ protein, tumor necrosis factor $(TNF){\alpha}$ mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of $NF-{\kappa}B$, $TNF{\alpha}$, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 $M{\Phi}$ against Escherichia coli. Conclusion: This study provides a novel insight into the regulation of $NF-{\kappa}B$ activity and lipid metabolism in $M{\Phi}$, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

• Clinical and Experimental Reproductive Medicine
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• 제44권3호
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• pp.152-158
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• 2017

Objective: To identify the associations between polymorphisms of the 3'-untranslated region (UTR) of methylenetetrahydrofolate reductase (MTHFR) gene, which codes for an important regulatory enzyme primarily involved in folate metabolism, and idiopathic recurrent pregnancy loss (RPL) in Korean women. Methods: The study population comprised 369 RPL patients and 228 controls. MTHFR 2572C > A, 4869C > G, 5488C > T, and 6685T > C 3'-UTR polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis or by TaqMan allelic discrimination assays. Natural killer cell proportions were determined by flow cytometry. Results: The MTHFR 2572-5488-6685 (A-C-T) haplotype had an adjusted odds ratio of 0.420 (95% confidence interval, 0.178-0.994; p= 0.048) for RPL. Analysis of variance revealed that MTHFR 4869C > G was associated with altered $CD56^+$ natural killer cell percentages (CC, $17.91%{\pm}8.04%$; CG, $12.67%{\pm}4.64%$; p= 0.024) and folate levels (CC, $12.01{\pm}7.18mg/mL$; CG, $22.15{\pm}26.25mg/mL$; p= 0.006). Conclusion: Variants in the 3'-UTR of MTHFR are potential biomarkers for RPL. However, these results should be validated in additional studies of ethnically diverse groups of patients.