• Journal of Microbiology and Biotechnology
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• 제26권5호
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• pp.854-866
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• 2016

The production cost of biodiesel from microalgae is still not competitive, compared with that of petroleum fuels. The genetic improvement of microalgal strains to increase triacylglycerol (TAG) accumulation is one way to reduce production costs. One of the most promising approaches is the isolation of starch-deficient mutants, which have been reported to successfully increase TAG yields. To date, such a stable mutant is not available in an oleaginous marine microalga, despite several advantages of using marine species for biodiesel production. Algae in the genus Dunaliella are known to tolerate high salt concentration and other environmental stresses. In addition, the cultivation processes for large-scale outdoor commercialization have been well established for this genus. In this study, Dunaliella tertiolecta was used to screen for starch-deficient mutants, using an iodine vapor-staining method. Four out of 20,016 UV-mutagenized strains showed a substantial reduction of starch content. A significantly higher TAG content, up to 3-fold of the wild-type level, was observed in three of the mutants upon induction by nitrogen depletion. The carotenoid production and growth characteristics of these mutants, under both normal and oxidative stress conditions, were not compromised, suggesting that these processes are not necessarily affected by starch deficiency. The results from this work open up new possibilities for exploring Dunaliella for biodiesel production.

• 생명과학회지
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• 제20권8호
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• pp.1173-1180
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• 2010

Salt stress가 벼 잎 내 광합성 효율과 ascorbate peroxidase (APX) 활성에 미치는 영향을 조사하였다. 염 농도가 증가(NaCl, 100-300 mM) 할수록 Fv/Fm 값이 감소한 반면, $H_2O_2$ 양과 APX 활성은 증가하였다. APX isoforms 중 APX 1 (stromal)은 300 mM NaCl 처리 시 활성이 거의 나타나지 않는 반면, chilling이나 drought 처리 시에는 변화가 없었다. 또한 gel 상에서의 서로 다른 APX isoforms의 활성이 유전자 발현에서도 확인이 되는지를 알아보기 위해 RT-PCR을 수행하였다. 구획별 APX isoforms의 RT-PCR 수행 결과, cytosolic/thylakoid bound APX 발현양은 증가한 반면, stromal APX 발현은 매우 감소하였다. 이러한 실험 결과는 salt에 의해 구획별로 APX 활성이 영향을 받음을 나타낸다.

• Journal of Plant Biotechnology
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• 제1권1호
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• pp.61-68
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• 1999

Rapid accumulation of reactive oxygen species (ROS) and their toxic reaction products with lipids and proteins significantly contributes to the damage of crop plants under biotic and abiotic stresses. We have identified several stress activated alfalfa genes, including the gene of the alfalfa ferritin and a novel NADPH-dependent aldose/aldehyde reductase enzyme. Transgenic tobacco plants that synthesize alfalfa ferritin in vegetative tissues-either in its processed form in chloroplast or in the cytoplasmic non-processed form-retained photosynthetic function upon free radical toxicity generated by paraquat treatment and exhibited tolerance to necrotic damage caused by viral and fungal infections. We propose that by sequestering intracellular iron involved in generation of the very reactive hydroxyl radicals through a Fenton reaction, ferritin protects plant cells from oxidative damage. Our preliminary results with the other stress-inducable alfalfa gene (a NADPH-dependent aldo-keto reductase) indicate, that the encoded enzyme may play role in the stress response of the plant cells. These studies reveal new pathways in plants that can contribute to the increased stress resistance with a potential use in crop improvement.

• 한국환경과학회지
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• 제23권1호
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• pp.89-96
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• 2014

Styela clava tunic is generated in large amounts as a waste from S. clava processing plants and causes environmental problem. Although biological activities of S. clava were reported by many investigators, study on S. clava tunic was little. In this study, therefore, tyrosinase inhibition and antioxidative activities of extracts from S. clava tunic using different solvent were investigated for recycling of the fishery waste. Among extraction methods tested, autoclaved extraction (25.7%) and hot water extraction (18.2%) appeared to be effective for extraction. The highest total phenolic content was 46.6 mg/g in autoclaved extract while the highest flavonoid content was 23.0 mg/g in chloroform extract. All extracts possessed tyrosinase inhibition activity and the inhibition activity was concentration-dependent. Inhibition concentration ($IC_{50}$) against tyrosinase activity was $0.36{\times}10^4$ mg/ml in ethanol extract, $0.11{\times}10^3$ mg/ml in acetone extract and 0.27 mg/ml in n-butanol extract. Among extracts tested, hot water and autoclaved extracts displayed higher antioxidative activity than organic solvent extracts. Therefore, our data suggest that extract from S. clava tunic may potential candidate for cosmetic product with whitening effect and medicine for diseases caused by various oxidative stresses.

• The Korean Journal of Physiology and Pharmacology
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• 제22권4호
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• pp.457-465
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• 2018

The expression of BCL-2 interacting cell death suppressor (BIS), an anti-stress or anti-apoptotic protein, has been shown to be regulated at the transcriptional level by heat shock factor 1 (HSF1) upon various stresses. Recently, HSF1 was also shown to bind to BIS, but the significance of these protein-protein interactions on HSF1 activity has not been fully defined. In the present study, we observed that complete depletion of BIS using a CRISPR/Cas9 system in A549 non-small cell lung cancer did not affect the induction of heat shock protein (HSP) 70 and HSP27 mRNAs under various stress conditions such as heat shock, proteotoxic stress, and oxidative stress. The lack of a functional association of BIS with HSF1 activity was also demonstrated by transient downregulation of BIS by siRNA in A549 and U87 glioblastoma cells. Endogenous BIS mRNA levels were significantly suppressed in BIS knockout (KO) A549 cells compared to BIS wild type (WT) A549 cells at the constitutive and inducible levels. The promoter activities of BIS and HSP70 as well as the degradation rate of BIS mRNA were not influenced by depletion of BIS. In addition, the expression levels of the mutant BIS construct, in which 14 bp were deleted as in BIS-KO A549 cells, were not different from those of the WT BIS construct, indicating that mRNA stability was not the mechanism for autoregulation of BIS. Our results suggested that BIS was not required for HSF1 activity, but was required for its own expression, which involved an HSF1-independent pathway.

• 한국자동차공학회논문집
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• 제13권5호
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• pp.152-162
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• 2005

Coolant rubber hoses for automobile radiators can be degraded and thus failed due to the influence of contacting stresses of air and coolant liquid under the thermal and mechanical loadings. In this study, test analysis was carried out for evaluating the degradation and failure mechanisms of coolant hose materials. Two kinds of EPDM rubber materials applicable to the hoses were adopted: commonly-used ethylene-propylene diene monomer(EPDM) rubbers and EPDM rubbers with high resistance against electro-chemical degradation (ECD). An increase of surface hardness and a large reduction of failure strain were shown due to the formation of oxidation layer for the specimens which had been kept in a high temperature air chamber. Coolant ageing effects took place only by an amount of pure thermal degradation. The specimens degraded by ECD test showed a swelling behavior and a considerable increase in weight on account of the penetration of coolant liquid into the skin and interior of the rubber specimens. The ECD induced material softening as well as drastic reduction in strength and failure strain. However EPDM rubbers designed for high resistance against ECD revealed a large improvement in reduction of failure strain and weight. This study finally established a procedure for reliability analysis and evaluation of the degradation and failure mechanisms of EPDM rubbers used in coolant hoses for automobile radiators.

• Mass Spectrometry Letters
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• 제1권1호
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• pp.25-28
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• 2010

The high-throughput identification and accurate quantification of proteins are essential strategies for exploring cellular functions and processes in quantitative proteomics. Stable isotope tagging is a key technique in quantitative proteomic research, accompanied by automated tandem mass spectrometry. For the differential proteome analysis of mouse neuronal cell lines, we used a multiplexed isobaric tagging method, in which a four-plex set of amine-reactive isobaric tags are available for peptide derivatization. Using the four-plex set of isobaric tag for relative and absolute quantitation (iTRAQ) reagents, we analyzed the differential proteome in several stroke time pathways (0, 4, and 8 h) after the mouse neuronal cells have been stressed using a glutamate oxidant. In order to obtain a list of the differentially expressed proteins, we selected those proteins which had apparently changed significantly during the stress test. With 95% of the peptides showing only a small variation in quantity before and after the test, we obtained a list of eight up-regulated and four down-regulated proteins for the stroke time pathways. To validate the iTRAQ approach, we studied the use of oxidant stresses for mouse neuronal cell samples that have shown differential proteome in several stroke time pathways (0, 4, and 8 h). Results suggest that histone H1 might be the key protein in the oxidative injury caused by glutamate-induced cytotoxicity in HT22 cells.

• Archives of Pharmacal Research
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• 제19권3호
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• pp.178-182
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• 1996

Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxidative stresses induced by 0.1 mM of copper ion $(Cu^{++})$ was studied. In aerobic culture condition, yeasts lacking MnSOD (mitochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared withwild type were observed under anaerobic condition. It was found that, under aerobic condition, the supplementation of 0.1 mM copper ioh:(Cu") into culture medium caused the remarkable increase of CuZnSOD but not so significant change in MnSOD. It was also observed that catalase activities appeared to be relatively high in the presence of copper ion in spite of the remarkable reduction of glutathion peroxidase in CuZnSOD-deficient yeasts, but the slight increments of catalase and glutathion peroxidase were detected in MnSOD-deficient strains. It implies that the lack of cytoplasmic SOD could be compensated mainly by catalase. However, these phenomena resulted in the significantincrease of cellular lipid peroxides content in CuZnSOD-deficient yeasts and the slight increment of lipid peroxides in MNSOD-deficient cells. In anaerobic cultivation supplementing copper ion, the cellular enzyme activities of catalase and glutathion peroxidase in SOD-deficient yeasts were slightly increased without any significant changes of lipid peroxides in cell membrane. It suggests that a little amount of free radicals generated by copper ion under anaerobic condition could be sufficiently overcome by catalase as well as glutathion peroxidase.dase.

• Molecules and Cells
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• 제40권7호
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• pp.503-514
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• 2017

Nicotinamide (NAM) plays essential roles in physiology through facilitating $NAD^+$ redox homeostasis. Importantly, at high doses, it protects cells under oxidative stresses, and has shown therapeutic effectiveness in a variety of disease conditions. In our previous studies, NAM lowered reactive oxygen species (ROS) levels and extended cellular life span in primary human cells. In the treated cells, levels of $NAD^+/NADH$ and SIRT1 activity increased, while mitochondrial content decreased through autophagy activation. The remaining mitochondria were marked with low superoxide levels and high membrane potentials (${\Delta}_{{\Psi}m}$); we posited that the treatment of NAM induced an activation of mitophagy that is selective for depolarized mitochondria, which produce high levels of ROS. However, evidence for the selective mitophagy that is mediated by SIRT1 has never been provided. This study sought to explain the mechanisms by which NAM lowers ROS levels and increases ${\Delta}_{{\Psi}m}$. Our results showed that NAM and SIRT1 activation exert quite different effects on mitochondrial physiology. Furthermore, the changes in ROS and ${\Delta}_{{\Psi}m}$ were not found to be mediated through autophagy or SIRT activation. Rather, NAM suppressed superoxide generation via a direct reduction of electron transport, and increased ${\Delta}_{{\Psi}m}$ via suppression of mitochondrial permeability transition pore formation. Our results dissected the effects of cellular $NAD^+$ redox modulation, and emphasized the importance of the $NAD^+/NADH$ ratio in the mitochondria as well as the cytosol in maintaining mitochondrial quality.

• Journal of Nutrition and Health
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• 제42권5호
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• pp.442-452
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• 2009

흡연은 신체 내 산화 스트레스를 유발할 뿐 아니라 체내 항산화 상태를 악화시킨다. 따라서 흡연으로 인해 유도되는 산화 스트레스를 줄여주기 위한 여러 다양한 식품영양학적인 시도들이 되어져 왔다. 프로폴리스는 꿀벌이 자신의 생존과 번식을 유지하기 위하여 여러 식물에서 뽑아낸 수지에 꿀벌 자신의 침과 효소 등을 혼합하여 만든 천연 물질이다. 본 연구에서는 프로폴리스가 산화 스트레스로 인해 나타난 흡연자의 DNA 손상을 회복시키어 항산화 영양 상태를 개선시키는지를 보고자 하여 placebo를 사용하는 double- blind cross-over 인체시험을 수행하였다. 흡연자에게 800 mg의 프로폴리스와 placebo를 4주 섭취시킨 후 2주 washout period를 가진 뒤 다시 군을 바꾸어 교차시험으로 4주간 섭취시킨 후, comet assay에 의한 DNA 손상도 및 신체 내 항산화 효소 수준, 총 항산화력 및 항산화 비타민 상태를 분석하였다. 처음 2주 동안의 고갈기간 후에 흡연자 29명 (평균나이: 34.38 ${\pm}$ 1.73세)을 프로폴리스군과 위약군의 두 군으로 나누어 하루에 프로폴리스 또는 위약을 4주 동안 공급하였고 2주 동안의 washout 기간을 가진 후에 교차실험을 위해 대상자의 군을 바꾸어 다시 4주 동안 프로폴리스와 위약을 공급하였다. Comet assay로 분석한 대상자의 임파구 DNA 손상정도는 프로폴리스를 섭취한 군이나 위약을 섭취한 군 간에 차이를 보이지 않았으며 총 항산화 영양상태도 두 군 간에 차이가 나타나지 않았다. 적혈구 항산화 효소인 catalase, glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) 활성도, 그리고 혈장 vitamin C와 tocopherol 수준도 프로폴리스군과 위약군 사이에 차이를 보이지 않았다. 따라서 흡연자에 있어서 프로폴리스의 항산화 효과를 평가하기 위해서는 앞으로 더 다양한 연구가 필요하리라고 생각된다.