• Archives of Pharmacal Research
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• 제28권7호
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• pp.839-847
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• 2005

The aim of this study was to provide the basis to further examine the mode of action of ethanol. Fluorescent probes reported to have different membrane mobilities were used to evaluate the effect of dimyristoylphosphatidylethanol (DMPEt) on the lateral and rotational mobilities of liposome lipid bilayers. An experimental procedure, based on the selective quenching of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1,3-di(1-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, was used. DMPEt increased the bulk lateral and rotational mobilities, and had a greater fluidizing effect on the outer than the inner monolayer. These effects of DMPEt on liposomes may be responsible for some, but not all, of the general anesthetic actions of ethanol.

• Molecules and Cells
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• 제21권2호
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• pp.229-236
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• 2006

Gaegurin 4 (GGN4), a novel peptide isolated from the skin of a Korean frog, Rana rugosa, has broad spectrum antimicrobial activity. A number of amphipathic peptides closely related to GGN4 undergo a coil to helix transition with concomitant oligomerization in lipid membranes or membrane-mimicking environments. Despite intensive study of their secondary structures, the oligomeric states of the peptides before and after the transition are not well understood. To clarify the structural basis of its antibiotic action, we used analytical ultracentrifugation to define the aggregation state of GGN4 in water, ethyl alcohol, and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). The maximum size of GGN4 in 15% HFIP corresponded to a decamer, whereas it was monomeric in buffer. The oligomeric transition is accompanied by a cooperative 9 nm blue-shift of maximum fluorescence emission and a large secondary structure change from an almost random coil to an ${\alpha}$-helical structure. GGN4 induces pores in lipid membranes and, using electrophysiological methods, we estimated the diameter of the pores to be exceed $7.3{\AA}$, which suggests that the minimal oligomer structure responsible is a pentamer.

• 한국산학기술학회논문지
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• 제12권4호
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• pp.1770-1774
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• 2011

본 논문은 리포포린에 의해 디아실글리세리드(DAG)가 Manduca sexta 유충 지방체로 운반되는 과정을 조사하였다. $[^3H]$-DAG 표지 리포포린($[^3H]$-DAG-Lp)을 시간별로 유충 지방체와 배양하여 지방체로 운반되는 DAG의 방사능을 결정하였다. $[^3H]$-DAG-Lp와 지방체를 배양하면 지방체에 DAG가 축적되며 이의 일부는 지방체에서 트리아실글리세리드(TAG)로 전환되는 것을 알 수 있었다. Suramin과 표지되지 않은 리포포린(unlabeled Lp)의 존재 하에서는 지방체로 운반되는 DAG가 억제되는 데, 이는 DAG를 운반하는 데 리포포린 수용체가 관여한다는 사실을 알 수 있었다. Suramin의 효과는 다소 복잡하지만 리포포린 수용체에 결합하여 막의 특성을 변화시켜 DAG 운반속도에 영향을 주는 것 같다. 지질 운반과정이 수용체-매개 내포작용이라는 사실을 조사하기 위하여 내포작용 억제자인 ammonium chloride와 chloroquine을 처리하였다. 그 결과 리포포린과 지방체사이에서의 지질 운반 기작이 수용체-매개 과정이라는 사실을 보여준다.

• 한국응용과학기술학회지
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• 제39권5호
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• pp.623-631
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• 2022

실험 계획법을 활용하여 세라마이드가 고함량으로 함유된 수화 액정형 베시클의 입자 사이즈에 영향을 미치는 인자를 분석하고 혼합물 조성의 최적화를 시도하였다. 베시클 입자 사이즈에 영향을 주는 주요 변수로 제조 온도, 에탄올 양, 초음파 시간을 각각 정하고, 다구찌 방법의 S/N비 산출과 ANOVA 분석을 통해, 이들 변수들이 입자 사이즈에 미치는 영향을 살펴보았다. 또한 혼합물 실험 계획법의 심플렉스 중심 설계에 따라, 베시클 막을 구성하는 세 지질 성분, 인지질(HPC), 콜레스테롤(Chol), 세라마이드(Cer)의 혼합 조성 실험을 수행하였다. 실험에서 얻어진 데이터로부터 회귀분석을 실시하여 모델식을 구하고, 베시클 입자 사이즈를 최소로 만들기 위한 세 지질 성분의 최적 혼합 조성은 HPC(0.6), Chol(0.1), Cer(0.3)으로 구해졌다.

• 대한의용생체공학회:의공학회지
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• 제45권1호
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• pp.20-25
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• 2024

Extracellular vesicles (EVs) are nano-sized lipid bilayer vesicles released by cells. EVs act as messengers for cell-to-cell communication. Inside, it contains various substances that show biological activity, such as proteins, lipids, nucleic acids, and metabolites. The study of EVs extracted from terrestrial organisms and stem cells on inflammatory environments and tissue regeneration have been actively conducted. However, marine organisms-derived EVs are limited. Therefore, we have extracted EVs from sea urchins belonging to the Echinoderm group with their excellent regenerative ability. First, we extracted extracellular matrix (ECM) from sea urchin gonads treated with hypotonic buffer, followed by collagenase treatment, and filtration to collect ECM-bounded EVs. The size of sea urchin gonad-derived EVs (UGEVs) is about 20-100 nm and has a round shape. The protein content was higher after EVs burst than before, which is evidence that proteins are contained inside. In addition, proteins of various sizes are distributed inside. PKH-26 was combined with UGEVs, which means that UGEVs have a lipid membrane. PHK-26-labeled UGEVs were successfully uptaken by cells. UGEVs can be confirmed to have the same characteristics as traditional EVs. Finally, it was confirmed that Schwann cells were not toxic by increasing proliferation after treatment.

• 공업화학
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• 제7권2호
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• pp.252-260
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• 1996

막의 기능설계를 하기 위하여 DPPC(dipalmitoylphosphatidylcholine)인 지질로 제조한 liposome과 DPPC와 DTAB(dodecyltrimethylammonium bromide)를 혼합하여 liposome을 제조하여 그 구조변화를 조사하였다. 막 구조의 변화는 CF(carboxylfluorescein)를 이용하여 DPPC의 상전이 온도($t_c=41^{\circ}C$) 이상과 이하에서 측정하여 평가하였다. DPPC liposome에 함유된 CF의 유출에 의한 형광강도는 $45^{\circ}C$에서 증가되었지만 $20^{\circ}C$에서는 그 변화는 관측되지 않았다. 동등한 조건에서 DPPC/DTAB liposome에서 CF 유출에 의한 형광강도의 변화는 DPPC liposome에 비하여 큰 것이 관측되었다. 이 결과는 DPPC/DTAB liposome이 불규칙한 배열을 하고 있다는 것을 시사한다. 한편 $Ca^{2+}$ 존재하에서 DPPC liposome과 DTAB/DPPC liposome에서 Quin 2의 형광강도는 $45^{\circ}C$에서 현저히 증가되었지만 $20^{\circ}C$에서는 거의 변화하지 않았다. 그리고 형광강도의 변화는 DPPC/DTAB liposome보다 DPPC liposome 쪽이 더 크게 나타났다. 이것은 DPPC/DTAB liposome의 구조가 DPPC liposome보다 더 안정하다는 것을 의미한다. 또한 형광물질인 ANS를 이용하여 막 표면의 양상을 조사한 결과 DPPC liposome과 DPPC/DTAB liposome에서 ANS의 형광강도는 $45^{\circ}C$와 $20^{\circ}C$에서 다른 양상을 보였다. 이 양상은 상전이 온도의 이상과 이하에서 DPPC liposome과 DPPC/DTAB liposome의 막유동성을 의미하고 그 변화량은 DPPC liposome이 DPPC/DTAB liposome보다 더 컸다. 또한 DSC에 의해 측정된 상전이 온도는 DPPC liposome의 경우 $41^{\circ}C$이었으며 DPPC/DTAB liposome의 경우에는 $32^{\circ}C$이었다. 이상의 결과로 DPPC/DTAB liposome의 분자배열 상태가 불규칙한 것을 의미한다. 그러므로 DPPC/DTAB liposome은 불규칙하게 배열되어 있지만 안정된 구조를 갖고 있다고 생각된다.

• 대한의생명과학회지
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• 제23권1호
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• pp.17-24
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• 2017

Estrogens are effective neuroprotectants in vivo and in vitro. To obtain a better insight into the molecular mechanisms of action of neuroprotection by $17{\beta}-estradiol$ (E2), we examined the differential effects of E2 on the fluidity of synaptosomal plasma membrane vesicles (SPMV) isolated from rat cerebral cortex. Intramolecular excimerization of 1,3-di(1-pyrenyl)-propane (Py-3-Py) was used to investigate the effects of E2 on the bulk and annular lateral diffusion of the SPMV. In addition, we examined the effects of E2 on the rotational diffusion of individual leaflet of SPMV exploiting selective quenching of outer monolayer 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence by trinitrophenyl groups. The $F{\ddot{o}}rster$ distance $R_0$ value for the tryptophan-Py-3-Py donor-acceptor pair was $26.9{\AA}$. E2 increased the lateral mobility of both bulk and annular lipids in SPMV in a dose-dependent manner, but a larger effect on bulk lipids was observed. Although E2 decreased the anisotropy of DPH in SPMV, E2 had a greater fluidizing effect on the outer leaflet compared to the inner leaflet. These results suggest that E2 selectively fluidizes the more fluid regions within SPMV. It is highly probable that E2 mostly fluidizes the bulk lipids, away from either annular lipids or lipid rafts, in the outer leaflet of SPMV. This selective fluidization may be one of the nongenomic mechanisms of neuroprotection by E2.

• 약학회지
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• 제51권4호
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• pp.285-290
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• 2007

The oxidative stress causes many diseases like cancer, aging, cardiovascular disease, degenerative neurological disorders (Parkinson’s disease, and Alzheimer's disease) by damage of cell membrane, protein deformation, and damage of DNA due to the oxidation of lipid of cell membrane, protein of tissue or enzyme, carbohydrate, and DNA. It is caused by the reactive oxygen species (ROS) that is produced in the metabolic process of oxygen in cell. The superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in cell systemize the antioxidative enzymes to control the oxidative stress. In this research, it is measured that the survival rate of cell by the typical isoflavonoid of daidzein or genistein, activity of antioxidative enzyme, and ROS level, in order to study the effect of isoflavonoid over the ROS production in cell and antioxidative system. As the similar action of the isoflavonoid with the estrogen is examined, women are encouraged to get bean. In view of this trend, it is very important to find out a combination medicine that lowers the oxidative stress caused by the daidzein in the ovarian cell. In the combined treatment of the typical antioxidant of vitamin C to oxidative stress which induced by daidzein recover the control level particularly lowering the ROS in cell by 30%. However, it made no effect in the combined treatment with genistein. Therefore, the research took the combination effect of daidzein with vitamin C in order to check it effect over the antioxidative system. In conclusion, it was disclosed that the oxidative stress caused by daidzein is related to the lowering activity of SOD, and the specific combination effect of daidzein with vitamin C is related to the recovery of SOD activity.

• 환경생물
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• 제28권2호
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• pp.101-107
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• 2010

Metal ions are essential to life. However, some metals such as mercury are harmful, even when present at trace amounts. Toxicity of mercury arises mainly from its oxidizing properties. Ionizing radiation (IR) is an active tool for destruction of cancer cells and diagnosis of diseases, etc. IR induces DNA double strand breaks in the nucleus, In addition, it causes lipid peroxidation, ceramide generation, and protein oxidation in the membrane, cytoplasm and nucleus. Yeasts have been a commonly used material in biological research. In yeasts, the physiological response to changing environmental conditions is controlled by the cell types. Growth rate, mutation and environmental conditions affect cell size and shape distributions. In this work, the effect of IR and mercury chloride (II) on the morphology of yeast cells were investigated. Saccharomyces cerevisiae cells were treated with IR, mercury chloride (II) and IR combined with mercury chloride (II). Non-treated cells were used as a control group. Morphological changes were observed by a scanning electron microscope (SEM). The half-lethal condition from the previous experimental results was used to the IR combined with mercury. Yeast cells were exposed to 400 and 800 Gy at dose rates of 400Gy $hr^{-1}$ or 800 Gy $hr^{-1}$, respectively. Yeast cells were treated with 0.05 to 0.15 mM mercury chloride (II). Oxidative stress can damage cellular membranes through a lipidic peroxidation. This effect was detected in this work, after treatment of IR and mercury chloride (II). The cell morphology was modified more at high doses of IR and high concentrations of mercury chloride(II). IR and mercury chloride (II) were of the oxidative stress. Cell morphology was modified differently according to the way of oxidative stress treatment. Moreover, morphological changes in the cell membrane were more observable in the frequently stress treated cells than the temporarily stress treated cells.

• BMB Reports
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• 제32권6호
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• pp.561-566
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• 1999

CA(1-8)-ME(1-12) [CA-ME], composed of cecropin A(1-8) and melittin(1-12), is a synthetic antimicrobial peptide having potent antibacterial and antitumor activities with minimal hemolytic activity. In order to investigate the effects of the flexible hinge sequence, Gly-Ile-Gly, of CA-ME on antibiotic activity, CA-ME and three analogues, CA-ME1, CA-ME2, and CA-ME3, were synthesized. The Gly-Ile-Gly sequence of Ca-ME was deleted in CA-ME1 and replaced with Pro and Gly-Pro-Gly in CA-ME2 and CA-ME3, respectively. CA-ME1 and CA-ME3 showed a significant decrease in antitumor activity and phospholipid vesicle-disrupting ability. However, CA-ME2 showed similar antitumor and vesicle-disrupting activities, as compared with CA-ME. These results suggest that the flexibility or ${\beta}$-turn induced by Gly-Ile-Gly or Pro in the central part of CA-ME may be important in the electrostatic interaction of the N-terminus cationic ${\alpha}$-helical region with the cell membrane surface and the hydrophobic interaction of the C-terminus amphipathic ${\alpha}$-helical region with the hydrophobic acyl chains in the cell membrane. CA-ME3 exhibited lower antitumor and vesicle-disrupting activities than CA-ME and CA-ME2. This result suggests that the excessive ${\beta}$-turn structure caused by the Gly-Pro-Gly sequence in CA-ME3 seems to interrupt ion channel/pore formation in the lipid bilayer. We concluded that the appropriate flexibility or bilayer. We concluded that the appropriate flexibility or ${\beta}$-turn structure provided by the central hinge is responsible for the effective antibiotic activity of the antimicrobial peptides with the helix-hinge-helix structure.