• The Korean Journal of Physiology and Pharmacology
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• v.7 no.3
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• pp.119-124
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• 2003

To elucidate the molecular mechanism of pharmacological action of local anesthetics, we studied membrane actions of tetracaine, bupivacaine, lidocaine, prilocaine and procaine. Fluorescence polarization of n-(9-anthroyloxy)stearic acid (n-AS) was used to examine the effects of these local anesthetics on differential rotational mobility of different positions of the number of synaptosomal plasma membrane vesicle (SPMV) phospholipid carbon atoms. The four membrane components differed with respect to 3, 6, 9 and 16-(9-anthroyloxy)stearic acid (3-AS, 6-AS, 9-AS and 16-AP) probes, indicating that differences in the membrane fluidity might be present. Degrees of the rotational mobility of 3-AS, 6-AS, 9-AS and 16-AP were different depending on depth of hydrocarbon interior. In a dose-dependentmanner, tetracaine, bupivacaine, lidocaine, prilocaine and procaine decreased anisotropy of 3-AS, 6-AS, 9-AS and 16-AP in the hydrocarbon interior of the SPMV. These results indicate that local anesthetics have significant disordering effects on hydrocarbon interior of the SPMV, thus affecting the transport of $Na^+$ and $K^+$ in nerve membranes and leading to anesthetic action.

• Journal of Pharmaceutical Investigation
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• v.24 no.3
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• pp.131-137
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• 1994

Drug delivery system by the use of red blood cells was established to sustain the release of drugs in the circulatory system by the intravenous injection. The entrapment method by the preswelling technique was re-examined and evaluated for searching the new entrapping conditions without hemolysis. The addition of 4 volume of $0.6{\times}\;hank's$ balanced salt solution (HBSS) into 1 volume of 50% red blood cells suspension did not induce the hemolysis and change the hematocrit level in this experimental condition (within 15 min). Most of daunorubicin could be entrapped into red blood cells within 15 min. While the intracellular adenosine triphosphate (ATP) level followed by the entrapment was reduced to 86% of normal ATP level, the membrane fluidity and the shape factor of red blood cells were not altered. The release rate of daunorubicin from red blood cells was affected by the hemolysis under this condition. To maintain the intracellular ATP in red blood cells, the new reaction buffer was made With the addition of ATP and sodium pyruvate during the entrapment procedure because the hemolysis during the release test would reflect the loss of intracellular ATP that might result in the decrease of the viability in vivo. The addition of ATP raised the intracellular ATP level, which protect the hemolysis during the release test.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.154-164
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• 2005

Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.

• Archives of Pharmacal Research
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• v.15 no.4
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• pp.298-303
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• 1992

Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex. The effects of barbiturates on the rotational relaxation time of 1.6-diphenyl-1, 3, 5-hexatriene (DPH) in intact SPMV and model membranes of total lipids (SPMVTL) and phosphlipids (SPMVPL) extracted from SPMV were examined. Barbiturates decreased the rotational relaxation time of DPH in intact SPMV in a dose-dependent manner. In contrast, they did not affect the rotational relaxation time of DPH in SPMVTL and even dose-dependently increased the rotational relaxation time of DPH in SPMVPL.

• Journal of Nutrition and Health
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• v.28 no.8
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• pp.706-716
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• 1995

This study was undertaken to investigate the influence of age and dietary linolenic acid content and the linolenic acid/linoleic acid (LAN/LA) ratio on the brain lipid composition and membrane-bound enzyme, acetylcholinesterase(AchE) activities. AchE was selected as a test case for the relationship between cell lipid composition and cell membrane function. The male rats were fed diets with 0.2, 0.4, 0.6 of LNA/LA ratio within 8% LNA(H-LNA) or 4% LNA(L-LAN) of total fatty acid content for different feeding period(1, 4, 12 month). The fats used s source were sesame oil, perilla oil, soybean oil and beef tallow. The AchE activity of brain crude synaptosomal fraction was reduced with advancing age, showing 20-30% reduction in 12M compared with 1 M, and the P/C ratio was reduced in old rats. In 1 and 4 monthed rats, AchE activites was higher in H-LAN-0.2 and L-LNA-0.2 and 0.4 group. In accordance with rising of AchE activities was higher in H-LNA-0.2 and L-LNA-0.2 and 0.4 group. In accordance with rising of AchE activities, the PC/PE ratio increasedin those groups. Paricularly in L-LNA, the PC/PE ratio increased as the AchE activites for decline of membrane fluidity with increasing cholesterol and decreasing P/C ratio when rats were old. Also, AchE activity increaed with increasing PC/PE ratio which depended on the dietary LNA/LA ratio within each LNA content. Therefore, it is concluded that the lipid composition of cell membrane influenced the AchE activiteis, which was mediated by aging and the modification of dietary LNA/LA ratio.

• Journal of Microbiology and Biotechnology
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• v.30 no.5
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• pp.739-748
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• 2020

In this study, a method of heat adaptation was implemented in an attempt to increase the upper thermal threshold of two Streptococcus thermophilus found in South Korea and identified the alterations in membrane fatty acid composition to adaptive response to heat. In order to develop heat tolerant lactic acid bacteria, heat treatment was continuously applied to bacteria by increasing temperature from 60℃ until the point that no surviving cell was detected. Our results indicated significant increase in heat tolerance of heat-adapted strains compared to the wild type (WT) strains. In particular, the survival ratio of basically low heat-tolerant strain increased even more. In addition, the strains with improved heat tolerance acquired cross protection, which improved their survival ratio in acid, bile salts and osmotic conditions. A relation between heat tolerance and membrane fatty acid composition was identified. As a result of heat adaptation, the ratio of unsaturated to saturated fatty acids (UFA/SFA) and C18:1 relative concentration were decreased. C6:0 in only heat-adapted strains and C22:0 in only the naturally high heat tolerant strain were detected. These results support the hypothesis, that the consequent increase of SFA ratio is a cellular response to environmental stresses such as high temperatures, and it is able to protect the cells from acid, bile salts and osmotic conditions via cross protection. This study demonstrated that the increase in heat tolerance can be utilized as a mean to improve bacterial tolerance against various environmental stresses.

• The Korean Journal of Pharmacology
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• v.29 no.1
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• pp.149-156
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• 1993

Intramolecular excimerization of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to examine the effects of ethanol on the rate and range of lateral diffusion of bulk bilayer structures of plasma membrane vesicles isolated from cultured mouse myeloma cell line Sp2/0-Ag14 (Sp2/0-PMV). In a concentration-dependent manner, ethanol increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in the Sp2/0-PMV and decreased the anisotropy (r), limiting anisotropy $(r_{\infty})$, and order parameter (S) of DPH in the Sp2/0-PMV. This indicates that ethanol increased both the lateral and rotational diffusion of the probes in the Sp2/0-PMV. Selective quenching of DPH by trinitrophenyl groups was utilized to examine the transbilayer asymmetric rotational diffusion of the Sp2/0-PMV. The anisotropy (r), limiting anisotropy $(r_{\infty})$, and order parameter (S) of DPH in the inner monolayer were 0.022, 0.029, and 0.063, respectively, greater than calculated for the outer monolayer of the Sp2/0-PMV. Selective quenching of DPH by trinitrophenyl groups was also utilized to examine the transbilayer asymmetric effects of ethanol on the range of rotational diffusion of the Sp2/0-PMV. Ethanol had a greater fluidizing effect on the outer monolayer as compared to the inner monolayer of the Sp2/0-PMV. It has been proven that ethanol exhibits a selective rather than nonselective fluidizing effect within transbilayer domains of the Sp2/0-PMV.

• Reproductive and Developmental Biology
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• v.40 no.2
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• pp.15-22
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• 2016

As a one of unsaturated fatty acid, polyunsaturated fatty acids (PUFAs) have multiple actions: as precursor of prostaglandins (PGs), steroid hormone synthesis and energy production in animal reproduction. PUFAs, which include omega-3 (n-3) and omega-6 (n-6), are derived from the diet and changed by diet, species, breed and season. The plasma membrane of spermatozoa in mammals contain various PUFAs. These composition of PUFAs regulate the membrane fluidity and cause lipid peroxidation via generation of reactive oxygen species (ROS). Induced lipid peroxidation by ROS decreased viability and motility of spermatozoa, and it is reduced by addition of antioxidant and low concentration of PUFAs. Because oocytes of animal have a high lipid components, process of oocyte maturation and embryo development are influenced by PUFAs. In in vitro study, oocyte maturation, embryo development, intracellular cAMP and MAPK activity were increased by treatment of n-3 ${\alpha}$-linolenic acid (ALA) during maturation, whereas n-6 linoleic acid (LA) negatively influenced. Also, inhibition of fatty acid metabolism in oocyte influenced blastocyst formation of cattle. PGs are synthesized from PUFAs and various PUFAs influence PGs via regulation of PG-endoperoxide synthase (PTGS). Steroid hormone synthesis from cholesterol is regulated by expression of steroid acute regulator (StAR) protein and mRNA. Exogenous n-3 and n-6 PUFAs altered sex hormone in animal through stimulate or inhibit StAR activity. Because PUFAs altered PG and steroid hormone synthesis, follicular development was influenced by PUFAs. This effect of unsaturated fatty acid could provide information for improvement of reproductive ability in animals.

• BMB Reports
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• v.33 no.6
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• pp.541-547
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• 2000

Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to evaluate the effects of chlorpromazine HCl on the range of the rotational mobility of bulk bilayer structure of the synaptosomal plasma membrane vesicles (SPMV) isolated from a bovine brain. In a dose-dependent manner, chlorpromazine HCl increased the anisotropy (r), limiting anisotropy ($r_{\infty}$) and order parameter (S) of DPH in the membranes. Cationic 1-[4-(trimethylammonio)-phenyl]-6-phenylhexa-1,3,5-hexatriene (TMA-DPH) and anionic 3-[p-(6-phenyl)-1,3,5-hexatrienyl]-phenylpropionic acid (PRO-DPH) were utilized to examine the range of transbilayer asymmetric rotational mobility of the neuronal membranes. The anisotropy (r) of TMA-DPH in the inner monolayer was 0.034 greater than the value of PRO-DPH in the outer monolayer of the membranes. Both cationic TMA-DPH and anionic PRO-DPH were also used to examine the transbilayer asymmetric effects of chlorpromazine HCl on the range of rotational mobility of the membranes. Chlorpromazine HCl have a decreasing effects on the rotational mobility of the bulk bilayer structures and have a greater decreasing effect on the mobility of the inner monolayer as compared to the outer monolayer of the membranes. It has been proven that chlorpromazine HCl exhibit a selective rather than nonselective fluidizing effect within the transbilayer domains of the SPMV.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.8
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• pp.1328-1336
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• 2003

This study was done to investigate the effects of n-3, n-6 fatty acid and d-limonene on the hepatic membrane lipid composition, protein kinase C (PKC) and glutathione S-transferase (GST) activities in experimental rat hepatocarcinogenesis. Sprague-Dawley female rats were fed with two different types of dietary oil for 20 weeks. Corn oil (CO) and sardine oil (SO) were used at 15% by weight as a source of n-6 and n-3 fatty acid, respectively. One week after feeding, rats were intraperitoneally injected twice with a dose of diethylnitrosamine (DEN, 50 mg/kg body weight) and after 1 week 0.05% phenobarbital (PB) was provided with drinking water. Membrane fractional lipid composition showed that the content of cholesterol was higher in 50 group than CO group and also significantly decreased by d-limonene. The content of phospholipid was increased by carcinogen treatment but not affected by dietary oils or d-limonene. Membrane C/PL molar ratio was significantly decreased by d-limonene or carcinogen treatment in 50 groups but not in CO groups. Fatty acid composition was changed by dietary oils but not by carcinogen treatment or d-limonene. Cytosolic PKC activity was not significantly different by dietary oils, d-limonene or carcinogen treatment. However, membrane PKC activity was significantly increased by carcinogen treatment and decreased by d-limonene. Cytosolic GST activity was affected by d-limonene or carcinogen treatment in all dietary groups. These data indicate that dietary oils, d-limonene and carcinogen treatment can not change much membrane phospholipid composition. But membrane C/PL molar ratio was changed by carcinogen treatment and d -limonene although the effect was different between dietary oils. Therefore, it is suggested that different dietary oils and d-limonene can somewhat modulate the changes of membrane fluidity and activities of membrane bound enzymes like membrane associated PKC during carcinogenesis.