• 한국산업보건학회지
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• 제19권3호
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• pp.261-269
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• 2009

To evaluate the effect of antioxidant on the cytotoxicity induced by oxidative stress of reactive oxygen species (ROS) in cultured human skin melanocytes, colorimeric assay of XTT and tyrosinase activity assay were adopted after human skin melanocytes were preincubated for 2 hours in the media containing various concentrations of superoxide dismutase (SOD) before the treatment of hydrogen peroxide. Light microscopic study was carried out in same cultures. The results of this study were as follows 1. Cell viability of human skin melanocytes was significantly decreased by 30 and $40{\mu}M$ of hydrogen peroxide($H_2O_2$), respectively. 2. XTT50 was determined at $30{\mu}M$ after human skin melanocytes were treated with $10{\sim}40{\mu}M$ of hydrogen peroxide for 6 hours. 3. The cell viability of cultured human skin melanocytes pretreated with SOD was increased than that of cultured human skin melanocytes treated with $H_2O_2$ dose-dependently. 4. In tyrosinase activity of human skin melanocytes, the cell treated with SOD showed brown stain compared with $H_2O_2$ treated cells, dark stain. 5. In light microscopy, cultured human skin melanocytes exposed to $H_2O_2$ showed morphological changes such as the decreased cell number and cytoplasmic processes, compared with control. 6. In light microscopy, cultured human skin melanocytes pretreated with SOD showed the increase of cell number and cytoplasmic processes compared with $H_2O_2-treated$ group. From these results, it is suggested that oxidative stress of ROS such as $H_2O_2$ has cytotoxicity by showing the decreased cell viability, the increased tyrosinase activity and mophological changes of the decreased cell number and cytoplasmic processes. While, antioxidant like SOD was effective in the prevention of oxidative stress-mediated cytotoxicity by the increased cell viability, decreased tyrosinase activity and the protection of degenerative morphological changes in cultured human skin melanocytes.

• 대한수의학회지
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• 제14권1호
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• pp.77-90
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• 1974

Hemolytic reaction of normal fresh chicken serum on sheep erythrocytes was studied and the following experimental results were obtained and summarized. 1. Chicken sera, 258 (78%) out of 344 samples showed hemolytic activity on sheep erythrocytes. 2. Distribution of a different hemolytic titer of chicken sera was not dependent to sex and age difference of test chicken. 3. Hemolytic activity of serum component obtained from normal fresh chicken was heat inactivated at $56^{\circ}C$. 30 minutes heating. 4. The most enhanced hemolytic activity of chicken serum on sheep erythrocytes was observed at the incubation temperature of $46^{\circ}C$. 5. The most effective pH for the hemolytic reaction of chicken serum on sheep erythrocytes was observed at 7.0, and pH 6.0 or 8.5 resulted less or no hemolysis. 6. Hemolytic reaction of chicken serum and sheep erythrocytes required Mg⧻ and Ca⧻ ions as, co-factor, and the former was required more compared to the latter. 7. Hemolytic activity of chicken serum was observed in ChC 2, 4 fraction but not in ChC 1, 3, ChC 3, 4, ChC 1, 2, 4 and ChC 1, 2, 3 fractions. 8. In electron micrography, morphological changes of sheep erythrocyte membrane by normal chicken serum was similar to that of immune hemolysis: that was, the hemolytic hole was circular and it was surrounded with a white ring. 9. Electron micrography of morphological changes on sheep erythrocyte membrane indicated that the size of hemolytic hole and white ring were functional to the chicken serum concentration used and reaction time.

• Journal of Nutrition and Health
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• 제35권10호
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• pp.1015-1022
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• 2002

This study was conducted to examine the effects of dietary xylooligosaccharides on hepatic HMG-CoA reductase activity and morphological exchange of liver in rats fed high fat diet. Sprague-Dawley male rats weighing 100 $\pm$ 10 g were randomly divided into four groups, two normal diets and two high fat diets containing 1% cholesterol and 10% lard. Two normal diets were classified into a basal diet (normal group) and 10% xylooligosaccharide diet (NX group). The high fat diet groups were classified into a HF group without xylooligosaccharides diet and HFX group supplemented 10% xylooligosacchride diet. Experimental diets were fed ad libidum to the rats for 4 weeks and then they were sacrificed. The body weight of high fat diet (HF group) was increased more than that of normal group, but it was significantly decreased by xylooligosacchrides supplementation. The food intake was not significantly different among the all groups. The weight of liver, small intestine and cecum of all xylooligosaccharide supplemented groups were significantly heavier than those of normal and HF groups. The activity of hepatic HMG-CoA reductase, a rate limiting enzyme of cholesterol biosynthesis, in xylooligosaccharide supplemented groups was higher than that of HF group. Light micrographs revealed that the structures of hepatocytes in xylooligosaccharide supplemented groups were preserved well, compared to HF group. The xylooligosaccharide supplementation exerted a lipid-lowering action by decreasing cholesterol and triglycerides contents in hepatic tissue. In conclusion, the activity of hepatic HMG-CoA reductase and damage of liver in rats fed high fat diets were improved by dietary xylooligosaccharides.

• 치위생과학회지
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• 제20권4호
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• pp.213-220
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• 2020

Background: The leaves of Perilla frutescens, commonly called perilla and used for food in Korea, contain components with a variety of biological effects and potential therapeutic applications. The purpose of this study was to identify the components of 70% ethanol extracted Perilla frutescens (EEPF) and determine its inhibitory effects on oral microbial activity and production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharides (LPS)-stimulated Raw264.7 macrophages, consequently, to confirm the possibility of using EEPF as a functional component for improving the oral environment and preventing inflammation. Methods: One kg of P. frutescens leaves was extracted with 70% ethanol and dried at -70℃. EEPF was analyzed using high-performance liquid chromatography analysis, and antimicrobial activity against oral microorganisms was revealed using the disk diffusion test. Cell viability was elucidated using a methylthiazolydiphenyl-tetrazolium bromide assay, and the effect of EEPF on LPS-induced morphological variation was confirmed through microscopic observation. The effect of EEPF on LPS-induced production of pro-inflammatory mediators, NO and PGE2 was confirmed by the NO assay and PGE2 enzyme-linked immunosorbent assay. Results: The main component of EEPF was rosemarinic acid, and EEPF showed weak anti-bacterial and anti-fungal effects against microorganisms living in the oral cavity. EEPF did not show toxicity to Raw264.7 macrophages and had inhibitory effects on the morphological variations and production of pro-inflammatory mediators, NO and PGE2 in LPS-stimulated Raw264.7 macrophages. Conclusion: EEPF can be used as a functional material for improving the oral environment through the control of oral microorganisms and for modulating inflammation by inhibiting the production of inflammatory mediators.

• Toxicological Research
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• 제8권2호
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• pp.285-302
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• 1992

Human and rat hepatocytes were isolated by nonperfusion method and cultured for longer than 5 days. Human liver biopsy sample and rat liver were used as hepatocyte source. Several physical and chemical factors which were influencing on hepatocyte isolation procedure were examined and a batch isolation procedure was established for small size sample of rat liver. Isolated hepatocytes showed normal morphlologica characteristics in microscopy and electron microscopical examinations and a morphologica response to phalloidin. Isolated cells were cultured as a monolayer and proven to have intact morphological characteristics for longer than 15 days. Because human liver sample is harder and tighter compared with rat liver, a standard procedure for rat hepatocytes was slightly modified to reduce mechanical damage. Similarly with rat hepatocytes, isolated human hepatocytes showed a normal morphological characteristics and could be cultured for longer than 15days. Human and rat hepatocytes were examined on their functional integrities including cytochrome-P450 related enzyme activity and it's inducibility, hormonal inducibility of AIB uptake and TAT activity, albumin synthesis, DNA synthesis, cellular protein maintenance. In all parameters used in the present study, human and rat hepatocytes showed normal functional characteristics.

• Journal of Plant Biology
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• 제30권4호
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• pp.257-266
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• 1987

Investigation on the ability of nitrogen usage by N2-fixing Nostoc muscorum and cultured tobacco cells when they were associately cultured on nitrogen-free media was carried out. Also, effect of polyamines on the associated cultured condition was carried out. In addition, morphological changes of N. muscorum and cultured tobacco cells in associate culture were observed to detect the possibility of induction of nitrogen fixing ability on cultued plant cells. The activity of nitrogenase increased markedly when N. muscorum was grown exclusively on nitrogen-free media. When N. muscorum was cultured associately with cultured tobacco cells on nitrogen-free media containing polyamines, high activity was detected in 10-4 M spermine treated group. Investigation on the change of polyamine amounts showed two times increase in spermidine and eight times increase in spermine on a associate culture. These effects of associated culture were shown through morphological change such as dense loclization of N. muscorum around the cultured tobacco cells as well as inside the cells. These results indicate the viability of N. muscorum in cultured tobacco cells and possible induction of nitrogen fixation ability by symbiosis.

• 한국균학회지
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• 제37권2호
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• pp.181-188
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• 2009

A Gram-positive, rod-shaped bacteria named DY5 was isolated from a peat sample collected from Daeam mountain in Korea. The culture filtrate of the bacterial isolate DY5 showed a broad spectrum of antifungal activity on various crop pathogenic fungi such as Trichoderma koningii, Fusarium oxysporum, Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Rhizoctonia solani AG-1(IA) For the identification of the DY5, morphological, biochemical, API 50 CHB test, analysis of fatty acid and molecular phylogenetic approaches were performed. The DY5 was found to be a member of the genus Paenibacillus on the basis of morphological and biochemical analysis. The 16S rRNA of DY5 showed high similarity(98%) with Paenibacillus polymyxa. On the basis of these results, the DY5 was identified as Paenibacillus polymyxa. Antifungal substance of the DY5 would be mild alkaline proteine molecule. The DY5 seems to have a great potential to be a biocontrol agent against various crop pathogens.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1237-1245
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• 1997

Interspecific fusants were made from the cells of two strains of Lactobacillus genus, a streptomycin resistant Lactobacillus sp. JC-7 and a kanamycin resistant L. acidophilus 88. The morphological and physiological properties of the fusants were examined by determining bacteriocin productivity, acid-producing activity, ability of carbohydrates utilization and three important enzyme activities. The fusants produced a bacteriocin against indicator strains and fusant No. 1, 4 exhibited a larger inhibition zone compared to that of L. acidophilus 88. $\beta$-Galactosidase, phospho-$\beta$-galactosidase, lipase activities and resistance to NaCl of Lactobacillus sp. JC-7 were better than those of L. acidophilus 88. Fusant No. 3 and 7 exhibited excellent lipase activities. Protease activity and acid productivity of L. acidophilus 88 were better than those of Lactobacillus sp. JC-7. Proteasse activities of all fusants were higher than those of parental strains, and expecially fusant No. 5 and 7 exhibited excellent proteolysis ability.

• The Korean Journal of Physiology and Pharmacology
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• 제24권3호
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• pp.223-232
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• 2020

Sesamin, a lipid-soluble lignin originally isolated from sesame seeds, which induces cancer cell apoptosis and autophagy. In the present study, has been reported that sesamin induces apoptosis via several pathways in human lung cancer cells. However, whether mitophagy is involved in sesamin induced lung cancer cell apotosis remains unclear. This study, the anticancer activity of sesamin in lung cancer was studied by reactive oxygen species (ROS) and mitophagy. A549 cells were treated with sesamin, and cell viability, migration ability, and cell cycle were assessed using the CCK8 assay, scratch-wound test, and flow cytometry, respectively. ROS levels, mitochondrial membrane potential, and apoptosis were examined by flow cytometric detection of DCFH-DA fluorescence and by using JC-1 and TUNEL assays. The results indicated that sesamin treatment inhibited the cell viability and migration ability of A549 cells and induced G₀/G₁ phase arrest. Furthermore, sesamin induced an increase in ROS levels, a reduction in mitochondrial membrane potential, and apoptosis accompanied by an increase in cleaved caspase-3 and cleaved caspase-9. Additionally, sesamin triggered mitophagy and increased the expression of PINK1 and translocation of Parkin from the cytoplasm to the mitochondria. However, the antioxidant N-acetyl-L-cysteine clearly reduced the oxidative stress and mitophagy induced by sesamin. Furthermore, we found that cyclosporine A (an inhibitor of mitophagy) decreased the inhibitory effect of sesamin on A549 cell viability. Collectively, our data indicate that sesamin exerts lethal effects on lung cancer cells through the induction of ROS-mediated mitophagy and mitochondrial apoptosis.

• Molecules and Cells
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• 제40권7호
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• pp.495-502
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• 2017

The $5-HT_6R$ has been considered as an attractive therapeutic target in the brain due to its exclusive expression in the brain. However, the mechanistic linkage between $5-HT_6Rs$ and brain functions remains poorly understood. Here, we examined the effects of $5-HT_6R$-mediated cell morphological changes using immunocytochemistry, Western blot, and live-cell imaging assays. Our results showed that the activation of $5-HT_6Rs$ caused morphological changes and increased cell surface area in HEK293 cells expressing $5-HT_6Rs$. Treatment with 5-HT specifically increased RhoA-GTP activity without affecting other Rho family proteins, such as Rac1 and Cdc42. Furthermore, live-cell imaging in hippocampal neurons revealed that activation of $5-HT_6Rs$ using a selective agonist, ST1936, increased the density and size of dendritic protrusions along with the activation of RhoA-GTP activity and that both effects were blocked by pretreatment with a selective $5-HT_6R$ antagonist, SB258585. Taken together, our results show that $5-HT_6R$ plays an important role in the regulation of cell morphology via a RhoA-dependent pathway in mammalian cell lines and primary neurons.