• Applied Chemistry for Engineering
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• v.29 no.4
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• pp.452-460
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• 2018

This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.625-634
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• 2011

Green tea seed coat (GTSC) was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether (PE), ethyl acetate (EtOAC) and butanol (BuOH). The EtOAC fraction showed the highest level in total phenol contents and the lowest level in nitric oxide (NO) production in LPS-stimulated RAW264.7 macrophage cell. Thus, this study was carried out to investigate the anti-inflammatory and its mechanisms of GTSC EtOAC fraction in LPS-stimulated RAW264.7 macrophage cell. GTSC EtOAC fraction contained EGC ($1146.48{\pm}11.01\;{\mu}g/g$), tannic acid ($966.99{\pm}32.24\;{\mu}g/g$), EC ($70.88{\pm}4.39\;{\mu}g/g$), gallic acid ($947.61{\pm}1.03\;{\mu}g/g$), caffeic acid ($37.69{\pm}1.46\;{\mu}g/g$), ECG ($35.46{\pm}3.19\;{\mu}g/g$), and EGCG ($15.53{\pm}0.09\;{\mu}g/g$) when analyzed by HPLC. NO production was significantly (p<0.05) suppressed in a dose-dependent manner with an $IC_{50}$ of $80.11\;{\mu}g$/mL. Also prostaglandin $E_2$ level was also inhibited in a dose-dependent manner. Moreover, iNOS protein expression was suppressed in dose-dependent manner but COX-2 gene expression was not affected. Total antioxidant capacity and glutathione (GSH) levels were enhanced more than the LPS-control. Expressions of antioxidative enzymes including catalase, GSH-reductase and Mn-SOD were elevated compared to LPS-control. Nuclear p65 level was decreased in the GTSC EtOAC fraction in a dose-dependent manner. These results indicate that GTSC EtOAC fraction inhibit oxidative stress and inflammatory responses through elevated GSH levels, antioxidative enzymes expressions and suppression of iNOS expression via NF-${\kappa}B$ down-regulation.

• Journal of Life Science
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• v.25 no.7
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• pp.733-739
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• 2015

Microalgae are a renewable natural resource that requires only sunlight, carbon dioxide, phosphorus, and nitrogen for rapid growth. They produce a broad variety of basic chemical substances―such as vitamins, fatty acids and carotenoids―that have high added value potential for the pharmaceutical and food industries. The aim of this study was to develop axenic culture and to establish a cell growth assay for microalgae. A further experiment was carried out to determine the yield of astaxanthin derived from microalgae. The axenic culture was developed using a mixture of antibiotics [ampicillin (100 ${\mu}g/ml$), streptomycin (10 ${\mu}g/ml$), chloramphenicol (10 ${\mu}g/ml$), penicillin (10 ${\mu}g/ml$), neomycin (50 ${\mu}g/ml$), gentamycin (50 ${\mu}g/ml$), kanamycin (10 ${\mu}g/ml$), and nystatin (1.5 ${\mu}g/ml$)] and then used to extract a variety of useful components from the microalgae. The optimal concentration for the antibiotic mixture was 1-3 percent. A spectrophotometric cell growth assay was also established. Astaxanthin was extracted from Haematococus lacustris with a yield of $1.9{\times}10^{-3}{\mu}g/l$ per 1 ml of culture medium. In conclusion, the axenic culture method developed here allows extraction of high-quality astaxanthin and other useful components from microalgae.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.1
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• pp.163-170
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• 2006

Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.7
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• pp.960-965
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• 2010

To develop Corni Fructus as a cancer preventive food material, the in vitro cytotoxicities and in vivo antitumor activities of various concentrations of 80% Corni Fructus ethanol extract (CFEE) were investigated using sarcoma-180 cancer cell. Viability was decreased and cell death rate was increased in both dose- and time-dependent manners in cells treated with CFEE at 10, 100, 300, and $500\;{\mu}g/mL$ concentrations for 24, 48, and 72 hr. Proliferation was also inhibited more than 60% in cells treated with CFEE at the $100\;{\mu}g/mL$ concentration for 48 hr. In addition, the morphology of cells treated with CFEE at the 100 and $500\;{\mu}g/mL$ concentrations was distorted with shrunken cell masses and lower cell numbers compared to the control cells. In the cells treated with CFEE, the formation of apoptotic bodies and nuclear condensation were observed in dose dependent manners. CFEE also increased DNA fragmentation values at the 100 and $500\;{\mu}g/mL$ concentrations. The apoptosis induced by CFEE was connected to the proteolytic activation of caspase-3. When CFEE was administered at 100 and 300 mg/kg, ip, for 7 consecutive days in mice inoculated with sarcoma-180 cancer cell, the life span of the mice was found to be longer than that of the control mice that did not receive the extract. These results suggest that Corni Fructus may be used as a potential cancer preventive food material.

• Applied Microscopy
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• v.30 no.3
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• pp.265-272
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• 2000

In this study, the brachial ganglion of Octopus minor was investigated with light microscope and electron microscope,andthefollowingresultswereobtained. The brachial ganglions of the octopus, round in shapes , are located under each of suckers. Their sizes are proportional to those of the suckers. A brachial ganglion of round shape consists of cortex and medulla. In cortex, nerve cells exist collectively while neuropiles in medulla. Three kinds of nerve cells (large, middle, and small neurons) are found in the cluster of nerve cells. The small one is a round cell of about $0.9{\mu}m$ in diameter while the middle and large ones are an elliptical cell of $1.6\times1.3{\mu}m$ and an ovoid cell of $2.8{\mu}m$ in diameter, respectively. All of those cells look light due to their low electron densities , in which cell organelle are not well developed. It was also observed that the middle neurons are surrounded by median electron-dense neuroglial cells of pyramidal shapes and about $0.6\times0.4{\mu}m$ in sizes. In the neuropiles of medulla, dendrites and axons of various sizes make a complex net. They contain four kinds of chemical synaptic vesicles-electron-dense synaptic vesicle of 100 nm in diameter, median electron-dense synaptic vesicle of 90 nm in diameter, electron-dense cored synaptic vesicle of 90 nm in diameter, and electron-lucent synaptic vesicle of 50 nm in diameter.

• The Journal of Advanced Prosthodontics
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• v.6 no.5
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• pp.406-414
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• 2014

PURPOSE. The objective of this study was to investigate the biologic effects of enamel matrix derivative (EMD) with different concentrations on cell viability and the genetic expression of human gingival fibroblasts (HGF) to zirconia surfaces. MATERIALS AND METHODS. Immortalized human gingival fibroblasts (HGF) were cultured (1) without EMD, (2) with EMD $25{\mu}g/mL$, and (3) with EMD $100{\mu}g/mL$ on zirconia discs. MTT assay was performed to evaluate the cell proliferation activity and SEM was carried out to examine the cellular morphology and attachment. The mRNA expression of collagen type I, osteopontin, fibronectin, and TGF-${\beta}1$ was evaluated with the real-time polymerase chain reaction (RT-PCR). RESULTS. From MTT assay, HGF showed more proliferation in EMD $25{\mu}g/mL$ group than control and EMD $100{\mu}g/mL$ group (P<.05). HGFs showed more flattened cellular morphology on the experimental groups than on the control group after 4h culture and more cellular attachments were observed on EMD $25{\mu}g/mL$ group and EMD $100{\mu}g/mL$ group after 24h culture. After 48h of culture, cellular attachment was similar in all groups. The mRNA expression of type I collagen increased in a concentration dependent manner. The genetic expression of osteopontin, fibronectin, and TGF-${\beta}1$ was increased at EMD $100{\mu}g/mL$. However, the mRNA expression of proteins associated with cellular attachment was decreased at EMD $25{\mu}g/mL$. CONCLUSION. Through this short term culture of HGF on zirconium discs, we conclude that EMD affects the proliferation, attachment, and cell morphology of HGF cells. Also, EMD stimulates production of extracellular matrix collagen, osteopontin, and TGF-${\beta}1$ in high concentration levels. CLINICAL RELEVANCE. With the use of EMD, protective barrier between attached gingiva and transmucosal zirconia abutment may be enhanced leading to final esthetic results with implants.

• Journal of Life Science
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• v.24 no.4
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• pp.337-342
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• 2014

Resveratrol (RSV), a natural polyphenolic compound, is a modulator for cell division and cell migration, and has diverse beneficial properties. Angiogenin (ANG) and vascular endothelial growth factor (VEGF) are considered to be important mechanisms for cell proliferation, angiogenesis, the formation of tubular structures, and migration. In this study, we investigated whether RSV has a migratory effect in HeLa cells. When cells were treated with $0{\sim}50{\mu}M$ of RSV for 24 hr, the expression of ANG and VEGF was significantly increased in a dose dependent manner measured by real-time PCR. Similarly, we performed time dependent experiments for $50{\mu}M$ RSV treated cells and identified the optimized time at 24 hr. The increased expression in RSV treated cells was confirmed by Western blot analysis. To examine the toxic effects of RSV at the determined conditions, MTT assays were performed. The viabilities were unchanged for $0{\sim}50{\mu}M$ RSV treated cells, while they decreased at $100{\mu}M$ RSV. To examine the effect of migration in RSV treated cells, we performed a wound-healing assay. The migratory rates were significantly enhanced in the RSV treated group. In this study, we found that RSV induces an increase in the expression of migration factors ANG, VEGF, and enhances cell migration for the determined conditions.

• Applied Microscopy
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• v.12 no.2
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• pp.55-68
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• 1982

The study on the nerve cells in the pars intercerebralis(IP) of 5-day-old cabbage butterfly Pieris rapae L. was performed to observe their ultrastructures and classify them on the basis. of the differences in size, shape and relative distribution cf cell organelles. The brain-subesophageal ganglion complex was fixed in 1% paraformaldehyde-1% gluaraldehyde mixture and embedded in araldite mixture. The transverse thin sections of IP were stained with uranyl acetate and lead citrate and examined by Hitachi 500 and ]EM 100B electron microscope. Five distinct types. of nerve cells are recognized and are arbitrarily designated as Type I, Type II Type III, Type IV and Type V. Type I neurone: These neurones are neurosecretory cells. Several neurosecretory cells are. recognized in the pars intercerebralis. They are roughly round or peach-shaped cells measuring $13{\sim}25{\mu}m$ in diameter. The rounded nucleus shows about $5{\sim}10{\mu}m$ in diameter. The chromatin is predominantly diffused with only occasional dense patches. The perikaryon contains numerous. mitochondria, free polyribosomes and neurosecretory granules. The neurosecretory granules are relatively uniform in electron density, and each one is about $100{\sim}400{\mu}m$ in diameter and surrounded by a single membrane. The granules are also observed mostly as in groups. In one group of neurones the cisternae of endoplasmic reticulum are distended or in other group of neurones are not distended. Golgi saccules are slightly dilated at their lateral extremities and contains. frequenty dense rounded materials. Type II neurone: Thes have the largest soma in the pars intercerebralis about $30{\sim}35{\mu}m$ in diameter. They also show roughly polygonal in shape. The nucleus is elongated or sickle-shaped. The chromatin is mainly in the euchromatin form. The perikarya in these cells are well populated with populated with free ribosomes and contain numerous mitochondria and Golgi bodies. The cisternae of granular endoplasmic reticulum are also well distributed. Type III neurone: They are oval or spindle-shaped and also medium-sized. neurones approximately $15{\sim}17{\mu}m$ in length. The nucleus is oval or slightly elongated in shape and $8{\sim}9{\mu}m$ in length. The chromatin occurs in diffused form. The cytoplasm contains many filamentous or oval mitochondria. The perikaryon has also numerous free polyribosomes and cisternae of granular endoplasmic reticulum. Type VI neurone: They are roughly polygonal in shape probably due to the close approximation of the adjacent cells. The soma is about $7{\sim}8{\mu}m$ in diameter. The nucleus is round or oval in shape and $5.0{\sim}5.8{\mu}m$ in diameter. The necleus also occupies a large proprion of the cell body. The perikaryon is well populated with free ribosomes and contains several mitochondria and cistenae of granular endoplasmic reticulum. Type V neurone: These neurones are similar to Type VI neurones in various respects such as cell size and cell inclusion, but they differ from Type IV neurones in shape. The soma is oval or slightly elongated. The cell body contains several filamentous and oval mitochondria.

• Korean Journal of Food Science and Technology
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• v.41 no.1
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• pp.87-92
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• 2009

The antioxidant properties and protective effects of Inula britannica on ${H_2}{O_2}$-induced SH-SY5Y neuroblastoma cell damage were investigated. A series of solvent fractions, including hexane(Fr.H), petroleum ether, chloroform, ethyl acetate(Fr.EA), and water fraction(Fr.W), were prepared from the 70% methanol extracts of Inula britannica. Fr.W had the highest total contents of phenolics and flavonoids, followed by Fr.EA. The antioxidant properties of the fractions were also evaluated by analyzing their scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH) radicals, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, and nitric oxide. Fr.W showed the strongest activities in all assays. The concentrations of Fr.W that resulted in 50% reductions of the DPPH and ABTS radicals were 20.7 ${\mu}g$/mL and 39.4 ${\mu}g$/mL, respectively. Fr.W showed the weakest cytotoxic activities on the SH-SY5Y cells, whereas it effectively protected ${H_2}{O_2}$-induced cell death, increasing cell survival by 35.0-77.0% at a concentration range of 62.5-250 ${\mu}g$/mL. In this range, Fr.W also significantly decreased intracellular ROS levels by 34-39%. Overall, the antioxidant properties of Inula britannica can contribute to rescuring neuronal cells from oxidative stress-induced cell injury.