• International Journal of Oral Biology
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• v.34 no.1
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• pp.7-14
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• 2009

Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.2 s.51
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• pp.147-152
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• 2005

As a part of searching tot the natural components which inhibit the skin aging and wrinkle formation, the $80\%$ methanolie extracts of 121 species of traditional herbal medicines used to treat lung and skin disease were investigated for their in vitro anti-oxidative activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals, and inhibitory activity against elastase. We selected 9 kinds of the traditional herbal medicines showing inhibitory activities of winkle formation. The effective concentrations of 9 candidates for anti-wrinke/skin firming activity was less than 0.1 mg/mL, and there is no toxicity to cell viability at these concentrations. Through analysis of human skin primary patch test data, the traditional herbal medicines represented non-irritant materials. We suggest that these 9 candidates with ability to help anti-wrinkle/skin firming may be useful for functional cosmetic materials.

• Food Science and Preservation
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• v.17 no.1
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• pp.58-65
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• 2010

We investigated the effect of Chun ma (Gastrodiae rhizoma) concentrated extract on the quality of mixed beverages. Chun ma beverages prepared using different concentrated extracts were divided into four groups: GCE 5 ($5^{\circ}$ Brix concentrated extracts) GCE 10 ($10^{\circ}$ Brix concentrated extracts) GCE 15 ($15^{\circ}$ Brix concentrated extracts) and GCE 20 ($20^{\circ}$ Brix concentrated extracts). The pH values ranged from a low of 4.37 in GCE 5 to a high of 4.68 in GCE 20. Soluble solid levels in GCE 20 ($19.6^{\circ}$ Brix) were higher than in the other samples. The b (yellowness) scores and the total phenolic contents of all samples increased with increasing extract concentration. The highest total phenolic contents were seen in GCE 20 samples at 232.23 mg%. Samples did not differ markedly in antiradical activity (75.07-76.00% DPPH inhibition). Free sugar levels in GCE 20 samples and organic acid concentrations of GCE 15 samples were higher than those of other preparations. Free amino acid and mineral contents of all samples increased with increasing extract concentration. The levels of free amino acids were in the order Glu > Gly > Ser > Arg > Hylys, and the Glu content was 249.15 ug/100 g for GCE-20 samples and 61 ug/100 g in GCE-5 products. The mineral contents of all samples were in the order K > Na > Mg > Ca. Higher scores for color, flavor, and overall acceptability were found in GCE 5 products compared with other extracts. These results indicate that Chun ma beverage can be prepared in various ways, as commercially desired, with reference to the above characteristics of Chun ma materials.

• Fisheries and Aquatic Sciences
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• v.20 no.7
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• pp.12.1-12.7
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• 2017

Soft tunic syndrome (STS) is a protozoal disease caused by Azumiobodo hoyamushi in the edible ascidian Halocynthia roretzi. Previous studies have proven that combined formalin-hydrogen peroxide ($H_2O_2$) bath is effective in reducing STS progress and mortality. To secure target animal safety for field applications, toxicity of the treatment needs to be evaluated. Healthy ascidians were bathed for 1 week, 1 h a day at various bathing concentrations. Bathing with 5- and 10-fold optimum concentration caused 100% mortality of ascidians, whereas mortality by 0.5- to 2.0-fold solutions was not different from that of control. Of the oxidative damage parameters, MDA levels did not change after 0.5- and 1.0-fold bathing. However, free radical scavenging ability and reducing power were significantly decreased even with the lower-than-optimal 0.5-fold concentration. Glycogen content tended to increase with 1-fold bathing without statistical significance. All changes induced by the 2-fold bathing were completely or partially restored to control levels 48 h post-bathing. Free amino acid analysis revealed a concentration-dependent decline in aspartic acid and cysteine levels. In contrast, alanine and valine levels increased after the 2-fold bath treatment. These data indicate that the currently established effective disinfectant regimen against the parasitic pathogen is generally safe, and the biochemical changes observed are transient, lasting approximately 48 h at most. Low levels of formalin and $H_2O_2$ were detectable 1 h post-bathing; however, the compounds were completely undetectable after 48 h of bathing. Formalin-$H_2O_2$ bathing is effective against STS; however, reasonable care is required in the treatment to avoid unwanted toxicity. Drug residues do not present a concern for consumer safety.

• Food Science and Preservation
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• v.19 no.2
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• pp.263-270
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• 2012

The chemical components and anti-oxidant activities of black currant were investigated. The pH, soluble solid and total acidity values were 3.36, 15.11 $^{\circ}Brix$, and 1.65%, respectively. The Hunter L, a, and b values were 18.20, 5.13, and 1.08, respectively. The proximate compositions were as follows; moisture, 77.64%; nitrogen free extract, 17.41%; crude fiber, 3.08%; crude protein, 1.28%; crude ash, 0.31%; and crude lipid, 0.28%, respectively. The mineral elements were K (177.36 mg/100 g), P (54.74 mg/100 g), and Ca (26.45 mg/100 g). The free sugar components were glucose (7.71%) and fructose (5.88%). The amino acid contents of the black currant were very rich in glutamic acid (105.73 mg/100 g) and deficient in cystine (5.29 mg/100 g). The ascorbic acid and total phenolic contents were 112.19 mg/100 g and 34.48 mg GAE/g, respectively. The ABTS and DPPH radical scavenging activity levels were 99.48% and 89.03% at the 10 and 1.25 mg/mL concentrations. The reducing power and FRAP of the black currant were dose-dependent. Thus, black currant can be an effective source of functional food substances, i.e., natural anti-oxidants.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.5
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• pp.670-674
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• 2013

The free radical theory of aging suggests that oxidative damage caused by free radicals plays a key role in normal aging. We measured the anti-oxidant activity of acorns and asked whether it can modulate the aging process in Caenorhabditis elegans. Different concentrations of acorn powder were added to culture medium, followed by the monitoring of fertility and survival under oxidative stress. The anti-oxidant activity of 500 mg/L of acorn powder exhibited significant increases in the resistance to oxidative stress in vivo. Acorn powder also significantly extended both the mean and maximum lifespan of C. elegans (the mean lifespan was increased up to 22.4%). The fertility assay indicates the lifespan extension from acorn does not accompany a reduced reproduction, which is common in long-lived mutants. These findings indicate that acorn has a strong antioxidant activity and can induce longevity without the trade-off of reduced reproduction in C. elegans.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.47-55
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• 2011

The solvent extracts of Aruncus dioicus var. kamtschaticus Hara, which were extracted by using several solvents with different polarities, were performed to investigate the antioxidant activities, whitening effect and antimicrobial activity. The content of total polyphenol of fractions from Aruncus dioicus var. kamtschaticus Hara extract showed the highest value ($335.88{\pm}2.26$ mg/g GAE) on ethyl acetate fraction. The ethyl acetate and n-butanol fractions were 0.06 mg/mL and 0.25 mg/mL as $IC_{50}$ values on DPPH radical scavenging, and $99.16{\pm}0.09%$ and $89.29{\pm}0.64%$ on ABTS radical scavenging activity, respectively. Also, reducing power and FRAP value were significantly higher on ethyl acetate fraction. The SOD like activity showed $80.76{\pm}0.61%$ on ethyl acetate and $72.34{\pm}0.79%$ on n-butanol. Tyrosinase inhibition activities (at 5 mg/mL) were $59.08{\pm}0.98%$ on ethyl acetate fraction. The chloroform fraction showed the strongest antimicrobial activities against B. cereus (14 mm), B. subtilis (12.5 mm), S. aureus (10.8 mm), E. coli (20.7 mm) at 0.1 mg/disc and the inhibition zone diameter of ethyl acetate fraction was 17.2 mm against E. coli at 0.5 mg/disc. The minimum inhibitory concentrations (MIC) of chloroform fraction against B. cereus and E. coli were 50 and $25{\mu}g$/mL, respectively. From these results, it is suggested that ethyl acetate and n-butanol fractions of Aruncus dioicus var. kamtschaticus Hara could be used as functional material for food additive ingredient and chloroform fraction could be suitable for the development of a food preservative.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.4
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• pp.361-373
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• 2015

This study was carried out to evaluate the antioxidant and antibacterial activities of Glycyrriza uralensis Fisher (Jecheon, Korea) extracts obtained by various extraction conditions (85% ethanol, heating temperatures and times), and to establish the optimal extraction condition of G. uralensis for the application as cosmetic ingredients. The extracts obtained under different conditions were concentrated and made in the powdered (sample-1) and were the crude extract solutions without concentration (sample-2). The antioxidant effects were determined by free radical scavenging activity ($FSC_{50}$), ROS scavenging activity ($OSC_{50}$), and cellular protective effects. Antibacterial activity was determined by minimum inhibitory concentration (MIC) on human skin flora. DPPH free radical scavenging activity of sample-1 ($100{\mu}g/mL$) was 10% higher in group extracted for 6 h than 12 h, but sample-2 didn't show any significant differences. The extraction yield extracted with same temperature for 12 h was 2.6 times higher than 6 h, but total flavonoid content was 1.1 times higher. These results indicated that total flavonoid content hardly increased with increasing extraction time. Free radical scavenging activity, ROS scavenging activity and cellular protective effects were not dependent on the yield of extraction, but total flavonoid content of extraction. Antibacterial activity on three skin flora (S. aureus, B. subtilis, P. acnes)of sample-1 in different extraction conditions were evaluated on same concentration, and the group extracted at 25 and $40^{\circ}C$ showed 16 times higher than methyl paraben ($2,500{\mu}g/mL$). In conclusion, 85% ethanol extracts of G. uralensis extracted at $40^{\circ}C$ for 6 h showed the highest antioxidant and antibacterial activity. These results indicate that the extraction condition is important to be optimized by comprehensive evaluation of extraction yield with various conditions, yield of active component, and activity test with concentrations, and activity of 100% extract, for manufacturing process of products.

• Korean Journal of Medicinal Crop Science
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• v.15 no.4
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• pp.296-303
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• 2007

Acer tegmentosum (Acereaceae) has been used a source of traditional medicines for the treatment of hepatic disorders in Korea. This research was conducted to determine biofunctional activities of A. tegmentosum stem extract and to identify its bioactive components. Methanolic extract from A. tegmentosum stem was partitioned by using organic solvents, including n-hexane, ethyl acetate, n-butanol, and water. Two compounds were isolated by using an ODS column chromatography from ethyl acetate soluble fraction shown to the strongest antioxidant activity ($RC_{50}=3.15\;{\mu}g/m{\ell}$) among the fractions. The isolated compounds were analyzed by $^1H$ and $^{13}C$ NMR, IR, UV/VIS, MS spectrum data and identified as catechin, ${\rho}-Hydroxyphenethyl$ alcohol $1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$. The compounds have shown strong antioxidant activity, with similar activity to BHA ($RC_{50}=2\;{\mu}g/m{\ell}$). Especially, ${\rho}-Hydroxyphenethyl$ alcohol 1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$ was shown strong anti-lipid peroxidative activity. However, the compounds were not shown antimicrobial activities. In antimicrobial activity assays, ethyl acetate soluble fraction was effective to bacterial inhibition, such as Escherichia coli and Klebsiella pneumonia, with minimum inhibitory concentrations in $125\;{\mu}g/m{\ell}$. Otherwise, antifungal activity against Candida albicans was shown in n-hexane soluble fraction exhibiting $63\;{\mu}g/m{\ell}$ of minimum inhibitory concentration. In anticomplementary activity assays, water soluble fraction was the most effective exhibiting 24% inhibitory activity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.2
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• pp.227-233
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• 2004

Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.