• Journal of Life Science
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• v.28 no.9
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• pp.1068-1075
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• 2018

Kaempferia parviflora, an herbaceous plant in the family Zingiberaceae, is popular in many tropical regions. It is called as black ginger or krachaidum in Thailand and Laos, and its raw or dried root have been used as spices and teas. The rhizomes also have been traditionally used to treat gastrointestinal disorders, ulcers, gout, dysentery, allergies and to improve physical work capacity. Recently, its anti-obesity, anti-oxidant, anti-inflammatory and blood clot-lysis activities were reported. In this study, the anti-thrombosis activity of black ginger was investigated, since improvement in blood fluidity leads to the prevention of various lifestyle-related diseases. The hot water and ethanol extract and their subsequent solvent fractions (hexane, ethylacetate, butanol fractions and water residue) were prepared, and their anti-coagulation and platelet aggregation inhibitory activities were determined, respectively. Among the black ginger extracts and their fractions, the ethylacetate fraction (EAF) of ethanol extract only showed significant extensions of blood coagulation time determined by thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (aPTT). At 5 mg/ml concentration, TT, PT and aPTT were extended to 1.22, 1.49 and >15-folds compared to non-treatment. The EAFs of ethanol and hot water extract showed strong inhibitions against collagen-induced platelet aggregations, which are comparable to inhibitions of aspirin. Also the EAFs from black ginger did not show any hemolysis activity against human RBC up to 0.5 mg/ml. Our results suggest that the EAF of black ginger has a potential as novel anti-coagulation and ant-platelet aggregation agent. This report provides the first evidence of anti-coagulation activity of black ginger.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.195-200
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• 2008

Saxifraga stolonifera (Saxifragaceae) is a perennial herbaceous plant growing in Korea, China, Japan and Russia. The aerial part has been used as herbal medicine for treatment of pneumonia, frostbite, inflammation and microbial infection. In this study, fresh juice and methanol extract were prepared from the aerial part of S. stolonifera, and their antimicrobial, antithrombin, and antioxidant activity were evaluated, respectively. The fresh juice showed weak antimicrobial activity against Proteus vulgaris, Escherichia coli O157:H7 and Candida albicans with ignorable DPPH scavenging activity. But, the methanol extract showed strong antioxidant activity ($IC_{50}$ of $37.5{\mu}g/mL$) with minor, broad-range antimicrobial activity. Antithrombin activities were not observed in fresh juice and extract, up to 1.5 mg/mL. Sequential organic solvent fractionation of methanol extract showed that $IC_{50}s$ of ethylacetate and the butanol fraction were 6.9 and $7.8{\mu}g/mL$, respectively, that is comparable with vitamin C or butylated hydroxytoluene. Analysis of component in extract and fractionates suggested that the antioxidants in fractions are diverse and the active substances have glycosylated phenolic structure. Our results suggest that the aerial part of S. stolonifera could be used as the natural source of potential antioxidant.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.661-676
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• 2000

Gingivitis and periodontitis are infectious diseases in that microorganisms are the primary extrinsic cause of the diseases. the occurrence of gingivitis has been associated clearly with the presence of microorganisms at the disease site, and the histologic nature of the tissue involved is indicative of an inflammatory response induced by microorganisms. additional evidence for the microbial etiology of periodontal disease is that numerous antimicrobial agents are effective in reducing plaque accumulation and periodontal diseases. the purpose of this article is to analyze the antimicrobial effects of Pulsatilla koreana. Well-dried Pulsatilla koreana purchased from herbs distributor was ground and extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH). we have then applied each solution to the bacteria samples(Bacteroides forsythus, Streptococcus mutans, Streptococcus sanguis, Porphylomonas gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, Prevotella intermedia, Actinomyces viscosus, Prevotella nigrescens , Rothia dentocariosa, Fusobacterium nucleatum, Pseudomonas aeruginosa, Staphylococcus aureus) collected from several organizations. To conduct susceptibility test(Kirby-Bauer method), plate contained each periodontopathic bacteria is spread extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH) and to measure the minimum inhibition concentration(MIC) of the bacteria against the solutions to ultimately determine antimicrobial effects of the solutions, insert bacteria sample into $20{\mu\ell}/{m\ell}$, $10{\mu\ell}/{m\ell}$, $5{\mu\ell}/{m\ell}$, $2.5{\mu\ell}/{m\ell}$ of each solution and control group(not contained solution) 1. Solution extracted into methanol did not show clear zone against all bacteria samples. Only P.nigrescens, S. mutans and S. sanguis in solution extracted into ethylacetate, S. mutans and S. anguis in solutions extracted into chlorform and Butyl alcohol showed clear zone against all bacteria samples. Solution extracted into Butyl alcohol showed clear zone against 13 types of bacteria, excluding P. gingivalis. 2. In Solution extracted into methanol, the bacteria samples grew in the highest concentrated plate, showing minimal variation from control group. 3. In Solution extracted into Butyl alcohol, S. aureus, P. intermedia, E. corrodens, A. actinomycetemcomitans, B. forsythus, P. gingivalis et al. showed decreased growth in the highest concentrated plate. P. auruginosa, R. dentocariosa, A. viscosus, P. nigrescens, S. mutans et al. showed decreased growth at MIC $20{\mu\ell}/{m\ell}$ and S. sanguis showed decreased growth at MIC $10{\mu\ell}/{m\ell}$. 4. By analyzing the MIC level through considering the results from Kirby-Bauer method, Solution extracted into methanol did not reveal any antimicrobial effects and Solution extracted into Butyl alcohol showed the highest antimicrobial effects In conclusion, it can be used the extracts of Pulsatilla koreana as wide spectrum antimicrobial agent.

• Journal of Life Science
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• v.24 no.5
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• pp.515-521
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• 2014

The oriental traditional medicine, Aruncus dioicus var kamtschaticus (ADK) is used for hemostasis (blood stopping) and the promotion of blood circulation. Recently, the demands of the aerial part of ADK as edible mountain herbs are rapidly increased due to its unique fragrance and bioactivity. In this study, to evaluate the anti-thrombosis activity of ADK, ethanol extract and organic solvent fractions were prepared from aerial parts of ADK, and their anticoagulation and anti-platelet aggregation activities were determined. In an anticoagulation activity assay, the ethanol extract of ADK increased the thrombin time, prothrombin time, and activated partial thromboplastin time (aPTT) 1.4-2.3 times at a concentration of 5 mg/ml. Among the fractions, the ethylacetate fraction showed strong inhibitory effects against blood clotting factors, as shown in an extension of the aPTT. In contrast, the butanol fraction strongly promoted blood clotting. In an anti-platelet aggregation assay, the activity of the ethanol extract was comparable to that of aspirin, a commercial anti-platelet aggregation agent, and the butanol fraction showed 2-fold higher aggregation inhibitory activity than aspirin. The aforementioned ethanol extract and active fractions have ignorable hemolytic activity against human red blood cells up to a concentration of 0.5 mg/ml. Considering the high content of total polyphenol, total flavonoid, and total sugar of the ethylacetate and butanol fractions, the purified active substances have potential as safe and novel anti-thrombosis agents. This report provides the first evidence of anti-thrombosis activity of ADK.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.144-151
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• 2012

In the course of this study aimed at the development of functional food ingredients from seaweeds, the in vitro antimicrobial activities of methanol extracts prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta) were determined against food-borne diseases and pathogenic microorganisms including multi-drug resistant (MDR) Pseudomonas sp. and Candida sp. Based on disc-diffusion assays at 500 g/disc concentration of the methanol extracts, Ishige okamurai, I. foliacea, Sargassum confusum, and S. yamade exhibited strong antibacterial activities in a broad-spectrum, except against Pseudomonas aeruginosa. In addition to the latter four seaweeds, Ecklonia stolonifera, E. cava and Eisenia bicyclis also demonstrated antifungal activity against C. albicans. Among these 8 selected seaweeds, I. okamurai, I. foliacea, and S. yamade exhibited strong hemolytic activity (55-93%) at 500 g/ml against human RBC. Organic solvent sequential fractions using hexane, ethylacetate and butanol, and water residues were prepared from the 8 selected seaweeds and their anti-Candida sp. activities were further determined. The ethylacetate and butanol fraction of I. okamurai, and the hexane fraction of I. foliacea demonstrated antifungal activity against MDR-pathogenic Candida sp. Although the solvent fractions had no activity against MDR-Pseudomonas sp., our results suggest that seaweeds, especially Ishige okamurai, I. foliacea, S. confusum, and S. yamade could be developed as broad-spectrum antimicrobial ingredients.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.168-174
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• 2011

In order to develop safe and economic novel antifungal agents, we prepared 73 methanol extracts from medicinal and edible herbs and examined their 365 solvent fractions using n-hexane, methylene chloride, ethylacetate, butanol and water residue based on the sequential organic solvent fraction method. When using the various fractions in the screening step for antifungal activity, we discovered ethylacetate fraction of Morus alba L., methylene chloride fraction of Pimpinella brachycarpa (MCPB), and n-hexane fraction of Salvia miltiorrhiza Bunge, which all have activities in methanol extracts, as potential sources of antifungal agents. Amongst these, the antifungal activity of P. brachycarpa has not to date been reported on. In addition, the mycelial growth inhibition and spore germination inhibition activities of MCPB against A. niger were confirmed by disc-diffusion assay in a 10 day culture. The MIC and MFC of MCPB were determined as 0.25 and 0.5 mg/ml, respectively. The MCPB has no hemolytic activity against human RBC at 0.5 mg/ml and glycoside-flavonoids are theorized to be active constituents. These results suggest that MCPB has a prominent antifungal activity and that the application of sequential organic solvent fractions, instead of simple natural product extracts, is useful in the screening process of novel bioactive substances.

• Food Science and Preservation
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• v.19 no.1
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• pp.110-115
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• 2012

$Epimedium$ $koreanum$ Nakai is a wild medicinal plant commonly consumed in South Korea due to its beneficial health effects. In this study, the antimutagenic and immunological activities of $E.$ $koreanum$ Nakai extracts were investigated for their use in food. In the immunomodulating activity, the effects of $E.$ $koreanum$ Nakai on the B cell (Rhamos) and T cell (Molt-4) were investigated. The results showed that the growth and viability of the B and T cells were increased and activated more in the ethylacetate (1.35 and 1.48 times) and water fraction (1.30 and 1.40 times), respectively. In the Ames test, none of the fractions produced a mutagenic effect on $Salmonella$. $typhimurium$ TA98 and TA 100. The ethylacetate fraction showed a strong antimutagenic effect (98%) on and a high butanol fraction (84%) of B(${\alpha}$)P in $S.$ $typhimurium$ TA98 and TA100, respectively. In 4-nitroquinoline-1-oxide (4NQO), all the solvent fractions showed an over 70% antimutagenic effect, except for the chloroform extract. Especially, ethylacetate and butanol showed strong inhibition of the mutagenic effects (80 and 90%) on 4NQO in $S.$ $typhimurium$ TA98 and TA100, respectively. These results provide preliminary data for the development of $E.$ $koreanum$ Nakai as an edible food material.

• Journal of Korea Foresty Energy
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• v.20 no.2
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• pp.9-19
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• 2001

This study was carried out to investigate natural insectifuge materials from tree extractives in order to substitute for organic synthetic insecticides for food wrapping corrugated board. Tree samples were collected, extracted, fractionated with hexane, $CH_2Cl_2$, ethylacetate(EtOAc) and $H_2O$, and then freezed dried for further study. EtOAc or $H_2O$ fractions were chromatographed on a Sephadex LH-20 column for isolation and purification, and the isolated compounds were characterized by spectroscopic tools such as NMR and MS. Crude extractives of EtOAc and $H_2O$ fractions were added to the printing ink for corrugated board with the concentration of 2% or 3% based on the weight of the ink, then the prepared ink was printed on the corrugated board to be used for evasion test using larva of indian meal moth(Plodia interpunctella(Hubner)). Robtin, dihydrorobinetin and leucorobinetinidin were isolated from the wood extractives of black locust(Robinia pseudoacacia) and the bark of poplar(Populus alba $\times$ glandulosa) contained many kinds of compounds such as (+)-catechin, naringenin, aromadendrin, eriodictyol, sakuranetin and its glycoside, taxifolin, neosaturanin, salireposide, p-coumaric acid and aesculin. Much of (+)-catechin was isolated from the bark extractives of willow(Salix koreensis) in addition to (+)-gallocatechin and p-coumaric acid and the bark of weeping willow(Salix babylonica) also contained (+)-catechin, (+)-gallocatechin, dihydromyricetin and myricetin.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1391-1398
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• 2005

The anti-thrombic properties of the oriental herbal medicine Daejowhan(DJW, 大造丸) which consists of 11 kinds of herbs (indicated as ratio) of Rehmanniae Radix 24%, Hominis Placenta 5%, Testudinis Carapax 9%, Eucommiae Cortex 9%, Asparagi Radix 9%, Phellodendri Cortex 9%, Achyranthis Radix 7%, Liriopis Tuber 7%, Angelicae Sinensis Radix 7%, Ginseng Radix 5% and Schizandrae Fructus 3% were investigated. The water extracts from DJW inhibited Platelet-activating factor(PAF) induced platelet aggregation. DJW was extracted with methanol and further fractionated by ethylacetate. A 70% methanol extract showed a strong inhibition against PAF-induced aggregation in vitro and in vivo assays. The ethylacetate soluble fraction was shown to have inhibitory effect on PAF-induced platelet aggregation in vitro assay. The ethylacetate soluble fraction specially protected against the lethality of PAF, while verapamil did not afford any protection. These results indicate that the water extracts and alcoholic-fractions inhibit the action of PAF in vivo by an antagonistic effect on PAF, so that it may be useful in treating disorders caused by PAF, such as acute allergy, inflammation, asthma, gastrointestinal ulceration, toxic shock and so forth. DJW was investigated regarding its assumed anti-thrombic action on human platelets which was deduced from its ability to suppress Arachidonic acid(AA)-induced aggregation, exocytosis of ATP, and inhibition of Cyclooxygenase(COX) and Thromboxane synthase(TXS) activity. The latter two effects were estimated from the generation of Prostaglandin $E_2(PGE_2)$ and Thromboxane $A_2(TXA_2)$ respectively. Exogenously applied AA ($100{\mu}mol/{\ell}$) provoked a $89\%$ aggregation of platelets, the release of 14 pmol ATP, and the formation of either 225 pg $TXA_2$ or 45 pg $PGE_2$, each parameter being related to 106 platelets. An application of DJW 5 min before AA dose-dependently diminished aggregation, ATP-release and the synthesis of $TXA_2$ and $PGE_2$ with $IC_{50}$ values of 74, 108, 65, $72{\mu}g/m{\ell}$, respectively. The similarity of the $IC_{50}$ values suggest an inhibition of COX by DJW as primary target, thus suppressing the generation of $TXA_2$ which induces aggregation of platelets and exocytosis of ATP by its binding on $TXA_2$-receptors.

• Korean Journal of Food Science and Technology
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• v.30 no.1
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• pp.230-236
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• 1998

We have screened total 32 herbal drugs to find the highest activity against human cariogenic enzyme, glucosyltransferase (GTase) from the extracts of Magnoliae bark. The extracts were separated into three phases, i. e. water, n-butanol and ethylacetate according to their solvent polarity. Among them, ethylacetate fraction had approximately more than 70% of total activities, and the active principle was further isolated by prep. HPLC following silicagel column chromatography to yield single compound as white powder. The chemical structure of the compound was finally elucidated to be 4,4'-dihydroxy-3,3'-dimethoxylignan from the spectral data of FAB-MS. $^1H-\;and\;^{13}C-NMR$ spectrometries. The compound was also shown to have relatively strong antibacterial activity against ten types of cariogenic oral bacteria and one kind of Actinomyces sp.