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        In Vitro Real-Time Interactions of Cranberry Constituents with Immobilized Fructosyltransferase

        Mark Feldman,Ervin I. Weiss,Itzhak Ofek,Moshe Shemesh,Doron Steinberg 한국식품영양과학회 2010 Journal of medicinal food Vol.13 No.5

        Cranberry has been proposed as an anti-biofilm agent that does not kill bacteria, but rather prevents the pathogen from survival in the host. This can be achieved by inhibiting the function of virulent factors essential for the pathogen to persist in a host environment. The oral bacterial enzyme fructosyltransferase (FTF) plays an important role in the pathogenesis of dental diseases. The real-time interaction of cranberry nondialyzable material (NDM) with immobilized FTF was investigated using the surface plasmon resonance (SPR) technique. To determine its binding efficiency, NDM at concentrations between 0μg/mL and 200μg/mL was applied onto the immobilized FTF. The effect of NDM or other polyphenols, myricetin, and epicatechin on FTF enzymatic activity was evaluated by applying the above compounds and sucrose onto immobilized FTF. Salivary amylase was applied with NDM onto immobilized FTF to explore the effect on NDM–FTF interaction. Our results show that NDM firmly attaches to immobilized FTF in a dose-dependent manner, whereas the presence of salivary amylase reduced this binding interaction. Using nonlinear regression we calculated that the affinity constant of NDM applied alone (106 M−1) was fivefold higher than NDM in the presence of amylase (0.2×106 M−1). At 200μg/mL, NDM, introduced together with sucrose, inhibited the activity of immobilized FTF by 63% within minutes, in comparison with the control (sucrose alone). The effect of NDM was sustained even after it was washed off the immobilized FTF. Myricetin also strongly inhibited FTF activity, whereas epicatechin was less effective. The real-time SPR observation suggests that one of the anti-biofilm modes of action of NDM is an immediate and irreversible inhibitory effect on the activity of immobilized FTF, which is due to a strong binding affinity to the immobilized enzyme

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        Garlic Allicin as a Potential Agent for Controlling Oral Pathogens

        Gilad Bachrach,Areen Jamil,Ronit Naor,Golan Tal,Zvi Ludmer,Doron Steinberg 한국식품영양과학회 2011 Journal of medicinal food Vol.14 No.11

        Garlic has been used medicinally throughout human history. Allicin is considered the most therapeutic constituent of garlic. This study tested the antimicrobial activity of garlic allicin on oral pathogens associated with dental caries and periodontitis. Allicin was found effective against all the tested bacteria. The broth dilution method revealed that planktonic growth of the cariogenic, gram-positive species Streptococcus mutans, S. sobrinus, and Actinomyces oris was inhibited by an allicin concentration of 600 μg/mL or higher. Planktonic growth of the tested gram-negative periopathogenic species Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum was inhibited by a minimum allicin concentration of 300 μg/mL. Porphyromonas gingivalis, an anaerobic, gram-negative pathogen and the bacterium most associated with chronic periodontitis, demonstrated the lowest sensitivity to allicin (2,400 μg/mL). Gel zymography and the synthetic chromogenic substrate N_α-benzoyl-L-arginine 4-nitroanilide hydrochloride demonstrated that allicin inhibits the proteases of P. gingivalis, including the arginine and lysine gingipains known as major virulence factors of this organism. A gingipain-inactivated mutant demonstrated high sensitivity to allicin (<300 μg/mL), revealing that gingipains confer resistance to allicin. Live/dead staining followed by analysis with confocal laser scanning microscopy revealed that allicin was bactericidal to S. mutans grown in mature biofilms. However, this bactericidal effect was reduced as biofilm depth increased. In conclusion, these results support the traditional medicinal use of garlic and suggest the use of allicin for alleviating dental diseases.

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