• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.2
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• pp.293-298
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• 1995

Antimicrobial effect of tea extracts from green tea(steamed, roasted), oolong tea and black tea was investigated. Minimal inhibitory concentration(MIC) of tea extraxcts against 9 well known strains of foodborne pathogenic bacteria such as the Gram positive and Gram negative bacteria was determined at 37$^{\circ}C$. Significant antimicrobial activity was observed in the steamed green tea and the roasted green tea of the water-soluble fraciton, and the steamed green tea of the methanol-soluble fraction, and the steamed green tea, roasted green tea and the oolong tea of the crude catechin fraction. The MIC of these extracts against B. subtillis were 700$\mu\textrm{g}$/ml, 500$\mu\textrm{g}$/ml and 120 $\mu\textrm{g}$/ml, respectively. The crude catechin fraction possessed greater antimicrobial activity than did the other fractions. Among tea extracts, extracts of steamed green tea, roasted green tea and oolong tea showed higher antimicrobial activity than them of black tea. The MIC of the crude catechin fraction obtained from tea extracts against Gram-positive bacteria such as M. Iuteus, B. subtillis and S. mutans were 30~50$\mu\textrm{g}$/ml, 120~240$\mu\textrm{g}$/ml and 120~180$\mu\textrm{g}$/ml, and against Gram-negative bacteri such as e. aerogenes and V. parahaemolyticus were 50~60$\mu\textrm{g}$/ml and 60~70$\mu\textrm{g}$/ml in the broth medium, respectively. Especially, the MIC to Streptococcus mutans which has known as a causative bacteria of a decayed tooth were 120$\mu\textrm{g}$/ml, 140$\mu\textrm{g}$/ml, 180$\mu\textrm{g}$/ml and above 1,000$\mu\textrm{g}$/ml in steamed green tea, roasted green tea, oolong tea and black tea, respectively. Tea extracts had strong growth inhibition activity against foodborne pathogenic and dental bacteria.

• Parasites, Hosts and Diseases
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• v.53 no.2
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• pp.147-154
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• 2015

Various constituents in clinical specimens, particularly feces, can inhibit the PCR assay and lead to false-negative results. To ensure that negative results of a diagnostic PCR assay are true, it should be properly monitored by an inhibition control. In this study, a cloning vector harboring a modified target DNA sequence (${\approx}375bp$) was constructed to be used as a competitive internal amplification control (IAC) for a conventional PCR assay that detects ${\approx}550bp$ of the Cryptosporidium oocyst wall protein (COWP) gene sequence in human feces. Modification of the native PCR target was carried out using a new approach comprising inverse PCR and restriction digestion techniques. IAC was included in the assay, with the estimated optimum concentration of 1 fg per reaction, as duplex PCR. When applied on fecal samples spiked with variable oocysts counts, ${\approx}2$ oocysts were theoretically enough for detection. When applied on 25 Cryptosporidium-positive fecal samples of various infection intensities, both targets were clearly detected with minimal competition noticed in 2-3 samples. Importantly, both the analytical and the diagnostic sensitivities of the PCR assay were not altered with integration of IAC into the reactions. When tried on 180 randomly collected fecal samples, 159 were Cryptosporidium-negatives. Although the native target DNA was absent, the IAC amplicon was obviously detected on gel of all the Cryptosporidium-negative samples. These results imply that running of the diagnostic PCR, inspired with the previously developed DNA extraction protocol and the constructed IAC, represents a useful tool for Cryptosporidium detection in human feces.

• Korean journal of food and cookery science
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• v.25 no.4
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• pp.496-505
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• 2009

The influence of roasting time on antibacterial and antioxidative effects of methanol and water coffee extracts was investigated. Extract yield differed with roasting time. The maximum yield of methanol extract was 20.02% and 24.00% at respective roasting times of 12 and 20 min. The maximum yield of water extracts was 2.70% and 18.58% at 5 and 25 min roasting time, respectively. Antibacterial effects of each extract were determined by the classical minimal inhibitory concentration (MIC) paper disc diffusion method. Methanol extracts of different coffee samples inhibited growth of various strains except Escherichia coli. Extracts obtained following roasting times of 12, 14, 16, 20, and 25 min in particular displayed the most potent activity against Staphylococcus aureus. Among these extracts, that obtained from 12 min roasted coffee samples produced a MIC of $16.125{\mu}g$/mL against S. aureus. Water extracts applied at $1,000{\mu}g$/mL were growth inhibitory except against Salmonella choleraesuis and Prevotella intermedia. However, growth inhibition by water extracts was weak, with inhibitory zones of only 6-8 mm diameter produced. Determinations of free radical elimination for the different coffee extracts using 1,1-diphenyl-2-picrylhydrazyl were compared with ascorbic acid and butylated hydroxytoluene positive controls. Methanol and water extracts of different coffee samples ($100{\mu}g$/mL) showed $67.1{\sim}92.3%$ and $66.4{\sim}93.3%$ radical scavenging activity, respectively. However, longer roasting time (especially >20 min) tended to somewhat lower free radical elimination using both extracts. Total phenol in different coffee samples measured by the Folin-Denis method revealed the highest level of phenol contents with non-roasted coffee, whereas phenol content differed with different roasting time, ranging from $87.{\sim}126.5\;mg/g$ in methanol extracts. In water extracts, the phenol content was maximum at 8 min roasting time, whereas in other samples the content was varied from $95.0{\sim}199.1\;mg/g$.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.450-455
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• 1997

The preparation of Bacillus coagulans is used as a therapeutics for human intestinal disorders. However, the bacterium in the preparation is very susceptible to rifampic in and fluoroquinolones. When the preparation is taken with rifampicin or fluoroquinolones, its therapeutic effect can not be expected. So B. coagulans RFR17 resistant to rifampicin was obtained by treating the parent B. coagulans with N-methyl-N'-nitro-N-nitrosoguanidine. B. coagulans OFR17 was produced by serial passage of B. coagulans RFR17 on agar with 2-fold minimal inhibitory concentration of ofloxacin or ciprofloxacin. B. coagulans OFR17 was resistant to fluoroquinolones up to 16~64 fold higher than that for the original strain. B. coagulans OFR17 also exhibited identical characteristics with the parent strain when they were tested for lactic acid production and growth inhibition of E. coli MB4-01 and Shigella sonnei MB4-10411. From in vitro test, it was also identified that rifampicin and ofloxacin are not inactivated by certain factors of B. coagulans OFR17. Conclusively, B. coagulans OFR17 can be regarded as a promising strain which can be developed as the preparation for the treatment of the intestinal disorders of the tuberculosis patients under rifampicin and ofloxacin therapy.

• Journal of Korean Medicine Rehabilitation
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• v.31 no.1
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• pp.47-57
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• 2021

Objectives This study was conducted to confirm the possibility of Clostridium difficile infection (CDI) treatment through natural herbal medicines. Methods After screening a total of 77 herbal medicines through the paper disc agar diffusion method, we selected the herbal medicines that showed a effectiveness compared to the positive control vancomycin. Afterwards, drugs that showed inhibitory effects compared to C. difficile without inhibition of Bifidobacterium bifidum and Lactobacillus plantarum, known as beneficial bacteria, were selected and minimal inhibitory concentration (MIC) was confirmed by applying the Broth microdilution method. Results The Coptidis Rhizoma, well known for its antimicrobial effect, was found to have antimicrobial effects on C. difficile, but also had inhibitory effects on the beneficial bacterium B. bifidum. 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus had antimicrobial effects on C. difficile without inhibiting the beneficial bacteria B. bifidum and L. plantarum. The MIC values of 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus were found to be 10 mg/mL, 20 mg/mL and 5 mg/mL, respectively. Conclusions Crataegi fructus, Corni fructus and Mume fructus were identified as candidate medicines for C. difficile. Further researchs will need to be done in vivo, and to find an optimal extraction method accompanied by economic evaluation.

• Journal of Society of Preventive Korean Medicine
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• v.4 no.2
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• pp.235-241
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• 2000

This study was carried out to evaluate cytotoxic effects of Poncirus trifoliata Raf. extract on lymphocytic leukemia tumor (L1210) cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay The comparison of Ic50 Values of Poncirus trifoliata Raf. extract in L1210 cell lines showed that their susceptibility to these fractons decreased in the following order: adriamycin > Fr.4> Fr. 6> Fr. 5> Fr. 3> Fr. 1> Fr. 2 by the MTT assay. In order to develop an antumicrobial agent, Poncirus trifoliata Raf. was extracted wit ethanol, and then it was fractionated with several mobile phase. The antitumor activities of fractions of the ethanol soluble extract was investigated. The minimal inhibitory concentrations (MIC) of fractions of the ethanol soluble extract of Poncirus trifoliata Raf. against microorganisms were also examined. Antimicrobial activities of ampicillin and ketoconazole as references were compared to those of fractions of the ethanol soluble extract of Poncirus trifoliata Raf. The antimicrobial activities of all fractions from the extract had growth inhibition activities against gram-positive bacteria, gram-negative bacteria and fungi $(MIC\;>\;200{\mu}g/ml)$. These results suggest that fraction 4 of the ethanol soluble extract of Poncirus trifoliata Raf. possessed the most antitumorous agent.

• Journal of Food Hygiene and Safety
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• v.35 no.4
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• pp.382-390
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• 2020

This study was carried out to evaluate the antimicrobial effect of red ginseng (Panax ginseng C.A. Meyer) against several foodborne pathogens including Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus niger. The antimicrobial effect was determined by agar diffusion method using red ginseng extract, crude saponin and non-water-soluble fractions. Red ginseng extract showed antimicrobial effect against S. aureus, but not C. albicans or A. niger. The extract showed anti-bacterial activity at concentration above 30% against S. aureus, which cause both food poisoning and atophic dermatitis. Crude saponin showed antibacterial activity above 7.5% against the bacterium. However, the ginsenosides purified from crude saponin showed no antimicrobial activities at 100-200 ㎍/mL. To investigate the mode of growth inhibition, red ginseng extract and crude saponin were added to 0.85% NaCl solution containing S. aureus and then incubated at 35℃ for 12 h. The results showed that viable cells were rapidly reduced in above 10% concentration of red ginseng extract and above 2% of crude saponin, respectively. However, the crude saponin and red ginseng extract did not inhibit the bacterial cells completely at those same concentrations. On the other hand, whereas all non-water-soluble fractions showed inhibition zones above 10 mm against S. aureus, they showed no inhibition effects against E. coli, C. albicans or A. niger. The methanol fraction-1 (MF-1) showed the highest antibacterial activity against S. aureus, and the MIC (minimal inhibitory concentration) was 0.625 mg/mL. These results suggest that red ginseng extract, crude saponin and non-water-soluble fractions show selective antibacterial activity against S. aureus, and non-water-soluble fractions might be used as natural antibacterial agents.

• Journal of Yeungnam Medical Science
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• v.9 no.2
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• pp.396-406
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• 1992

Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.1
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• pp.65-80
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• 2014

Objectives: The object of this study was to observe the in vitro antibacterial effects of Gagam-seopyoungjeon aqueous extracts (GGSYJ) against Gardnerella vaginalis and the possible synergic combination effects with clindamycin. Methods: Antibacterial activities against Gardnerella vaginalis of GGSYJ were detected using minimal inhibition concentration (MIC), and the effects on the bacterial growth curve were also monitored at MIC and MIC${\times}$2 levels. The combination effects of GGSYJ with clindamycin were observed by checkboard microtiter assay, and the effects of bacterial growth curve treated with GGSYJ MIC+clindamycin MIC, 1/2 MIC and 1/4 MIC, respectively. The effects on the bacterial invasion and intracellular killing of GGSYJ were also observed using human vaginal epithelial (VK2) and murine macrophage (Raw264.7) cells with combination effects with clindamycin after treatment of GGSYJ MIC+clindamycin 1/2 MIC, 1/4 MIC and 1/6 MIC, respectively. Results: The MIC of clindamycin and GGSYJ against Gardnerella vaginalis were detected as $0.012{\pm}0.006$ (0.004~0.016)${\mu}g/ml$ and $1.016{\pm}0.524$ (0.391~1.563) mg/ml, respectively. Clindamycin and GGSYJ were also showed marked dosage-dependent inhibition of bacterial growth, and significant decreases of viable cells were detected in clindamycin MIC+GGSYJ MIC and clindamycin 1/2 MIC+GGSYJ MIC treatment as compared with each of single clindamycin MIC and GGSYJ MIC treatments. And significant decreases of intraepithelial and intra-macrophage viable bacteria numbers were detected in clindamycin 1/2 MIC+GGSYJ 1/2 MIC and clindamycin 1/4 MIC+GGSYJ 1/2 MIC treatment as compared with each of single clindamycin GGSYJ 1/2 MIC treatments, respectively. Conclusions: GGSYJ showed slight antibacterial effects against Gardnerella vaginalis, but they showed dosage-dependent inhibitory effects on the bacterial growth and VK2 epithelial invasions of bacteria with favorable accelerating effects of intracellular killing activities of macrophages. In addition, combination of GGSYJ also increased the inhibitory effects of clindamycin on the epithelial invasions of Gardnerella vaginalis and intracellular killing activities of macrophages against Gardnerella vaginalis as 2-fold higher as compared with clindamycin single treatment, respectively. Therefore, we expected that the clinical dosages of clindamycin can be reduced as 1/2 levels as combination with GGSYJ.

• Journal of Life Science
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• v.20 no.2
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• pp.269-274
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• 2010

An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.