• 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.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.428-436
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• 2016

The emergence of antibiotic-resistant bacteria has been increased and become a public health concern worldwide. Many bacterial infections can be sequentially treated with different types of antibiotics. Thus, this study was designed to evaluate the changes in survival, antibiotic susceptibility, mutant frequency, ${\beta}$-lactamase activity, biofilm formation, and gene expression in Klebsiella pneumoniae after exposure to sequential antibiotic treatments of ciprofloxacin and meropenem. Treatments include control (CON; no addition), 1/2 MIC ciprofloxacin addition (1/2CIP), 2 MIC ciprofloxacin addition (2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC ciprofloxacin (16 h-incubation) (1/2CIP-1/2MER-2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC meropenem (16 h-incubation) (1/2CIP-1/2MER-2MER), and initial 1/2 MIC ciprofloxacin addition followed by 2 MIC ciprofloxacin(8 h-incubation) and 2 MIC meropenem(16 h-incubation) (1/2CIP-2CIP-2MER). No growth of K. pneumoniae was observed for the 2CIP throughout the incubation period. The numbers of planktonic cells varied with the treatments (7~10 log CFU/ml), while those of biofilm cells were not significantly different among treatments after 24-h incubation, showing approximately 7 log CFU/ml. Among the sequential treatments, the least mutant frequency was observed at the 1/2CIP-1/2MER-2CIP (14%). Compared to the CON, 1/2CIP-2CIP-2MER decreased the sensitivity of K. pneumoniae to piperacillin, cefotaxime, and nalidixic acid. The highest ${\beta}$-lactamase activity was 22 nmol/min/ml for 1/2CIP-1/2MER-2CIP, while the least ${\beta}$-lactamase activity was 6 nmol/min/ml for 1/2CIP-2CIP-2MER. The relative expression levels of multidrug efflux pump-related genes (acrA, acrB, and ramA) were increased more than 2-fold in K. pneumoniae exposed to 1/2CIP-1/2MER-2MER and 1/2CIP-2CIP-2MER. The results suggest that the sequential antibiotic treatments could change the antibiotic resistance profiles in K. pneumoniae.

• 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.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.147-154
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• 1997

국내에서 많이 사용되고 있는 제2세대 quinolone 항생제인 norfloxacin(NFX)에 대한 약역학적인 특성을 구명하기 위하여 국내에서 분리된 동물유래 병원성 세균에 대하여 시험관내에서 실험을 수행하여 다음과 같은 결과를 얻었다. 즉, E coli(n=89) 대한 NFX의 $MIC_{50}$와 $MIC_{90}$는 공히 0.02g/ml이었으며, Streptococcus spp.(n=36)에 대한 NFX의 $MIC_{50}$는 2g/ml 그리고 $MIC_{90}$는 4g/ml로 나타났다. Salmonella spp.(n=56)에 대한 NFX의 $MIC_{50}$와 $MIC_{90}$ 모두 0.2g/ml로 강한 항균력을 보였으며, Streptococcus spp.(n=24)에 대한 NFX의 $MIC_{50}$는 2g/ml 그리고 $MIC_{90}$가 4g/ml로 나타났다. Bacillus spp.(n=34)는 NFX의 $MIC_{50}$와 $MIC_{90}$는 모두 0.4g/ml으로서 대부분의 병원성 세균에 대해서 $MIC_{50}$와 $MIC_{90}$치가 동일하든지 또는 매우 비슷한 수치를 보여주었다. 그러나 NFX는 혐기성세균인 Clostridium spp.(n=34)에 대해서는 항균력이 매우 낮았다. 현재 수의임상에서 항균제 병용요법이 많이 응용되고 있는 것을 고려하여 NFX와 다른 항생물질간의 분획억제농도 (FICs)를 E coli 88ac을 시험균주로 하여 실험한 결과, NFX와 colistin과 병용할 때 FIC 값이 0.38로서 상승작용을 그리고 gentamicin, trimethoprim, amikacin, penicillin 및 tylosin과의 병용시 FIC 값이 각각 0.52, 0.56, 0.63, 1.00 및 1.02로서 상가작용을 보여주었으며, tetracyclin과의 병용시의 FIC값은 1.49로서 길항작용을 나타냄을 알 수 있었다. 한편 실제 항균제의 임상적용시 매우 주요한 요소인 항균활성후 저농도유효성(PAE)을 알아보기 위하여 E coli AB1157을 시험균주로 측정한 결과 PAE은 0.90~1.02 시간 그리고 S aureus R-209에 대해서는 PAE가 1.58~1.99 시간으로서 그람음성균 및 그람양성균 모두에 대해서 긴 PAE를 갖고 있음을 알 수 있었다.

• Biomedical Science Letters
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• v.22 no.4
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• pp.174-183
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• 2016

In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.2
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• pp.1-12
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• 2011

Objectives: The object of this study was to observe the in vitro antibacterial effects of Chungdae-tang aqueous extracts, traditionally used for treating various gynecological diseases including vaginitis in Korea against Gardnerella vaginalis, and combination effects of Chungdae-tang extracts with Clindamycin were also monitored in this study. Methods: Antibacterial activities against Gardnerella vaginalis of Chungdae-tang aqueous extracts were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at MIC and MIC${\times}$2 levels. The combination effects of Chungdae-tang aqueous extracts with Clindamycin were observed by Checkerboard microtiter assay, and the effects of bacterial growth curve treated with or Chungdae-tang aqueous extracts MIC+Clindamycin MIC, 1/2MIC and 1/4MIC, respectively. In the present study, Gardnerella vaginalis were incubated under $37^{\circ}C$, 10% $CO_2$; and bacterial growth curves were calculated at 24, 48, 72, 96 and 120hrs after incubations. Results: MIC of Chungdae-tang aqueous extracts against Gardnerella vaginalis were detected as $3.906{\pm}2.344$(0.782~6.250) mg/$m\ell$, respectively. MIC of Clindamycin was detected as $0.010{\pm}0.006$(0.004~0.016) ${\mu}g/m\ell$ at same conditions. In addition, Clindamycin and Chungdae-tang aqueous extracts also showed marked dosage-dependent inhibition of bacterial growth, and more dramatical inhibitions were detected in Clindamycin+Chungdae-tang aqueous extracts MIC treatment as compared with each of single Clindamycin MIC and Chungdae-tang aqueous extracts MIC treatments, respectively. In addition, quite similar inhibitory effects on bacterial growth were detected in Clindamycin 1/4 MIC+Chungdae-tang aqueous extracts MIC treatment as compared with single Clindamycin MIC treatment in the present study. FIC index in combination of Chungdae-tang and Clindamycin were detected as $0.775{\pm}0.285$ (0.500~1.250) at Checkerboard microtiter assay. Conclusions: The results obtained in this study suggest that Chungdae-tang aqueous extracts showed antibacterial effects against Gardnerella vaginalis, and it also showed dosage-dependent inhibitory effects on the bacterial growth. In addition, combination treatment of Chungdae-tang aqueous extract with Clindamycin showed more potent inhibitory effects on the growth of Gardnerella vaginalis with FIC index $0.775{\pm}0.285$(0.500~1.250), respectively. It means, the combination of Chungdae-tang aqueous extract with Clindamycin is partially synergistic effects. It, therefore, is expected that effective dosages of Clindamycin will be reduced to 1/4 or over 1/4 levels as combination with Chungdae-tang extracts, respectively.

• BMB Reports
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• v.43 no.2
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• pp.91-96
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• 2010

The function of macrophage inhibitory cytokine-1 (MIC-1) in cancer remains controversial, and its signaling pathways remain poorly understood. In this study, we demonstrate that MIC-1 induces the transactivation of EGFR, ErbB2, and ErbB3 through the activation of c-Src in SK-BR-3 breast cells. MIC-1 induced significant phosphorylation of EGFR at Tyr845, ErbB2 at Tyr877, and ErbB3 at Tyr1289 as well as Akt and p38, Erk1/2, and JNK mitogen-activated protein kinases (MAPKs). Treatment of SK-BR-3 cells with MIC-1 increased the phosphorylation level of Src at Tyr416, and induced invasiveness of those cells. Inhibition of c-Src activity resulted in the complete abolition of MIC-1-induced phosphorylation of the EGFR, ErbB2, and ErbB3, as well as invasiveness and matrix metalloproteinase (MMP)-9 expression in SK-BR-3 cells. Collectively, these results show that MIC-1 may participate in the malignant progression of certain cancer cells through the activation of c-Src, which in turn may transactivate ErbB-family receptors.

• International Journal of Oral Biology
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• v.39 no.2
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• pp.115-120
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• 2014

Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of a microorganism. It has been reported that sub-MIC of antibiotics may result in morphological alterations along with biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of periodontal pathogens after treatment with sub-MIC antibiotics. Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis were used in this study. The MIC for amoxicillin, doxycycline, metronidazole, penicillin, and tetracycline were determined by broth dilution method. The bacterial morphology was observed with bright field microscope after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans and F. nucleatum were increased after incubation with metronidazole; penicillin and amoxicillin. P. gingivalis were increased after incubating with metronidazole and penicillin. However, F. nucleatum showed decreased length after incubation with doxycycline and tetracycline. In this study, we observed that sub-MIC antibiotics can affect the morphology of periodontal pathogens.

• Journal of Pharmaceutical Investigation
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• v.8 no.1
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• pp.1-5
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• 1978

Drug interaction of a new antibiotic, methampicillin lysinate (MAL) with nine drugs were investigated using four species of gram positive and gram negative bacteria. The experimental results were as follows: 1. MIC of MAL were found to be decreased against E. coil when combined with mefenamic acid, probenecid, aluminium hydroxide gel or corticosteroids. The other drugs did not affect MIC of MAL against the same bacteria. 2. MIC of MAL were found to be increased against Staphylococcus aureus ATCC 6538-P, 9441 when combined with mefenamic acid, aluminum hydroxide gel or dexamethasone acetate. The other drugs did not affect MIC of MAL against the same bacteria. 3. MIC of MAL were found to be increased against Shigella dysenteriae when either of the nine drugs was combined. 4. MIC of MAL were found to be increased approximately 2.5 times when combined with Streptokinase-Streptodornase or hydrocortisone and to be decreased approximately 2 times when combined with probenecid or dexamethasone against Salmonella typhi(type 2). It seems the other drugs do not affect the MIC of MAL against the same bacteria.

• Mycobiology
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• v.31 no.2
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• pp.81-85
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• 2003

Antifungal properties of methyl 2-benzimidazole carbamate(BMC) were investigated using 16 fungi. Cytotoxicity test of BMC revealed that the morphology of HeLa cells was considerably deformed even at the concentrations as low as 0.1 ppm. Minimum inhibitory concentration(MIC) values of BMC for 7 fungi among the 16 tested ones were lower than $1.95{\times}10^{-4}{\mu}g/ml$, while Aspergillus flavus showed an MIC value higher than 1.0 ${\mu}g/ml$. Tolerance induction against BMC was successful only for Paecilomyces farinosus LAR10, contrary to the expectation that tolerance would be induced for the fungi having high MIC values such as Aspergillus niger ATCC 9642 and A. flavus ATCC 9643. Spore germination of A. niger ATCC 9642 was suppressed by BMC. However the mycelial growth of the fungus once germinated was not retarded at all by BMC up to 8 MIC. Addition of lanosterol provided a remedy for the reduced germination rate of A. niger ATCC 9642 spores.