• Journal of Ginseng Research
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• v.41 no.2
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• pp.180-187
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• 2017

Background: The incidence of halitosis has a prevalence of 22-50% throughout the world and is generally caused by anaerobic oral microorganisms, such as Fusobacterium nucleatum, Clostridium perfringens, and Porphyromonas gingivalis. Previous investigations on the structure-activity relationships of ginsenosides have led to contrasting results. Particularly, the antibacterial activity of less polar ginsenosides against halitosis-related bacteria has not been reported. Methods: Crude saponins extracted from the Panax quinquefolius leaf-stem (AGS) were treated at $130^{\circ}C$ for 3 h to obtain heat-transformed saponins (HTS). Five ginsenoside-enriched fractions (HTS-1, HTS-2, HTS-3, HTS-4, and HTS-5) and less polar ginsenosides were separated by HP-20 resin absorption and HPLC, and the antimicrobial activity and mechanism were investigated. Results: HPLC with diode-array detection analysis revealed that heat treatment induced an extensive conversion of polar ginsenosides (-Rg1/Re, -Rc, -Rb2, and -Rd) to less polar compounds (-Rg2, -Rg3, -Rg6, -F4, -Rg5, and -Rk1). The antimicrobial assays showed that HTS, HTS-3, and HTS-4 were effective at inhibiting the growth of F. nucleatum, C. perfringens, and P. gingivalis. Ginsenosides-Rg5 showed the best antimicrobial activity against the three bacteria, with the lowest values of minimum inhibitory concentration and minimum bactericidal concentration. One major reason for this result is that less polar ginsenosides can more easily damage membrane integrity. Conclusion: The results indicated that the less polar ginsenoside-enriched fraction from heat transformation can be used as an antibacterial agent to control halitosis.

• Journal of Veterinary Science
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• v.22 no.2
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• pp.18.1-18.12
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• 2021

Background: We previously elucidated the protective mechanism of Korean red ginseng oil (RGO) against Brucella abortus infection, and our phytochemical analysis revealed that palmitic acid (PA) was an abundant component of RGO. Consequently, we investigated the contribution of PA against B. abortus. Objectives: We aimed to investigate the efficacy of PA against B. abortus infection using a murine cell line and a murine model. Methods: Cell viability, bactericidal, internalization, and intracellular replication, western blot, nitric oxide (NO), and superoxide (O₂^-) analyses and flow cytometry were performed to determine the effects of PA on the progression of B. abortus infection in macrophages. Flow cytometry for cytokine analysis of serum samples and bacterial counts from the spleens were performed to determine the effect of PA in a mouse model. Results: PA did not affect the growth of B. abortus. PA treatment in macrophages did not change B. abortus uptake but it did attenuate the intracellular survivability of B. abortus. Incubation of cells with PA resulted in a modest increase in sirtuin 1 (SIRT1) expression. Compared to control cells, reduced nitrite accumulation, augmented O₂^-, and enhanced pro-inflammatory cytokine production were observed in PA-treated B. abortus-infected cells. Mice orally treated with PA displayed a decreased serum interleukin-10 level and enhanced bacterial resistance. Conclusions: Our results suggest that PA participates in the control of B. abortus within murine macrophages, and the in vivo study results confirm its efficacy against the infection. However, further investigations are encouraged to completely characterize the mechanisms involved in the inhibition of B. abortus infection by fatty acids.

• Journal of Pharmacopuncture
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• v.22 no.1
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• pp.49-54
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• 2019

Objective: Antibiotic resistance is a global health problem and threatens health of societies. These problems have led to a search for alternative approaches such as combination therapy. The aim of the present study was to investigate the effect of caffeine and omeprazole in combination with gentamicin or ciprofloxacin against standard and clinically resistant isolates of Staphylococcus aureus and Escherichia coli. Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of different agents against bacterial strains were determined. The interaction of non- antibiotic drugs with gentamicin and ciprofloxacin was studied in vitro using a checkerboard method and calculating fraction inhibitory concentration index (FICI). Verapamil as efflux pump inhibitor was used to evaluate the possible mechanism of bacterial resistance to antibiotics. Results: The MIC and MBC values of gentamicin against bacterial strains were in the range of $20-80{\mu}g/ml$ and $40-200{\mu}g/ml$, respectively. Caffeine and omeprazole had no intrinsic inhibitory activity against tested microorganisms. However, upon combination of caffeine with antibiotics, the synergistic effects were observed. Verapamil was able to reduce the MIC values of gentamicin (4 folds) only in some bacterial strains. Conclusion: These findings indicated that caffeine was effective in removing bacterial infection caused by S. aureus and E. coli. The relevant mechanisms of antibiotic resistance were not related to the drug efflux.

• Journal of Oral Medicine and Pain
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• v.32 no.2
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• pp.137-150
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• 2007

Trees emit phytoncide into atmosphere to protect them from predation. Phytoncide from different trees has its own unique fragrance that is referred to as forest bath. Phytoncide, which is essential oil of trees, has microbicidal, insecticidal, acaricidal, and deodorizing effect. The present study was performed to examine the effect of phytoncide on Porphyromonas gingivalis, which is one of the most important causative agents of periodontitis and halitosis. P. gingivalis 2561 was incubated with or without phytoncide extracted from Hinoki (Chamaecyparis obtusa Sieb. et Zucc.; Japanese cypress) and then changes were observed in its cell viability, antibiotic sensitivity, morphology, and biochemical/molecular biological pattern. The results were as follows: 1. The phytoncide appeared to have a strong antibacterial effect on P. gingivalis. MIC of phytoncide for the bacterium was determined to be 0.008%. The antibacterial effect was attributed to bactericidal activity against P. gingivalis. It almost completely suppressed the bacterial cell viability (>99.9%) at the concentration of 0.01%, which is the MBC for the bacterium. 2. The phytoncide failed to enhance the bacterial susceptibility to ampicillin, cefotaxime, penicillin, and tetracycline but did increase the susceptibility to amoxicillin. 3. Numbers of electron dense granules, ghost cell, and vesicles increased with increasing concentration of the phytoncide, 4. RT-PCR analysis revealed that expression of superoxide dismutase was increased in the bacterium incubated with the phytoncide. 5. No distinct difference in protein profile between the bacterium incubated with or without the phytoncide was observed as determined by SDS-PAGE and immunoblot. Overall results suggest that the phytoncide is a strong antibacterial agent that has a bactericidal action against P. gingivalis. The phytoncide does not seem to affect much the profile of the major outer membrane proteins but interferes with antioxidant activity of the bacterium. Along with this, yet unknown mechanism may cause changes in cell morphology and eventually cell death.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.2
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• pp.83-88
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• 2004

The purpose of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Large unilamellar vesicles (OPGTL) were prepared with total lipids extracted from cultured Porphyromonas gingivalis outer membranes (OPG). The anthroyloxy probes were located at a graded series of depths inside a membrane, depending on its substitution position (n) in the aliphatic chain. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine effects of chlorhexidine digluconate on differential rotational mobility, while changing the probes' substitution position (n) in the membrane phospholipids aliphatic chain. Magnitude of the rotational mobility of the intact six membrane components differed depending on the substitution position in the descending order of 16-(9-anthroyloxy)palmitic acid (16-AP), 12, 9, 6, 3 and 2-(9-anthroyloxy)stearic acid (12-AS, 9-AS, 6-AS, 3-AS and 2-AS). Chlorhexidine digluconate increased in a dose-dependent manner the rate of rotational mobility of hydrocarbon interior of the OPGTL prepared with total lipids extracted from cultured OPG, but decreased the mobility of membrane interface of the OPGTL. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

• Journal of Veterinary Clinics
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• v.28 no.6
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• pp.576-580
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• 2011

Silver has potent antibacterial activity against a variety of bacteria while maintaining low toxicity in mammalian cells. This study was conducted to investigate the possible mechanism underlying the bactericidal effects of silver ions against bovine mastitis pathogens using electron microscopy. We used two different bacterial strains, Escherichia coli and Staphylococcus aureus, which are primarily responsible for the majority of bovine mastitis cases. Interaction between the bacteria and silver ions (50 ${\mu}g/mL$, 2 hours) were studied using energy-filtering transmission electron microscopy (EFTEM). EFTEM images showed that E. coli and S. aureus cells treated with the silver ions had distorted plasma membranes, silver ions attached to the outer membranes, scattered electron-light material, and leakage of cell contents from disrupted cell membranes.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.1
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• pp.80-91
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• 2003

Mutans streptococci, especially S. mutans and S. sobrinus strongly implicated in pathogenesis of dental caries, the major cause of tooth loss in children. Use of an antibacterial agent controlling dental caries has been rationalized. The present study was performed to observe the antibacterial effect of inorganic polyphosphates (polyP) on S. mutans and S. sobrinus. S. mutans GS5 and S. sobrinus 6715 were grown in brain-heart infusion broth with or without polyP. Minimal inhibitory concentration (MIC) of polyP for S. mutans GS5 was determined to be 0.08% and that for S. sobrius 6715 was 0.17%. PolyP 15 added to the growing culture of S. mutans GS5 and S. sobrinus 6715 at their exponential phase was as effective in inhibiting the growth of S. mutans GS5 and S. sobrinus 6715 as polyP added at the very beginning of the culture. More than 85% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. mutans GS5 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. And more than 99.9% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. sobrinus 6715 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. Intracellular nucleotide release from S. mutans CS5 and S. sobrinus 6715 was increased in the presence of polyP 15 for 5h but was not really reversed by the addition of divalent cations like $Ca^{++}\;and\;Mg^{++}$. The majority of the cells appeared to be atypical in their shape, demonstrating accumulation of highly electron-dense granules and ghost cells. The overall results suggest that polyP have a strong bactericidal activity against S. mutans and S. sobrinus in which lysis in relation to chelation may not play the major role but unknown mechanism that possibly affects the viability of the bacterium may be involved. PolyP may be used as an agent for prevention of dental caries.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.723-727
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• 2007

Resistance to metronidazole in Helicobacter pylori results from inactivation of rdxA and frxA, the chromosomal genes for a nitroreductase that normally converts metronidazole from prodrug to bactericidal agent. Two types of metronidazole susceptible strains had been found distinguishable by their apparent levels of frxA expression. Most common in the populations we had studied were strains that required only rdxA inactivation to become resistant to moderate levels of metronidazole(type I strains). The second strain type required inactivation of both frxA and rdxA to become resistance to metronidazole(type II strains): this was linked to a relatively high level of frxA gene transcription in the type II strains. The fdxA gene regulated fdxA as well as rdxA gene. Thus, to study the function of fdxA as a regulatory gene we constructed a null mutant of fdxA in H. pylori genome and identified over-and under-expressed proteins by fdxA using two-dimensional(2-D) electrophoresis and MALDI-TOP-MS. There were four over-expressed proteins in fdxA mutant; nifU-like protein(HP0221), frxA(HP0642), nonheme ferritin(HP0653), and hypothetical protein(HP0902). Three under-expressed proteins were also identified in fdxA mutant, including 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (HP0089), (3R)-hydroxymyristoyl ACP dehydratase(HP1376), and thioredoxin(HP1458).

• Journal of Life Science
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• v.33 no.2
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• pp.158-168
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• 2023

Natural products have been used to mitigate the effects of cancer and infectious diseases, as they feature diverse bioactivities, such as antioxidant, antibacterial, anti-inflammatory, and immunomodulatory effects. Here, we chose 10 natural products that are well-known as pulmonary enhancers and investigated their bactericidal effects on Streptococcus pneumoniae. In the disk diffusion assay, the growth of S. pneumoniae was significantly regulated by G. fructus treatment regardless of extraction method used. We first adopted spraying as a novel delivery method for G. fructus. Interestingly, mice exposed to G. fructus three times a day for 2 weeks were resistant to S. pneumoniae intranasal infection (shown both through body weight loss and survival rates compared to the control group). Moreover, we confirmed that exposure to G. fructus regulated the colonization of the bacteria despite the sustained inflammation in the lung after exposure to S. pneumoniae, indicating that migrated inflammatory immune cells may involve a host defense mechanism against pulmonary infectious diseases. While a similar number of granulocytes (CD11b⁺Ly6C⁺Ly6G⁺), neutrophils (CD11b⁺Ly6C^intLy6G⁺), and monocytes (CD11b⁺Ly6C^intLy6G^-) were found between groups, a significantly increased number of alveolar macrophages (CD11b⁺CD11c^hiF4/80⁺) was detected in BAL fluids of mice pre-exposed to G. fructus at 5 days after S. pneumonia infection. Taken together, our data suggest that this usage of G. fructus can induce protective immunity against bacterial infection, indicating that facial spray may be helpful in enhancing the defense mechanism against pulmonary inflammation and in evaluating the efficacy of natural products as immune enhancers against respiratory diseases.

• The Journal of Engineering Research
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• v.3 no.1
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• pp.189-197
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• 1998

Hyeroxyapatite(HAp) which has good biocompatibility was made by Wet Chemical Process. The surface of Ti-6Al-4V, coated with HAp by lon Beam Assisted Deposition (IBAD), was treated with 5ppm, 10ppm, 20ppm, and 100ppm of $AgNO_3$ solution. In this Ag impregnation process, $Ca^{2+}$ of HAp was substituted with $Ag^+$ of $AgNO_3$. In this study, the antimicrobial effect and biocompatibility of Ti-6Al-4V alloy which was coated with Ag-HAp were examined. The antimicrobial test was carried out with two kinds of bacteria(P. Aeruginosa, S. Epidermidis), which are highly infectious in a transplanting operation of implant materials. As a result of the test, it was observed that Ti-6Al-4V alloy which was treated by 20ppm of $AgNO_3$ solution has good biocompatibility. In order to observe the antimicrobial mechanism of $Ag^+$, E. coli which is the most common bacterium was treated by Ag-HAp. Then cell morphology of E. coli was observed by the transmission electron microscope(TEM). The destruction of cell wall and cytoplasm of E. coil were observed. A black spot appeared in the cytoplasm was analyzed by energy dispersive analysis X-ray (EDAX) and it showed a small amount of $Ag^+$. Thus, it was proved that $Ag^+$ destroys bacteria effectively and Ti-6Al-4V alloy which was impregnate with Ag ion show antimicrobial effect on infection bacteria.