• Journal of Oral Medicine and Pain
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• v.47 no.1
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• pp.10-26
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• 2022

Purpose: The purpose of this study was to investigate whether various saliva collection methods affect the observed salivary microbiome and whether microbiomes of stimulated and unstimulated saliva and plaque differ in richness and diversity. Methods: Seven sampling methods for unstimulated saliva, stimulated saliva, and plaque samples were applied to six orally and systemically healthy participants. Bacterial 16S ribosomal RNA genes of 10 major oral bacterial species, namely, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Streptococcus mitis, Streptococcus sobrinus, and Lactobacillus casei, were analyzed by real-time polymerase chain reaction. We comprehensively examined the dependence of the amount of bacterial ribosomal DNA (rDNA), bacterial-community composition, and relative abundance of each species on sample collection methods. Results: There were significant differences in the bacterial rDNA copy number depending on the collection method in three species: F. nucleatum, P. nigrescens, and S. mitis. The species with the highest richness was S. mitis, with the range from 89.31% to 100.00%, followed by F. nucleatum, P. nigrescens, T. denticola, T. forsythia, and P. intermedia, and the sum of the proportions of the remaining five species was less than 1%. The species with the lowest observed richness was P. gingivalis (<0.1%). The Shannon diversity index was the highest in unstimulated saliva collected with a funnel (4.449). The Shannon diversity index was higher in plaque samples (3.623) than in unstimulated (3.171) and stimulated (3.129) saliva and in mouthwash saliva samples (2.061). Conclusions: The oral microbial profile of saliva samples can be affected by sample collection methods, and saliva differs from plaque in the microbiome. An easy and rapid technique for saliva collection is desirable; however, observed microbial-community composition may more accurately reflect the actual microbiome when unstimulated saliva is assayed.

• Korean Journal of Pharmacognosy
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• v.54 no.1
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• pp.27-37
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• 2023

This research was conducted to investigate the comprehensive effects of methanol extract of Coptidis rhizoma (MECR) against oral pathogen. We studied the antibacterial, anti-biofilm, anti-gingipain and anti-inflammatory activity of MECR. The minimum bactericidal concentration (MBC) of MECR was 100 ㎍/mL against several oral pathogens. The formation of biofilm of Streptococcus mutans was reduced to 8.93~24.12% in the presence of 25 ㎍/mL of MECR. The gingipain activity of Porphyromonas gingivalis were reduced to 3.91~6.23% in case of Kgp and 5.73~7.78% in case of Rgp in the presence of 10 mg/mL of MECR. The expression of fadA mRNA, virulence factor of Fusobacterium nucleatum (F. nucleatum) was 3 folds decreased in the presence of 25 ㎍/mL of MECR. In case of YD-38 cells challenged with F. nucleatum, RQ values of IL-8 and IL-6 were reduced about 12 folds and 5.45 folds in the presence of 2 ㎍/mL of MECR. In case of RAW 264.7 murine cell challenged with F. nucleatum, RQ values of IL-1β and IL-6 were 2.52 folds and 2.55 folds reduced in the presences of 2 ㎍/mL of MECR. Conclusively, MECR showed potent antibacterial and anti-inflammatory effects against oral pathogenic bacteria.

• Tuberculosis and Respiratory Diseases
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• v.86 no.4
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• pp.294-303
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• 2023

Background: The human lung serves as a niche for a unique and dynamic bacterial community related to the development and aggravation of multiple respiratory diseases. Therefore, identifying the microbiome status is crucial to maintaining the microecological balance and maximizing the therapeutic effect on lung diseases. Therefore, we investigated the histological type-based differences in the lung microbiomes of patients with lung cancer. Methods: We performed 16S rRNA sequencing to evaluate the respiratory tract microbiome present in bronchoalveolar lavage fluid. Patients with non-small cell lung cancer were stratified based on two main subtypes of lung cancer: adenocarcinoma and squamous cell carcinoma (SqCC). Results: Among the 84 patients analyzed, 64 (76.2%) had adenocarcinoma, and 20 (23.8%) had SqCC. The α- and β-diversities showed significant differences between the two groups (p=0.004 for Chao1, p=0.001 for Simpson index, and p=0.011 for PERMANOVA). Actinomyces graevenitzii was dominant in the SqCC group (linear discriminant analysis [LDA] score, 2.46); the populations of Haemophilus parainfluenza (LDA score, 4.08), Neisseria subflava (LDA score, 4.07), Porphyromonas endodontalis (LDA score, 3.88), and Fusobacterium nucleatum (LDA score, 3.72) were significantly higher in the adenocarcinoma group. Conclusion: Microbiome diversity is crucial for maintaining homeostasis in the lung environment, and dysbiosis may be related to the development and prognosis of lung cancer. The mortality rate was high, and the microbiome was not diverse in SqCC. Further large-scale studies are required to investigate the role of the microbiome in the development of different lung cancer types.

• Journal of dental hygiene science
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• v.24 no.2
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• pp.97-106
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• 2024

Background: Periodontitis and peri-implantitis are diseases caused by pathogenic microorganisms that cause tissue damage and alveolar bone destruction resulting in the loss of teeth and implants. Due to the biological differences in the tissues surrounding the implants, peri-implantitis progresses more rapidly and intensely than periodontitis, underscoring the importance of understanding the characteristics and interactions of pathogenic bacteria. This study aimed to quantitatively analyze the pathogenic microorganisms associated with periodontitis and peri-implantitis in Korean patients and evaluate the correlation between these bacteria. Methods: A total of 98 (52 males and 46 females) were randomly selected and classified into three groups (healthy group [HG]=25; periodontitis group [PG]=31; and peri-implantitis group [PIG]=42). The relative expression levels of 11 pathogenic microorganisms collected from the gingival sulcus fluid were determined using multiplex real-time polymerase chain reaction. Results: Eikenella corrodens, Fusobacterium nucleatum, and Prevotella nigrescens were highly prevalent in the HG, PG, and PIG patients. The results of the relative quantitative analysis of microorganisms showed that all bacteria belonging to the green, orange, and red complexes were significantly more abundant in the PG and PIG than in the HG (p<0.05). Porphyromonas gingivalis in the red complex showed a positive correlation with all microorganisms in the orange complex (p<0.05). Campylobacter rectus in the orange complex showed a significant positive correlation with all microorganisms in the red complex, and with F. nucleatum, P. nigrescens, Prevotella intermedia, and Eubacterium nodatum (p<0.05). Conclusion: P. gingivalis, C. rectus, and F. nucleatum exhibit strong interactions. Removing these bacteria can block complex formation and enhance the prevention and treatment of periodontitis and peri-implantitis.

• Journal of the korean academy of Pediatric Dentistry
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• v.36 no.2
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• pp.237-244
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• 2009

When the symptom of periapical infection is not released by mechanical instrumentation. anti-microbial agents including antibiosis become necessary in order to remove microorganisms from the root canal. Since anti-microbial agents of natural origins are currently popular, more natural remedies are being sought out. As it turns out, it is well known isothiocyanates (ITCs) in horseradish root extract have anti-microbial activity from many studies. In this research, anti-microbial effects of horseradish root extract and chlorhexidine, a typical anti-microbial agent, were investigated and compared against two kinds of obligate anaerobes. Fusobacterium nucleatum and Prevotella nigrescens, that are often discovered in infected root canal, and Clostridium perfringens, which is resistant to antibiotics and frequently used as a control strain for antibacterial studies 1. The MIC and MBC of horseradish root extract were ranged from 87 to 470 ppm and from 156 to 625 ppm against three kinds of obligate anaerobes, respectively. Horseradish root extract showed the strongest anti-bacterial activity (MBC, 156 ppm) against F. nucleatum and also showed anti-bacterial activity against antibiotic resistant obligate anaerobes. C. perfringens. 2. The MIC and MBC of chlorhexidine were ranged from 3.12 to 6.25 ppm and 10.94 ppm against three kinds of obligate anaerobes, respectively. 3. The MIC with 87-470 ppm of horseradish root exact has the same growth inhibiting effect as the one of 3.12-6.25 ppm of chlorhexidine. Likewise, the MBC with 156-625 ppm of horseradish has the similar bactericidal effect as 10.94 ppm of chlorhexidine.

• Journal of Oral Medicine and Pain
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• v.33 no.2
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• pp.159-166
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• 2008

An ideal anti-bacterial medication for oral infection requires selective effect on pathogens causing dental caries and periodontal disease but not on normal flora. In addition, it should be less toxic for human and even for environment. This study was to seek such a natural anti-bacterial medication and thus anti-bacterial effect of Hamamelis virginiana was evaluated. Many recent researches on the anti-bacterial effect of natural plant extract and essential oil have reported that natural products can be used as medication for prevention and restrainment of dental caries, halitosis and periodontitis. It has been also reported that Hamamelis virginiana has anti-bacterial effect on Porphyromonas gingivalis, Fusobacterium nucleatum, Capnocytophaga gingivalis, Veilonella parvula, Eikenella corrodens, Peprostreptococcus micros, and Actinomyces odontolyticus. This study evaluated anti-bacterial effect of Hamamelis virginiana on Streptoccoccus mutans, Haemophylus actinomycetemcomitans, and Klebsiella pneumoniae to expand its anti-bacterial effect on other important oral pathogens and eventually to develop its oral care products or apply to clinical purpose. In this study, anti-bacterial tests for antibiotic disk susceptibility, minimal inhibitory concentration and minimal bactericidal concentration were performed to evaluate anti-bacterial effect of Hamamelis virginiana against Streptoccoccus mutans, Haemophylus actinomycetemcomitans, and Klebsiella pneumoniae. The results showed that Hamamelis virginiana has anti-bacterial effect on all pathogen strains tested in this study and furthermore Hamamelis virginiana possesses bactericidal effect other than bacteriostatic effect on Streptoccoccus mutans, Haemophylus actinomycetemcomitans, Klebsiella pneumoniae. This study indicates that a natural anti-bacterial medication for oral diseases can be developed using Hamamelis virginiana.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.3
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• pp.457-468
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• 2006

It is widely known that individuals with mental retardation (MR) and Down's syndrome (DS) often develop early onset periodontal diseases. In this study, the prevalence of periodontopathic bacteria in MR persons and DS patients was compared with normal persons. Plaque index and gingival index were measured. Five periodontopathic bacteria, Actinobacillus actinomycetemcomitans. Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum were surveyed in subgingival plaque samples by the polymerase chain reaction. Results : 1. Plaque index and gingival index were higher in MR persons group and DS patients group than normal persons group (p<0.05). 2. The prevalence of periodontopathic bacteria in normal persons group were lower than that of MR persons group and DS. Significant differences were observed in the prevalence of P. gingivalis, T. denticola and A. actinomycetemcomitans(P<0.05). 3. Prevalence of P. gingivalis(5.9%) at age 8-10 was lower than other ages in normal persons group, and its prevalence increased with age Prevalence of P. gingivalis, T. denticola and A. actinomycetemcomitans at MR persons group and DS patients group were higher than those of same ages of normal persons group. 4. Plaque index was associated with T. denticola and gingival index was associated with T. denticola and A. actinomycetemcomitans(P<0.05). These results suggested that plaque index, gingival index and prevalence of periodontopathic pathogens, especially P. gingivalis, T. denticola and A actinomycetemcomitans in DS patients group and MR persons group are higher than those of normal persons group.

• 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 Microbiology and Biotechnology
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• v.29 no.12
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• pp.1947-1956
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• 2019

The gut microbiome influences the health and well-being of dogs. However, little is known about the impact of breed on the fecal microbiome composition in dogs. Therefore, we aimed to investigate the differences in the fecal microbiome in three breeds of dog fed and housed under the same conditions, namely eight Maltese (8.0 ± 0.1 years), eight Miniature Schnauzer (8.0 ± 0.0 years), and nine Poodle dogs (8.0 ± 0.0 years). Fresh fecal samples were collected from the dogs and used to extract metagenomic DNA. The composition of the fecal microbiome was evaluated by 16S rRNA gene amplicon sequencing on the MiSeq platform. A total of 840,501 sequences were obtained from the 25 fecal samples and classified as Firmicutes (32.3-97.3% of the total sequences), Bacteroidetes (0.1-62.6%), Actinobacteria (0.2-14.7%), Fusobacteria (0.0-5.7%), and Proteobacteria (0.0-5.1%). The relative abundance of Firmicutes was significantly lower in the Maltese dog breed than that in the other two breeds, while that of Fusobacteria was significantly higher in the Maltese than in the Miniature Schnauzer breed. At the genus level, the relative abundance of Streptococcus, Fusobacterium, Turicibacter, Succinivibrio, and Anaerobiospirillum differed significantly among the three dog breeds. These genera had no correlation with age, diet, sex, body weight, vaccination history, or parasite protection history. Within a breed, some of these genera had a correlation with at least one blood chemistry value. This study indicates that the composition of the fecal microbiome in dogs is affected by breed.

• Journal of Periodontal and Implant Science
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• v.44 no.2
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.