Receptor activator of NF-kB ligand (RANKL) is known to play a major role in bone metabolism and the immune system, and its recombinant form has been expressed in bacterial systems for research since the last two decades. However, most of these recombinant forms are used after purification or directly using living cells. Here, there were cell extracts of recombinant Lactococcus lactis expressing mouse RANKL (mRANKL) used to evaluate its biological activity in mice. Mice were divided into three groups that were fed phosphate-buffered saline (PBS), wild-type L. lactis IL1403 (WT_CE), and recombinant L. lactis expressing mRANKL (mRANKL_CE). The small intestinal transcriptome and fecal microbiome were then profiled. The biological activity of mRANKL_CE was confirmed by studying RANK-RANKL signaling in vitro and in vivo. For small intestinal transcriptome, differentially expressed genes (DEGs) were identified in the mRANKL_CE group, and no DEGs were found in the WT_CE group. In the PBS vs. mRANKL_CE gene enrichment analysis, upregulated genes were enriched for heat shock protein binding, regulation of bone resorption, and calcium ion binding. In the gut microbiome analysis, there were no critical changes among the three groups. However, Lactobacillus and Sphingomonas were more abundant in the mRANKL_CE group than in the other two groups. Our results indicate that cell extracts of mRANKL_CE can play an effective role without a significant impact on the intestine. This strategy may be useful for the development of protein drugs.
Objectives: Dental caries is one of the most prevalent human diseases worldwide. The disease initiates with bacterial adherence to the tooth surface followed by the formation of dental plaques. Mutans streptococci and Candida albicans are principal oral microorganisms involved in the initiation and development of dental caries. Phytochemicals have been shown to possess promising antimicrobial properties against a wide range of microorganisms and can be used for the prevention and treatment of dental caries. Herein, we reviewed literature on plants that are traditionally used for their antimicrobial properties or possess promising anticariogenic activity. We selected aerial parts of Scrophularia striata (S. striata) and galls of Quercus infectoria (Q. infectoria) and investigated their antimicrobial activity against cariogenic microorganisms. Methods: Water soluble fractions were obtained from hydroalcoholic extracts of S. striata and Q. infectoria and their antimicrobial activity against Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), and Candida albicans (C. albicans) was evaluated separately and in combination. The extracts were then used for preparing an herbal mouthwash whose stability and tannic acid content were evaluated over 60 days. Results: Q. infectoria gall extract possesses efficient antimicrobial activity that was synergistically enhanced in the presence of S. striata extract. Mouthwash prepared using these extracts showed desirable organoleptic characteristics, antimicrobial activity, and stability. Conclusion: Extracts of S. striata and Q. infectoria galls can be used together for preparing dental products with effective anticariogenic properties. Our study highlights the importance of extensive pharmacological investigations when using herbal products alone or in combination with other chemical substances.
Alisha Wehdnesday Bernardo Reyes;Heejin Kim;Tran Xuan Ngoc Huy;Trang Thi Nguyen;Wongi Min;Hu Jang Lee;Jin Hur;John Hwa Lee;Suk Kim
Journal of Microbiology and Biotechnology
/
v.33
no.4
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pp.441-448
/
2023
Brucellosis is a contagious zoonotic disease that infects millions of people annually with hundreds of millions more being exposed. It is caused by Brucella, a highly infectious bacterial species capable of infecting humans with an estimated dose of 10-100 organisms. Sirtuin 1 (SIRT1) has been reported to contribute to prevention of viral diseases as well as a chronic infection caused by Mycobacterium bovis. Here, we investigated the role of SIRT1 in the establishment of Brucella abortus infection in both in vitro and in vivo systems using the reported SIRT1 activators resveratrol (RES), piceatannol (PIC), and ginsenoside Rg3 (Rg3). In RAW264.7 cells, SIRT1 activators did not alter the adherence of Brucella or Salmonella Typhimurium. However, reduced uptake of Brucella was observed in cells treated with PIC and Rg3, and survival of Brucella within the cells was only observed to decrease in cells that were treated with Rg3, while PIC treatment reduced the intracellular survival of Salmonella. SIRT1 treatment in mice via oral route resulted in augmented Brucella resistance for PIC and Rg3, but not RES. PIC treatment favors Th2 immune response despite reduced serum pro-inflammatory cytokine production, while Rg3-treated mice displayed high IL-12 and IFN-γ serum production. Overall, our findings encourage further investigation into the complete mechanisms of action of the different SIRT1 activators used as well as their potential benefit as an effective alternative approach against intracellular and extracellular pathogens.
Yeonhoo Jung;Moonseok Jang;Rahye Kang;Wanghui Lee;Seongjun Park
Journal of Veterinary Clinics
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v.41
no.1
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pp.49-53
/
2024
A two-year-old, spayed female, 22.5 kg Pungsan was referred with chronic crusts and erosion on the nose. A referring veterinarian prescribed an anti-inflammatory dose (0.5-1 mg/kg/day) of oral glucocorticoids for 5 months, but skin lesions showed no meaningful improvement. A dermatological evaluation revealed a crust, depigmentation, erosion, and erythematous lesion over the nasal planum with a loss of the normal cobblestone texture. Also, firm and multifocal plaques over the thigh, groin, axilla, and dorsum were detected. A cytology examination on the nose, thigh, groin, axilla, and dorsum revealed moderate neutrophilic inflammation and bacterial infection. Abdominal radiography and ultrasonography revealed subcutaneous calcified materials along the thigh, groin, axilla, and dorsum. Calcinosis cutis was suspected because of the adverse effect of previous prolonged corticosteroid therapy. A histopathology examination of the nose lesion revealed moderate to severe degenerative or apoptotic changes of the basal layer and lymphoplasmacytic interface dermatitis. Facial discoid lupus erythematosus (FDLE) was diagnosed based on the history and the clinical, cytological, and histopathological results. Minocycline (7 mg/kg PO q 12 h) and niacinamide (500 mg/dog PO q 12 h) were prescribed as initial treatment. Glucocorticoids were not administered due to the presence of calcinosis cutis induced by previous corticosteroid treatment. After 6 weeks of treatment, the clinical signs on the nose were mildly improved. At this time, topical 1% pimecrolimus cream (twice daily) was initiated, while minocycline and niacinamide were continued at the same dose. The nasal planum markedly improved after 6 weeks of additional treatment, hence minocycline and niacinamide were prescribed for an additional 2 weeks and stopped, and the patient was continued solely on topical pimecrolimus. The dog's skin lesion has been maintained in clinical remission with topical 1% pimecrolimus twice daily for more than 5 months.
Industrial advancements have resulted in food culture development, followed by increased seafood consumption and large-scale seafood farming, which has been accompanied by an increased prevalence of fish disease. The antibiotic oxytetracycline (OTC) is commonly used to prevent and treat bacterial diseases in fish. However, overuse of OTC had led to negative aspects. In view of this, we conducted a research with regard to aspects of remnants on olive flounder skin, liver, and muscle through dipping treatment and oral feeding of OTC and analyzed the results with bioassay and HPLC quantitative analyses. The dipping treatment was carried out once with 25 g/ton/hr of OTC, and the oral treatment with 62.5 mg/kg body weight/7 days. The results underwent a bioassay analysis. The dipping group reacted only on the skin right after dipping, while the oral feeding group responded on the skin for 77 days after feeding and on the muscle for 14 days. In the dipping group, the HPLC quantitative analysis revealed remnants in the skin on the 37th day and on the 13th day in the liver group. No remnants were found in the muscle, even immediately after dipping. In the oral feeding group, there was a high concentration (1.07 mg/kg) of remnant in the skin, even on the 77th day. 0.56 mg/kg in the liver, even a small amount, and no remnant in the muscle on the 42nd day. To sum up, the results suggest that it will not be harmful to our body to observe the OTC withdrawal period of 40 days with the muscle because OTC will hardly remain on it. When using olive flounder for sashimi, the skin and liver should not be used for broth, as the quantity of OTC residue is several times higher than that found in muscle. As previous studies reported that the concentration of remnants gradually decreased with heating, so it was likely to lessen, depending on the cooking temperature.
Water supplied through dental unit waterlines (DUWLs) has been shown to contain high number of bacteria. To reduce the contamination of DUWLs, it is essential to develop effective disinfectants. It is, however, difficulty to obtain proper DUWL samples for studies. The purpose of this study was to establish a simple laboratory model for reproducing DUWL biofilms. The bacteria obtained from DUWLs were cultured in R2A liquid medium for 10 days, and then stored at $-70^{\circ}C$. This stock was inoculated into R2A liquid medium and incubated in batch mode. After 5 days of culturing, it was inoculated into the biofilm formation model developed in this study. Our biofilm formation model comprised of a beaker containing R2A liquid medium and five glass rods attached to DUWL polyurethane tubing. Biofilm was allowed to form on the stir plate and the medium was replaced every 2 days. After 4 days of biofilm formation in the laboratory model, biofilm thickness, morphological characteristics and distribution of the composing bacteria were examined by confocal laser microscopy and scanning electron microscopy. The mean of biofilm accumulation was $4.68{\times}10^4$ colony forming unit/$cm^2$ and its thickness was $10{\sim}14{\mu}m$. In our laboratory model, thick bacterial lumps were observed in some parts of the tubing. To test the suitability of this biofilm model system, the effectiveness of disinfectants such as sodium hypochlorite, hydrogen peroxide, and chlorhexidine, was examined by their application to the biofilm formed in our model. Lower concentrations of disinfectants were less effective in reducing the count of bacteria constituting the biofilm. These results showed that our DUWL biofilm laboratory model was appropriate for comparison of disinfectant effects. Our laboratory model is expected to be useful for various other purposes in further studies.
Kim, Yeowon;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
Journal of the korean academy of Pediatric Dentistry
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v.47
no.4
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pp.446-453
/
2020
The purpose of this study was to evaluate the effect of Indocyanine Green (ICG) and near-infrared (NIR) diode laser on Streptococcus mutans biofilms depending on ICG concentrations. S. mutans biofilms were formed on a Hydroxyapatite disk, and 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solutions dissolved in sterile distilled water and a NIR diode laser having a power of 300 mW and a wavelength of 808 nm were applied to the biofilms. The temperature changes of the biofilm surface according to the concentrations of the ICG solution were measured using a 1-channel thermocouple thermometer. Compared to the control group, in the groups with only the 3.0, 4.0, 5.0 mg/mL ICG solution application, and in the groups with the 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solution application and light irradiation, a statistically significant decrease in the bacterial counts were observed. The temperature increase according to the concentration of the ICG solutions was 9.53℃, 10.43℃, 11.40℃, 12.10℃, 12.67℃, and 13.63℃ in ICG solutions of 0.5, 1.0, 2.0, 3.0, 4.0, and 5.0 mg/mL respectively. This study presents the potential for clinical application of ICG and NIR diode lasers as a new method for preventing dental caries.
The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.
Objective: The purpose of this study was to compare the bacterial flora at the peri-implant sulcus of the orthodontic mini-implant placed in the alveolar mucosa with the bacterial flora at the adjacent healthy gingival sulcus. Methods: Two plaque samples from 7 patients were collected by inserting paper points into the sulcus between the mini-implant and ligature wire connected to the mini-implant head and inflamed alveolar mucosa, and from the gingival sulcus of a healthy tooth adjacent to the mini-implant. Results: Using 16S rDNA clone library, the 24 kinds of bacteria including Haemophilus aphrophilus, Sphingomonas species, Capnocytophaga species, Prevotella melaninogenica, Lachnospiraceae species, Porphyromonas species, Neisseria flava were identified only from the sulcus around the mini-implant. These bacteria constituted only 9.2% of total clones, and the bacteria identified from both the sulcus around mini-implants and the gingival sulcus constituted 80.4% of total clones. Of these bacteria, clones of Prevotella species, Atopobium rimae, Veillonella species, Streptococcus intermedius/constellatus, Streptococcus salivarius were more frequently isolated from the peri-implant sulcus. Conclusion: This study suggests that a broad epidemiological study is needed to find causative bacteria which induce inflammation from the peri-implant sulcus.
Kim, Dong-Hyeok;Lim, Jung-Ju;Lee, Jin-Ju;Jung, Won-Chul;Shin, Hyeon-Jin;Lee, Hu-Jang;Kim, Gon-Sup;Kim, Suk
Korean Journal of Environmental Agriculture
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v.27
no.2
/
pp.156-162
/
2008
Salmonellosis is a major bacterial zoonosis that causes a variety of disease syndromes, self-limited enteritis to fatal infection in animals and food-borne infection and typhoid fever in humans. Recently, the emergence of multidrug resistant strains of Salmonella spp. causes more serious problems in environment and public health. The present study was investigated the antibacterial effect of Houttuynia cordata ethanol extract(HCEE) for murine salmonellosis. In the cytotoxic effect of HCEE on RAW 264.7 cells, there was no detectable effect with any concentrations between 25 and 100 ${\mu}g/ml$ after 8 h incubation. The bacteriocidal effect of HCEE was not showed on a Salmonella enterica serovar Typhimurium(S. typhimurium). HCEE makes morphological change of the RAW 264.7 cells, and there was significant decreased bacterial uptake and intracellular replication within Salmonella infected cells. And further nitric oxide(NO) production of Salmonella infected RAW 264.7 cells with HCEE was decreased comparing to RAW 264.7 cells without HCEE until 8 h post infection. Oral administration of HCEE showed a therapeutic effect for S. typhimurium infected BALB/c mice. The mortality of HCEE treated mouse was 80% until 12 days, while that of HCEE untreated mouse was 100 % until 8 days after lethal dose of S. typhimurium infection. These data suggested that HCEE has a potency treatment for intracellular replicative pathogen including salmonellosis, brucellosis, tuberculosis, listeriosis etc., and the application of HCEE makes new strategies for safety medicine development without antibiotic resistance bacterial appearance and residue problem in food and solves the public health problem from antibiotic mis- and over use.
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