• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.98-105
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• 1999

Intestinal bacteria comprise one-third of the contents of the large intestine in humans. Their interactions with the gastrointestinal immune system induce characteristic immunological responses which stimulate or suppress the host's defense system. RAW 264.7 murine cell line was used as a macrophage model to assess the effects of the exposure to the isolated human intestinal bacteria, Bacteroides, Bifidobacterium, Eubacterium, Streptococcus, and E. coli, on NO (nitric oxide), $H_2O_2$(hydrogen peroxide), and cytokines IL (interleukin)-6 and TNF (tumor necrosis factor)-a production. RAW 264.7 cells were cultured in the presence of heat-killed bacteria for 24 h at concentrations of 0-$50\mu$g/ml. Our results showed that Bacteroides and E. coli stimulated IL-6, TNF-$\alpha$, NO, and $H_2O_2$production at high levels even at $1\mu$g/ml, whereas Bifidobacterium, Eubacterium, and Streptococcus showed a low level of stimulation at $1\mu$g/ml, and a gradual increase as the cell concentration increased up to $50\mu$g/ml. This result suggests that gram-negative Bacteroides and E. coli are better able to stimulate macrophage than gram-positive Bifidobacterium, Streptococcus, and Eubacterium. The in vitro approaches employed here should be useful in further characterization of the effects of intestinal bacteria on gastrointestinal and systemic immunity.

• KSBB Journal
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• v.11 no.4
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• pp.420-430
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• 1996

One-step immuno-chromatography assay system for heat-killed Salmonella typhimurium antigens was developed. Three major components used were a glass fiber membrane (placed at the bottom of the system) with an antibody (specific to the analyse, detection antibody)-gold conjugate deposited in a dry state on the surface, a nitrocellulose membrane (middle) with an antibody (also, specific to the analyse but recognized different epitome: capture antibody) and anti-detection antibody immobilized in spatially separated areas, and a cellulose membrane (top) as absorption pad. These membranes were partially superimposed such that a wicking of aqueous solution containing sample can continuously take place through membranes. Variables that affected the system performance were the concentration of capture antibody, the location on the membrane, inert protein used for blocking of the membrane and for carrying the sample, and the concentration of the gold conjugate. Under optimal conditions, within 15 minutes after absorption of a sample solution from the bottom of the system antigen-antibody complexes of sandwich type were formed on the membrane surface area with immobilized capture antibody and a color signal was generated in proportion to the analyse concentration. The minimum do tection limit of the analyse was $1{\times}106$ Salmonella cells/mL.

• Journal of Microbiology
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• v.42 no.2
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• pp.117-125
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• 2004

Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to >1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, rang-ing from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92％ (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-${\alpha}$) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypep-tide reactant was eluted from the gel and was found to cause dose dependent stimulation of the pro-ductions of IL-8 and TNF-${\alpha}$. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.201-207
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• 2016

The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.

• ALGAE
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• v.32 no.4
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• pp.309-324
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• 2017

Recently, the heterotrophic nanoflagellate Katablepharis japonica has been reported to feed on diverse red-tide species and contribute to the decline of red tides. However, if there are effective predators feeding on K. japonica, its effect on red tide dynamics may be reduced. To investigate potential effective protist predators of K. japonica, feeding by the engulfment-feeding heterotrophic dinoflagellates (HTDs) Oxyrrhis marina, Gyrodinium dominans, Gyrodinium moestrupii, Polykrikos kofoidii, and Noctiluca scintillans, the peduncle-feeding HTDs Luciella masanensis and Pfiesteria piscicida, the pallium-feeding HTD Oblea rotunda, and the naked ciliates Strombidium sp. (approximately $20{\mu}m$ in cell length), Pelagostrobilidium sp., and Miamiensis sp. on K. japonica was explored. We found that none of these heterotrophic protists fed on actively swimming cells of K. japonica. However, O. marina, G. dominans, L. masanensis, and P. piscicida were able to feed on heat-killed K. japonica. Thus, actively swimming behavior of K. japonica may affect feeding by these heterotrophic protists on K. japonica. To the contrary, K. japonica was able to feed on O. marina, P. kofoidii, O. rotunda, Miamiensis sp., Pelagostrobilidium sp., and Strombidium sp. However, the specific growth rates of O. marina did not differ significantly among nine different K. japonica concentrations. Thus, K. japonica may not affect growth of O. marina. Our findings suggest that the effect of predation by heterotrophic protists on K. japonica might be negligible, and thus, the effect of grazing by K. japonica on populations of red-tide species may not be reduced by mortality due to predation by protists.

• Journal of Korean Society of Water and Wastewater
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• v.23 no.2
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• pp.199-205
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• 2009

It is very important to differentiate of DNA derived from live or dead bacteria within mixed microbial communities in aquatic environments. Ethidium monoazide (EMA) is a DNA intercalating agent and the treatment of EMA with strong visible light cleaves the genomic DNA of bacteria. In dead bacterial cells, EMA intercalates into the genomic DNA, induces the cleavage of DNA, and inhibits the PCR amplification. In this study, we developed the EMA-PCR and EMA real-time PCR to detect the DNA derived from viable Escherichia coli (E.coli) in mixed cultures of live and dead E.coli. The treatment of EMA, $50{\mu}g/mL$, and 650 W visible halogen light exposure for 2 minutes cleaved the genomic DNA derived from heat killed E.coli but did not those of live E.coli. EMA-PCR could detect the DNA from live E.coli in mixed culture samples of live and dead E.coli at various ratio and there was no DNA amplification in only dead E.coli cultures. Similar results were observed in EMA real-time PCR. Further studies are needed to develop various EMA-PCR methods to detect viable waterborne pathogens such as Helicobacter pylori, Giardia lamblia, and so on.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.999-1010
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• 2021

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment. They are highly toxigenic and carcinogenic. Probiotic bacteria isolated from fermented foods were tested to check their ability to degrade and/or detoxify PAHs. Five probiotic bacteria with distinct morphologies were isolated from a mixture of 26 fermented foods co-cultured with benzo(a)pyrene (BaP) containing Bushnell Haas minimal broth. Among them, B. velezensis (PMC10) significantly reduced the abundance of BaP in the broth. PMC10 completely degraded BaP presented at a lower concentration in broth culture. B. velezensis also showed a clear zone of degradation on a BaP-coated Bushnell Haas agar plate. Gene expression profiling showed significant increases of PAH ring-hydroxylating dioxygenases and 4-hydroxybenzoate 3-monooxygenase genes in B. velezensis in response to BaP treatment. In addtion, both live and heat-killed B. velezensis removed BaP and naphthalene (Nap) from phosphate buffer solution. Live B. velezensis did not show any cytotoxicity to macrophage or human dermal fibroblast cells. Live-cell and cell-free supernatant of B. velezensis showed potential anti-inflammatory effects. Cell-free supernatant and extract of B. velezensis also showed free radical scavenging effects. These results highlight the prospective ability of B. velezensis to biodegrade and remove toxic PAHs from the human body and suggest that the biodegradation of BaP might be regulated by ring-hydroxylating dioxygenase-initiated metabolic pathway.

• KSBB Journal
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• v.25 no.4
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• pp.303-310
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• 2010

Probiotics beneficially affect the health of the host via various mechanisms in the intestine. Recent developments in probiotic products have mainly been made to maximize probiotic effects in human. In this regard, probiotic products containing doubly coated or encapsulated cells, multi-species probiotics, or high viable cell number (1010 viable cells/gram or more) have been developed and are already available in the market. Until now, the majority of probiotics contain live cells but little attention has been paid to other alternative products such as heat-killed cell or bacteriocin-containing ones, which could have broad applications due to advantages over live cell-based probiotics, such as safety and stability. In addition, genetically engineered lactic acid bacteria could be of great importance in the field of alimentary health if they are carefully designed for biological safety. Although a number of probiotics are marketed by claiming health benefits, regulations for health claims will be more stringent. Therefore sufficient scientific and clinical evidences supporting the safety and efficacy of the potential probiotic strain will be required by the regulatory authority for a health claim, which thus may have a huge impact on the future probiotic market.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.1
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• pp.67-73
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• 2011

Dental bacteria can cause gum diseases, i.e. gingivitis and periodontitis, by inducing inflammation in human gingiva. Therefore, the most effective way to prevent and treat gum diseases is the control of the inflammatory reactions induced by dental bacteria. Almost all present dental care products contain anti-bacterial agents to eliminate dental bacteria. However, recent studies report that even heat-killed dental bacteria can induce the inflammation responses in oral cells. Therefore, the method using anti-bacterial agents should be improved for better anti-inflammatory effect and the effective natural anti-inflammatory substances need to be found. In addition, the mechanisms of gingival inflammation should be elucidated. In this study, we tried to find out the mechanism of the gingival inflammation and effective natural anti-inflammatory substances with human gingival epithelial cells and Prevotella intermedia which is well known as a typical dental bacteria inducing gingivitis and periodontitis. In results, Prevotell intermedia initiated the gingival inflammation response by stimulating gingival epithelial cells to release an inflammatory cytokine, IL-8. Furthermore, the inflammation by Prevotella intermedia is related to COX-2, AP-1, and TNF-${\alpha}$ pathways. Green tea extract could effectively suppress the inflammatory responses induced by Prevotella intermedia. We find out the effective natural substance for the improvement of gum diseases by studying the mechanism of the gingival inflammation induced by dental bacteria.

• Journal of Life Science
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• v.26 no.8
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• pp.970-975
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• 2016

Acne vulgaris is a common chronic skin disorder that affects millions of people. The pathogenesis of acne has been known to be closely associated with the bacterium Propionibacterium acnes. Here we investigated the anti-acne activity of Lactobacillus plantarum K-1 BR by observing the expressions of proinflammatory cytokines, TNF-α, IFN-γ and IL-8, of human keratinocytes. When we applied heat-killed P. acnes to HaCaT cells, the inflammatory cytokines were induced by two- to four-fold compared to the normal control. When the bacteriocin, purified from L. plantarum K-1 BR, was pretreated to the HaCaT cells, the expression levels of TNF-α and IFN-γ stimulated by P. acnes significantly decreased to 25% and 30% of the induced levels, respectively. The IL-8 levels also significantly decreased with the concentration dependent manner of the bacteriocin. These results suggest that the bacteriocin from L. plantarum K-1 BR could reduce the expression levels of inflammatory cytokines and thus may relieve inflammations caused by acne.