• Restorative Dentistry and Endodontics
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• 제26권4호
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• pp.285-295
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• 2001

The objective of this study was to investigate the inhibitory effect of taurine and alendronate on the osteoclast differentiation. Osteoblasts and bone marrow cells from 1-2 day old mouse were co-cultured in 10% fetal bovine serum - minimal essential media (FBS-MEM). Osteoclast differentiation was induced by adding the sonicated extracts of Porphyromonas gingivalis (P.gingivalis). Osteoclasts were identified using tartrate resistant acid phosphotase staining (TRAP). Alendronate of 10$^{-7}$, 10$^{-6}$, 10$^{-5}$M and taurine of 500, 1000, 1500$\mu\textrm{g}$/ml were added respectively. The cytotoxic effects of alendronate and taurine were examined using MTT(3-(4,5-dimethylthiazol -2-yl-2,5-diphenyltetrazo- lium bromide) method. After culturing with the sonicated extracts of P.gingivalis, the amounts of IL-6 in the culture supernatant were measured and compared using the ELISA method. The results were as follows : 1. Osteoclasts were differentiated at the concentration of 0.01~0.1$\mu\textrm{g}$/ml sonicated extracts of P.gingivalis. (P<0.05). 2. Alendronate inhibited osteoclasts differentiation at the concentration of 10$^{-5}$ M when the concentration of sonicated extracts of P.gingivalis was 0.01$\mu\textrm{g}$/ml. 3. Taurine inhibited osteoclasts differentiation at the concentration of 1500$\mu\textrm{g}$/ml when the concentration of sonicated extracts of P.gingivalis 0.01$\mu\textrm{g}$/ml. 4. In cytotoxic test (MTT test), no cytotoxic effect was evident in all concentrations of alendronate and taurine. 5. Taurine (10$^{-5}$M) and alendronate(1500$\mu\textrm{g}$/ml) did not change the amounts of IL-6 induced by sonicated extracts of P.gingivalis significantly.

• Restorative Dentistry and Endodontics
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• 제29권5호
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• pp.479-484
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• 2004

감염근관내 spirochetes의 존재 유무에 대한 논란이 많았으나 최근 PCR을 사용한 세균검출 실험에서 Treponema denticola 균주가 감염근관의 50%이상의 경우에서 발견됨에 따라 이 세균이 치수 및 치근단 질환의 병인과정에 관여하는 지에 대한 관심이 높아지고 있다. 이와 관련하여 Shenker 등이 T. denticola의 sonicated extract에서 순수분리된 면역억제 단백질 (immunoinhibitory protein)이 T 임파구의 proliferation을 방해함을 보고한 바 있다. 하지만 이 세균성 단백질이 T 임파구의 기능에 어떤 영향을 미치는 지에 대한 연구는 부족한 실정이다. 따라서 T. denticola의 면역억제 단백질이 처리되기 전과 후의 T 세포에서 분비되는 cytokine Interleukine-2와 Interleukine-4의 발현 정도를 비교하여 그 작용기전을 밝히는 것이 본 연구의 목적이다. Treponema denticola LL2513를 혐기성 상태에서 TYGVS 배지에 배양한 다음 PBS 세척과 lyophilize 과정을 거친 후 sonication을 시행한다. 이 과정을 거쳐 추출된 상층액이 T. denticola의 면역억제 단백질인 sonicated extract이다. 실험을 위해 건강한 혈액 공여자로부터 T 세포를 Buoyant density 방법으로 추출해 낸 다음 24-well plate에 100만개의 세포를 주입한 다음 Group 1에는 2% FBS의 medium만으로 배양하고, Group 2에는 100 $\mu$l의 PHA로만 증식 자극을 하였고, Group 3에는 PHA처리 전 sonicated extract로 T세포를 자극하였다. 72시간 동안 배양한 다음 상층액을 추출하여 ELISA assay를 사용하여 IL-2와 IL-4의 발현정도를 측정하였다. PHA로 자극받은 Group 2에서는 IL-2와 IL-4가 대조군인 Group 1에서보다 높은 수준으로 발현되었다. 하지만 $12.5{\;}\mu\textrm{g}/ml$의 T. denticola sonicated 추출물로 전처리한 Group 3에서는 IL-2와 IL-4의 수준이 유의성있게 억제되어 발현되었다 (p < 0.05). 이러한 결과를 통하여 T. denticola에서 추출된 면역억제 단백질이 Th1과 Th2의 cytokine 분비 기능을 억제하는 것으로 확인 되었으며 이 기전이 감염 근관에서 발견되는 T. denticola의 치수 및 치근단 질환에 대한 병인기전과 관련이 있는 것으로 사료된다.

• 한국축산식품학회지
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• 제40권6호
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• pp.1044-1054
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• 2020

This study was performed to investigate the effects of lotus leaf hot water extracts treatment on the quality and stability of eggs using impregnation treatment through ultrasonication during storage. A total of 480 eggs were categorized into four treatment groups (n=30 each)-non-treated (CON), soaked for 30 min in lotus leaf hot water extracts without ultrasonication (T1), sonicated in distilled water (T2), and sonicated in lotus leaf hot water extracts (T3)-and stored for 15 d at 30℃. The egg weight, Haugh unit (HU), egg grade, albumen height, yolk color, eggshell thickness, eggshell breaking strength, and weight loss were measured for egg quality assessment. 2-Thiobarbituric acid reactive substance (TBARS) and volatile basic nitrogen (VBN) contents were measured as stability indicators. Additionally, total phenolic contents (TPC), total flavonoid contents (TFC), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were evaluated. The HU, egg grade, albumen height, and yolk color of T3 were significantly higher than those of CON (p<0.05). No significant differences in eggshell thickness and eggshell breaking strength are observed among the groups. The weight loss of T3 was significantly lower than that of the other groups during storage (p<0.05). The application of lotus leaf hot water extracts also significantly reduced TBARS and VBN (p<0.05). The TPC, TFC, and DPPH radical scavenging activity of T3 were significantly higher than those of the other groups (p<0.05). These results suggest that lotus leaf hot water extracts may be useful as a natural ingredient for improving the quality and stability of eggs during storage.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1186-1194
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• 2022

The intake of probiotic lactic acid bacteria not only promotes digestion through the microbiome regulated host intestinal metabolism but also improves diseases such as irritable bowel syndrome and inflammatory bowel disease, and suppresses pathogenic harmful bacteria. This investigation aimed to evaluate the immunomodulatory effects in intestinal epithelial cells and to study the clinical efficacy of the selected the Bifidobacterium breve and Bifidobacterium longum groups. The physiological and biochemical properties were characterized, and immunomodulatory activity was measured against pathogenic bacteria. In order to find out the mechanism of inflammatory action of the eight viable and sonicated Bifidobacterium spp., we tried to confirm the changes in the pro-inflammatory cytokines (TNF-α, interleukin (IL)-6, IL-12) and anti-inflammatory cytokine (IL-10), and chemokines, (monocyte chemoattractant protein-1, IL-8) and inflammatory enzymatic mediator (nitric oxide) against Enterococcus faecalis ATCC 29212 infection in Caco-2 cells and RAW 264.7 cells. The clinical efficacy of the selected B. breve and B. longum group was studied as a probiotic adjuvant for acute diarrhea in children by oral administration. The results showed significant immunomodulatory effects on the expression levels of TNF-α, IL-6, IL-12, MCP-1, IL-8 and NO, in sonicated Bifidobacterium extracts and viable bifidobacteria. Moreover, each of the Bifidobacterium strains was found to react more specifically to different cytokines. However, treatment with sonicated Bifidobacterium extracts showed a more significant effect compared to treatment with the viable bacteria. We suggest that probiotics functions should be subdivided according to individual characteristics, and that personalized probiotics should be designed to address individual applications.

• 한국작물학회지
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• 제58권2호
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• pp.119-127
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• 2013

The interaction between Agrobacterium and soybean has been studied at the transcriptome level but not at the metabolic level. However, it is necessary to investigate the difference in metabolites between susceptible and non-susceptible cultivars for high efficiency transformation. We investigated the difference in metabolites from sonicated soybean cotyledons of Korean cultivars and Bert cultivar. To identify difference in metabolites, sonicated extracts were analysed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The soybean cultivars were classified by susceptibility using green fluorescent protein expression. We found a difference in metabolites between the high susceptible and low susceptible cultivars. The FT-ICR/MS experimental m/z data of different metabolites were compared with theoretical m/z in KNApSAcK database. The candidate list was made using KNApSAcK and focused on phenolic compounds. These candidate metabolites are speculated to influence factors in the interaction. This list of candidates may be useful to investigate the interaction between Agrobacterium and plants to increase transformation efficiency.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.275-288
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• 1995

The inhibitory effect of cell wall extracts of streptococci, have been investigated to know host-parasite relationship or pathogenesis of abscess formation. Streptococci isolated from the infected root canals were sonicated to get cell wall extracts which have been known as one of the factors of pyogenesis. Lymphocytes separated by density gradient were stimulated with phytohemagglutinin and exposed to cell wall extracts of Streptococcus sanguis, S. mitis, S. uberis, S. mutans (ATCC 10449), and S. faecalis (ATCC 19433). [$^3H$]-thymidine uptake of lymphocytes was analyzed with scintillation counter and lactate dehyrogenase (LD) activity was measured with autochemistry analyzer. S. faeealis had the strongest inhibitory effect. beginning at $100\;{\mu}g/ml$ concentration of sonic extracts. S. sanguis and S. mitis had inhibitory effect at $300\;{\mu}g/ml$, while S. uberis and S. mutans showed no inhibitory, effect on DNA syntheis even at $300\;{\mu}g/ml$. Each streptococci showed different inhibitory effect on the DNA synthesis of lymphocytes, which finding indicated wide spectrum of susceptibility of lymphocytes according to streptococcus spp. There were no significant difference of LD activities between control and each streptococcal extracts. Streptococcal sonic extracts did not affect the morphological findings or number of colonies activated lymphocytes. These finding suggested the inhibitory effect of sonic extract of streptococci to lymphocytes could be detected by DNA synthesis inhibition, not by cellular membrane damage.

• Restorative Dentistry and Endodontics
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• 제25권4호
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• pp.606-618
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• 2000

Dental pulp infection is most commonly caused by extensive dental caries, and some bacterial species invade root canals; bacterial components and products are thought to be associated with the pathogenesis of periapical periodontitis. A principle driving force behind pulpal disease response appears to lie in the host immune system's to bacteria and their products. We examined the production of interleukin $1{\beta}$ (IL-$1{\beta}$) and tumor necrosis factor ${\alpha}$(TNF-${\alpha}$) from human peripheral mononuclear cells, lymphocytes and monocytes stimulated by heat-killed Acitnobacillus actinomycetemcomitans (ATCC 29523), Porphyromonas gingivalis (ATCC 33277) and Prevotella intermedia (ATCC 25611), and also by their sonicated bacterial extracts (SBE), respectively. The effects of three strains of heat-killed bacteria and their SBEs on the morphology of cultured blood cell lines HL-60 (KCLB 10240) and J774A.1 (KCLB 40067) were observed under the inverted microscope. Ultrastructural changes of J774A.1 exposed to heat-killed P. intermedia and its SBE were investigated using transmission electron microscopy. Production of IL-$1{\beta}$ was reduced in human peripheral mononuclear cells after stimulation by sonic bacterial extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. Heat-killed and sonic extract of P. gingivalis inhibited the production of TNF-${\alpha}$ in peripheral mononuclear cells. Production of TNF-${\alpha}$ was inhibited in peripheral monocytes after stimulation by sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. HL-60 and J 774A.1 cells showed granular degeneration after treatment with heat-killed and sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia Chromatin margination and shrinkage were observed in 774A.1 treated with heat-killed P. intermedia. Cell wall structure and organelles were destroyed and vacuoles were formed in cytoplasm in J774A.1 treated with P. intermedia sonic extract. These results suggest that A actinomycetemcomitans, P gingivalis and P intermedia may have an important role in the formation and progression of pulpal diseases via both modulation of production of IL-$1{\beta}$ and TNF-${\alpha}$ from blood mononuclear cells and cytopathic effects.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.740-744
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• 2005

Bifidobacterium has been previously shown to potentiate immune function, which was mediated through the stimulation of cytokine production by macrophage. This study was performed to further characterize the effective component of Bifidobacterium by measuring the level of interleukin (IL)-6 cytokine using the RAW 264.7 murine cell line as a macrophage model. RAW 264.7 cells were cultured for 24 h in the presence of whole cells (WCs), cell walls (CWs), and cell-free extracts (CFEs) from various strains of Bifidobacterium and other lactic acid bacteria at various concentrations. The most effective component was different depending on the strains and the concentrations used. When tested with each cell fraction from Bifidobacterium sp. BGN4, heat treatment of the cell fractions lowered the production of IL-6. Synergistic effect was obtained, especially when CWs and CFEs were combined. Sonicated WCs stimulated IL-6 production more than intact WCs. The in vitro approaches employed here should be useful in further characterization of the effects of Bifidobacterium on gastrointestinal and systemic immunity.

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2001년도 추계학술대회(제116회) 및 13회 Workshop 제3회 한ㆍ일 치과보존학회 공동학술대회 초록집
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• pp.566.2-566
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• 2001

The objective of this study was to investigate the ability of alendronate and taurine in inhibiting in vitro osteoclast differentiation induced by bacteria. Whole cell sonicates of P. gingivalis were used as an osteoclast-stimulating factor in a mouse coculture system and differentiated osteoclasts were confirmed by tartrate-resistant acid phosphatase (TRAP) staining. Alendronate at the concentrations of 10-7, 10-6, and 10-5 M, and taurine at the concentrations of 4mM, 8mM, and 12mM were used.(omitted)

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.621-621
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• 2003

I. Objectives In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2(IL-2), Interleukin-4(IL-4) and Transforming growth $factor-{\beta}1(TGF-{\beta}1)$ in human lymphocytes. II. Materials and methods Enterococcus faecalis(ATCC29212) strains were used in this study. Strains were grown in 1-liter cultures in 85% N2-10% H2-5% $CO_2$chamber for 3 days at $37^{\circ}C$. The medium used was 3.7% brain heart infusion broth. Bacterial cells harvested from 1-liter cultures were washed, suspended in 20ml of phosphate-buffered saline(PBS). Suspensions of bacterial cells were disrupted by sonication on ice for 5 min. Protein concentration was determined by the Bicinchoninic acid(BCA) protein assay.(omitted)