• Title/Summary/Keyword: Sonicated extracts

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THE INHIBITORY EFFECT OF TAURINE AND ALENDRONATE ON THE OSTEOCLAST DIFFERENTIATION MEDIATED BY SONICATED EXTRACTS OF PORPHYROMONAS GINGIVALIS IN VITRO. (Porphyromonas gingivalis 분쇄액으로 유도된 파골세포의 분화에 미치는 Taurine과 Alendronate의 효과)

  • Park, Ju-Hyun;Kum, Kee-Yeon;Lee, Jung-Hyun;Yu, Jung-Yun;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.26 no.4
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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.

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The Effect of Treponema denticola immunoinhibitory protein on cytokine expression in T cells (Treponema denticola 면역억제 단백질이 T 세포의 cytokine 발현에 미치는 영향)

  • Lee Sang-Yup;Shon Won-Jun;Lee WooCheol;Baek Seung-Ho;Bae Kwang-Shik;Lim SungSam
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.479-484
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    • 2004
  • Immunoinhibitory protein extracted from sonicated Treponema denticola have been shown to suppress cell cycle progression of human lymphocytes. To study in detail about the effect of this microorganism on the function of lymphocytes. we investigated the levels of Interleukin-2 (IL-2) and Interleukin-4 (IL-4) production by T lymphocytes before and after the addition of $12.5{\;}\mu\textrm{g}/ml$ T. denticola sonicated extracts. In this study. levels of IL-2 and IL-4 produced from T cells pretreated with sonicated extracts were evaluated by using the quantitative sandwich enzyme immunoassay technique. In response to phytohemagglutinin (PHA) stimulation. T cell produced increased levels of IL-2 and IL-4. However. the expressions of both cytokines were significantly inhibited when PHA activated-T cells were pre-exposed to sonicated T. denticola extracts (p < 0.05). These findings suggest that the T. denticola sonicated extracts induced-immunosuppression in Th1 and Th2 cell functions could be a part of the pathogenic mechanism of the endodontic failure associated with this microorganism.

Effects of Lotus (Nelumbo nucifera) Leaf Hot Water Extracts on the Quality and Stability of Eggs using Ultrasonication Treatment during Storage

  • Lee, Jihye;Seo, Han Geuk;Lee, Chi-Ho
    • Food Science of Animal Resources
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    • v.40 no.6
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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.

Immunomodulatory Effects of Bifidobacterium spp. and Use of Bifidobacterium breve and Bifidobacterium longum on Acute Diarrhea in Children

  • Choi, Yae Jin;Shin, Seon-Hee;Shin, Hea Soon
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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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.

Relationship of Transformation Efficiency and Metabolites Induced in Korean Soybean Cotyledons Treated with Sonication

  • Song, Kitae;Yim, Won Cheol;Jung, Gun-Ho;Kim, Sun Lim;Kwon, Young-Up;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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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.

THE INHIBITORY EFFECT OF STREPTOCOCCAL CELL WALL EXTRACTS ON STIMULATION OF LYMPHOCYTES (연쇄구균의 세포벽 단백질 추출물이 림프구 활성의 억제에 미치는 영향)

  • Sang, Hyoung-Sook;Jeong, Hee-Il;Oh, Se-Hong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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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.

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EFFECTS OF HEAT-KILLED AND SONIC EXTRACTS OF MICROORGANISM ON CULTURED CELLS (세균액 및 세균단백질 추출물이 배양 세포에 미치는 영향)

  • Yu, Young-Dae;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.25 no.4
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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.

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Effect of Bifidobacterium Cell Fractions on IL-6 Production in RAW 264.7 Macrophage Cells

  • Lee, Byung-Hee;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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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.

The effect of taurine and alendronate on the osteoclast differentiated by the sonicated extracts of Porphyromonas Gingivalis in vitro

  • Kim, Hyung-Su;Lee, Seung-Jong;Lee, Chan-Young;Kum, Kee-Yeon
    • Proceedings of the KACD Conference
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    • 2001.11a
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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)

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Effects of Enterococcus faecalis sonicated extracts on IL-2, IL-4 and $TGF-{\beta}1$ production from human lymphocytes

  • Kim, Hyeon-Sik;Lee, Woo-Cheol;Lim, Sung-Sam
    • Proceedings of the KACD Conference
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    • 2003.11a
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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)

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