• Title/Summary/Keyword: bacterial cell growth

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Kinetic Analysis of CpG-Induced Mouse B Cell Growth and Ig Production

  • Kim, Young-Ha;Lee, Sang-Hoon;Yoo, Yung-Choon;Lee, Jung-Lim;Park, Jong-Hwan;Park, Seok-Rae
    • IMMUNE NETWORK
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    • v.12 no.3
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    • pp.89-95
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    • 2012
  • Immune cells express toll-like receptors (TLRs) and respond to molecular patterns of various pathogens. CpG motif in bacterial DNA activates innate and acquired immune systems through binding to TLR9 of immune cells. Several studies reported that CpG can directly regulate B cell activation, differentiation, and Ig production. However, the role of CpG in B cell growth and Ig production is not fully understood. In this study, we analyzed the effect of CpG on the kinetics of mouse B cell viability, proliferation, and Igs production. Overall, CpG enhanced mouse B cell growth and production of Igs in a dose-dependent manner. Unlike LPS, 100 nM CpG (high dose) did not support TGF-${\beta}1$-induced IgA and IgG2b production. Moreover, 100 nM CpG treatment abrogated either LPS-induced IgM or LPS/TGF-${\beta}1$-induced IgA and IgG2b production, although B cell growth was enhanced by CpG under the same culture conditions. We subsequently found that 10 nM CpG (low dose) is sufficient for B cell growth. Again, 10 nM CpG did not support TGF-${\beta}1$-induced IgA production but, interestingly enough, supported RA-induced IgA production. Further, 10 nM CpG, unlike 100 nM, neither abrogated the LPS/TGF-${\beta}1$- nor the LPS/RA-induced IgA production. Taken together, these results suggest that dose of CpG is critical in B cell growth and Igs production and the optimal dose of CpG cooperates with LPS in B cell activation and differentiation toward Igs production.

Substrate specificity of bacterial endoribonuclease toxins

  • Han, Yoontak;Lee, Eun-Jin
    • BMB Reports
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    • v.53 no.12
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    • pp.611-621
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    • 2020
  • Bacterial endoribonuclease toxins belong to a protein family that inhibits bacterial growth by degrading mRNA or rRNA sequences. The toxin genes are organized in pairs with its cognate antitoxins in the chromosome and thus the activities of the toxins are antagonized by antitoxin proteins or RNAs during active translation. In response to a variety of cellular stresses, the endoribonuclease toxins appear to be released from antitoxin molecules via proteolytic cleavage of antitoxin proteins or preferential degradation of antitoxin RNAs and cleave a diverse range of mRNA or rRNA sequences in a sequence-specific or codon-specific manner, resulting in various biological phenomena such as antibiotic tolerance and persister cell formation. Given that substrate specificity of each endoribonuclease toxin is determined by its structure and the composition of active site residues, we summarize the biology, structure, and substrate specificity of the updated bacterial endoribonuclease toxins.

In vivo anti-metastatic action of Ginseng Saponins is based on their intestinal bacterial metabolites after oral administration

  • Saiki, Ikuo
    • Journal of Ginseng Research
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    • v.31 no.1
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    • pp.1-13
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    • 2007
  • We found that the main bacterial metabolite M1 is an active component of orally administered protopanxadiol-type ginsenosides, and that the anti-metastatic effect by oral administration of ginsenosides may be primarily mediated through the inhibition of tumor invasion, migration and growth of tumor cells by their metabolite M1. Pharmacokinetic study after oral administration of ginsenoside Rb1 revealed that M1 was detected in serum for 24 h by HPLC analysis but Rb1 was not detected. M1, with anti-metastatic property, inhibited the proliferation of murine and human tumor cells in a time- and concentration-dependent manner in vitro, and also induced apoptotic cell death (the ladder fragmentation of the extracted DNA). The induction of apoptosis by M1 involved the up-regulation of the cyclin-dependent kinase(CDK) inhibitor $p27^{Kip1}$ as well as the down-regulation of a proto-oncogene product c-Myc and cyclin D1 in a time-dependent manner. Thus, M1 might cause the cell-cycle arrest (G1 phase arrest) in honor cells through the up/down-regulation of these cell-growth related molecules, and consequently induce apoptosis. The nucleosomal distribution of fluorescence-labeled M1 suggests that the modification of these molecules is induced by transcriptional regulation. Tumor-induced angiogenesis (neovascularization) is one of the most important events concerning tumor growth and metastasis. Neovascularization toward and into tumor is a crucial step for the delivery of nutrition and oxygen to tumors, and also functions as the metastatic pathway to distant organs. M1 inhibited the tube-like formation of hepatic sinusoidal endothelial (HSE) cells induced by the conditioned medium of colon 26-L5 cells in a concentration-dependent manner. However, M1 at the concentrations used in this study did not affect the growth of HSE cells in vitro.

THE EFFECTS OF HONOKIOL AND MAGNOLOL ON THE ANTIMICROBIAL, BACTERIAL COLLAGENASE ACTIVITY, CYTOTOXICITY AND CYTOKINE PRODUCTION (Magnolol과 Honokiol이 항균, 교원질 분해효소, 세포독성 및 Cytokine생산에 미치는 영향)

  • Jang, Beom-Seok;Son, Seong-Heai;Chung, Chong-Pyoung;Bae, Ki-Hwan
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.145-158
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    • 1993
  • The oral microbiota such as P. gingivalis, P. intermedia and A. actinomycetemcomitans play a primary role in the initiation and progression of the periodontal disease. The purpose of this study was to evaluate the antimicrobial effects and inhibitory effects of honokiol and magnolol on the bacterial collagenase activity, cytotoxicity and cytokine production of periodontopathic microorganisms. The antimicrobial activities of honokiol and magnolol was evaluted with minimum inhibition concentration. Honokiol was more active than magnolol, but less than chlorhexidine on antimicrobial activity. The inhibitory effects of magnolol and honokiol on the collagenolytic activity and cytotoxicity were evaluated using a Collagenokit CLN-100 and rapid colorimetric assay (MTT method) for cellular growth and survival of gingival fibroblast and periodontalligament cell and $[^3H]-thymidine$ incorporation for the gingival epithelial cell. The inhibitory effects on the collagenolytic activity was the highest in chlorhexidine, and the lowest in magnolol. Magnolol had the lowest cytotoxic effect and chlorhexidine had the highest. The inhibitory effects on cytokine production was evaluated using $interleukin-1{\beta}$ ELISA kit (Cistron Biotech.), IL-6, $TNF-{\alpha}$ ELISA kit (Genzyme) and inhibitory effects were higher than bacterial LPS and there is no difference among the honokiol, magnolol and chlorhexidine. From these results, the antimicrobial and antienzymatic activities of honokiol and magnolol were seemed to inhibit bacterial growth and enzyme activities with lesser cytotoxic activities. Therefore, it was suggested that honokiol and magnolol are very effective antimicrobial agents on periodontal pathogens.

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Effect of GlycinecinA on the Control of Bacterial Leaf Spot of Red Pepper and Bacterial Leaf Blight of Rice

  • Jeon, Yong-Ho;Moonjae Cho;Cho, Yong-Sup;Ingyu Hwang
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.249-256
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    • 2001
  • Xanthomonas axonopodis pv. glycines 8ra produces a bacteriocin called glycinecinA, which specifically inhibits the growth of bacteria belonging to Xanthomonas species. GlycinecinA was produced by culturing Escherichia coli DH5 containing biosynthetic genes for glycinecinA, and was tested for its control effect against X. vesicatoria on red pepper and X. oryzae pv. oryzae on rice. The bacteriocin activity was much higher in the cell extract than in the supernatant. It reached a maximum level at the stationary phase, ws maintained up to 2 months at room temperature and approximately 10 months at $4^{\circ}$. The optimum concentration of glycinecinA for the control in the greenhouse and in the field was 12,800 AU/ml. In this study, the activity of glycinecinA on rice and red pepper leaves continued for 7-8 days, during which the pathogen populations remained at low levels. Bacterial leaf spot of red pepper and bacterial leaf blight of rice were significantly reduced by the bacteriocin treatments. The control efficacy was as high as, or even higher than, the chemical treatment of copper hydroxide. These results suggest that the bacteriocin is a potential control agent for bacterial diseases.

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Fermentation of a Potential Biocontrol Agent, Bacillus amyloliquefaciens SKU-78 Strain (풋마름병균의 길항세균 Bacillus amyloliquefaciens SKU-78의 대량 배양 조건 확립)

  • Kim, Shin-Duk;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.84-86
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    • 2014
  • Mass production of biocontrol agent is an essential step for its commercial use. Media composition and culture conditions for production of Bacillus amyloliquefaciens SKU-78, a potential biocontrol agent against bacterial wilts, were optimized by a flask culture. Low cost media combining nitrogen and carbon sources were tested. Maximum cell growth (> $2{\times}10^9$ CFU/ml) was obtained in a medium of 5% soy flour combined with 3% corn starch after 24 h cultivation. The optimum initial pH, temperature and shaking speed was 5.5, $30^{\circ}C$ and 150-250 rpm, respectively. Fermentation of SKU-78 was scaled up in 30 L fermenter and the profiles of cell density, pH, dissolved oxygen and spore formation were recorded. After 8 h lag phase, exponential growth occurred and reached at maximum viable cell number ($1.2{\times}10^{11}$ CFU/ml) after 20 h. The SKU-78 strain grown in a low cost medium exhibited the high suppression of bacterial wilts. The results indicate that SKU-78 strain can be produced in a low cost medium and provide a basis for scaling up to industrial level.

Evaluation on Antimicrobial Activity of Psoraleae semen Extract Controlling the Growth of Gram-Positive Bacteria

  • Kim, Sejeong;Lee, Soomin;Lee, Heeyoung;Ha, Jimyeong;Lee, Jeeyeon;Choi, Yukyung;Oh, Hyemin;Hong, Joonbae;Yoon, Yohan;Choi, Kyoung-Hee
    • Food Science of Animal Resources
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    • v.37 no.4
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    • pp.502-510
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    • 2017
  • This study investigated bacterial growth-inhibitory effect of 69 therapeutic herbal plants extracts on 9 bacterial strains using a disc diffusion assay. Especially, the antimicrobial activity of Psoraleae semen, which showed different activity on pathogenic Gram-positive and Gram-negative bacteria, was evaluated by MIC (minimal inhibition concentration) and biofilm formation assay. The effect of Psoraleae semen extract on bacterial cell membranes was examined by measurement of protein leakage (optical density at 280 nm) and scanning electron microscope (SEM). No clear zone was formed on discs containing Gram-negative bacteria, but Gram-positive bacteria exhibited clear zones. The MICs of Psoraleae semen extract were $8{\mu}g/mL$ for Streptococcus mutans, and $16{\mu}g/mL$ for Enterococci and Staphylococcus aureus. In addition, biofilm formation was inhibited at concentration $8-16{\mu}g/mL$. Protein leakage values and SEM images revealed that cell membranes of Gram-positive bacteria were impaired following exposure to the extract. Further, the extract inhibited the growth of Listeria monocytogenes in sausages. These results indicate that Psoraleae semen extract could be utilized as a natural antimicrobial agent against Gram-positive bacteria.

Anti-Oomycete Activity and Pepper Root Colonization of Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 against Phytophthora capsici

  • Elena, Volynchikova;Ki Deok, Kim
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.123-135
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    • 2023
  • Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO4 solution (untreated control). In diluted (nutrient-deficient) V8 juice broth, the tested strain populations were maintained at >108 cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 106 cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.

Artificial Induction of Environmental Mammary Stress by Temperature and Micro-organism Causing Mastitis and Modulation of Mammary Growth by Adenosine, IGF-I and Prolatin In Vitro (In Vitro내 유선조직에의 인위적인 온도 및 유방염 발생 미생물에 의한 환경스트레스 유기와 Adenosine, IGF-I 및 Prolactin에 의한 성장조절작용)

  • 정석근;장병배;이창수;박춘근;홍병주;여인서
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.325-333
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    • 1997
  • Recent evidence indicates that growth factors modulate response of mammary epithelial cells to environmental stress. The objective of this study was to examine the cellular and biochemical responses of mammary tissue to environmental stress caused by artificial mastitis. For experimental a, pp.oach, toxins of most mastitis causing organisms(Staph. aureus or Strep. agalactiae) and heat stress(42$^{\circ}C$) were artificially exposed to mammary tissue. Effects of these environmental stresses on cell growth, cell death and heat shock protein synthesis were examined. Lactating mammary tissure were cultured under basal medium(DMEM) su, pp.emented with insulin(10$\mu\textrm{g}$/ml) and aldosterone(1$\mu\textrm{g}$/ml). All treatment groups in heat stress at 42$^{\circ}C$ incubation significantly decreased DNA synthesis rates in comparison with those at 39$^{\circ}C$(P<0.05), however, these decreased DNAa synthesis rates were recovered by addition of adenosine(10$\mu$M) and IGFI(10ng/ml). Similar results were obtained when tissue growth rates were measured by DNA content/tissue. Strep. agalactiae toxin did not significantly decreased DNA content/tissue in comparison with no treatment of bacterial toxin with or without heat stress, however, tended to decrease DNA contents/tissue without heat stress. In the fluorography analysis, heat stress(42$^{\circ}C$ incubation) slightly increased 35S-methoionine labelled 70kd protein synthesis. These results indicate that environmental stress caused by artificial mastitis slightly decreased mammary growth or mammary size, however, these results could be recovered by addition of adenosine and IGF-I.

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Bacterial determinants involved in the induction of systemic resistance ana plant growth promotion in tobacco by Pseudomonas chlororaphis O6.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.101.2-102
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    • 2003
  • The ability of P. chlororaphis O6 to induce resistance to Erwinia carotovora subsp. carotovara SCCI and to promote growth in tobacco was demonstrated in microtiter assays on plants pre-inoculated at the root level with the bacteria before challenge with the leaf pathogen. To identify th bacterial determinants involved in induced systemic resistance and plant growth promotion, cell culture of O6 grown in King's medium B was fractionated with organic solvents and purified using various columns. in vivo and in vitro assays with samples from successive fractionation steps of the O6 supernatant led to the conclusion that antibacterial compounds were observed in aqueous layer, and to the isolation of fractions containing metabolites that retained most of the resistance-inducing activity (70:30, methanol:water) and the plant growth promotion (80:20 and 90:10, methanol:water) after ODS column chromatography. Although these molecules remain to be purified further and structurally characterized, its isolation is an addition to the range of determinants from plant growth-promoting rhizobacteria known to stimulate plant defence.

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