• Title/Summary/Keyword: intracellular survival

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Quorum-Sensing Mechanisms in Bacterial Communities and Their Potential Applications (세균의 의사 소통(Quorum-Sensing) 기구와 그 잠재적 응용성)

  • Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.402-409
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    • 2006
  • Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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Effects of Chrysanthemum indicum L. Extract on the Function of Osteoblastic MC3T3-E1 Cells under Oxidative Stress Induced by Hydrogen PeroxideJee (감국(Chrysanthemum indicum L.) 추출물이 H2O2로 유도한 산화적 스트레스에서 MC3T3-E1 조골세포 기능에 미치는 영향)

  • Yun, Jee-Hye;Hwang, Eun-Sun;Kim, Gun-Hee
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.82-88
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    • 2012
  • Chrysanthemum indicum L. (Asteraceae) is a traditional herbal medicine that has been used for the treatment of inflammation, hypertension, and respiratory diseases due to its strong antagonistic activity against inflammatory cytokines. The effects of Chrysanthemum indicum L. Extract (CIE) for increasing cell growth, alkaline phosphatase (ALP) activity, and collagen content were totally inhibited, suggesting that the effect of CIE might be partly involved with estrogen activity. Furthermore, the protective effects of CIE on the response of osteoblasts to oxidative stress were evaluated. Osteoblastic MC3T3-E1 cells were incubated with hydrogen peroxide and/or CIE, and markers of osteoblast function and oxidative damage were examined. CIE significantly increased cell survival, ALP activity, and calcium deposition, and decreased the production of Reactive Oxygen Species (ROS) and Tumor Necrosis Factor-${\alpha}$ (TNF-${\alpha}$) in osteoblasts. Taken together, these results indicate that the enhancement of osteoblast function by CIE may prevent osteoporosis and inflammatory bone diseases.

Antioxidant Properties and Protective Effects of Inula britannica var. chinensis Regel on Oxidative Stress-induced Neuronal Cell Damage (금불초 추출물의 항산화 효과 및 산화 스트레스에 대한 신경세포 보호작용)

  • Lee, Na-Hyun;Hong, Jung-Il;Kim, Jin-Yung;Chiang, Mae-Hee
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.87-92
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    • 2009
  • The antioxidant properties and protective effects of Inula britannica on ${H_2}{O_2}$-induced SH-SY5Y neuroblastoma cell damage were investigated. A series of solvent fractions, including hexane(Fr.H), petroleum ether, chloroform, ethyl acetate(Fr.EA), and water fraction(Fr.W), were prepared from the 70% methanol extracts of Inula britannica. Fr.W had the highest total contents of phenolics and flavonoids, followed by Fr.EA. The antioxidant properties of the fractions were also evaluated by analyzing their scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH) radicals, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, and nitric oxide. Fr.W showed the strongest activities in all assays. The concentrations of Fr.W that resulted in 50% reductions of the DPPH and ABTS radicals were 20.7 ${\mu}g$/mL and 39.4 ${\mu}g$/mL, respectively. Fr.W showed the weakest cytotoxic activities on the SH-SY5Y cells, whereas it effectively protected ${H_2}{O_2}$-induced cell death, increasing cell survival by 35.0-77.0% at a concentration range of 62.5-250 ${\mu}g$/mL. In this range, Fr.W also significantly decreased intracellular ROS levels by 34-39%. Overall, the antioxidant properties of Inula britannica can contribute to rescuring neuronal cells from oxidative stress-induced cell injury.

Cloning of cDNA Encoding Putative Cellular Receptor Interacting with E2 protein of Hepatitis C Virus (C형 간염바이러스 E2 단백질에 결합하는 추정 세포수용체 cDNA의 클로닝)

  • 이성락;백재은;석대현;박세광;최인학
    • Journal of Life Science
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    • v.13 no.4
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    • pp.541-550
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    • 2003
  • E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

Effect of Water Temperature on Infectivity of the Parasitoid Amoebophrya sp. Infecting the Harmful Bloom-forming Dinoflagellate Akashiwo sanguinea (유해 적조생물 Akashiwo sanguinea를 감염시키는 포식성 기생생물 Amoebophrya sp.의 감염력에 대한 수온의 영향)

  • JUNG, YOUNGGYO;KIM, SUNJU
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.23 no.1
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    • pp.20-31
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    • 2018
  • Marine parasitoid Amoebophrya infects and kills various bloom-forming dinoflagellates and strongly influences the harmful algal bloom dynamics. We investigated the effect of temperature on survival, infectivity, generation time of the parasite from the parasitoid Amoebophrya sp. and the harmful dinoflagellate host Akashiwo sanguinea system. Temperature had a significant effect on the parasite generation time and infectivity. While the lower temperature ($15^{\circ}C$) arrested parasite intracellular development and infectivity, resulting in the longer generation time ($115{\pm}0.1h$), the higher temperatures ($25^{\circ}C$ and $20^{\circ}C$) accelerated the parasite development, with the generation times of $58{\pm}0.1h$ and $83{\pm}0.1h$, respectively. Parasite prevalence (percent of host infected) was $71.5{\pm}0.30%$, $54.3{\pm}1.68%$, and $29.6{\pm}1.42%$ at $25^{\circ}C$, $20^{\circ}C$, and $15^{\circ}C$, respectively. These results suggest that biological control by parasitism on A. sanguinea bloom would not be highly effective during low water temperature season. Further, water temperature would be an important factor of bottom-up controls for the host-parasite population dynamics.

The effects of $\beta$-1,3/1,6-linked glucan in the diet on immune responses of olive flounder, Paralichthys olivaceus by oral admimistration ($\beta$-1,3 / 1,6-linked glucan 첨가사료가 넙치의 비특이적 면역반응에 미치는 영향)

  • Won, Gyeong-Mi;Kim, Su-Mi;Park, Su-Il
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.29-38
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    • 2004
  • The effects of dietary $\beta$-1,3/1,6-linked glucan administration on nonspecific immune responses, hematology and disease resistance against Edwardsiella tarda in olive flounder, Paralichthys olivaceus were evaluated. Fish were fed the experimental diets supplemented with 0 %, 0.01 %, 0.05 %, 0.1 %, and 0.5 % of $\beta$-glucan to a commercial diet for 7 weeks. The test was performed in two ways such as fed on each $\beta$-glucan concentrations daily and every other week alternately. The body weight gain from the fish fed on daily the 0.05 % supplemented diet and fed on alternately the 0.1 % supplemented diet of $\beta$-glucan were significantly higher than the control. Both lysozyme activity and intracellular superoxide anion production of kidney phagocytes were higher in the all experimental groups than in the control. But there was no large difference in hematology among each group. The relative percent survival rate (RPS) after an artificial challenge with $4{\times}10^6$ cells of E. tarda per fish was higher than the control, except for that fed on daily the 0.01 % supplemented diet.

Remifentanil induces autophagy and prevents hydrogen peroxide-induced apoptosis in Cos-7 cells

  • Yoon, Ji-Young;Baek, Chul-Woo;Woo, Mi-Na;Kim, Eun-Jung;Yoon, Ji-Uk;Park, Chang-Hoon
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.16 no.3
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    • pp.175-184
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    • 2016
  • Background: This study investigated the effect of remifentanil pretreatment on Cos-7 cells exposed to oxidative stress, and the influence of remifentanil on intracellular autophagy and apoptotic cell death. Methods: Cells were divided into 4 groups: (1) Control: non-pretreated cells were incubated in normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$). (2) $H_2O_2$: non-pretreated cells were exposed to $H_2O_2$ for 24 h. (3) RPC+$H_2O_2$: cells pretreated with remifentanil were exposed to $H_2O_2$ for 24 h. (4) 3-MA+RPC+$H_2O_2$: cells pretreated with 3-Methyladenine (3-MA) and remifentanil were exposed to $H_2O_2$ for 24 h. We determined the cell viability of each group using an MTT assay. Hoechst staining and FACS analysis of Cos-7 cells were performed to observe the effect of remifentanil on apoptosis. Autophagy activation was determined by fluorescence microscopy, MDC staining, and AO staining. The expression of autophagy-related proteins was observed using western blotting. Results: Remifentanil pretreatment increased the viability of Cos-7 cells exposed to oxidative stress. Hoechst staining and FACS analysis revealed that oxidative stress-dependent apoptosis was suppressed by the pretreatment. Additionally, fluorescence microscopy showed that remifentanil pretreatment led to autophagy-induction in Cos-7 cells, and the expression of autophagy-related proteins was increased in the RPC+$H_2O_2$ group. Conclusions: The study showed that remifentanil pretreatment stimulated autophagy and increased viability in an oxidative stress model of Cos-7 cells. Therefore, we suggest that apoptosis was activated upon oxidative stress, and remifentanil preconditioning increased the survival rate of the cells by activating autophagy.

The enhancing effect of Acanthopanax sessiliflorus fruit extract on the antibacterial activity of porcine alveolar 3D4/31 macrophages via nuclear factor kappa B1 and lipid metabolism regulation

  • Hwang, Eunmi;Kim, Gye Won;Song, Ki Duk;Lee, Hak-Kyo;Kim, Sung-Jo
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1776-1788
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    • 2019
  • Objective: The demands for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). Methods: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells ($M{\Phi}$) were activated by phorbol 12-myristate 13-acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B ($NF-{\kappa}B$) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative polymerase chain reaction was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 $M{\Phi}$ cells was evaluated by the colony forming unit assay. Results: ASF upregulated the cell viability and growth rate of 3D4/31 $M{\Phi}$, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of $NF-{\kappa}B$ protein, tumor necrosis factor $(TNF){\alpha}$ mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of $NF-{\kappa}B$, $TNF{\alpha}$, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 $M{\Phi}$ against Escherichia coli. Conclusion: This study provides a novel insight into the regulation of $NF-{\kappa}B$ activity and lipid metabolism in $M{\Phi}$, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

Relative Association of Overhydration and Muscle Wasting with Mortality in Hemodialysis Patients: Assessment by Bioelectrical Impedance Analysis (혈액투석 환자에서 Bioelectrical Impedance Analysis를 활용하여 측정한 과수분량과 근육량 감소와 사망률의 상관관계)

  • Kim, Eunju;Seo, Sang Oh;Choi, Yu Bum;Lee, Mi Jung;Lee, Jeong Eun;Kim, Hyung Jong
    • The Korean Journal of Medicine
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    • v.93 no.6
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    • pp.548-555
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    • 2018
  • Background/Aims: Assessment of fluid status in hemodialysis patents is very important. Overhydration in hemodialysis is associated with generalized edema, cardiovascular complications, and hypertension. The aim of this study was to determine the factors correlated with mortality of hemodialysis patients, assessing body muscle mass and fluid status using bioelectrical impedance analysis (BIA). Methods: This study enrolled 93 patients who underwent hemodialysis between January 2010 and May 2015 at CHA Bundang Medical Center. Medical records of enrollees up to June 2017 were reviewed retrospectively. These included laboratory results (serum albumin, C-reactive protein [CRP], lipid profile, etc.) and BIA data (extracellular water, intracellular water, total body water, soft lean mass, fat free mass, skeletal muscle mass, etc.). Results: Eleven of 93 patients had expired by May 2017. Among the surviving subjects, mean age was younger, CRP levels were lower, albumin levels were higher, and extracellular water/total body water (ECW/TBW) ratios were lower than in the expired patient group. Kaplan-Meier survival analysis revealed that overhydration (ECW/TBW > 0.4) was associated with higher mortality. Conclusions: In hemodialysis patients, overhydration is an important factor in mortality, and BIA could be a reliable modality in its assessment. We suggest that, for hemodialysis patients, overhydration is more of a risk factor for mortality than is muscle wasting.