• Title/Summary/Keyword: Extracellular $K^+$

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Role of flgA for Flagellar Biosynthesis and Biofilm Formation of Campylobacter jejuni NCTC11168

  • Kim, Joo-Sung;Park, Changwon;Kim, Yun-Ji
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1871-1879
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    • 2015
  • The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

Fluvastatin inhibits advanced glycation end products-induced proliferation, migration, and extracellular matrix accumulation in vascular smooth muscle cells by targeting connective tissue growth factor

  • Hwang, Ae-Rang;Nam, Ju-Ock;Kang, Young Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.2
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    • pp.193-201
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    • 2018
  • Connective tissue growth factor (CTGF) is a novel fibrotic mediator, which is considered to mediate fibrosis through extracellular matrix (ECM) synthesis in diabetic cardiovascular complications. Statins have significant immunomodulatory effects and reduce vascular injury. We therefore examined whether fluvastatin has anti-fibrotic effects in vascular smooth muscle cells (VSMCs) and elucidated its putative transduction signals. We show that advanced glycation end products (AGEs) stimulated CTGF mRNA and protein expression in a time-dependent manner. AGE-induced CTGF expression was mediated via ERK1/2, JNK, and Egr-1 pathways, but not p38; consequently, cell proliferation and migration and ECM accumulation were regulated by CTGF signaling pathway. AGE-stimulated VSMC proliferation, migration, and ECM accumulation were blocked by fluvastatin. However, the inhibitory effect of fluvastatin was restored by administration of CTGF recombinant protein. AGE-induced VSMC proliferation was dependent on cell cycle arrest, thereby increasing G1/G0 phase. Fluvastatin repressed cell cycle regulatory genes cyclin D1 and Cdk4 and augmented cyclin-dependent kinase inhibitors p27 and p21 in AGE-induced VSMCs. Taken together, fluvastatin suppressed AGE-induced VSMC proliferation, migration, and ECM accumulation by targeting CTGF signaling mechanism. These findings might be evidence for CTGF as a potential therapeutic target in diabetic vasculature complication.

Protective actions of Rubus coreanus ethanol extract on collagenous extracellular matrix in ultraviolet-B irradiation-induced human dermal fibroblasts

  • Bae, Ji-Young;Lim, Soon-Sung;Choi, Jung-Suk;Kang, Young-Hee
    • Nutrition Research and Practice
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    • v.1 no.4
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    • pp.279-284
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    • 2007
  • Solar ultraviolet (UV) irradiation leads to distinct changes in the skin connective tissues by degradation of collagen, which is a major structural component in the extracellular matrix. UV irradiation induces the production of matrix metalloproteinases (MMP) capable of attacking native fibrillar collagen and responsible for inhibiting the construction of collagenous extracellular matrix. In this study, we attempted to investigate the protective actions of Rubus coreanus ethanol extract (RCE) on the MMP production and the consequent procollagen/collagen degradation in UV-B-irradiated human dermal fibroblasts. The analytical data showed that Rubus coreanus ethanol extract was mostly comprised of cyanidin 3-rutinoside. Pre-treatment of fibroblasts with this extract inhibited UV-B-induced production of MMP-1, MMP-8 and MMP-13 in dose-dependent manners. In addition, Western blot analysis and immunocytochemical staining assay revealed that RCE markedly augmented the cellular levels of procollagen/collagen declined in UV-B-exposed dermal fibroblasts. These results demonstrate that RCE blocks UV-B-induced increase of the collagen degradation by inhibiting MMP production. Thus, RCE may act as an agent inhibiting excessive dermal collagen degradation leading to the skin photoaging.

CLIP-domain serine proteases in Drosophila innate immunity

  • Jang, In-Hwan;Nam, Hyuck-Jin;Lee, Won-Jae
    • BMB Reports
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    • v.41 no.2
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    • pp.102-107
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    • 2008
  • Extracellular proteases play an important role in a wide range of host physiological events, such as food digestion, extracellular matrix degradation, coagulation and immunity. Among the large extracellular protease family, serine proteases that contain a "paper clip"-like domain and are therefore referred to as CLIP-domain serine protease (clip-SP), have been found to be involved in unique biological processes, such as immunity and development. Despite the increasing amount of biochemical information available regarding the structure and function of clip-SPs, their in vivo physiological significance is not well known due to a lack of genetic studies. Recently, Drosophila has been shown to be a powerful genetic model system for the dissection of biological functions of the clip-SPs at the organism level. Here, the current knowledge regarding Drosophila clip-SPs has been summarized and future research directions to evaluate the role that clip-SPs play in Drosophila immunity are discussed.

Endothelial Cell Seeding onto Extracellular Matrix for Development of Polyurethane Vascular Prosthesis (폴리우레탄 인공혈관을 위한 extracellular matrix 기질상의 내피세포이식)

  • Lee, Yoon-Shin;Park, Dong-Kook;Min, Byoung-Goo
    • Journal of Biomedical Engineering Research
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    • v.12 no.3
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    • pp.165-170
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    • 1991
  • Many experiments about endothelial cell seeding on artificial vessels were studied and conducted For this one or a combination of the extramatrix was used for the underlying matrix. But we used the whole ECM(extracellular matrix) that made excreated from flbroblasl. In thls study, we obtained human adult omental microvascular endothelium by collagenase digestion and used polyurthane sheets in order to make a new artificial vessel material. We cultured fibroblast on the polyurethane and gelatin - coated polyurethane. After confluent ingrowth we treated the polyure thane with triton in order to destroy the cytoskeleton and nucleus. We observed the preformed extra cellular matrix on the ployurethane and cultured the isolated microvascular endothelium. We also ok served the growth of microvascular endothelium on the polyurethane and gelatin. We conclude that the use of the whole ECM is promising fair as a new underying substrate for endothelial cell seeding on artificial vessels.

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Extracellular Matrix Metalloproteinase Inducer is Regulated Developmentally and Functionally in the Rat Submandibular Gland

  • Yoo, Hong-Il;Suh, Han-Young;Kim, Sun-Hun
    • International Journal of Oral Biology
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    • v.40 no.3
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    • pp.127-133
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    • 2015
  • The salivary gland undergoes complex process of growth and differentiation of the branching morphogenesis of ductal system during the prenatal and early postnatal periods which are regulated by various elements in the extracellular matrix. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule. In the present study, localization and expression of EMMPRIN in development and effects of chorda-lingual denervation and cyclosporine A (CsA) treatment on the EMMPRIN expression were investigated. Immunohistochemistry, RT-PCR and Western blot were used to determine expression level. Immunohistochemistry revealed that EMMPRIN was localized specifically in the cytoplasm of ductal cells, not acini of the submandibular gland all the postnatal periods. At prenatal day 18, when the formation of ducts was not definite, no immunoreactivity was observed. Both Western blot and RT-PCR analyses revealed that EMMPRIN expression was maintained up to postnatal day 7, decreased after postnatal day 10. The EMMPRIN expression was upregulated by the surgical denervation of the chorda-lingual nerve in the gland as well as by the CsA treatment. The present study suggests that EMMPRIN is a crucial molecule for maintaining physiological functions of the salivary gland.

Characterization of Extracellular Protease Secreted from Chryseobacterium sp. JK1 (Chryseobacterium sp. JK1이 분비하는 세포외 단백질분해효소 특성)

  • Lee, Yu-Kyong;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.78-82
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    • 2013
  • A novel Chryseobacterium sp. JK1 strain isolated from soil had been reported that this isolate produced large amount of extracellular protease at mesophilic temperature in previous study. The optimal temperature and pH of extracellular protease were $40^{\circ}C$ and 7.0, respectively, showing narrow range of optimal temperature and relatively broad activity from pH 6.0 to 9.0. In addition, the protease showed greatest activity against skim milk and lowest against bovine serum albumin (BSA). The protease strongly inhibited by ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA) or phenylmethylsulfonyl fluoride (PMSF), and addition of cation $Ag^+$ or $Cu^{2+}$, and slightly inhibited by $Al^{3+}$. No significant inhibition was found with pepstatin, and addition of cation, $K^+$, $Ca^{2+}$, $Na^+$, $Fe^{2+}$ or $Mg^{2+}$. On the contrary, protease was enhanced by addition of divalent cation $Mn^{2+}$ (5 mM). Zymography analysis of concentrated culture supernatant revealed two major bands at 67 and 145 kDa. These results suggest that Chryseobacterium sp. JK1 strain produced extracellular neutral serine proteases which could apply in food industry.

Effect of pH on the Vascular Tone and $^{45}Ca$ Uptake in the Aorta of Spontaneously Hypertensive Rats

  • Chang, Seok-Jong;Jeon, Byeong-Hwa;Kim, Se-Hoon;Kim, Hoe-Suk;Park, Hae-Kun
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.169-179
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    • 1994
  • The effect of extracellular and intracellular pH on vascular tone and $^{45}Ca$ uptake were investigated in aortic strips and dispersed single aortic smooth muscle cells of spontaneously hypertensive rats (SHR) and aged-matched Wistar-Kyoto rats (WKY). The contraction produced by a change of extracellular pH (pHo) in the range of $6.5{\sim}8.3$ was estimated by comparison with the level of vascular tone at pH 7.4. Contraction was induced below pHo 6.5 in WKY, pHo 7.1 in SHR, and over pHo 8.0 on both strains. The amplitude of contraction induced by high pHo (over pHo 7.7) was similar in SHR and WKY, but that induced by low pHo (below pHo 7.1) in SHR was greater than that in WKY. Either high pHo- or low pHo-induced contractions in WKY and SHR were not induced in the Ca-free Tyrode's solution and were induced by the addition of Ca. $^{45}Ca$ uptake increased progressively as pHo was increased from 6.8 to 8.1 in the single aortic smooth muscle cells of WKY and SHR. $NH_4Cl$ induced a gradually developing contraction in a dose-dependent manner $(5\;mM{\sim}30\;mM)$ and the removal of $NH_4Cl$ induced transient contraction was followed by profound relaxation in the aortic rings of both strains. The contractions induced by $NH_4Cl$ or by the removal of $NH_4Cl$ in SHR were significantly greater than that in WKY. These contractions were not induced in Ca-free Tyrode's solution. $^{45}Ca$ uptake was increased by $NH_4Cl$ (20 mM) and was not changed by the removal of $NH_4Cl$ (20 mM) in the aortic strips of WKY and SHR. As a summary of above results, the vascular tone of SHR was more sensitive to the change pHi and pHo than that of WKY. The contractions induced by change of extracellular or intracellular pH depended on extracellular Ca in the aorta of SHR nnd WKY. However, the Ca uptake was in accord with the changes of contraction but increase in contraction by low pH was not accompanied by an increase in Ca uptake in both strains.

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Adsorption of Heavy Metal onto the Extracellular Polysaccharide Produced by the Purple Nonsulfur Photosynthetic Bacteria Rhodopseudomonas sp. KH4 (홍색 비황 광합성 세균 Rhodopseudomonas sp. KH4의 Extracellular polysaccharide의 중금속 흡착)

  • Jeong, Jeong-Hwa;Seo, Pil-Soo;Kong, Sung-Ho;Lee, Jong-Yeol;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.326-331
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    • 2006
  • In the present study, we examined biosorption characteristics of heavy metals onto the extracellular polysaccharide (EPS) produced by the purple nonsulfur photosynthetic bacteria Rhodopseudomonas sp. KH4, which was isolated from a stream in Anyang, Kyonggi-Do. When Cd (100 mg/L) and Cu (100 mg/L) were added to EPS (1.0 g/L) in the optimal condition (Cd; pH 8, Cu; pH 5, $40^{\circ}C$), 84.2 mg/L of Cd and 70.0 mg/L of Cu were adsorbed within 30 min and 10 min, respectively. When 100 mg/L of Cd and Cu were present as mixture, 16.8 mg/L of Cd and 48.7 mg/L of Cu were adsorbed at $25^{\circ}C$, pH 5. The maximum adsorption capacity determined by fitting Langmuir isotherms model was suitable for describing the biosorption of Cd (76.9 mg/g) and Cu (67.1 mg/g) by EPS. The neutral monosaccharide in the EPS determined by GC consisted of arabinose (2.4%), glucose (7.1%) and mannose (90.5%).

Assessment of the Ability of Extracellular Enzyme Production in Hybrid Strains of Lentinula edodes by Chromogenic Reaction-based Plate Assay (발색반응 분석법을 이용한 표고 교배균주의 세포외효소 분비 능력 평가)

  • Kwon, Hyuk-Woo;Kim, Jun-Young;Ko, Han-Gyu;Park, Heung-Soo;Kim, Seong-Hwan
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.99-104
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    • 2011
  • Shiitake breeding requires the procedures of mating of two different parental strains and selection of hybrid strains that have good traits for the mushroom production. In this study, we tested the possibility of the use of chromogenic plate-based assay for extracellular enzyme production in order to assess and find good biochemical properties-possessed hybrid strains that were generated from genetic cross of the monokaryotic strains derived from two different parental strains of Lentinula edodes Sanjo-101ho and Sanjo-108ho. We observed that there was difference in the ability of producing ${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease among the monokaryotic strains. We could also comparatively assess that the ability of the seven extracellular enzymes production in the hybrid strains depended on the mating combination of the monokaryotic strains. Our results demonstrate that the assessment method for extracellular enzyme production using chromogenic plate assay could be usefully applied to the assessment of the hybrid strains derived from the breeding procedure of L. edodes.