• Title/Summary/Keyword: extracellular

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Influence of Extracellular Products from Haematococcus pluvialis on Growth and Bacteriocin Production by Three Species of Lactobacillus

  • Kim Jeong-Dong;Lee Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.849-854
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    • 2006
  • The effects of Haematococcus pluvialis extracellular products on microbial growth and bacteriocin production were investigated to improve bacteriocin synthesis during the growth cycle of Lactobacilli. Lactobacillus pentosus KJ-108, L. plantarum KJ-10311, and L. sakei KJ-2008 were cultured in MRS and enriched medium (ERM) with or without supplement of the extracellular products obtained from a late exponential phase culture of Haematococcus pluvialis in modified Bold's basal medium (MBBM). In both MRS and ERM, the extracellular products strongly enhanced the growth as well as the bacteriocin production of all the lactic acid bacteria tested. The enhancing effect was observed in ERM with pH adjusted at 5 and 6. In addition, some difference in growth effects with the extracellular products of H. pluvialis was observed between pH 5 and 6 in ERM, but no effect was observed in the minimal medium. The final biomass and the final concentration of bacteriocin activity were associated with the cell growth that was promoted by the extracellular products of H. pluvialis, and the enhanced cell growth of the three lactic acid bacterial strains induced the increase of the specific bacteriocin production. Therefore, bacteriocin production and activity were influenced by the addition of the extracellular products of H. pluvialis in the culture medium.

Extracellular vesicles as emerging intercellular communicasomes

  • Yoon, Yae Jin;Kim, Oh Youn;Gho, Yong Song
    • BMB Reports
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    • v.47 no.10
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    • pp.531-539
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    • 2014
  • All living cells release extracellular vesicles having pleiotropic functions in intercellular communication. Mammalian extracellular vesicles, also known as exosomes and microvesicles, are spherical bilayered proteolipids composed of various bioactive molecules, including RNAs, DNAs, proteins, and lipids. Extracellular vesicles directly and indirectly control a diverse range of biological processes by transferring membrane proteins, signaling molecules, mRNAs, and miRNAs, and activating receptors of recipient cells. The active interaction of extracellular vesicles with other cells regulates various physiological and pathological conditions, including cancer, infectious diseases, and neurodegenerative disorders. Recent developments in high-throughput proteomics, transcriptomics, and lipidomics tools have provided ample data on the common and specific components of various types of extracellular vesicles. These studies may contribute to the understanding of the molecular mechanism involved in vesicular cargo sorting and the biogenesis of extracellular vesicles, and, further, to the identification of disease-specific biomarkers. This review focuses on the components, functions, and therapeutic and diagnostic potential of extracellular vesicles under various pathophysiological conditions.

Purification and Characterization of Intracellular and Extracellular Inulase from Kluyveromyces marxianus (Kluyveromyces marxianus 가 생산하는 Intracellular 및 Extracellular Inulase 의 정제 및 특성비교)

  • Kim, Su-Il;Moon, Hang-Sik
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.169-178
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    • 1987
  • The extracellular and intracellular inulases from Kluyveromyces marxianus were purified and characterized. The maximum production of both inulases was achieved at stationary phase in a pH-controlled medium at pH 5 with yeast nitrogen base as organic nitrogen source. Each enzyme was concentrated by tannic acid precipitation and separated into two fractions by DEAF-cellulose chromatography. Electrophoretic analysis showed that the four fractions had three glycoprotein bards each. Only main glycoprotein band, however, had both inulase and invertase activities. There were no significant differences between two enzymes in the optimum pH and temperature. But the intracellular inulases had higher heat stability and less affinity toward inulin than the extracellular enzymes do. All the purified enzymes were considered to be exo-inulases using hydrolyzate analysis with TLC.

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Scant Extracellular NAD Cleaving Activity of Human Neutrophils is Down-Regulated by fMLP via FPRL1

  • Hasan, Md. Ashraful;Sultan, Md. Tipu;Ahn, Won-Gyun;Kim, Yeon-Ja;Jang, Ji-Hye;Hong, Chang-Won;Song, Dong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.6
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    • pp.497-502
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    • 2014
  • Extracellular nicotinamide adenine dinucleotide (NAD) cleaving activity of a particular cell type determines the rate of the degradation of extracellular NAD with formation of metabolites in the vicinity of the plasma membrane, which has important physiological consequences. It is yet to be elucidated whether intact human neutrophils have any extracellular NAD cleaving activity. In this study, with a simple fluorometric assay utilizing $1,N^6$-ethenoadenine dinucleotide (etheno-NAD) as the substrate, we have shown that intact peripheral human neutrophils have scant extracellular etheno-NAD cleaving activity, which is much less than that of mouse bone marrow neutrophils, mouse peripheral neutrophils, human monocytes and lymphocytes. With high performance liquid chromatography (HPLC), we have identified that ADP-ribose (ADPR) is the major extracellular metabolite of NAD degradation by intact human neutrophils. The scant extracellular etheno-NAD cleaving activity is decreased further by N-formyl-methionine-leucine-phenylalanine (fMLP), a chemoattractant for neutrophils. The fMLP-mediated decrease in the extracellular etheno-NAD cleaving activity is reversed by WRW4, a potent FPRL1 antagonist. These findings show that a much less extracellular etheno-NAD cleaving activity of intact human neutrophils compared to other immune cell types is down-regulated by fMLP via a low affinity fMLP receptor FPRL1.

Identification of Genes Modulated by High Extracellular Calcium in Coculture of Mouse Osteoblasts and Bone Marrow Cells by Oligo Chip Assay

  • Kim, Hyung-Keun;Song, Mi-Na;Jun, Ji-Hae;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.31 no.2
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    • pp.53-65
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    • 2006
  • Calcium concentration in the bone resorption lacunae is high and is in the mM concentration range. Both osteoblast and osteoclast have calcium sensing receptor in the cell surface, suggesting the regulatory role of high extracellular calcium in bone metabolism. In vitro, high extracellular calcium stimulated osteoclastogenesis in coculture of mouse osteoblasts and bone marrow cells. Therefore we examined the genes that were commonly regulated by both high extracellular calcium and $1,25(OH)_2vitaminD_3(VD3)$ by using mouse oligo 11 K gene chip. In the presence of 10 mM $[Ca^{2+}]e$ or 10 nM VD3, mouse calvarial osteoblasts and bone marrow cells were co-cultured for 4 days when tartrate resistant acid phosphatase-positive multinucleated cells start to appear. Of 11,000 genes examined, the genes commonly regulated both by high extracellular calcium and by VD3 were as follows; 1) the expression of genes which were osteoclast differentiation markers or were associated with osteoclastogenesis were up-regulated both by high extracellular calcium and by VD3; trap, mmp9, car2, ctsk, ckb, atp6b2, tm7sf4, rab7, 2) several chemokine and chemokine receptor genes such as sdf1, scya2, scyb5, scya6, scya8, scya9, and ccr1 were up-regulated both by high extracellular calcium and by VD3, 3) the genes such as mmp1b, mmp3 and c3 which possibly stimulate bone resorption by osteoclast, were commonly up-regulated, 4) the gene such as c1q and msr2 which were related with macrophage function, were commonly down-regulated, 5) the genes which possibly stimulate osteoblast differentiation and/or mineralization of extracellular matrix, were commonly down-regulated; slc8a1, admr, plod2, lox, fosb, 6) the genes which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were commonly up-regulated; s100a4, npr3, mme, 7) the genes such as calponin 1 and tgfbi which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were up-regulated by high extracellular calcium but were down-regulated by VD3. These results suggest that in coculture condition, both high extracellular calcium and VD3 commonly induce osteoclastogenesis but suppress osteoblast differentiation/mineralization by regulating the expression of related genes.

Extracellular Vesicles in Psychiatry Research in the Context of RDoC Criteria

  • Ilgin, Can;Topuzoglu, Ahmet
    • Psychiatry investigation
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    • v.15 no.11
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    • pp.1011-1018
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    • 2018
  • The analysis of extracellular vesicles has been accelerated because of the technological advancements in omics methods in recent decades. Extracellular vesicles provide multifaceted information regarding the functional status of the cells. This information would be critical in case of central nervous system cells, which are confined in a relatively sealed biological compartment. This obstacle is more dramatic in psychiatric disorders since their diagnosis primarily depend on the symptoms and signs of the patients. In this paper, we reviewed this rapidly advancing field by discussing definition of extracellular vesicles, their biogenesis and potential use as clinical biomarkers. Then we focused on their potential use in psychiatric disorders in the context of diagnosis and treatment of these disorders. Finally, we tried to combine the RDoC (Research Domain Criteria) with the use of extracellular vesicles in psychiatry research and practice. This review may offer new insights in both basic and translational research focusing on psychiatric disorders.

Extracellular Mechanisms of Neutrophils in Immune Cell Crosstalk

  • Sanjeeb Shrestha;Chang-Won Hong
    • IMMUNE NETWORK
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    • v.23 no.5
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    • pp.38.1-38.14
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    • 2023
  • Neutrophils are professional phagocytes that provide defense against invading pathogens through phagocytosis, degranulation, generation of ROS, and the formation of neutrophil extracellular traps (NETs). Although long been considered as short-lived effector cells with limited biosynthetic activity, recent studies have revealed that neutrophils actively communicate with other immune cells. Neutrophils employ various types of soluble mediators, including granules, cytokines, and chemokines, for crosstalk with immune cells. Additionally, ROS and NETs, major arsenals of neutrophils, are utilized for intercellular communication. Furthermore, extracellular vesicles play a crucial role as mediators of neutrophil crosstalk. In this review, we highlight the extracellular mechanisms of neutrophils and their roles in crosstalk with other cells.

Inverse Correlation between Extracellular DNase Activity and Biofilm Formation among Chicken-Derived Campylobacter Strains

  • Jung, Gi Hoon;Lim, Eun Seob;Woo, Min-Ah;Lee, Joo Young;Kim, Joo-Sung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1942-1951
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    • 2017
  • Campylobacter jejuni and Campylobacter coli are important foodborne pathogenic bacteria, particularly in poultry meat. In this study, the presence of extracellular DNase activity was investigated for biofilm-deficient Campylobacter strains versus biofilm-forming Campylobacter strains isolated from chickens, to understand the relationship between extracellular DNase activity and biofilm formation. A biofilm-forming reference strain, C. jejuni NCTC11168, was co-incubated with biofilm non-forming strains isolated from raw chickens or their supernatants. The biofilm non-forming strains or supernatants significantly prohibited the biofilm formation of C. jejuni NCTC11168. In addition, the strains degraded pre-formed biofilms of C. jejuni NCTC11168. Degradation of C. jejuni NCTC11168 biofilm was confirmed after treatment with the supernatant of the biofilm non-forming strain 2-1 by confocal laser scanning microscopy. Quantitative analysis of the biofilm matrix revealed reduction of extracellular DNA (16%) and proteins (8.7%) after treatment. Whereas the biofilm-forming strains C. jejuni Y23-5 and C. coli 34-3 isolated from raw chickens and the C. jejuni NCTC11168 reference strain showed no extracellular DNase activity against their own genomic DNA, most biofilm non-forming strains tested, including C. jejuni 2-1, C. coli 34-1, and C. jejuni 63-1, exhibited obvious extracellular DNase activities against their own or 11168 genomic DNA, except for one biofilm non-former, C. jejuni 22-1. Our results suggest that extracellular DNase activity is a common feature suppressing biofilm formation among biofilm non-forming C. jejuni or C. coli strains of chicken origin.

Expression and Purification of Extracellular Solute-Binding Protein (ESBP) in Escherichia coli, the Extracellular Protein Derived from Bifidobacterium longum KACC 91563

  • Song, Minyu;Kim, Hyaekang;Kwak, Woori;Park, Won Seo;Yoo, Jayeon;Kang, Han Byul;Kim, Jin-Hyoung;Kang, Sun-Moon;Van Ba, Hoa;Kim, Bu-Min;Oh, Mi-Hwa;Kim, Heebal;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.39 no.4
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    • pp.601-609
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    • 2019
  • Bifidobacterium longum KACC 91563 secretes family 5 extracellular solute-binding protein via extracellular vesicle. In our previous work, it was demonstrated that the protein effectively alleviated food allergy symptoms via mast cell specific apoptosis, and it has revealed a therapeutic potential of this protein in allergy treatment. In the present study, we cloned the gene encoding extracellular solute-binding protein of the strain into the histidine-tagged pET-28a(+) vector and transformed the resulting plasmid into the Escherichia coli strain BL21 (DE3). The histidine-tagged extracellular solute-binding protein expressed in the transformed cells was purified using Ni-NTA affinity column. To enhance the efficiency of the protein purification, three parameters were optimized; the host bacterial strain, the culturing and induction temperature, and the purification protocol. After the process, two liters of transformed culture produced 7.15 mg of the recombinant proteins. This is the first study describing the production of extracellular solute-binding protein of probiotic bacteria. Establishment of large-scale production strategy for the protein will further contribute to the development of functional foods and potential alternative treatments for allergies.

Physiochemical Properties of Extracellular Lipid Produced by Rhodotorula glutinis K-501 as a Biosurfactant (Rhodotorula glutinis K-501에 의해 생산된 세포외지질의 생체계면활성제로서 물리화학적 특성)

  • 박평규;채희정;김의용
    • KSBB Journal
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    • v.13 no.1
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    • pp.65-70
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    • 1998
  • The physiochemical properties of extracellular lipid produced by an oleaginous yeast, Rhodotorula glutinis K-501 were examined. From the analytical experiments, it was suggested that the extracellular lipid produced is glycolipidic compound. Critical micelle concentration and minimum surface tension of the extracellular lipid in aqueous solution were 89mg/L and 31dyne/cm, respectively. Surface tension was also constant throughout wide range of pH. The emulsifying abilities and dispersing power of the extracellular lipid were much greater than those of commercial surfactants such as Tween 80 and Triton X-100 by factors of 2-3 and 1.3, respectively.

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