• Journal of Microbiology
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• 제39권2호
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• pp.133-138
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• 2001

We investigated the antagonism of indigenous bacteria isolated from stressed mussels and their extracellular metabolites on the adult zebra mussel, Dreissena polymorpha. Selective bacterial isolates including Aeromonas media, A. salmonicida, A. veronii, and Shewanella putrefaciens, showed strong lethality against adult mussels and 100% mortality was observed within 5 days of incubation. Bacterial metabolites, fractionated and concentrated from stationary-phase culture supernatants of these bacterial isolates, displayed varying degrees of antagonistic effects on zebra mussels. Among the three size fractions examined, <5, 5-10, and >10 kDa, the mast lethal fraction seems to be >10 kDa for three of the four isolates tested. Further chemical analyses of these size fractions revealed that the predominant constituents were polysaccharides and proteins. No 2-keto-3-deoxyoctanoic acid (2-KDO), deoxyribonucleic acids (DNA) or uranic acid were detectable. Extraction of supernatants of two antagonistic isolates with polar solvent suggested that polar molecules are present in the active fraction. Our data suggest that extracellular metabolites produced by antagonistic bacteria are also involved in disease development in zebra mussels and elucidation of the mechanisms involved may offer a novel strategy for control of biofouling invertebrates.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1485-1490
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• 2006

In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1464-1472
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• 2016

The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.

• Journal of Microbiology
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• 제35권3호
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• pp.213-221
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• 1997

The secretion of the alkaliphilic Bacillus sp. S-1 extracellular pullulanase involves translocation across the cytoplasmic membrane of the Gram-positive bacterial cell envelope. Translocation of the intracellular pullulanase PUL-I, was traced to elucidate the mechanism and pathway of protein secretion from an alkaliphilic Bacillus sp. S-1. Pullulanase could be slowly bue quantitatively released into the medium during growth of the cells in medium contianing proteinase K. The released pullulanase lacked the N-terminal domain. The N-terminus is the sole membrane anchor in the pullulanase protein and was not affected by proteases, confirming that it is not exposed on the cell surface. Processing of a 180,000M$\_$r/ pullulanase to a 140,000M$\_$r/ polypeptide has been demonstrated in cell extracts using antibodies raised against 140,000M$\_$r/ extracellular form. Processing of the 180,000 M$\_$r/ protein occured during the preparation of extracts in an alkaline pH condition. A modified rapid extraction procedure suggested that the processing event also occured in vivo. Processing apparently increased the activity of pullulanase. The western blotting analysis with mouse anti-serum against 140-kDa extracellular pullulanase PUL-E showed that PUL-I is processed into PUL-X via intermediate form of PUL-E. Possible explanationa for the translocation are discussed.

• 대한의용생체공학회:의공학회지
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• 제40권4호
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• pp.137-142
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• 2019

Numerous efforts are being made to develop an ideal dermal filler that should be bio-compatibility, non-immunogenicity, long-lasting and biodegradable without a toxic secretion. Biomaterials of dermal fillers are hyaluronic acid filler, calcium filler, PMMA filler and collagen filler depending on the ingredient. Although hyaluronic acid (HA) is most widely used, it has shortages such as short shelf life and low mechanical strength compare to extracellular matrix (ECM). The cartilage ECM composed of collagen type II, proteoglycans, glycosaminoglycans (GAGs) and in a minor part with glycoproteins. In this study, we developed a cartilage ECM injectable filler capable of improving biocompatibility and longevity compared with hyaluronic acid (HA) fillers. The ECM hydrogel was cross-linked by the reaction of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) for mechanical enhancement. Prepared ECM filler was compared with cross-linked HA by butanediol diglycidyle ether (BDDE), which is the most widely used natural polymers for dermal filler. In the results, the articular cartilage ECM hydrogel has great potential as a dermal filler to improve the biophysical and biological performance.

• Translational and Clinical Pharmacology
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• 제26권3호
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• pp.103-110
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• 2018

The human microbiome is known to play an essential role in influencing host health. Extracellular vesicles (EVs) have also been reported to act on a variety of signaling pathways, distally transport cellular components such as proteins, lipids, and nucleic acid, and have immunomodulatory effects. Here we shall review the current understanding of the intersectionality of the human microbiome and EVs in the emerging field of microbiota-derived EVs and their pharmacological potential. Microbes secrete several classes of EVs: outer membrane vesicles (OMVs), membrane vesicles (MVs), and apoptotic bodies. EV biogenesis is unique to each cell and regulated by sophisticated signaling pathways. EVs are primarily composed of lipids, proteins, nucleic acids, and recent evidence suggests they may also carry metabolites. These components interact with host cells and control various cellular processes by transferring their constituents. The pharmacological potential of microbiome-derived EVs as vaccine candidates, biomarkers, and a smart drug delivery system is a promising area of future research. Therefore, it is necessary to elucidate in detail the mechanisms of microbiome-derived EV action in host health in a multi-disciplinary manner.

• 생약학회지
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• 제50권1호
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• pp.18-24
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• 2019

Andrographolide, the main compound of Andrographis paniculata (A. paniculata), shows various biological properties including anti-viral, anti-inflammatory, anti-diabetic, and hepatoprotective effects. Our previous study has shown that A. paniculata extract exerts antiaging effects by activation of stemness in epidermal stem cells (EpSCs). In this study, we investigated the effect of andrographolide as a main compound of A. paniculata on EpSCs and its mechnism of action using several in vitro assays. Andrographolide increased the proliferation of EpSCs and induced cell cycle progression. Additionally, andrographolide increased VEGF production and the expression of stem cell markers integrin ${\beta}1$ and p63. Furthermore, phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), S6 ribosomal protein (S6RP) and Akt were increased by andrographolide. Taken together, these results indicate that andrographolide-induced proliferation of EpSCs is mediated by the ERK1/2, Akt-dependent pathway with increased production of VEGF and upregulated stemness through integrin ${\beta}1$ and p63.

• 한국동물생명공학회지
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• 제37권1호
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• pp.2-12
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• 2022

Extracellular vesicles (EVs) are nanovesicles that carry bioactive cargoes of proteins, lipids, mRNAs, and miRNAs between living cells. Their role in cellular communication has gained the attention of several research reports globally in the last decade. EVs are critically involved in sperm functions, oocyte functions, fertilization, embryonic development, and pregnancy. The review summarizes the state-of-the-art of EVs research in the diagnostic and therapeutic (theranostic) potentials of the EVs during the pregnancy that might provide a solution for gestational disturbances such as implantation failure, maternal health problems, gestational diabetes, and preeclampsia. EVs can be found in all biological fluids of the fetus and the mother and would provide a non-invasive and excellent tool for diagnostic purposes. Moreover, we provide the current efforts in manufacturing and designing targeted therapeutics using synthetic and semi-synthetic nanovesicles mimicking the natural EVs for efficient drug delivery during pregnancy.

• BMB Reports
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• 제55권1호
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• pp.20-29
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• 2022

Stem cell-based therapy is a promising approach for treating a variety of disorders, including acute brain insults and neurodegenerative diseases. Stem cells such as mesenchymal stem cells (MSCs) secrete extracellular vesicles (EVs), circular membrane fragments (30 nm-1 ㎛) that are shed from the cell surface, carrying several therapeutic molecules such as proteins and microRNAs. Because EV-based therapy is superior to cell therapy in terms of scalable production, biodistribution, and safety profiles, it can be used to treat brain diseases as an alternative to stem cell therapy. This review presents evidences evaluating the role of stem cell-derived EVs in stroke, traumatic brain injury, and degenerative brain diseases, such as Alzheimer's disease and Parkinson' disease. In addition, stem cell-derived EVs have better profiles in biocompatibility, immunogenicity, and safety than those of small chemical and macromolecules. The advantages and disadvantages of EVs compared with other strategies are discussed. Even though EVs obtained from native stem cells have potential in the treatment of brain diseases, the successful clinical application is limited by the short half-life, limited targeting, rapid clearance after application, and insufficient payload. We discuss the strategies to enhance the efficacy of EV therapeutics. Finally, EV therapies have yet to be approved by the regulatory authorities. Major issues are discussed together with relevant advances in the clinical application of EV therapeutics.

• BMB Reports
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• 제55권2호
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• pp.65-71
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• 2022

Regenerative medicine is a research field that develops methods to restore damaged cell or tissue function by regeneration, repair or replacement. Stem cells are the raw material of the body that is ultimately used from the point of view of regenerative medicine, and stem cell therapy uses cells themselves or their derivatives to promote responses to diseases and dysfunctions, the ultimate goal of regenerative medicine. Stem cell-derived extracellular vesicles (EVs) are recognized as an attractive source because they can enrich exogenous microRNAs (miRNAs) by targeting pathological recipient cells for disease therapy and can overcome the obstacles faced by current cell therapy agents. However, there are some limitations that need to be addressed before using miRNA-enriched EVs derived from stem cells for multiplexed therapeutic targeting in many diseases. Here, we review various roles on miRNA-based stem cell EVs that can induce effective and stable functional improvement of stem cell-derived EVs. In addition, we introduce and review the implications of several miRNA-enriched EV therapies improved by multiplexed targeting in diseases involving the circulatory system and nervous system. This systemic review may offer potential roles for stem cell-derived therapeutics with multiplexed targeting.