• Title/Summary/Keyword: laser scanning confocal microscopy

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Membrane Perturbation Induced by Papiliocin Peptide, Derived from Papilio xuthus, in Candida albicans

  • Lee, June-Young;Hwang, Jae-Sam;Hwang, Bo-Mi;Kim, Jin-Kyoung;Kim, Seong-Ryul;Kim, Yang-Mee;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1185-1188
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    • 2010
  • Previously, papiliocin was isolated from the swallowtail butterfly Papilio xuthus and its antimicrobial activity was suggested. In this study, the antifungal mechanism of papiliocin against Candida albicans was investigated. Confocal laser scanning microscopy (CLSM) and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence analysis indicated that papiliocin disturbed the fungal plasma membrane. Moreover, the assessment of the release of FITC-dextran (FD) from liposomes further demonstrated that the antifungal mechanism of papiliocin could have originated from the pore-forming action and that the radius of the pores was presumed to be anywhere from 2.3 to 3.3 nm.

Direct membrane filtration of wastewater under very short hydraulic retention time

  • Yoon, Seong-Hoon
    • Advances in environmental research
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    • v.7 no.1
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    • pp.39-52
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    • 2018
  • Direct membrane filtration (DMF) of wastewater has many advantages over conventional biological wastewater treatment processes. DMF is not only compact, but potentially energy efficient due to the lack of biological aeration. It also produces more biosolids that can be used to produce methane gas through anaerobic digestion. Most of ammoniacal nitrogen in wastewater is preserved in effluent and is used as fertilizer when effluent is recycled for irrigation. In this study, a technical feasibility of DMF was explored. Organic and nitrogen removal efficiencies were compared between DMF and membrane bioreactor (MBR). Despite the extremely high F/V ratio, e.g., $14.4kg\;COD/m^3/d$, DMF provided very high COD removal efficiencies at ~93%. Soluble microbial products (SMP) and extracellular polymeric substances (EPS) were less in DMF sludge, but membrane fouling rate was far greater than in MBR. The diversity of microbial community in DMF appeared very narrow based on the morphological observation using optical microscope. On the contrary, highly diverse microbial community was observed in the MBR. Microorganisms tended to form jelly globs and attach on reactor wall in DMF. FT-IR study revealed that the biological globs were structurally supported by feather-like materials made of secondary amines. Confocal laser scanning microscopy (CLSM) study showed microorganisms mainly resided on the external surface of microbial globs rather than the internal spaces.

Bacterial Dynamics of Biofilm Development During Toluene Degradation by Burkholderia vietnamiensis G4 in a Gas Phase Membrane Bioreactor

  • Kumar, Amit;Dewulf, Jo;Wiele, Tom Van De;Langenhove, Herman Van
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.1028-1033
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    • 2009
  • In this study, the dynamics of living cells (LC) and dead cells (DC) in a laboratory-scale biofilm membrane bioreactor for waste gas treatment was examined. Toluene was used as a model pollutant. The bacterial cells were enumerated as fluoromicroscopic counts during a 140 operating day period using BacLight nucleic acid staining in combination with epifluorescence and confocal laser scanning microscopy (CSLM). Overall, five different phases could be distinguished during the biofilm development: (A) cell attachment, (B) pollutant limitation, (C) biofilm establishment and colonization, (D) colonized biofilm, and (E) biofilm erosion. The bioreactor was operated under different conditions by applying different pollutant concentrations. An optimum toluene removal of 89% was observed at a loading rate of 14.4 kg $m^{-3}d^{-1}$. A direct correlation between the biodegradation rate of the reactor and the dynamics of biofilm development could be demonstrated. This study shows the first description of biofilm development during gaseous toluene degradation in MBR.

Expression of Heat Shock Protein Protein 70 in Umbilical Vein Endothelial Cells Infected by Staphylococcus aureus

  • Chang, Hyun-Ah;Chang, Jun-Keun;Kim, Jong-Won;Kim, Mal-Nam
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.137-142
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    • 2000
  • Environmental stres is known to induce heat shock proteins (HSPs) in eukaryotic cells. However, the induction of HSPs in host cells by microbial infection has not yet been well explained. Staphylococcus aureus (S. aureus) is one of the major pathogens in the pathogenesis of endovascular diseases such as infective endocarditis. In this study, the synthesis of stress-inducible 70 kDa HSP was investigated in the endothelial cells (ECs) after 3 h to 20 h of incubation with S. aureus. The dffect of S. aureus infection on the expression of HSP70 in cultured ECs was analyzed using laser scanning confocal microscopy (LSCM). The increase of HSP70 expression in ECs infected by S. aureus ($10^4{\;}cfu/ml$) for 20 h was 1.1-fold higher than that in heat shock treated ECs and 2.2-fold higher than that in untreated cells. Heat shock is known to induce intranucleus HSP70 expression in mammalian cells, whereas the S. aureus infection induced perinuclear expression in ECs as observed by LSCM. Consequently, the expression of HSP70 in ECs plays an important role in the pathogenesis of endovascular infection.

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Changes in Caenorhabditis elegans Exposed to Vibrio parahaemolyticus

  • Durai, Sellegounder;Pandian, Shunmugiah Karutha;Balamurugan, Krishnaswamy
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1026-1035
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    • 2011
  • Vibrio parahaemolyticus, which owes its origin to the marine environment, is considered as one of the most common causes of infectious diarrhea worldwide. The present study investigated the pathogenicity of V. parahaemolyticus against the model organism, Caenorhabditis elegans. Infection in the host was localized with GFP-tagged V. parahaemolyticus using confocal laser scanning microscopy. The times required for causing infection, bacterial load in intestine, chemotactic response, and alteration in pharyngeal pumping were analyzed in the host system. In addition, the regulation of innate immune-related genes, lys-7, clec- 60, and clec-87, was analyzed using real-time PCR. The role of immune-responsible pmk-1 was studied using mutant strains. The pathogenicity of environmental strain CM2 isolated from the Gulf of Mannar, India was compared with that of a reference strain obtained from ATCC. The pathogen infected animals appeared to ward off infection by up-regulating candidate antimicrobial genes for a few hours after the exposure, before succumbing to the pathogen. For the first time, the pathogenicity of V. parahaemolyticus at both the physiological and molecular levels has been studied in detail using the model organism C. elegans.

Nuclear Localization Signal of Human Foamy Virus Integrase (인간 포미바이러스 인테그라제의 핵위치 신호)

  • Oh Soo-A;Kang Seung-Yi;Han Sung-Tae;An Dog-Gn;Shin Cha-Gyun
    • YAKHAK HOEJI
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    • v.50 no.2
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    • pp.93-98
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    • 2006
  • Human foamy virus (HFV) integrase mediates integration of viral c-DNA into cellular DNA. In this process, HFV prointegration complex (PIC) in which integrase is a key component moves to nuclei of the infected cells and leads to integration of viral DNA to the cellular genome, which is essential in viral life cycle. In general nuclear localization signals (NLS) have been suggested to be involved in localizing retroviral PIC to nuclei, but the mechanisms for nuclear localization of the HFV PIC remains unclear. To functionally identify the NLS of HFV integrase, various subdomains of the protein were expressed as GFP fusions and their subcellular locations were analyzed with confocal laser scanning microscopy. Wild type HFV integrase was karyophilic by targeting the fusion protein to nuclei of the COS-1 and 293T cells. Our results showed that strong NLS of HFV integrase was mapped to the C-terminal regions. In addition the karyophilic properties of N-terminal and central regions are not individually strong enough to direct localization of the fusion proteins to nuclei, but their cooperative activity for nuclear import was confirmed.

Use of Cationic PAM as a Surface Sizing Additive to Improve Paper Properties

  • Seo, Man-Seok;Lee, Hak-Lae;Youn, Hye-Jung
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2006.06b
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    • pp.245-250
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    • 2006
  • This study was focused on the use of cationic PAM (Polyacrylamide) as a surface sizing additive to improve the surface sizing properties of paper. Effects of the ionic property, viscosity and charge density of PAM on bending stiffness of surface sized papers were investigated. Use of cationic PAM as a surface sizing additive improved bending stiffness while addition of anionic PAM did not show any effect. Increase of starch holdout with the addition of cationic PAM was attributed as a prime reason of stiffness increase. Viscosity of PAM was one of the most important factors affecting surface sizing due to its influence on the interaction between cationic PAM and oxidized starch solution. Greater improvement of bending stiffness of paper was obtained when high charged PAM was used as an additive. The order of addition was found to have significant influence on the effect of additives since it influences the formation of network structure among starch, cationic PAM, and SA (styrene acrylic acid copolymer). Investigation on the penetration of starch solution was carried out with CLSM (Confocal Laser Scanning Microscopy), and it was shown that the addition of cationic PAM to oxidized starch solution made starch molecules stay on the paper surface rather than penetrating into the paper structure because of the electrostatic interaction between negatively charged fibers and positively charged cationic PAM.

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Bacterial Traits Involved in Colonization of Arabidopsis thaliana Roots by Bacillus amyloliquefaciens FZB42

  • Dietel, Kristin;Beator, Barbara;Budiharjo, Anto;Fan, Ben;Borriss, Rainer
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.59-66
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    • 2013
  • Colonization studies previously performed with a green-fluorescent-protein, GFP, labeled derivative of Bacillus amyloliquefaciens FZB42 revealed that the bacterium behaved different in colonizing surfaces of plant roots of different species (Fan et al., 2012). In order to extend these studies and to elucidate which genes are crucial for root colonization, we applied targeted mutant strains to Arabidopsis seedlings. The fates of root colonization in mutant strains impaired in synthesis of alternative sigma factors, non-ribosomal synthesis of lipopeptides and polyketides, biofilm formation, swarming motility, and plant growth promoting activity were analyzed by confocal laser scanning microscopy. Whilst the wild-type strain heavily colonized surfaces of root tips and lateral roots, the mutant strains were impaired in their ability to colonize root tips and most of them were unable to colonize lateral roots. Ability to colonize plant roots is not only dependent on the ability to form biofilms or swarming motility. Six mutants, deficient in abrB-, sigH-, sigD-, nrfA-, yusV and RBAM017410, but not affected in biofilm formation, displayed significantly reduced root colonization. The nrfA- and yusV-mutant strains colonized border cells and, partly, root surfaces but did not colonize root tips or lateral roots.

Anti-biofilm Activity of Xanthorrhizol Isolated from Curcuma xanthorrhiza Roxb. against Bacterial Biofilms Formed by Saliva and Artificial Multi-species Oral Strains

  • Yanti, Yanti;Rukayadi, Yaya;Lee, Kwan-Hyoung;Han, Sung-Hwa;Hwang, Jae-Kwan
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.556-560
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    • 2009
  • Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.

Galactosylated Chitosan (GC)-graft-Poly(vinyl pyrrolidone) (PVP) as Hepatocyte-Targeting DNA Carrier: In Vitro Transfection

  • Park, In-Kyu;Jiang, Hu-Lin;Cook, Seung-Eun;Cho, Myung-Haing;Kim, Su-Il;Jeong, Hwan-Jeong;Akaike, Toshihiro;Cho , Chong-Su
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1284-1289
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    • 2004
  • Galactosylated chitosan-graft-poly(vinyl pyrrolidone) (GCPVP) was synthesized and characterized for hepatocyte-targeting gene carrier. GCPVP itself as well as GCPVP/DNA complex had negligible cytotoxicity regardless of the concentration of GCPVP and the charge ratio, but GCPVP/DNA complex had slightly cytotoxic effect on HepG2 cells only in the case of the higher charge ratio and 20 mM of $Ca^{2+}$ concentration used. Through the confocal laser scanning microscopy, it is shown that the endocytosis by interaction between galactose ligands of GCPVP and ASGPR of the hepatocytes was the major route of transfection of GCPVP/F-plasmid complexes.