Soojin Shim

Silver-perfluorodecanethiolate complexes having superhydrophobic, antifouling, antibacterial properties

Silver-perfluorodecanethiolate complexes having superhydrophobic, antifouling, antibacterial properties were prepared by a reaction of silver nitrate with perfluorodecanethiol. When the silver nitrate to perfluorodecanethiol molar ratio was 1/2, silver-perfluorodecanethiolate complexes having hierarchical micro-/nano-sized wire shapes were obtained, and they showed superhydrophobic and antifouling properties. After UV irradiation, silver nanoparticles were generated on the wires and exhibited antibacterial properties.

Prevention of Pseudomonas aeruginosa adhesion by electric currents

The process of controlling bacterial adhesion using an electric current deserves attention because of its ease of automation and environmentally friendly nature. This study investigated the role of electric currents (negative, positive, alternating) for preventing adhesion of Pseudomonas aeruginosa and achieving bacterial inactivation. Indium tin oxide (ITO) film was used as a working electrode to observe adhesion and inactivation under electric polarization. Electric current types were classified into negative, positive, and alternating current. The working electrode acted as a cathode or anode by applying a negative or positive current, and an alternating current indicates that the negative current was combined sequentially with the positive current. The numbers of adhered cells were compared under a flow condition, and the in situ behavior of the bacterial cells and the extent of their inactivation were also investigated using time-lapse recording and live/dead staining, respectively. The application of a negative current prevented bacterial adhesion significantly (∼81% at 15.0 μA cm−2). The positive current did not significantly inhibit adhesion (<20% at 15.0 μA cm−2), compared to the nonpolarized case. The alternating current had a similar effect as the negative current on preventing bacterial adhesion, but it also exhibited bactericidal effects, making it the most suitable method for bacterial adhesion control.

Bacterial translational motion on the electrode surface under anodic electric field.

Application of an electric field (alternating or cathodic polarization) has been suggested as a possible mean of controlling biofilm development. Bacteria on an anodically polarized surface were shown to be active and highly motile when compared with a nonpolarized condition, but no quantitative information on bacterial motion has been reported. This study investigated the effects of environmental conditions (current density and ionic strength) on the translational motion of P. aeruginosa PAO1 cells under an anodic electric field using a quantitative tracking method. Bacterial displacement for 10 s was found to be approximately 1.2 μm, irrespective of wide-ranging current densities (7.5-30 μA/cm(2)). However, the local dynamics of bacterial communities differed under varied current densities. The distribution of bacterial displacement appeared to exhibit a more oscillating (subdiffusive) at high current density. At the same time, the number of bacteria with a circular trajectory (superdiffusive) decreased. Bacterial movement decreased with increased ionic strength of the media, because of strong electrostatic interactions. The motion of bacterial communities on an anodically polarized surface under various conditions is discussed, along with possible mechanisms. In addition, the control of biofilm growth was partly demonstrated by changing the motility of bacterial cells under anodic polarization.

Cytokines production and toll-like receptors expression in human leukemic monocyte cells, THP-1, stimulated with Brucella abortus cellular antigens

A zoonotic pathogen, Brucella spp. is the causative agent of brucellosis, which results in abortion and loss in milk production in domestic animals, and undulant fever, osteoarticular pain and splenomegaly in humans. Due to the capability of the bacteria to modulate the host cell functions and survive in macrophages, early detection and eradication of the intracellular bacteria has received significant attention. Moreover, understanding the immunological alterations in Brucella infection is crucial to help develop control measures. Cytokines and toll-like receptors (TLRs) are some of the major compounds that play important roles in modulating the innate immunity and acquired immunity in host after infection. In this study, therefore, human leukemic monocyte cells (THP-1 cells) were stimulated with five Brucella abortus cellular components: outer membrane protein 10 (OMP10), outer membrane protein 19 (OMP19), thiamine transporter substrate-binding protein (TbpA), arginase (RocF), and malate dehydrogenase (Mdh). Post stimulation, the cytokine productions and TLR expressions in the cells were evaluated at different time points (12 h and 24 h), and analyzed using ELISA and real time RT-PCR, respectively. In the production of cytokines, it was observed that the production of TNF-α and IL-6 was highly induced in THP-1 cells stimulated with five recombinant protein antigens. Also, TLR8 was induced in a time-dependent manner after stimulation with two recombinant proteins, rOMP19 and rMdh, until 24 h. These results suggest that the two B. abortus antigens, rOMP19 and rMdh, might be involved in TLR8 signaling pathway in THP-1 cells in a time-dependent manner. These two proteins are therefore potentially effective antigen candidates which would help to provide better understandings of the immune responses after Brucella infection.

Analysis of protein expression in Brucella abortus mutants with different growth rates by two-dimensional gel electrophoresis and LC-MS/MS peptide analysis

Brucella abortus is a bacterium that causes brucellosis and is the causative agent of worldwide zoonoses. Pathogenesis of the B. abortus infection is complicated, and several researchers have attempted to elucidate the infection mechanism of B. abortus. While several proteins have been revealed as pathogenic factors by previous researchers, the underlying mechanism of B. abortus infection is unresolved. In this study, we identified proteins showing different expression levels in B. abortus mutants with different biological characteristics that were generated by random insertion of a transposon. Five mutants were selected based on biological characteristics, in particular, their growth features. Total proteins of mutant and wild-type B. abortus were purified and subjected to two-dimensional gel electrophoresis. Thirty protein spots of each mutant with expression increases or decreases were selected; those with a change of more than 2-fold were compared with the wild-type. Selected spots underwent liquid chromatography tandem mass spectrometry for peptide analysis. DnaK and ClpB, involved in protein aggregation, increased. SecA and GAPDH, associated with energy metabolism, decreased in some mutants with a growth rate slower than that of the wild-type. Mutants with slower growth showed a decrease in energy metabolism-related proteins, while mutants with faster growth showed an increase in pathogenicity-related proteins.

Global gene-expression profiles of intracellular survival of the BruAb2_1031 gene mutated Brucella abortus in professional phagocytes, RAW 264.7 cells

BackgroundSince recognizing the interaction between Brucella and host cells is crucial to the elucidation of the infectious process, Brucella researches have prioritized the investigation of genes related to pathogenicity. To demonstrate the roles of Brucella genes, RAW 264.7 cells were infected with the Brucella abortus wild-type and mutant strains (generated using transposon mutagenesis), after which the different transcriptional responses of the infected cells were determined using microarray.ResultsFollowing infection, enhanced strategies for intracellular survival, such as down-regulation of genes associated with cytokine responses and apoptosis, were observed in RAW 264.7 cells infected with C3 mutant strain when compared to the transcriptional responses of wild-type infected cells. Using sequence analysis, we determined the mutation site of a C3 mutant strain as the ATP-binding cassette transporter permease (BruAb2_1031). These results were evidenced by an increased level of intracellular survival of the C3 mutant strain.ConclusionsCharacteristics of each mutant strain including bacterial growth rate, abilities to induce cytokine production in macrophages after infection, internalization, and levels of intracellular survival and replication, were investigated by performing RAW 264.7 cell infection experiments. Our results indicate that the BruAb2_1031 gene might be closely related with intracellular survival of B. abortus in RAW 264.7 cells.

Oscillational motion properties of bacteria and polystyrene particles on a positively polarized substrate surface.

The oscillational motion of bacteria and non-biological particles on a positively polarized substrate surface were investigated in this study using several bacterial species (Staphylococcus epidermidis ATCC12228 and Pseudomonas aeruginosa PA14) and polystyrene particles (modified with sulfate or carboxylate) that have different cell/particle size, surface potential, surface ionizable functional group, and surface appendage with respect to the mean square displacement (MSD) and motion trajectory. The attractive/repulsive interactions between the bacteria/particle and a positively polarized substrate surface are further discussed with the results of the motion analysis based on the extended Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. As our major findings, all the bacterial species and particles showed oscillational motion, a kind of sub-diffusive motion that is more limited than the Brownian motion of the suspended bacteria/particles, on a positively polarized substrate surface. However, the motion properties among the bacteria/particles were found to differ in motion radius and MSD. As the size and negative surface potential of the bacteria/particle got smaller, the oscillational motion became more active, which may result from a decrease in attractive interactions such as van der Waals interaction and electrostatic attractive interaction. In the case in which some surface functional group (e.g., sulfate group) contributed to the formation of a strong Lewis acid-base interaction, the oscillational motion was significantly reduced regardless of the surface potential of the particle. The bacterial surface appendages were found to have no influence in explaining motion differences between the bacteria and non-biological particle.

Control of Bacterial Adhesion and Biofilm Using Electric Field

The use of electric field has been studied as an alternative for biofilm control dominated by disinfectants and antibiotics. This technology would be advantageous in the environmental respect that biofilm can be controlled based on electron transfer, not using chemical disinfectants and antibiotics. Control mechanisms which were reported by earlier studies are organized as; (1) bacterial adhesion control by electrostatic repulsion at a negative current, (2) bacterial adhesion control using bacterial motion and (3) bacterial inactivation by direct oxidation at a positive current, (4) bioelectric effect leading to biofilm inactivation. In this review article, we summarized the technologies for biofilm control using electric field and provided some application examples from previous studies.