Fluid flow facilitates inward rectifier K+ current by convectively restoring [K+] at the cell membrane surface.

The inward rectifier Kir2.1 current (IKir2.1) was reported to be facilitated by fluid flow. However, the mechanism underlying this facilitation remains uncertain. We hypothesized that during K+ influx or efflux, [K+] adjacent to the outer mouth of the Kir2.1 channel might decrease or increase, respectively, compared with the average [K+] of the bulk extracellular solution, and that fluid flow could restore the original [K+] and result in the apparent facilitation of IKir2.1. We recorded the IKir2.1 in RBL-2H3 cells and HEK293T cells that were ectopically over-expressed with Kir2.1 channels by using the whole-cell patch-clamp technique. Fluid-flow application immediately increased the IKir2.1, which was not prevented by either the pretreatment with inhibitors of various protein kinases or the modulation of the cytoskeleton and caveolae. The magnitudes of the increases of IKir2.1 by fluid flow were driving force-dependent. Simulations performed using the Nernst-Planck mass equation indicated that [K+] near the membrane surface fell markedly below the average [K+] of the bulk extracellular solution during K+ influx, and, notably, that fluid flow restored the decreased [K+] at the cell surface in a flow rate-dependent manner. These results support the "convection-regulation hypothesis" and define a novel interpretation of fluid flow-induced modulation of ion channels.

The fabrication of protein chip based on surface plasmon resonance for detection of pathogens.

Protein chip based on surface plasmon resonance (SPR) was developed for detection of pathogens existing in contaminated environment, such as Escherichia coli O157:H7, Salmonella typhimurium, Legionella pneumophila, and Yersinia enterocolitica. Protein G was immobilized to endow the orientation of antibody molecules on the SPR surface. The pathogen binding of the protein chip was investigated by SPR spectroscopy. Consequently, it was found that the four kinds of pathogen could be selectively detected by using SPR-based protein chip.

Biofilm Formation of Staphylococcus aureus on Various Surfaces and Their Resistance to Chlorine Sanitizer.

UNLABELLED: This study investigated the effect of material types (polystyrene, polypropylene, glass, and stainless steel) and glucose addition on Staphylococcus aureus biofilm formation, and the relationship between biofilm formation measured by crystal violet (CV) staining and the number of biofilm cells determined by cell counts was studied. We also evaluated the efficacy of chlorine sanitizer on inhibiting various different types of S. aureus biofilms on the surface of stainless steel. Levels of biofilm formation of S. aureus were higher on hydrophilic surfaces (glass and stainless steel) than on hydrophobic surfaces (polypropylene and polystyrene). With the exception of biofilm formed on glass, the addition of glucose in broth significantly increased the biofilm formation of S. aureus on all surfaces and for all tested strains (P ≤ 0.05). The number of biofilm cells was not correlated with the biomass of the biofilms determined using the CV staining method. The efficacy of chlorine sanitizer against biofilm of S. aureus was not significantly different depending on types of biofilm (P > 0.05). Therefore, further studies are needed in order to determine an accurate method quantifying levels of bacterial biofilm and to evaluate the resistance of bacterial biofilm on the material surface. PRACTICAL APPLICATION: Biofilm formation of Staphylococcus aureus on the surface was different depending on the surface characteristics and S. aureus strains. There was low correlation between crystal violet staining method and viable counts technique for measuring levels of biofilm formation of S. aureus on the surfaces. These results could provide helpful information for finding and understanding the quantification method and resistance of bacterial biofilm on the surface.

Detection of insulin-antibody binding on a solid surface using imaging ellipsometry.

Imaging ellipsometry (IE) was used to detect the binding of insulin to its antibody on a solid surface. The modification of a gold surface with 11-mecaptoundecanoic acid (11-MUA), the adsorption of protein G, and antibody immobilization onto the protein G layer were confirmed by surface plasmon resonance. Ellipsometric images and ellipsometric angles of the surface antibody were acquired using the IE system by off-null ellipsometry. Ellipsometric images of antigen binding to the antibody were acquired, and their mean optical intensities estimated. Changes in mean optical intensity indicated that the detection range for insulin was from 10 ng/ml to 100 microg/ml.

Immunosensor for detection of Legionella pneumophila using surface plasmon resonance.

Immunosensor using surface plasmon resonance (SPR) onto self-assembled protein G layer was developed for the detection of Legionella pneumophila. A self-assembled protein G layer on gold (Au) surface was fabricated by adsorbing a mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol (molar ratio of 1:2) and the activation process for chemical binding between free amine (-NH(2)) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of self-assembled protein G layer on Au substrate and the binding of antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of self-assembled protein G layer on Au substrate and monoclonal antibody against L. pneumophila immobilized on protein G were performed by atomic force microscope (AFM). The immunosensor for detection of L. pneumophila using SPR was developed and its detection limit could find up to 10(5) cells/ml.

Inhibitory effects of UV treatment and a combination of UV and dry heat against pathogens on stainless steel and polypropylene surfaces.

Pathogens that contaminate the surfaces of food utensils may contribute to the occurrence of foodborne disease outbreaks. We investigated the efficacy of UV treatment combined with dry heat (50 °C) for inhibiting 5 foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus) on stainless steel and polypropylene surfaces in this study. We inoculated substrates with each of the 5 foodborne pathogens cultured on agar surface and then UV treatment alone or a combination of both UV and dry heat (50 °C) was applied for 30 min, 1 h, 2 h, and 3 h. The initial populations of the 5 pathogens before treatment were 8.02 to 9.18 and 8.73 to 9.16 log10 CFU/coupon on the surfaces of stainless steel and polypropylene coupons, respectively. UV treatments for 3 h significantly inhibited S. Typhimurium, L. monocytogenes, and S. aureus on the stainless steel by 3.06, 2.18, and 2.70 log10 CFU/coupon, and S. aureus on the polypropylene by 3.11 log10 CFU/coupon, respectively. The inhibitory effects of the combined UV and dry heat treatment (50 °C) increased as treatment time increased, yielding significant reductions in all samples treated for 3 h, with the exception of S. aureus on polypropylene. The reduction level of E. coli O157:H7 treated for 3 h on the surface of stainless steel and polypropylene treated was approximately 6.00 log10 CFU/coupon. These results indicate that combined UV and dry heat (50 °C) treatments may be effective for controlling microbial contamination on utensils and cooking equipment surfaces as well as in other related environments.

Resistance of pathogenic bacteria on the surface of stainless steel depending on attachment form and efficacy of chemical sanitizers.

Various bacteria including food spoilage bacteria and pathogens can form biofilms on different food processing surfaces, leading to potential food contamination or spoilage. Therefore, the survival of foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, Cronobacter sakazakii) in different forms (adhered cells, biofilm producing in TSB, biofilm producing at RH 100%) on the surface of stainless steel and stored at various relative humidities (RH 23%, 43%, 68%, 85%, and 100%) at room temperature for 5 days was investigated in this study. Additionally, the efficacy of chemical sanitizers (chlorine-based and alcohol-based commercial sanitizers) on inhibiting various types of biofilms of E. coli O157:H7 and S. aureus on the surface of stainless steel was investigated. The number of pathogens on the surface of stainless steel in TSB stored at 25°C for 7 days or RH 100% at 25°C for 7 days was significantly increased and resulted in the increase of 3 log(10) CFU/coupon after 1 day, and these levels were maintained for 7 days. When stainless steel coupons were stored at 25°C for 5 days, the number of pathogens on the surface of stainless steel was significantly reduced after storage at RH 23%, 43%, 68%, and 85%, but not at 100%. When the bacteria formed biofilms on the surface of stainless steel in TSB after 6 days, the results were similar to those of the attached form. However, levels of S. aureus and C. sakazakii biofilms were more slowly reduced after storage at RH 23%, 43%, 68%, and 85% for 5 days than were those of the other pathogens. Formation of biofilms stored at RH 100% for 5 days displayed the highest levels of resistance to inactivation. Treatment with the alcohol sanitizer was very effective at inactivating attached pathogens or biofilms on the surface of stainless steel. Reduction levels of alcohol sanitizer treatment ranged from 1.91 to 4.77 log and from 4.35 to 5.35 log CFU/coupon in E. coli O157:H7 and S. aureus, respectively. From these results, the survival of pathogens contaminating the surfaces of food processing substrates such as stainless steel varied depending on RH and attachment form. Also, alcohol-based sanitizers can be used as a potential method to remove microbial contamination on the surfaces of utensils, cooking equipment, and other related substrates regardless of the microbial attached form.

Pulpal blood flow measurement with ultrasound Doppler imaging.

INTRODUCTION: This study compared the difference in pulpal blood flow between vital and root-filled teeth by using ultrasound Doppler imaging. METHODS: To compare the difference in pulpal blood flow between vital and root-filled teeth, 11 patients (mean age, 32.06 years; 3 male, 8 female) who had undergone root canal treatment on the anterior tooth of the maxilla or mandible and had a vital contralateral tooth were examined. Pulpal blood flow measurements were performed on the vital and root canal-treated teeth by using ultrasound Doppler imaging. The parameters examined were the maximum linear velocity (Vas), average linear velocity (Vam), minimum linear velocity (Vakd), pulsation index (PI), and circulation resistance (RI), which are indicators of the pulpal blood flow. The differences between the vital and root-filled teeth were examined by using a paired t test at the 95% confidence interval. RESULTS: There were significant differences in the Vas, Vam, Vakd, and RI between the vital and root-filled teeth (P<.05). With the root-filled teeth, ultrasound Doppler imaging revealed a linear and nonpulsed waveform, whereas the vital teeth showed a pulsed waveform that is characteristic of an arteriole. CONCLUSIONS: Ultrasound Doppler imaging can detect pulpal blood flow in vital tooth through indicators such as Vas, Vam, Vakd, PI, and RI.