Reduction of Listeria monocytogenes on Green Peppers (Capsicum annuum L.) by Gaseous and Aqueous Chlorine Dioxide and Water Washing and Its Growth at 7°C†

2001 ◽  
Vol 64 (11) ◽  
pp. 1730-1738 ◽  
Author(s):  
Y. HAN ◽  
R. H. LINTON ◽  
S. S. NIELSEN ◽  
P. E. NELSON
Keyword(s):  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Membrane Surface ◽  
Fluorescein Isothiocyanate ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Green Pepper ◽  
Gas Treatment ◽  
Water Washing ◽  
Significant Difference

Reduction of Listeria monocytogenes Scott A on uninjured and injured surfaces of green peppers after 0.3- and 3-mg/liter gaseous and aqueous ClO2 treatment and water washing for 10 min at 20°C was studied. Growth of the L. monocytogenes untreated or treated with 0.6 mg/liter ClO2 gas for 30 min at 20°C on green peppers also was investigated. A membrane-surface-plating method was used for resuscitation and enumeration of L. monocytogenes treated with ClO2. The bacterial viability on pepper surfaces was visualized using confocal laser scanning microscopy (CLSM). Live and dead cells of L. monocytogenes were labeled with a fluorescein isothiocyanate-labeled antibody and propidium iodide, respectively. More than 6 log CFU/5 g L. monocytogenes on uninjured surfaces and about 3.5 log CFU/5 g on injured surfaces were inactivated by both 3-mg/liter and 0.6-mg/liter ClO2 gas treatments. The 3-mg/liter aqueous ClO2 treatment achieved 3.7- and 0.4-log reductions on uninjured and injured surfaces, respectively; whereas, water washing alone showed 1.4- and 0.4-log reductions, respectively. ClO2 gas treatment was the most effective in reducing L. monocytogenes on both uninjured and injured green pepper surfaces, when compared with aqueous ClO2 treatment and water washing. The significant difference (P < 0.05) between log reductions on uninjured and injured surfaces and the results from CLSM analysis suggested that injured surfaces protected more bacteria from sanitation treatments than did uninjured surfaces. Not only could L. monocytogenes grow on green pepper surfaces at 7°C, bacteria that survived the 0.6-mg/liter ClO2 gas treatment also could grow.

Use of confocal laser scanning microscopy and a computer model to understand ink cavitation and filamentation

TAPPI Journal ◽  
2010 ◽  
Vol 9 (10) ◽  
pp. 7-15
Author(s):  
HANNA KOIVULA ◽  
DOUGLAS BOUSFIELD ◽  
MARTTI TOIVAKKA
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Print Quality ◽  
Length Scale ◽  
Offset Printing ◽  
Significant Difference ◽  
Printing Speed

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.

scholarly journals Vacuolar-type H+-ATPase regulates cytoplasmic pH in Toxoplasma gondii tachyzoites

1998 ◽  
Vol 330 (2) ◽  
pp. 853-860 ◽  
Author(s):  
N. J. Silvia MORENO ◽  
Li ZHONG ◽  
Hong-Gang LU ◽  
Wanderley DE SOUZA ◽  
Marlene BENCHIMOL
Keyword(s):  
Plasma Membrane ◽  
Toxoplasma Gondii ◽  
Laser Scanning ◽  
Membrane Surface ◽  
Surface Proteins ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Dependent Manner ◽  
Cytoplasmic Ph ◽  
Bafilomycin A1

Cytoplasmic pH (pHi) regulation was studied in Toxoplasma gondii tachyzoites by using the fluorescent dye 2ʹ,7ʹ-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. Their mean baseline pHi (7.07±0.06; n = 5) was not significantly affected in the absence of extracellular Na+, K+ or HCO3- but was significantly decreased in a dose-dependent manner by low concentrations of N,Nʹ-dicyclohexylcarbodi-imide (DCCD), N-ethylmaleimide (NEM) or bafilomycin A1. Bafilomycin A1 also inhibited the recovery of tachyzoite pHi after an acid load with sodium propionate. Similar concentrations of DCCD, NEM and bafilomycin A1 produced depolarization of the plasma membrane potential as measured with bis-(1,3-diethylthiobarbituric)trimethineoxonol (bisoxonol), and DCCD prevented the hyperpolarization that accompanies acid extrusion after the addition of propionate, in agreement with the electrogenic nature of this pump. Confocal laser scanning microscopy indicated that, in addition to being located in cytoplasmic vacuoles, the vacuolar (V)-H+-ATPase of T. gondii tachyzoites is also located in the plasma membrane. Surface localization of the V-H+-ATPase was confirmed by experiments using biotinylation of cell surface proteins and immunoprecipitation with antibodies against V-H+-ATPases. Taken together, the results are consistent with the presence of a functional V-H+-ATPase in the plasma membrane of these intracellular parasites and with an important role of this enzyme in the regulation of pHi homoeostasis in these cells.

scholarly journals Micromorphological analysis and bond strength comparison of two adhesives for different degrees of dental fluorosis

2020 ◽  
Author(s):  
Shuangfeng Liu ◽  
Yanxia Zhu ◽  
Tana Gegen
Keyword(s):  
Shear Strength ◽  
Bond Strength ◽  
Bonding Strength ◽  
Laser Scanning ◽  
Dental Fluorosis ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Cohesive Failure ◽  
Bonding System ◽  
Significant Difference

Abstract The objective of this study was to analyze morphologically the all-etching bonding system and self-etching bonding system for enamel with different degrees of fluorosis and evaluate the bond strength of each system. Teeth that were indicated for extraction owing to orthodontic or periodontal problems were selected. According to Dean’s index and the Thylstrup-Fejerskov index, 180 extracted teeth were divided into three groups of mild, moderate, and severe dental fluorosis (DF), with 60 teeth in each group. The teeth in each group were randomly divided into two subgroups (n = 30), which were then subjected to the all-etching bonding system (Prime & Bond NT) and self-etching bonding system (SE-Bond). Each group of adhesives was used to bond Z350 universal resin (3M) to the etched dental enamel. Tensile and shear tests were conducted to determine the bond strength. Subsequently, the fractured specimens were investigated using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The Prime & Bond NT was statistically significant for the tensile and shear strength of enamel with mild fluorosis (P < 0.05) but did not exhibit a significant difference for moderate and severe DF (P > 0.05). The SE-Bond was not statistically significant for the tensile and shear strength of mild, moderate, or severe DF (P > 0.05). The SEM and CLSM results reveal that the mild fluorosis enamel crystals were relatively dense, and a small amount of resin remained. The moderate fluorosis enamel crystals were loosely arranged, and the gaps were widened. The severe fluorosis enamel crystals were irregularly arranged. The disorder was aggravated, and the dentinal orifice was exposed by partial enamel exfoliation. The bonding strength of mild fluorosis enamel with the Prime & Bond NT was better than that with the SE-Bond, and cohesive failure was the most common mode of failure. Because there was no difference in the bonding strength of the SE-Bond for different degrees of DF, we recommend the use of the all-etching adhesive system in the clinical treatment of teeth with mild fluorosis.

scholarly journals Primary Neurons and Differentiated NSC-34 Cells Are More Susceptible to Arginine-Rich ALS Dipeptide Repeat Protein-Associated Toxicity than Non-Differentiated NSC-34 and CHO Cells

2019 ◽  
Vol 20 (24) ◽  
pp. 6238 ◽  
Author(s):  
Anna L. Gill ◽  
Monica Z. Wang ◽  
Beth Levine ◽  
Alan Premasiri ◽  
Fernando G. Vieira
Keyword(s):  
Motor Neurons ◽  
Laser Scanning ◽  
Neuronal Cell ◽  
Fluorescein Isothiocyanate ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Primary Neurons ◽  
Repeat Protein ◽  
Neuronal Cell Lines ◽  
Dipeptide Repeat

A repeat expansion mutation in the C9orf72 gene is the most common known genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). In this study, using multiple cell-based assay systems, we reveal both increased dipeptide repeat protein (DRP) toxicity in primary neurons and in differentiated neuronal cell lines. Using flow cytometry and confocal laser scanning microscopy of cells treated with fluorescein isothiocyanate (FITC)-labeled DRPs, we confirm that poly-glycine-arginine (GR) and poly-proline-arginine (PR) DRPs entered cells more readily than poly-glycine-proline (GP) and poly-proline-alanine (PA) DRPs. Our findings suggest that the toxicity of C9-DRPs may be influenced by properties associated with differentiated and aging motor neurons. Further, our findings provide sensitive cell-based assay systems to test phenotypic rescue ability of potential interventions.

scholarly journals Evaluation of four final irrigation protocols for cleaning root canal walls

2020 ◽  
Vol 12 (1) ◽  
Author(s):  
Qiang Li ◽  
Qian Zhang ◽  
Xiaoying Zou ◽  
Lin Yue
Keyword(s):  
Root Canal ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Smear Layer ◽  
Confocal Laser ◽  
Bacterial Survival ◽  
Bacterial Inhibition ◽  
Layer Removal ◽  
Significant Difference ◽  
Canal Systems

Abstract The aim of this study was to compare the efficiency of four final irrigation protocols in smear layer removal and bacterial inhibition in root canal systems. Thirty roots inoculated with Enterococcus faecalis were prepared with ProTaper Universal files. The teeth were disinfected by conventional needle irrigation, sonic agitation using the EndoActivator device, passive ultrasonic irrigation, or an M3 Max file. Teeth with no root canal preparation served as blank controls for the establishment of the infection baseline. Teeth with preparation but no final irrigation served as a post-instrumentation baseline. After the final irrigation, the teeth were sectioned in half. One half of each tooth was examined by scanning electron microscopy (SEM) to assess smear layer removal using a five-point scale. The other half was examined by confocal laser scanning microscopy (CLSM) using the LIVE/DEAD BackLight bacterial viability kit to evaluate the depth of bacterial survival in dentinal tubules. SEM analysis revealed no significant difference in smear layer removal throughout the whole canal among the EA, PUI, and M3 Max groups (P > 0.05). CLSM revealed that PUI achieved the greatest bacterial inhibition depth in the coronal ((174.27 ± 31.63) μm), middle ((160.94 ± 37.77) μm), and apical ((119.53 ± 28.49) μm) thirds of the canal (all P < 0.05 vs. other groups). According to this comprehensive SEM and CLSM evaluation, PUI appears to have the best infection control ability in root canal systems.

scholarly journals Enhanced Bactericidal Efficacy of NaOCl at pH 12 Followed by Acidified NaOCl at pH 6.5 on Enterococcus faecalis Biofilm

2020 ◽  
Vol 10 (17) ◽  
pp. 6096
Author(s):  
Ronald Wigler ◽  
Shlomo Matalon ◽  
Tomer Goldberger ◽  
Anat Or Lerner ◽  
Anda Kfir
Keyword(s):  
Contact Time ◽  
Laser Scanning ◽  
Saline Solution ◽  
Volume Ratio ◽  
Bactericidal Effect ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Red Fluorescence ◽  
Bactericidal Efficacy ◽  
Significant Difference

This study aimed to determine the bactericidal efficacy of sequential use of NaOCl pH 12 followed by acidified NaOCl pH 6.5, and compare it to that of either of these NaOCl solutions alone. E. faecalis biofilm was grown on standardized dentine specimens for four weeks. The specimens were randomly divided into four groups: (A) 4 min exposure to 0.9% saline solution (control); (B) 4 min exposure to 4% NaOCl pH 12; (C) 4 min exposure to 4% NaOCl pH 6.5; and (D) 2 min exposure to 4% NaOCl pH 12 followed by 2 min exposure to 4% NaOCl pH 6.5. The bactericidal activity was evaluated after the 4 min of contact time using confocal laser scanning microscopy. The volume ratio of red fluorescence to green and red fluorescence indicated the proportion of dead cells in the biofilm. The percent of dead cells in the saline solution group was significantly lower than those in the other groups. There was no significant difference between NaOCl pH 12 compared to NaOCl pH 6.5. The sequential use of NaOCl pH 12 followed by pH 6.5 significantly increased the percent of dead cells compared to both the samples exposed to either NaOCl pH 12 or pH 6.5. These results show that sequential irrigation protocol had a stronger bactericidal effect than the commonly used NaOCl pH 12.

Effects of Bacteriophage P100 at Different Concentrations on the Structural Parameters of Listeria monocytogenes Biofilms

2018 ◽  
Vol 81 (12) ◽  
pp. 2040-2044 ◽  
Author(s):  
CRISTINA RODRÍGUEZ-MELCÓN ◽  
ROSA CAPITA ◽  
CAMINO GARCÍA-FERNÁNDEZ ◽  
CARLOS ALONSO-CALLEJA
Keyword(s):  
Image Analysis ◽  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Digital Image Analysis ◽  
Surface Coverage ◽  
Structural Parameters ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Foodborne Diseases ◽  
Average Thickness

ABSTRACT Because listeriosis is one of the deadliest foodborne diseases, controlling and eradicating Listeria monocytogenes biofilms is a serious challenge for food safety. Biofilms (24 h old) formed on polystyrene by a L. monocytogenes strain of food origin were exposed for a further 24 h to 12 different concentrations (from 100 to 1011 PFU/mL) of the bacteriophage P100 (Listex P100). The structural parameters of biofilms were studied by using confocal laser scanning microscopy and digital image analysis. The biovolume in the observation field (14,121 μm2) of control (untreated) biofilms was 237,333.1 ± 2,692.6 μm3. The biomass of treated biofilms ranged from 164.7 ± 89.0 μm3 (biofilms exposed to 1010 PFU/mL) to 231,170.5 ± 15,142.0 μm3 (100 PFU/mL). The lowest biomass was achieved after treatment with 108 PFU/mL, with no further decrease in biovolume when higher phage concentrations were used. A strong (P &lt; 0.001) correlation was found between phage concentration (log units) and biovolume (−0.965), surface coverage (−0.939), roughness (0.976), maximum thickness (−0.853), and average thickness (−0.965). Findings from this research suggest that bacteriophage P100 at concentrations equal to or greater than 8 log PFU/mL successfully removes L. monocytogenes biofilms from polystyrene surfaces.

Bacterial Colonization in the Marginal Region of Ceramic Restorations: Effects of Different Cement Removal Methods and Polishing

2016 ◽  
Vol 41 (6) ◽  
pp. 642-654 ◽  
Author(s):  
SMB Pereira ◽  
LC Anami ◽  
CA Pereira ◽  
ROA Souza ◽  
KZ Kantorski ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Bacterial Colonization ◽  
Resin Cement ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Significant Difference ◽  
Ceramic Blocks ◽  
Cement Removal ◽  
Ceramic Restorations ◽  
Marginal Region

SUMMARY This study evaluated the effects of excess cement removal techniques, with or without subsequent polishing, on biofilm formation and micromorphology in the marginal region of the tooth/restoration. From bovine teeth, 96 dentin blocks (4 × 8 × 2 mm) were produced, molded, and reproduced in type IV gypsum, on which 96 pressed ceramic blocks (Vita PM9, Vita Zahnfabrik; 4 × 8 × 2 mm) were produced via the lost wax technique. The dentin blocks and their respective ceramic blocks were cemented with a self-adhesive resin cement (RelyX U200, 3M ESPE), and cement excess was removed from the margin using four different techniques, followed or not by polishing with silicone rubber tips: MBr, removal with microbrush and photoactivation; MBr-Pol, MBr + polishing; Br, removal with brush and photoactivation; Br-Pol, Br + polishing; Photo-Expl, 5 seconds of initial photoactivation, removal with explorer, and final curing; Photo-Expl-Pol, Photo-Expl + polishing; Photo-SB, 5 seconds of initial photoactivation, removal with scalpel, and final curing; and Photo-SB-Pol, Photo-SB + polishing. After 24 hours, the roughness in the marginal region was analyzed using a profilometer (three measurements on each sample). Micromorphological analyses of the region were performed by stereomicroscope and scanning electron microscopy (SEM). Then the samples were contaminated with sucrose broth standardized suspension with Streptococcus mutans, Staphylococcus aureus, and Candida albicans and incubated for a period of 48 hours. The samples were quantitatively analyzed for bacterial adherence in the marginal region by confocal laser scanning microscopy and counting of colony-forming units (CFUs/mL) and qualitatively analyzed using SEM. Roughness data (Ra) were submitted to two-way analysis of variance, Tukey test at a confidence level of 95%, and Student t-tests. CFU, biomass, and biothickness data were analyzed by Kruskal-Wallis, Mann-Whitney, and Dunn tests. The removing technique statistically influenced Ra (MBr, p=0.0019; Br, p=0.002; Photo-Expl, p=0.0262; Photo-SB, p=0.0196) when comparing the polished and unpolished groups. The MBr and MBr-Pol technique differed significantly for CFU/mL values (p=0.010). There was no significant difference in the amounts of biomass and biothickness comparing polished and unpolished groups and when all groups were compared (p&gt;0.05). Different morphological patterns were observed (more regular surface for polished groups). We conclude that margin polishing after cementation of feldspar/pressed ceramic restorations is decisive for achieving smoother surfaces, as the excess cement around the edges can increase the surface roughness in these areas, influencing bacterial adhesion.

scholarly journals Macropinocytosis as a Mechanism of Entry into Primary Human Urethral Epithelial Cells by Neisseria gonorrhoeae

2000 ◽  
Vol 68 (3) ◽  
pp. 1696-1699 ◽  
Author(s):  
Michael K. Zenni ◽  
Peter C. Giardina ◽  
Hillery A. Harvey ◽  
Jianqiang Shao ◽  
Margaret R. Ketterer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Laser Scanning ◽  
Actin Polymerization ◽  
Kinase Inhibitors ◽  
Fluorescein Isothiocyanate ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Transmission Electron ◽  
Microscopy Analysis ◽  
Phosphoinositide 3 Kinase

ABSTRACT Gonococcal entry into primary human urethral epithelial cells (HUEC) can occur by macropinocytosis. Scanning and transmission electron microscopy revealed lamellipodia surrounding gonococci, and confocal laser scanning microscopy analysis showed organisms colocalized with M r 70,000 fluorescein isothiocyanate-labeled dextran within the cells. Phosphoinositide 3-kinase inhibitors and an actin polymerization inhibitor prevented macropinocytic entry of gonococci into HUEC.

scholarly journals Influence of Light-curing Parameters on Biofilm Development and Flexural Strength of a Silorane-based Composite

2016 ◽  
Vol 41 (2) ◽  
pp. 219-227 ◽  
Author(s):  
E Brambilla ◽  
A Ionescu ◽  
G Cazzaniga ◽  
M Ottobelli
Keyword(s):  
Flexural Strength ◽  
Laser Scanning ◽  
Testing Machine ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Curing Time ◽  
Two Samples ◽  
Significant Difference ◽  
Light Curing ◽  
Biofilm Development

SUMMARYObjectives: The aim of this study was to evaluate the differences in biological and mechanical performances of a silorane-based and a methacrylate-based composite. Another aim was to assess the influence of light-curing time and light-curing intensity on in vitro biofilm formation and flexural strength of the two tested composites.Methods: Experiment 1: 432 specimens obtained from a silorane-based composite and from a standard methacrylate-based composite were divided into six groups and light-cured for 10, 20, 30, 40, 60, or 80 seconds, using one of two light-curing intensities, 400 mW/cm2 or 800 mW/cm2. At 24 hours, a monospecific Streptococcus mutans biofilm adherent to the surfaces of the samples was obtained. Then, a colorimetric technique (MTT assay) was used to evaluate the adherent viable biomass. Two samples per group were observed using confocal laser scanning microscopy. Analysis of variance (ANOVA) and Tukey tests were used to analyze the results (p&lt;0.05). Experiment 2: 192 bar-shaped specimens were obtained and light-cured as in the previous experiment. A three-point bend test using a universal testing machine was performed to obtain flexural strength values. ANOVA and Tukey tests were used to analyze the results (p&lt;0.05).Results: In experiment 1, a highly significant difference (p&lt;0.0001) in biofilm development was shown between silorane-based and methacrylate-based composites. In fact, the silorane-based composite exhibited better biological performance. Significant differences were also found between the two light-curing intensities (p&lt;0.018) and for curing times (p&lt;0.0001): silorane-based composite light-cured for 80 seconds at 800 mW/cm2 light-curing intensity showed the lowest biofilm development. In experiment 2, a significant difference in flexural strength (p&lt;0.0318) was only found between the different composites. Nevertheless, both resin composites showed flexural strength values in accordance with International Organization for Standardization guidelines even after 10 seconds of light-curing time.Conclusions: Silorane-based composite was less prone to biofilm development compared with a methacrylate-based composite. Acceptable flexural strength values for both composites were obtained after 10 seconds of light-curing time.

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