scholarly journals Inhibition of Hydrophobic Protein-MediatedCandida albicans Attachment to Endothelial Cells during Physiologic Shear Flow

2001 ◽  
Vol 69 (5) ◽  
pp. 2815-2820 ◽  
Author(s):  
Pati M. Glee ◽  
Jim E. Cutler ◽  
Evelyn E. Benson ◽  
Robert F. Bargatze ◽  
Kevin C. Hazen
Keyword(s):  
Endothelial Cells ◽  
Shear Flow ◽  
Cell Surface ◽  
Protective Efficacy ◽  
Cell Surface Hydrophobicity ◽  
Surface Hydrophobicity ◽  
Host Cells ◽  
Yeast Cells ◽  
Cytokine Activation ◽  
Endothelial Monolayers

ABSTRACT Adhesion interactions during hematogenous dissemination ofCandida albicans likely involve a complex array of host and fungal factors. Possible C. albicans factors include changes in cell surface hydrophobicity and exposed antigens that have been shown in static adhesion assays to influence attachment events. We used a novel in vitro shear analysis system to investigate host-pathogen interactions and the role of fungal cell surface hydrophobicity in adhesion events with human endothelial cells under simulated physiologic shear. Endothelial monolayers were grown in capillary tubes and tested with and without interleukin-1β activation in buffered medium containing human serum. Hydrophobic and hydrophilic stationary-phase C. albicans yeast cells were infused into the system under shear flow and found to adhere with widely varying efficiencies. The average number of adherent foci was determined from multiple fields, sampled via video microscopy, between 8 and 12 min after infusion. Hydrophobic C. albicans cells demonstrated significantly more heterotypic binding events (Candida-endothelial cell) and greater homotypic binding events (Candida-Candida) than hydrophilic yeast cells. Cytokine activation of the endothelium significantly increased binding by hydrophobic C. albicans compared to unactivated host cells. Preincubation of hydrophobic yeast cells with a monoclonal antibody against hydrophobic cell wall proteins significantly blocked adhesion interactions with the endothelial monolayers. Because the antibody also blocks C. albicans binding to laminin and fibronectin, results suggest that vascular adhesion events with endothelial cells and exposed extracellular matrix may be blocked during C. albicans dissemination. Future studies will address the protective efficacy of blocking or redirecting blood-borne fungal cells to favor host defense mechanisms.

scholarly journals New Assay for Measuring Cell Surface Hydrophobicities of Candida dubliniensis andCandida albicans

2001 ◽  
Vol 8 (3) ◽  
pp. 585-587 ◽  
Author(s):  
M. A. Jabra-Rizk ◽  
W. A. Falkler ◽  
W. G. Merz ◽  
T. F. Meiller
Keyword(s):  
Cell Surface ◽  
Anaerobic Bacterium ◽  
Hydrophobic Interactions ◽  
Cell Surface Hydrophobicity ◽  
Fusobacterium Nucleatum ◽  
Surface Hydrophobicity ◽  
Yeast Cells ◽  
Pathogenic Yeast ◽  
Yeast Cell Surface ◽  
The Many

ABSTRACT Hydrophobic interactions, based on cell surface hydrophobicity (CSH), are among the many and varied mechanisms of adherence deployed by the pathogenic yeast Candida albicans. Recently it was shown that, unlike C. albicans, C. dubliniensisis a species that exhibits an outer fibrillar layer consistent with constant CSH. Previously, C. dubliniensis grown at 25 or 37°C was shown to coaggregate with the oral anaerobic bacteriumFusobacterium nucleatum. C. albicans, however, demonstrated similar coaggregation only when hydrophobic or grown at 25°C. This observation implied that coaggregation of Candida cells with F. nucleatum is associated with a hydrophobic yeast cell surface. To test this hypothesis, 42 C. albicans and 40 C. dubliniensis clinical isolates, including a C. albicans hydrophobic variant, were grown at 25 and 37°C and tested with the established hydrophobicity microsphere assay, which determines CSH levels based on the number of microspheres attached to the yeast cells. The coaggregation assay was performed in parallel experiments. All C. dubliniensis isolates grown at either temperature, hydrophobic 25°C-grown C. albicans isolates, and the C. albicans hydrophobic variant, unlike the 37°C-hydrophilic C. albicans isolates, exhibited hydrophobic CSH levels with the microsphere assay and simultaneously showed maximum, 4+, coaggregation with F. nucleatum. The parallel results obtained for C. dubliniensis using both assays support the use of the CoAg assay both as a rapid assay to determine CSH and to differentiate between C. dubliniensisand C. albicans.

Hydrophobic cell wall protein glycosylation by the pathogenic fungus Candida albicans

1994 ◽  
Vol 40 (4) ◽  
pp. 266-272 ◽  
Author(s):  
Kevin C. Hazen ◽  
Pati M. Glee
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Cell Surface ◽  
Molecular Mass ◽  
Pathogenic Fungus ◽  
Cell Surface Hydrophobicity ◽  
Surface Hydrophobicity ◽  
Protein Glycosylation ◽  
Yeast Cells ◽  
Cell Wall Proteins

Cell surface hydrophobicity influences adhesion and virulence of the opportunistic fungal pathogen Candida albicans. Previous studies have shown that cell surface hydrophobicity is due to specific proteins that are exposed on hydrophobic cells but are masked by long fibrils on hydrophilic cells. This observation suggests that hydrophobic cell wall proteins may contain little or no mannosylation. In the present study, the glycosylation levels of three hydrophobic cell wall proteins (molecular mass range between 36 and 40 kDa) derived from yeast cells were examined. One hydrophilic protein (90 kDa) was also tested. Various endoglycosidases (endoglycosidase F – N-glycosidase F, O-glycosidase, β-mannosidase, N-glycosidase F), an exoglycosidase (α-mannosidase), and trifluoromethane sulfonic acid were used to deglycosylate the proteins. All four proteins were reactive to the lectin concanavalin A, demonstrating that they were mannoproteins. However, gel electrophoresis of the control and treated proteins revealed that mannosyl groups of hydrophobic proteins were less than 2 kDa in size, while the mannosyl group of the hydrophilic protein had a molecular mass of approximately 20 kDa. These results suggest that unlike many hydrophilic proteins, hydrophobic proteins may have low levels of glycosylation. Changes in glycosylation may determine exposure of hydrophobic protein regions at the cell surface.Key words: Candida albicans, cell wall, mannoproteins, hydrophobicity, fibrils.

scholarly journals Analysis of Functional Domains of theEnterococcus faecalis Pheromone-Induced Surface Protein Aggregation Substance

2001 ◽  
Vol 183 (19) ◽  
pp. 5659-5667 ◽  
Author(s):  
C. M. Waters ◽  
G. M. Dunny
Keyword(s):  
Cell Surface ◽  
High Efficiency ◽  
Cell Surface Hydrophobicity ◽  
Surface Protein ◽  
Cell Aggregation ◽  
Surface Hydrophobicity ◽  
Host Cells ◽  
Functional Domains ◽  
Aggregation Substance ◽  
C Terminus

ABSTRACT Pheromone-inducible aggregation substance (AS) proteins ofEnterococcus faecalis are essential for high-efficiency conjugation of the sex pheromone plasmids and also serve as virulence factors during host infection. A number of different functions have been attributed to AS in addition to bacterial cell aggregation, including adhesion to host cells, adhesion to fibrin, increased cell surface hydrophobicity, resistance to killing by polymorphonuclear leukocytes and macrophages, and increased vegetation size in an experimental endocarditis model. Relatively little information is available regarding the structure-activity relationship of AS. To identify functional domains, a library of 23 nonpolar 31-amino-acid insertions was constructed in Asc10, the AS encoded by the plasmid pCF10, using the transposons TnlacZ/in and TnphoA/in. Analysis of these insertions revealed a domain necessary for donor-recipient aggregation that extends further into the amino terminus of the protein than previously reported. In addition, insertions in the C terminus of the protein also reduced aggregation. As expected, the ability to aggregate correlates with efficient plasmid transfer. The results also indicated that an increase in cell surface hydrophobicity resulting from AS expression is not sufficient to mediate bacterial aggregation.

scholarly journals Evaluation of Relative Yeast Cell Surface Hydrophobicity Measured by Flow Cytometry

2005 ◽  
Vol 13 (1) ◽  
pp. 43-48 ◽  
Author(s):  
Lisa Colling ◽  
Richard N. Carter ◽  
Michael Essmann ◽  
Bryan Larsen
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Binding Capacity ◽  
Cell Surface Hydrophobicity ◽  
Fungal Growth ◽  
Surface Hydrophobicity ◽  
Growth Conditions ◽  
Yeast Cells ◽  
Appreciable Effect ◽  
Polystyrene Microspheres

Objective:To develop an efficient method for evaluating cell surface hydrophobicity and to apply the method to demonstrate the effects of fungal growth conditions on cell surface properties.Methods:Yeast isolates were suspended in phosphate-buffered saline and mixed with deep blue-dyed polystyrene microspheres. Flow cytometry was used to detect the degree of microsphere binding to yeast cells. Different strains of yeast were compared for intrinsic microsphere binding activity and changes in growth conditions were invoked to modify the relative surface hydrophobicity.Results:Commercially available blue-dyed polystyrene microspheres showed strong fluorescence in the FL3 channel, whereas yeast cells did not show appreciable FL3 fluorescence. Microspheres and yeast were generally distinguishable on the basis of size revealed by forward light scatter. This method showed a wide variation in intrinsic cell surface hydrophobicity amongCandida albicansstrains. Likewise, variation in hydrophobicity of non-albicans yeast species was observed. Growth on solid media, incubation at 25°C, or 250 mg/dl glucose concentration increased hydrophobicity compared with growth in liquid media, incubation at 37°C, or 50 mg/dl glucose, respectively. Growth in1×10−9M estradiol had no appreciable effect on hydrophobicity.Conclusions:Stained latex microspheres fluoresced in the FL3 channel of the flow cytometer and bound to yeast cells to an extent related to the surface hydrophobicity of the yeast. Binding detected by flow cytometry showed that clinical yeast isolates varied in intrinsic binding capacity and this binding ability was altered by different growth conditions. The implications for virulence regulation among yeast isolates are discussed.

Autecology of scum producing bacteria

1998 ◽  
Vol 37 (4-5) ◽  
pp. 527-530 ◽  
Author(s):  
Hilde Lemmer ◽  
George Lind ◽  
Margit Schade ◽  
Birgit Ziegelmayer
Keyword(s):  
Wastewater Treatment ◽  
Cell Surface ◽  
Wastewater Treatment Plants ◽  
Cell Surface Hydrophobicity ◽  
Surface Hydrophobicity ◽  
Residence Times ◽  
Loading Conditions ◽  
Emulsifying Activity ◽  
Substrate Preferences

Non-filamentous hydrophobic scum bacteria were isolated from scumming wastewater treatment plants (WWTP) by means of adhesion to hydrocarbons. They were characterized with respect to taxonomy, substrate preferences, cell surface hydrophobicity, and emulsification capability. Their role during flotation events is discussed. Rhodococci are selected by hydrolysable substrates and contribute to flotation both by cell surface hydrophobicity and emulsifying activity at long mean cell residence times (MCRT). Saprophytic Acinetobacter strains are able to promote flotation by hydrophobicity and producing emulsifying agents under conditions when hydrophobic substrates are predominant. Hydrogenophaga and Acidovorax species as well as members of the Cytophaga/Flavobacterium group are prone to proliferate under low loading conditions and contribute to flotation mainly by emulsification.

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