172 research outputs found
Surface modifying substances that reduce apparent yeast cell hydrophobicity.
OBJECTIVE: To determine whether several topical compounds and other chemical entities are able to diminish the surface hydrophobicity of yeast cells. METHOD: Hydrophobicity of yeast cells was determined by binding styrene microspheres to the surface of untreated yeast or yeast pre-incubated with various substances with potential for cell surface modification. The degree of microsphere adherence to yeast cells was measured by flow cytometry. RESULTS: A significant reduction in cell surface hydrophobicity was observed when yeast was incubated in protein-containing media. Other compounds that effectively reduced microsphere binding were various formulations of K-Y and heparin. Divalent cations (Ca+ + , Mg+ + , Zn+ + , Cu + + ) were also potent inhibitors of microsphere adherence. It was possible to remove substances contributing to microsphere binding by chemical extraction of the yeast. Yeast having reduced microsphere binding activity also showed diminished binding of concanavalin A. CONCLUSIONS: Several commercially available compounds were able to block binding of styrene microspheres to yeast. Some of the binding activity appeared to be attributable to mannose-containing surface components. These findings have implications for formulating therapeutic products that might block yeast binding to tissues
Tetracycline Effects on Candida Albicans Virulence Factors
Object. To determine if tetracycline, previously reported to increase the probability of developing symptomatic vaginal yeast infections, has a direct effect on Candida albicans growth or induction of virulent phenotypes. Method. In vitro, clinical isolates of yeast were cultivated with sublethal concentrations of tetracycline and yeast cell counts, hyphal formation, drug efflux pump activity, biofilm production, and hemolysin production were determined by previously reported methods.
Results. Tetracycline concentrations above 150 μg/mL inhibited Candida albicans, but at submicrogram/mL, a modest growth increase during the early hours of the growth curve was observed. Tetracycline did not inhibit hyphal formation at sublethal concentrations. Hypha formation appeared augmented by exposure to tetracycline in the presence of chemically defined medium and especially in the presence of human serum. Efflux pump CDR1 was upregulated and a nonsignificant trend toward increased biofilm formation was noted. Conclusion. Tetracycline appears to have a small growth enhancing effect and may influence virulence through augmentation of hypha formation, and a modest effect on drug efflux and biofilm formation, although tetracycline did not affect hemolysin. It is not clear if the magnitude of the effect is sufficient to attribute vaginitis following tetracycline treatment to direct action of tetracycline on yeast
Adherence and Blocking of Candida Albicans to Cultured Vaginal Epithelial Cells: Treatments to Decrease Adherence
Background. Pathogenesis of mucosal microorganisms depends on adherence to the tissues they colonize and infect. For Candida albicans, cell surface hydrophobicity may play a significant role in tissue binding ability. Methods. A continuous cell line of vaginal epithelial cells (VEC) was grown in keratinocyte serum-free medium (KSFM) with supplements and harvested by trypsinization. VEC were combined with yeast cells to evaluate adherence and inhibition of adherence. In this experimental setup, yeast stained with fluorescein isothiocyanate were allowed to attach to VEC and the resulting fluorescent VEC were detected by flow cytometry. Results. VEC were cultured and examined daily after plating and showed morphology similar to basal epithelial cells. Culture media supplemented with estradiol showed increased VEC proliferation initially (first 24 h) but cell morphology was not altered. Fluorescinated Candida cells bound effectively to the cultured VEC. Using fresh cells exposed to various preparations of K-Y, we showed that all formulations of the product reduced Candida binding to VEC by 25% to 50%. While VEC were generally harvested for use in experiments when they were near confluent growth, we allowed some cultures to grow beyond that point and discovered that cells allowed to become overgrown or stressed appeared to bind yeast cells more effectively. Conclusion. Flow cytometry is a useful method for evaluating binding of stained yeast cells to cultured VEC and has demonstrated that commercially available products have the ability to interfere with the process of yeast adherence to epithelial cells
Candidiasis During Pregnancy May Result From Isogenic Commensal Strains
Objective: Our laboratory previously demonstrated that asymptomatic vaginal colonization during pregnancy is a factor predisposing patients to subsequent symptomatic vulvovaginal candidiasis. It is unknown whether symptoms result from strain replacement or a change in host relationship to the original colonizing strain. This study was undertaken to determine whether Candida albicans isolates from asymptomatic women could be responsible for subsequent symptomatic vaginitis. Methods: We retained isolates of C. albicans from women followed longitudinally through pregnancy, and identified six pairs of cultures from women who were colonized without symptoms and who later became symptomatic (average time 14 weeks). We used a random amplification of polymorphic DNA (RAPD) analysis to determine whether isolates from our study patients were genetically similar or dissimilar. Results: Analysis of these pairs of yeast strains by RAPD revealed that five of the six women had symptoms apparently due to the same yeast strain that was found initially as a commensal strain. To increase the power of these observations, we also performed RAPD analysis on six randomly selected yeast strains from other women in this study who had not become symptomatic to determine whether any of these unrelated strains matched strains from those women who became symptomatic. Conclusion: Symptomatic yeast vaginitis is usually due to strains of C. albicans already carried in the lower genital tract, underscoring the need to understand regulation of growth and virulence of the organism in vivo
Evaluation of relative yeast cell surface hydrophobicity measured by flow cytometry.
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 among Candida albicans strains. Likewise, variation in hydrophobicity of non-albicans yeast species was observed. Growth on solid media, incubation at 25 degrees C, or 250 mg/dl glucose concentration increased hydrophobicity compared with growth in liquid media, incubation at 37 degrees C, or 50 mg/dl glucose, respectively. Growth in 1 x 10(-9) M 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
Expertise and Trust-Aware Social Web Service Recommendation
With the increasing number of Web services, the personalized recommendation of Web services has become more and more important. Fortunately, the social network popularity nowadays brings a good alternative for social recommendation to avoid the data sparsity problem that is not treated very well in the collaborative filtering approach. Since the social network provides a big data about the users, the trust concept has become necessary to filter this abundance and to foster the successful interactions between the users. In this paper, we firstly propose a trusted friend detection mechanism in a social network. The dynamic of the users’ interactions over time and the similarity of their interests have been considered. Secondly, we propose a Web service social recommendation mechanism which considers the expertise of the trusted friends according to their past invocation histories and the active user’s query. The experiments of each mechanism produced satisfactory results
Growth of a vortex polycrystal in type II superconductors
We discuss the formation of a vortex polycrystal in type II superconductors
from the competition between pinning and elastic forces. We compute the elastic
energy of a deformed grain boundary, that is strongly non-local, and obtain the
depinning stress for weak and strong pinning. Our estimates for the grain size
dependence on the magnetic field strength are in good agreement with previous
experiments on NbMo. Finally, we discuss the effect of thermal noise on grain
growth.Comment: 4 pages, 2 figure
Whole-Sample Mapping of Cancerous and Benign Tissue Properties
Structural and mechanical differences between cancerous and healthy tissue
give rise to variations in macroscopic properties such as visual appearance and
elastic modulus that show promise as signatures for early cancer detection.
Atomic force microscopy (AFM) has been used to measure significant differences
in stiffness between cancerous and healthy cells owing to its high force
sensitivity and spatial resolution, however due to absorption and scattering of
light, it is often challenging to accurately locate where AFM measurements have
been made on a bulk tissue sample. In this paper we describe an image
registration method that localizes AFM elastic stiffness measurements with
high-resolution images of haematoxylin and eosin (H\&E)-stained tissue to
within 1.5 microns. Color RGB images are segmented into three structure types
(lumen, cells and stroma) by a neural network classifier trained on
ground-truth pixel data obtained through k-means clustering in HSV color space.
Using the localized stiffness maps and corresponding structural information, a
whole-sample stiffness map is generated with a region matching and
interpolation algorithm that associates similar structures with measured
stiffness values. We present results showing significant differences in
stiffness between healthy and cancerous liver tissue and discuss potential
applications of this technique.Comment: Accepted at MICCAI201
Simulations of HIV capsid protein dimerization reveal the effect of chemistry and topography on the mechanism of hydrophobic protein association
Recent work has shown that the hydrophobic protein surfaces in aqueous
solution sit near a drying transition. The tendency for these surfaces to expel
water from their vicinity leads to self assembly of macromolecular complexes.
In this article we show with a realistic model for a biologically pertinent
system how this phenomenon appears at the molecular level. We focus on the
association of the C-terminal domain (CA-C) of the human immunodeficiency virus
(HIV) capsid protein. By combining all-atom simulations with specialized
sampling techniques we measure the water density distribution during the
approach of two CA-C proteins as a function of separation and amino acid
sequence in the interfacial region. The simulations demonstrate that CA-C
protein-protein interactions sit at the edge of a dewetting transition and that
this mesoscopic manifestation of the underlying liquid-vapor phase transition
can be readily manipulated by biology or protein engineering to significantly
affect association behavior. While the wild type protein remains wet until
contact, we identify a set of in silico mutations, in which three hydrophilic
amino acids are replaced with nonpolar residues, that leads to dewetting prior
to association. The existence of dewetting depends on the size and relative
locations of substituted residues separated by nm length scales, indicating
long range cooperativity and a sensitivity to surface topography. These
observations identify important details which are missing from descriptions of
protein association based on buried hydrophobic surface area
Binding of the human nucleotide excision repair proteins XPA and XPC/HR23B to the 5R-thymine glycol lesion and structure of the cis-(5R,6S) thymine glycol epimer in the 5′-GTgG-3′ sequence: destabilization of two base pairs at the lesion site
The 5R thymine glycol (5R-Tg) DNA lesion exists as a mixture of cis-(5R,6S) and trans-(5R,6R) epimers; these modulate base excision repair. We examine the 7:3 cis-(5R,6S):trans-(5R,6R) mixture of epimers paired opposite adenine in the 5′-GTgG-3′ sequence with regard to nucleotide excision repair. Human XPA recognizes the lesion comparably to the C8-dG acetylaminoflourene (AAF) adduct, whereas XPC/HR23B recognition of Tg is superior. 5R-Tg is processed by the Escherichia coli UvrA and UvrABC proteins less efficiently than the C8-dG AAF adduct. For the cis-(5R, 6S) epimer Tg and A are inserted into the helix, remaining in the Watson–Crick alignment. The Tg N3H imine and A N6 amine protons undergo increased solvent exchange. Stacking between Tg and the 3′-neighbor G•C base pair is disrupted. The solvent accessible surface and T2 relaxation of Tg increases. Molecular dynamics calculations predict that the axial conformation of the Tg CH3 group is favored; propeller twisting of the Tg•A pair and hydrogen bonding between Tg OH6 and the N7 atom of the 3′-neighbor guanine alleviate steric clash with the 5′-neighbor base pair. Tg also destabilizes the 5′-neighbor G•C base pair. This may facilitate flipping both base pairs from the helix, enabling XPC/HR23B recognition prior to recruitment of XPA
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