scholarly journals Effect of surface treatments of titanium on amphotericin B-treated Candida albicans persister cells

Mycoses ◽  
2011 ◽  
Vol 54 (3) ◽  
pp. 189-194 ◽  
Author(s):  
C. S. P. Tsang ◽  
D. Y. K. Tang
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Surface Treatments ◽  
Persister Cells ◽  
Effect Of Surface

scholarly journals Inhibition of Nucleic Acid Biosynthesis Makes Little Difference to Formation of Amphotericin B-Tolerant Persisters in Candida albicans Biofilm

2014 ◽  
Vol 59 (3) ◽  
pp. 1627-1633 ◽  
Author(s):  
Jing Sun ◽  
Xiaohua Liu ◽  
Guangshui Jiang ◽  
Qingguo Qi
Keyword(s):  
Candida Albicans ◽  
Nucleic Acid ◽  
Amphotericin B ◽  
Total Population ◽  
Acid Synthesis ◽  
Persister Cells ◽  
Content Type ◽  
Nucleic Acid Biosynthesis ◽  
Low Levels ◽  
Small Subpopulation

ABSTRACTCandida albicanspersisters constitute a small subpopulation of biofilm cells and play a major role in recalcitrant chronic candidiasis; however, the mechanism underlying persister formation remains unclear. Persisters are often described as dormant, multidrug-tolerant, nongrowing cells. Persister cells are difficult to isolate and study not only due to their low levels inC. albicansbiofilms but also due to their transient, reversible phenotype. In this study, we tried to induce persister formation by inducingC. albicanscells into a dormant state.C. albicanscells were pretreated with 5-fluorocytosine (planktonic cells, 0.8 μg ml−1; biofilm cells, 1 μg ml−1) for 6 h at 37°C, which inhibits nucleic acid and protein synthesis. Biofilms and planktonic cultures of eightC. albicansstrains were surveyed for persisters after amphotericin B treatment (100 μg ml−1for 24 h) and CFU assay. None of the planktonic cultures, with or without 5-fluorocytosine pretreatment, contained persisters. Persister cells were found in biofilms of all testedC. albicansstrains, representing approximately 0.01 to 1.93% of the total population. However, the persister levels were not significantly increased inC. albicansbiofilms pretreated with 5-fluorocytosine. These results suggest that inhibition of nucleic acid synthesis did not seem to increase the formation of amphotericin B-tolerant persisters inC. albicansbiofilms.

scholarly journals Patients with Long-Term Oral Carriage Harbor High-Persister Mutants of Candida albicans

2009 ◽  
Vol 54 (1) ◽  
pp. 39-44 ◽  
Author(s):  
Michael D. LaFleur ◽  
Qingguo Qi ◽  
Kim Lewis
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Antimicrobial Agents ◽  
Oral Candidiasis ◽  
Wild Type ◽  
Persister Cells ◽  
Microtiter Plates ◽  
High Concentrations

ABSTRACT Fungal biofilms produce a small number of persister cells which can tolerate high concentrations of fungicidal agents. Persisters form upon attachment to a surface, an important step in the pathogenesis of Candida strains. The periodic application of antimicrobial agents may select for strains with increased levels of persister cells. In order to test this possibility, 150 isolates of Candida albicans and C. glabrata were obtained from cancer patients who were at high risk for the development of oral candidiasis and who had been treated with topical chlorhexidine once a day. Persister levels were measured by exposing biofilms growing in the wells of microtiter plates to high concentrations of amphotericin B and plating for survivors. The persister levels of the isolates varied from 0.2 to 9%, and strains isolated from patients with long-term carriage had high levels of persisters. High-persister strains were isolated from every patient with Candida carriage of more than 8 consecutive weeks but from no patients with transient carriage. All of the high-persister isolates had an amphotericin B MIC that was the same as that for the wild type, indicating that these strains were drug-tolerant rather than drug-resistant mutants. Biofilms of the majority of high-persister strains also showed an increased tolerance to chlorhexidine and had the same MIC for this antimicrobial as the wild type. This study suggests that persister cells are clinically relevant, and antimicrobial therapy selects for high-persister strains in vivo. The drug tolerance of persisters may be a critical but overlooked component responsible for antimicrobial drug failure and relapsing infections.

scholarly journals Candida albicans Biofilms Produce Antifungal-Tolerant Persister Cells

2006 ◽  
Vol 50 (11) ◽  
pp. 3839-3846 ◽  
Author(s):  
Michael D. LaFleur ◽  
Carol A. Kumamoto ◽  
Kim Lewis
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Biofilm Formation ◽  
Fungal Pathogens ◽  
Wild Type ◽  
Persister Cells ◽  
Dormant Cells ◽  
Multidrug Tolerance ◽  
Candida Albicans Biofilms

ABSTRACT Fungal pathogens form biofilms that are highly recalcitrant to antimicrobial therapy. The expression of multidrug resistance pumps in young biofilms has been linked to increased resistance to azoles, but this mechanism does not seem to underlie the resistance of mature biofilms that is a model of in vivo infection. The mechanism of drug resistance of mature biofilms remains largely unknown. We report that biofilms formed by the major human pathogen Candida albicans exhibited a strikingly biphasic killing pattern in response to two microbicidal agents, amphotericin B, a polyene antifungal, and chlorhexidine, an antiseptic, indicating that a subpopulation of highly tolerant cells, termed persisters, existed. The extent of killing with a combination of amphotericin B and chlorhexidine was similar to that observed with individually added antimicrobials. Thus, surviving persisters form a multidrug-tolerant subpopulation. Interestingly, surviving C. albicans persisters were detected only in biofilms and not in exponentially growing or stationary-phase planktonic populations. Reinoculation of cells that survived killing of the biofilm by amphotericin B produced a new biofilm with a new subpopulation of persisters. This suggests that C. albicans persisters are not mutants but phenotypic variants of the wild type. Using a stain for dead cells, rare dark cells were visible in a biofilm after amphotericin B treatment, and a bright and a dim population were physically sorted from this biofilm. Only the dim cells produced colonies, showing that this method allows the isolation of yeast persisters. Given that persisters formed only in biofilms, mutants defective in biofilm formation were examined for tolerance of amphotericin B. All of the known mutants affected in biofilm formation were able to produce normal levels of persisters. This finding indicates that attachment rather than formation of a complex biofilm architecture initiates persister formation. Bacteria produce multidrug-tolerant persister cells in both planktonic and biofilm populations, and it appears that yeasts and bacteria have evolved analogous strategies that assign the function of survival to a small part of the population. In bacteria, persisters are dormant cells. It remains to be seen whether attachment initiates dormancy that leads to the formation of fungal persisters. This study suggests that persisters may be largely responsible for the multidrug tolerance of fungal biofilms.

scholarly journals Absence of Amphotericin B-Tolerant Persister Cells in Biofilms of Some Candida Species

2008 ◽  
Vol 52 (5) ◽  
pp. 1884-1887 ◽  
Author(s):  
Rawya S. Al-Dhaheri ◽  
L. Julia Douglas
Keyword(s):  
Drug Resistance ◽  
Candida Albicans ◽  
Amphotericin B ◽  
Candida Glabrata ◽  
Candida Parapsilosis ◽  
Candida Krusei ◽  
Cell Populations ◽  
Planktonic Cells ◽  
Persister Cells ◽  
Tolerant Cell

ABSTRACT Biofilms and planktonic cells of five Candida species were surveyed for the presence of persister (drug-tolerant) cell populations after exposure to amphotericin B. None of the planktonic cultures (exponential or stationary phase) contained persister cells. However, persisters were found in biofilms of one of two strains of Candida albicans tested and in biofilms of Candida krusei and Candida parapsilosis, but not in biofilms of Candida glabrata or Candida tropicalis. These results suggest that persister cells cannot solely account for drug resistance in Candida biofilms.

scholarly journals Sublethal Injury and Resuscitation of Candida albicans after Amphotericin B Treatment

2003 ◽  
Vol 47 (4) ◽  
pp. 1200-1206 ◽  
Author(s):  
Robert S. Liao ◽  
Robert P. Rennie ◽  
James A. Talbot
Keyword(s):  
Cell Death ◽  
Candida Albicans ◽  
Amphotericin B ◽  
Antifungal Agents ◽  
Sybr Green ◽  
Sequential Treatment ◽  
Tetrazolium Salt ◽  
Fungal Cell ◽  
Sublethal Injury

ABSTRACT Amphotericin B treatment was previously shown to inhibit Candida albicans reproduction and reduce the fluorescence of vitality-specific dyes without causing a corresponding increase in the fluorescence of the mortality-specific dyes bis-(1,3-dibutylbarbituric acid)trimethine oxonol and SYBR Green Ι. In the present study, we have confirmed these results and have shown that the numbers of CFU are reduced by 99.9% by treatment with 0.5 μg of amphotericin B per ml for 10 h at 35°C. This reduction was not due to fungal cell death. First, the level of reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide increased in the presence of concentrations of amphotericin B that caused greater than 90% reductions in the numbers of CFU. Second, fungal cells treated with amphotericin B at a concentration of 0.5 μg/ml were resuscitated by further incubation at 22°C for 15 h in the continued presence of amphotericin B. Third, recovery of the ability to replicate was prevented by sequential treatment with 20 μg of miconazole per ml, which also increased the fluorescence of mortality-specific dyes to near the maximal levels achieved with 0.9 μg of amphotericin B per ml. Sequential treatment with fluconazole and flucytosine did not increase the levels of staining with the mortality-specific dyes. Itraconazole was less effective than ketoconazole, which was less effective than miconazole. The practice of equating the loss of the capacity of C. albicans to form colonies with fungal cell death may give incorrect results in assays with amphotericin B, and the results of assays with caution with other antifungal agents that are lipophilic or that possess significant postantifungal effects may need to be interpreted.

scholarly journals Effect of surface treatments on push-out bond strength of calcium silicate-based cements to fiber posts

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Amr M. Elnaghy ◽  
Ayman Mandorah ◽  
Ali H. Hassan ◽  
Alaa Elshazli ◽  
Shaymaa Elsaka
Keyword(s):  
Bond Strength ◽  
Surface Treatment ◽  
Glass Fiber ◽  
Calcium Silicate ◽  
Surface Treatments ◽  
The Other ◽  
Fiber Posts ◽  
Glass Fiber Posts ◽  
Push Out ◽  
Effect Of Surface

Abstract Background To evaluate the effect of surface treatments on the push-out bond strength of Biodentine (BD) and white mineral trioxide aggregate (WMTA) to fiber posts. Methods Two brands of fiber posts were used: Reblida post; RP and RelyX post; RX. Each type of post (n = 80/group) was divided into four groups (n = 20/group) and exposed to surface treatment as follows: Control (no treatment), sandblasting (SB), hydrofluoric acid (HF), and TiF4 4 wt/v%. Each group was further subdivided into two subgroups (n = 10/subgroup) based on the type of CSCs used as follows: Subgroup A: BD and Subgroup B: WMTA. Push-out bond strength of BD and WMTA to glass fiber posts was assessed. Data were statistically analyzed using three-way ANOVA and Tukey’s test. A Weibull analysis was performed on the push-out bond strength data. Results BD showed higher bond strength than WMTA (P < 0.001). The push-out bond strength for posts treated with TiF4 4 wt/v% showed greater bond strength than the other surface treatments (P < 0.05). The BD/RP-TiF4 4 wt/v% showed the greater characteristic bond strength (σ0) (15.93) compared with the other groups. Surface treatments modified the surface topography of glass fiber posts. Conclusions The BD/RP-TiF4 4 wt/v% showed greater bond strength compared with the other groups. The TiF4 4 wt/v% surface treatment enhanced the bond strength of BD and WMTA to glass fiber posts than the other treatments. Surface treatment of fiber post with TiF4 4 wt/v% could be used to improve the bond strength with calcium silicate-based cements.

scholarly journals Dectin-2-Targeted Antifungal Liposomes Exhibit Enhanced Efficacy

mSphere ◽  
2019 ◽  
Vol 4 (5) ◽  
Author(s):  
Suresh Ambati ◽  
Emma C. Ellis ◽  
Jianfeng Lin ◽  
Xiaorong Lin ◽  
Zachary A. Lewis ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Aspergillus Fumigatus ◽  
Effective Dose ◽  
Cryptococcus Neoformans ◽  
Amphotericin B ◽  
Antifungal Drugs ◽  
Extracellular Matrices ◽  
Content Type ◽  
Order Of Magnitude ◽  
Life Threatening

ABSTRACT Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus cause life-threatening candidiasis, cryptococcosis, and aspergillosis, resulting in several hundred thousand deaths annually. The patients at the greatest risk of developing these life-threatening invasive fungal infections have weakened immune systems. The vulnerable population is increasing due to rising numbers of immunocompromised individuals as a result of HIV infection or immunosuppressed individuals receiving anticancer therapies and/or stem cell or organ transplants. While patients are treated with antifungals such as amphotericin B, all antifungals have serious limitations due to lack of sufficient fungicidal effect and/or host toxicity. Even with treatment, 1-year survival rates are low. We explored methods of increasing drug effectiveness by designing fungicide-loaded liposomes specifically targeted to fungal cells. Most pathogenic fungi are encased in cell walls and exopolysaccharide matrices rich in mannans. Dectin-2 is a mammalian innate immune membrane receptor that binds as a dimer to mannans and signals fungal infection. We coated amphotericin-loaded liposomes with monomers of Dectin-2’s mannan-binding domain, sDectin-2. sDectin monomers were free to float in the lipid membrane and form dimers that bind mannan substrates. sDectin-2-coated liposomes bound orders of magnitude more efficiently to the extracellular matrices of several developmental stages of C. albicans, C. neoformans, and A. fumigatus than untargeted control liposomes. Dectin-2-coated amphotericin B-loaded liposomes reduced the growth and viability of all three species more than an order of magnitude more efficiently than untargeted control liposomes and dramatically decreased the effective dose. Future efforts focus on examining pan-antifungal targeted liposomal drugs in animal models of fungal diseases. IMPORTANCE Invasive fungal diseases caused by Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus have mortality rates ranging from 10 to 95%. Individual patient costs may exceed $100,000 in the United States. All antifungals in current use have serious limitations due to host toxicity and/or insufficient fungal cell killing that results in recurrent infections. Few new antifungal drugs have been introduced in the last 2 decades. Hence, there is a critical need for improved antifungal therapeutics. By targeting antifungal-loaded liposomes to α-mannans in the extracellular matrices secreted by these fungi, we dramatically reduced the effective dose of drug. Dectin-2-coated liposomes loaded with amphotericin B bound 50- to 150-fold more strongly to C. albicans, C. neoformans, and A. fumigatus than untargeted liposomes and killed these fungi more than an order of magnitude more efficiently. Targeting drug-loaded liposomes specifically to fungal cells has the potential to greatly enhance the efficacy of most antifungal drugs.

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