Postantifungal Effect Research Articles

In vitro pharmacodynamic characteristics of flucytosine determined by time-kill methods☆

Two Candida albicans isolates, three non- albicans Candida isolates ( Candida glabrata, Candida krusei, and Candida tropicalis), and one Cryptococcus neoformans isolate were evaluated by time-kill methods to characterize the relationship of flucytosine concentrations to antifungal activity and the duration of the post-antifungal effect (PAE). Against Candida and Cryptococcus isolates, flucytosine at concentrations > 1 × MIC exhibited fungistatic (≤99% reduction in CFU) activity over a 24-h time-period. The rate and extent of fungistatic activity of flucytosine against all isolates was generally not increased when 5-FC concentrations exceeded 4 × MIC. A notable PAE was detected for flucytosine against both Candida and Cryptococcus species that persisted 2 to 4 h. These in vitro data suggest that flucytosine is predominately a concentration-independent fungistatic agent at clinically achieved serum concentrations. This pharmacodynamic characteristic coupled with the persistent PAE and the relatively long half-life of flucytosine in humans (>5 h), suggests lower daily dosing may possible without loss of antifungal efficacy.

Influence of pH and concentration on the postantifungal effect and on the effects of sub-MIC concentrations of 4 antifungal agents on previously treated Candida spp.

This study investigates the impact of different pH values (5.5 and 7.4) on the postantifungal effect (PAFE) and the effect of sub-MIC concentrations (1/4 x MIC) on C. albicans and C. glabrata in the PAFE stage (PAFSE). The PAFE stage was induced by a 1.5 h pretreatment with different doses (1, 4 and 8 x MIC) of 4 antifungal agents. An increase in the pH and/or an increase in the dose of the antimycotic prolonged the duration of the PAFE induced by amphotericin B or 5-fluorocytosine and the PAFSE induced by all 4 antifungal agents in both species. 5-Fluorocytosine and amphotericin B (except for treatment with 1 x MIC at pH 5.5) induced significant PAFEs (0.5-3.0 h and 1.4-4.8 h, respectively), which were increased (to 0.9-3.2 h and 0.8-3.4 h, respectively) by posterior (PLEASE EXPLAIN WHAT YOU MEAN BY THE WORD "POSTERIOR" HERE) exposure to 1/4 X MIC of the respective antifungal agent. Although ketoconazole and fluconazole were not able to induce significant PAFEs, posterior exposure to 1/4 x MIC of each of these 2 azoles led to significant PAFSEs of up to 2.6 h in both yeast species when the concentrations and pH were high enough.

In vitropost-antifungal effect (PAFE) elicited by chlorhexidine gluconate on oral isolates ofCandida albicans

The post-antifungal effect (PAFE) is defined as the suppression of growth that persists following brief exposure of yeasts and other fungi to antimycotics and subsequent removal of the drug. There is no data available on the PAFE of chlorhexidine gluconate on oral isolates of Candida albicans . As chlorhexidine gluconate is by far the commonest antiseptic mouth wash prescribed in dentistry, the main aim of this investigation was to measure the PAFE of 10 oral isolates of C. albicans following limited exposure (30 min) to sub-therapeutic concentrations of this antiseptic. The PAFE was determined as the difference in time (h) required for the growth of the drug-free control and the drug-exposed test cultures to increase to 0.05 absorbance level at 520 nm following removal of the antifungal agent. The mean duration of PAFE elicited by 0.005, 0.0025 and 0.00125% chlorhexidine gluconate was 9.91( ± 1.43) h, 1.65 ( ± 0.21) h and 0.67 ( ± 0.12) h, respectively. These findings, reported here for the first time, indicate that even a brief exposure to sub-therapeutic and sub-lethal levels of chlorhexidine gluconate may suppress the growth of oral C. albicans isolates, thereby minimising the pathogenic potential of this opportunistic fungus. Keywords: Candida albicans , chlorhexidine gluconate, post-antifungal effect.

The in vitro post-antifungal effect of nystatin on Candida species of oral origin.

The post-antifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of yeasts to antimycotics and subsequent removal of the drug. Although limited data are available on the PAFE of nystatin on oral isolates of C albicans, there is no information on non-albicans Candida species. As nystatin is the commonest antifungal agent prescribed in dentistry, the main aim of this investigation was to measure the PAFE of oral isolates of Candida belonging to six different species (five isolates each of C. albicans, C. tropicalis, C. krusei, C. parapsilosis, C. glabrata and C. guilliermondii) following limited exposure (1 h) to nystatin. The yeasts were examined for the presence of the PAFE after 1 h exposure to the minimum inhibitory concentration (MIC) of nystatin. The PAFE was determined as the difference in time (h) required for the growth of the drug-free control and the drug-exposed test cultures to increase to the 0.05 absorbance level following removal of the antifungal agent. The mean duration of nystatin-elicited PAFE was lowest for C. albicans (6.85 h) and greatest for C. parapsilosis (15.17 h), while C. krusei (11.58 h), C. tropicalis (12.73 h), C. glabrata (8.51 h), and C. guilliermondii (8.68 h) elicited intermediate values. These findings clarify another intriguing possibility for the persistent, chronic recurrence of oral C. albicans infections despite apparently adequate antifungal drug regimens. The significant variations in nystatin-induced PAFE amongst non-albicans species may also have clinical implications, in terms of nystatin regimens used in the management of these fungal infections.

Activity of Voriconazole: Post-antifungal Effect, Effects of Low Concentrations and of Pretreatment on the Susceptibility of Candida albicans to Leucocytes

This study examined: (i) the post-antifungal effect (PAFE) of Voriconazole (UK 109,496) on Candida albicans, at 2 concentrations (MIC and 4 x MIC) in the presence or absence of 10% human serum; (ii) the activity of low concentrations of the drug (1/4 x MIC) on yeasts that had previously been exposed to Voriconazole (PAFSE) with or without 10% human serum; and (iii) the effect of Voriconazole pretreatment on the fungicidal activity of leucocytes and serum against C. albicans (PALE). Two concentrations (0.25 and 1 mg/l) of Voriconazole induced no PAFE against C. albicans between -4.3 and -1.4 h, but when the assays were performed in the presence of serum, positive and concentration-dependent PAFEs were obtained (0.2-4.1 h). Pretreated yeasts were more susceptible than untreated yeasts to low concentrations (0.0625 mg/l) of Voriconazole, so the drug showed positive PAFSE that was dependent on the concentration used in pretreatment without serum (0.3-1.9 h) or with 10% human serum (0.5-2.5 h). Pretreatment of the growing C. albicans cells with Voriconazole (0.25 mg/l) increased their vulnerability to killing by leucocytes during the last 2 h (p < 0.05), leading to PALE of 2 h. The results suggest that these effects might be used to evaluate the in vivo activity of an antifungal agent. The sum of the durations of these effects (PAFE, PAFSES and PALE) cause a considerable delay in yeast growth in treated cultures compared with control cultures.

In vitro post-antifungal effect (PAFE) elicited by chlorhexidine gluconate on oral isolates of Candida albicans

In vitro post-antifungal effect (PAFE) elicited by chlorhexidine gluconate on oral isolates of Candida albicans

The postantifungal effect (PAFE) of antimycotics on oral C. albicans isolates and its impact on candidal adhesion.

Postantifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of yeasts to antimycotics and subsequent removal of the drug. As there are no data on the PAFE of oral C. albicans isolates the main aim of this investigation was to measure the PAFE of 10 oral isolates of C. albicans following limited exposure (1 h) to five antifungal drugs, including nystatin which has not been previously used in PAFE assays. A secondary aim of the study was to evaluate the biological significance of PAFE, using a nystatin pre-exposed isolate of C. albicans and observing its adherence to denture acrylic surfaces, during the PAFE period. A total of 10 oral isolates of C. albicans were examined for the presence of the PAFE after 1 h exposure to five antifungal drugs, nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole. PAFE was automatically assessed with the help of a Spectramax machine which utilizes the principle of periodic turbidometric assessment of growth rates at a given temperature over a given period. The data thus collected are automatically processed in a graphic format as a computer printout. The PAFE was determined as the difference in time (h) required for growth of the drug-free control and the drug-exposed test cultures to increase to 0.05 absorbance level following removal of the antifungal agent (by repeated washing). The adhesion of the single isolate to denture acrylic following limited exposure to nystatin was assessed by a previously described in vitro adhesion assay. Significant PAFE were observed for nystatin, amphotericin-B and 5-fluorocytosine. A marginal PAFE was observed for ketoconazole and little or none for fluconazole. The mean duration of the PAFE of nystatin, amphotericin-B, 5-fluorocytosine, ketoconazole and fluconazole were 2.89 (+/- 0.27) h, 2.83 (+/- 0.23) h, 3.18 (+/- 0.31) h, 0.65 (+/- 0.11) h and 0.16 (+/- 0.06) h, respectively. The mean percentage reduction of adhesion of oral C. albicans BU47204 to denture acrylic during the PAFE period following exposure to nystatin for 10, 30, 50, 70 and 90 min was 9.12%, 61.73%, 65.99%, 82.16% and 83.14%, respectively. These in vitro findings imply that even a short period of exposure to antifungals may result in modulation of the growth and the virulent attributes of C. albicans, which however is largely dictated by the antimycotic agent in question. Whether such mechanisms operate in vivo needs to be clarified by further studies.

Influence of human serum on the postantifungal effect of four antifungal agents on Candida albicans.

This study evaluates the influence of both fresh and heated human serum on the postantifungal effect (PAFE) induced by different concentrations of amphotericin B (AmB), 5-fluorocytosine (5-Fc), Ketoconazole (Kz) and fluconazole (Flu) on two strains of Candida albicans. The concentrations were selected in harmony with the pharmacokinetic properties and toxicity of the drugs. Without serum there was no delay in the growth of yeast cultures pretreated with Kz or Flu, leading to negative PAFEs, however with AmB and 5-Fc the PAFEs were positive. When assays were made in the presence of 10% fresh human serum, the duration of the PAFEs increased with all drugs tested, and those induced by azolic agents became positive. In the presence of 10% human serum heated at 56 degrees C for 30 min, the PAFEs of the antifungal agents were similar to those obtained in the absence of serum. Our results suggest that fresh serum positively influenced PAFE which may be an important factor in determining the dosing regimen for infection by yeasts.

Postantifungal effect and effects of low concentrations of amphotericin B and fluconazole on previously treated Candida albicans.

The objective of the study was to assess (a) the postantifungal effect (PAFE) of amphotericin B (AmB) and fluconazole (Flu) on two Candida albicans strains, and (b) the effect of low concentrations of AmB or Flu on yeasts previously exposed to AmB or Flu, respectively, in the presence or absence of 10% human serum. In the absence of serum, AmB exerted a positive effect (at 0.5-5.0 h) depending on the concentration and the strain used. Flu, however, produced negative effects (at -1.2 to -0.1 h). When the experiment was conducted in the presence of serum, the duration of all PAFEs increased significantly, especially those induced by Flu that became positive. Pretreated yeasts were more susceptible than untreated controls to the antifungal activity of low concentrations of the drugs. Growth delay was more pronounced with Flu (up to 5.2 h), whereas the greatest decrease in log10 at 12 h was slightly more pronounced for AmB. A knowledge of PAFEs and the effects of low drug concentrations on pretreated yeasts and the effects of serum on these is important in order to gain more insight into the in vivo activity of the study drugs.

The Postantifungal Effect: Synergistic Interactions of Cispentacin and Pradimicin (BMY-28864) with Flucytosine against Candida albicans.

A turbidometric method was used to evaluate Candida albicans yeast cell growth and to quantitate the in vitro postantifungal effect (PAFE) after exposure to various concentrations of cispentacin and pradimicin (BMY-28864), alone and in combination with flucytosine, for 2hr at 30°C and 37°C. Initial MIC and FIC (fractional inhibitory concentration) assays were made to determine approximate drug concentrations to use in the PAFE studies. The PAFE was calculated by the difference in time (hr) required for growth of the control and test cultures to reach the 0.5 absorbance level following removal of the drug by dilution. A synergistic PAFE was evidenced with both cispentacin and pradimicin (BMY-28864) when combined with flucytosine. Optimal PAFEs ranging from 5.7 to 18.5hr were induced by concentrations of cispentacin (0.078 to 0.313μ/ml) and flucytosine (0.049-0.098μg/ml) and from 3.3 to 11.7hr with pradimicin (BMY-28864) (0.078 to 0.313μg/ml) and flucytosine (0.049-0.098μg/ml). The PAFEs persisted for 1.9 to 4.8hr (cispentacin+flucytosine) and for 0.3 to 2.9hr (pradimicin-BMY-28864+flucytosine) longer than those achieved with either agent alone.

Synergistic postantifungal effect of flucytosine and fluconazole on Candida albicans.

The in vitro efficacy of flucytosine and fluconazole, separately and in combination, with respect to induction of a postantifungal effect (PAFE) on Candida albicans was studied. PAFE refers to the persistent suppression of fungal cell growth following a short period of exposure to an antifungal agent. A turbidometric method was used to measure cell growth and to quantitate the PAFE following exposure of C. albicans yeast cells to different concentrations of the two agents for 2 h. The PAFE was determined by the difference in time (h) required for growth of the control and test cultures to increase to the 0.5 absorbance level following removal of the drug by dilution. Minimum (MIC) and fractional inhibitory concentration determinations were made and the data used for selecting the concentrations used in the PAFE evaluations. A synergistic interaction of the two drugs at concentrations well below their individual MICs was evidenced. Flucytosine:fluconazole ratios of 1:16-1:32 at concentrations ranging from 0.024-0.098 micrograms ml-1 and from 0.78-1.56 micrograms ml-1, with flucytosine and fluconazole, respectively, induced PAFEs which persisted for 2.5 h longer than those achieved when each of the two agents was assayed separately.

The postantifungal effect of 5-fluorocytosine on Candida albicans.

The in-vitro postantifungal effect (PAFE) of 5-fluorocytosine for Candida albicans for short periods of time was investigated. Yeast cells were exposed for 0.5, 1 or 2 h to a range of concentrations (0.1-3.2 mg/L) of 5-fluorocytosine. The PAFE was quantitated by determinations of the number of colony forming units at hourly intervals (0-10 h) after removal of the drug by dilution. The length of the PAFE was dependent upon the concentration of 5-fluorocytosine and the duration of exposure. An exposure time of 0.5 h resulted in PAFE's ranging from 0 to 4.2 h. Exposure times of 1 and 2 h resulted in longer PAFEs and in many instances suppression of cell growth was seen for the entire evaluation period (up to ten hours).

Comparative Studies on the Postantifungal Effect of Cispentacin and Pradimicin (BMY-28864) on Candida albicans.

Comparative Studies on the Postantifungal Effect of Cispentacin and Pradimicin (BMY-28864) on Candida albicans.

In vitro comparative evaluations of the postantifungal effect: synergistic interaction between flucytosine and fluconazole against Candida albicans

In vitro comparative evaluations were performed to study the efficacy of combinations of flucytosine and fluconazole in producing a postantifungal effect (PAFE) on Candida albicans. Initial studies were done to determine MIC, FIC (fractional inhibitory concentration) and optimal PAFE parameters. A turbidometric method was used to measure yeast cell growth following exposure to different concentrations of the two drugs for periods of 0.5, 1 or 2 h at temperatures of 30 degrees C and 37 degrees C. The PAFE was determined by the difference in time (h) required for growth of the control and test cultures to reach the 0.5 absorbance level following removal of the drug by dilution. Ten strains of C. albicans were then assayed (30 degrees C; 2 h exposure time) and a synergistic PAFE was evidenced with the two drugs at concentrations well below their individual MICs. PAFEs ranging from 3.8 to 10.5 h, which persisted for 1.2-2.5 h longer than those achieved with either agent separately, were evidenced when flucytosine and fluconazole were combined (flucytosine: fluconazole ratios of 1:16-1:32) at concentrations ranging from 0.024 to 0.098 micrograms ml-1 and 0.78 to 1.56 micrograms ml-1 respectively. The concentrations of each agent required to produce an optimal PAFE varied according to the C. albicans strain being assayed.

Turbidometric characterization of the postantifungal effect: comparative studies with amphotericin B, 5‐fluorocytosine and miconazole on Candida albicans

The phenomenon of persistent suppression of Candida albicans yeast cell growth after short drug exposures (postantifungal effect, PAFE) was determined by performing comparative studies using different concentrations of 5-fluorocytosine (5-FC), amphotericin B (AMPH) and miconazole (MCZ). An in vitro turbidometric method was used to measure cell growth and to quantitate the PAFE after removal of the drug by dilution following exposure to C. albicans yeast cells for 0.5 h, 1 h or 2 h. The PAFE was determined by the difference in time (h) required for growth of the control and test cultures to increase to the 0.5 absorbance level following removal of the antifungal agent. A PAFE was demonstrated with each agent and generally the length of the PAFE was dependent upon the concentration of the drug and the time of exposure. An exposure time of 0.5 h resulted in PAFEs ranging from 0.6 to 16.7 h with 5-FC, 0 to 16.5 h with AMPH and 0.1 to 14.1 h with MCZ. In most instances exposure of the cells to each drug for 1 or 2 h resulted in slightly longer PAFEs, respectively. Longer PAFEs were induced with lower concentrations of 5-FC as compared to AMPH and MCZ. The data from such PAFE assays may be useful for determining in vivo treatment regimens, since longer PAFEs may allow for intermittent dosing instead of continuous drug administration.