In vitro analysis of flufenamic acid activity against Candida albicans biofilms
Open accessAbstract
In a recent high-throughput screen against specific Candida albicans drug targets, several compounds that exhibited non-specific antifungal activity were identified, including the non-steroidal anti-inflammatory drug flufenamic acid (FFA). This study sought to determine the effect of different doses of FFA, alone or in combination with fixed concentrations of the standard antifungal agents amphotericin B (AmB), caspofungin (CAS) or fluconazole (FLU), for the prevention and treatment of C. albicans biofilms. Biofilms were formed in a 96-well microplate followed by evaluation of antifungal activity using the XTT assay. FFA concentrations of ≥512 mg/L demonstrated >80% prevention of biofilm formation. FFA concentrations of 1024 mg/L demonstrated >85% reduction of mature biofilms. When FFA (≥8 mg/L) was used in combination with FLU (32 mg/L), antifungal activity increased to 99% for the prevention of biofilm formation. Similarly, when a FFA concentration of ≥8 mg/L was used in combination with either AmB (0.25 mg/L) or CAS (0.125 mg/L), antifungal activity also increased up to 99% for the prevention of biofilm formation. The inhibitory effect of FFA on C. albicans biofilms has not been reported previously, therefore these findings suggest that FFA in combination with traditional antifungals might be useful for the treatment and prevention of C. albicans biofilms.
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Written from the paper’s full text.
TL;DR
Flufenamic acid (FFA) alone inhibited Candida albicans biofilm formation and mature biofilms at high concentrations, and low FFA concentrations enhanced the activity of fluconazole, amphotericin B, and caspofungin in vitro. The authors propose FFA as a possible adjunctive antifungal or lock therapy, but note that mechanism and in vivo relevance remain unresolved.
Key findings
- FFA alone prevented biofilm formation by >80% at ≥512 mg/L and reduced mature biofilms by >85% at 1024 mg/L; sMIC50 values were 128–512 mg/L for prevention and >256 mg/L for mature biofilms, with echinocandin-resistant strains 42379 and 53264 showing sMIC50 of 1024 mg/L for mature biofilms.
- FFA at ≥8 mg/L combined with fluconazole (32 mg/L) increased prevention activity to 99%; FFA at ≥8 mg/L combined with amphotericin B (0.25 mg/L) or caspofungin (0.125 mg/L) also increased prevention activity up to 99%.
- For mature biofilms, fluconazole plus FFA was significantly active at FFA ≥16 mg/L; amphotericin B plus FFA at 128–1024 mg/L reached up to 99% reduction; caspofungin plus FFA showed enhanced activity at 32, 512, and 1024 mg/L but a paradoxical effect at 64–256 mg/L with low-dose caspofungin.
- Light microscopy showed reduced biofilm density with FFA and, at 512 mg/L, predominantly pseudohyphal cells after treatment of mature biofilms or yeast-form cells for prevention.
Methods
- In vitro 96-well static biofilm model with XTT reduction assay; six C. albicans strains: four wild-type (ATCC 10231, ATCC 14053, ATCC 24433, SC5314) and two clinically derived echinocandin-resistant strains (42379, 53264).
- Biofilms were formed for 24 h at 37 °C and then treated for 24 h with FFA (2–1024 mg/L) alone or in combination with fixed fluconazole (32 mg/L), amphotericin B (0.25 mg/L), or caspofungin (0.125 mg/L); prevention assays co-incubated cells with FFA.
- Experiments were performed three times independently, each in quadruplicate; metabolic activity was compared by one-way ANOVA with Tukey post-test, significance at P < 0.05; light microscopy was also used.
Limitations
- The paper states that further studies of FFA’s effects on fungal cell biology are needed to support the hypothesized mechanism, and that molecular mechanisms for amphotericin B combined with NSAIDs against C. albicans remain undefined.
- The paper notes that systemic FFA alone would not achieve levels effective against Candida biofilms, though high-dose FFA could be considered for antifungal lock therapy.
- Evident from the text: all data are from in vitro static biofilm assays, with no in vivo or clinical validation; combination experiments were performed only with strain SC5314.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
0 of 17 cited sources checked · 0 supported · 0 flagged
- Findingown data
The echinocandin-resistant strains 42379 and 53264 showed an sMIC50 of 1024 mg/L for treatment of mature biofilms.
“Interestingly, the echinocandin-resistant strains 42379 and 53264 showed an sMIC50 of 1024 mg/L (the highest concentration of FFA tested) for treatment of mature biofilms.”
Quote in the paperNot a substantial claim - Findingown data
FFA concentration of 1024 mg/L demonstrated >85% reduction of mature biofilms.
“FFA concentrations of 1024 mg/L demonstrated >85% reduction of mature biofilms.”
Quote in the paperSubstantial claim - Findingown data
FLU alone showed minimal activity against mature biofilms.
“There was substantial inhibition of biofilm formation (50–99% decrease in biofilm metabolic activity) when FLU (32 mg/L) was used alone against formation of biofilms (prevention) (Fig. 2a); however, as expected, FLU alone showed minimal activity against mature biofilms (Fig. 3a).”
Quote in the paperSubstantial claim - Findingown data
For treatment of mature biofilms, sMIC50 values were >256 mg/L.
“For the prevention of biofilm formation, sMIC50 values (sMIC that inhibits 50% of biofilm growth) varied from 128 mg/L to 512 mg/L; for treatment of mature biofilms, sMICs values were higher than those observed for the prevention of biofilm formation, with sMIC50 values of >256 mg/L.”
Quote in the paperSubstantial claim - Findingown data
When AmB was used in combination with serially increasing concentrations of FFA, a progressive reduction of biofilm formation (prevention) was achieved at FFA concentrations of 8-1024 mg/L.
“When AmB was used in combination with serially increasing concentrations of FFA, a progressive reduction of biofilm formation (i.e. prevention) was achieved at concentrations of 8–1024 mg/L of FFA (Fig. 2b).”
Quote in the paperSubstantial claim
- Findingown data
For treatment of mature biofilms, a statistically significant reduction was observed when FLU was combined with FFA at concentrations of ≥16 mg/mL.
“For treatment of mature biofilms, a statistically significant reduction was observed when FLU was combined with FFA at concentrations of ≥16 mg/mL (Fig. 3a).”
Quote in the paperSubstantial claim - Findingown data
FFA concentrations of 512 mg/L and 1024 mg/L markedly reduced mature biofilm metabolic activity by ≥85% compared with lower concentrations.
“For the treatment of mature biofilms, a significant reduction in metabolic activity was observed at concentrations of 128 mg/L, and FFA concentrations of 512 mg/L and 1024 mg/L markedly reduced biofilm metabolic activity (≥85%) compared with lower concentrations (Fig. 1b).”
Quote in the paperSubstantial claim - Findingown data
For treatment of biofilms, the combination of lower concentrations of FFA (16, 8, 4 and 2 mg/L) with AmB did not result in statistically significant differences in antifungal activity compared with AmB alone, meeting the criteria of an indifferent effect.
“For treatment of biofilms, the combination of lower concentrations of FFA (16, 8, 4 and 2 mg/L) with AmB did not result in statistically significant differences in antifungal activity compared with the effect of AmB alone, meeting the criteria of an indifferent effect (Fig. 3b).”
Quote in the paperSubstantial claim - Findingown data
For treatment of mature biofilms, AmB combined with FFA at concentrations of 128-1024 mg/L resulted in a maximum effect up to 99% reduction.
“For treatment of mature biofilms, AmB combined with FFA at concentrations of 32 mg/L and 64 mg/L resulted in a 75% decrease, and a maximum effect (up to 99% reduction) was observed at FFA concentrations of 128–1024 mg/L (Fig. 3b), meeting the criteria of enhanced activity.”
Quote in the paperSubstantial claim - Findingown data
FFA concentrations of ≥16 mg/L caused a significant decrease (P < 0.05) in biofilm metabolic activity compared with the untreated control for prevention of biofilm formation.
“For the prevention of biofilm formation, FFA concentrations of ≥16 mg/L caused a significant decrease (P < 0.05) in biofilm metabolic activity compared with the untreated control.”
Quote in the paperSubstantial claim - Findingown data
For the prevention of biofilm formation across six C. albicans strains, sessile minimum inhibitory concentration (sMIC50) values varied from 128 mg/L to 512 mg/L.
“For the prevention of biofilm formation, sMIC50 values (sMIC that inhibits 50% of biofilm growth) varied from 128 mg/L to 512 mg/L; for treatment of mature biofilms, sMICs values were higher than those observed for the prevention of biofilm formation, with sMIC50 values of >256 mg/L.”
Quote in the paperSubstantial claim - Findingown data
FLU alone at 32 mg/L caused substantial inhibition of biofilm formation (50-99% decrease in biofilm metabolic activity).
“There was substantial inhibition of biofilm formation (50–99% decrease in biofilm metabolic activity) when FLU (32 mg/L) was used alone against formation of biofilms (prevention) (Fig. 2a); however, as expected, FLU alone showed minimal activity against mature biofilms (Fig. 3a).”
Quote in the paperSubstantial claim - Findingown data
When biofilms were treated with high concentrations (512 mg/L) of FFA, many biofilm cells appeared predominantly pseudohyphal after treatment of mature biofilms or yeast form for prevention of biofilm formation.
“Interestingly, when biofilms were treated with high concentrations (512 mg/L) of FFA, many of the biofilm cells appeared to be predominantly pseudohyphal (after treatment of mature biofilms) or yeast form (for prevention of biofilm formation).”
Quote in the paperNot a substantial claim - Findingown data
Compared with the untreated control, no significant differences in metabolic activity were observed using lower concentrations of FFA (8, 4 and 2 mg/L).
“Compared with the untreated control, no significant differences in metabolic activity were observed using lower concentrations of FFA (8, 4 and 2 mg/L).”
Quote in the paperNot a substantial claim - Findingown data
A clear visual difference in biofilm density was observed at FFA concentrations of 16, 128, 256 and 512 mg/L for the prevention and treatment of biofilms.
“A clear visual difference in biofilm density was observed at concentrations of 16, 128, 256 and 512 mg/L for the prevention and treatment of biofilms (Fig. 1c).”
Quote in the paperNot a substantial claim - Findingown data
For treatment of mature biofilms, AmB combined with FFA at concentrations of 32 mg/L and 64 mg/L resulted in a 75% decrease.
“For treatment of mature biofilms, AmB combined with FFA at concentrations of 32 mg/L and 64 mg/L resulted in a 75% decrease, and a maximum effect (up to 99% reduction) was observed at FFA concentrations of 128–1024 mg/L (Fig. 3b), meeting the criteria of enhanced activity.”
Quote in the paperSubstantial claim - Findingown data
FFA concentration of 128 mg/L caused a significant reduction in metabolic activity for treatment of mature biofilms.
“For the treatment of mature biofilms, a significant reduction in metabolic activity was observed at concentrations of 128 mg/L, and FFA concentrations of 512 mg/L and 1024 mg/L markedly reduced biofilm metabolic activity (≥85%) compared with lower concentrations (Fig. 1b).”
Quote in the paperSubstantial claim - Findingown data
FFA concentrations of 512 mg/L and 1024 mg/L achieved ≥80% reduction in biofilm metabolic activity for prevention of biofilm formation.
“FFA concentrations of ≥512 mg/L demonstrated >80% prevention of biofilm formation.”
- Also stated in: § 3.1. Effects of flufenamic acid on Candida albicans biofilms · plain
“A dramatic reduction (≥80%) in biofilm metabolic activity was achieved at concentrations of 512 mg/L and 1024 mg/L (Fig. 1a).”
Quote in the paperSubstantial claim - Also stated in: § 3.1. Effects of flufenamic acid on Candida albicans biofilms · plain
- Findingown data
The combination of FFA with FLU significantly enhanced the activity of either agent against biofilm formation and mature biofilms.
“In distinct contrast, the combination of FFA with FLU significantly enhanced the activity of either agent against biofilm formation and mature biofilms.”
Quote in the paperSubstantial claim - Findingown data
Statistically significant inhibition of biofilm formation began when FLU was combined with ≥8 mg/L of FFA.
“Statistically significant inhibition of biofilm formation began when FLU was combined with ≥8 mg/L of FFA (Fig. 2a).”
Quote in the paperSubstantial claim - Findingown data
For treatment of mature biofilms, a paradoxical effect was observed when FFA at concentrations of 64, 128 and 256 mg/L was combined with low-dose CAS.
“An enhanced effect of the combination of FFA with CAS was observed for prevention of biofilm formation; surprisingly, for treatment of mature biofilms a paradoxical effect (defined as a reduced fungicidal effect at high concentrations) [14] was observed when increased concentrations of FFA (64–256 mg/L) were combined with low-dose CAS (0.125 mg/L).”
Quote in the paperSubstantial claim - Findingown data
For the treatment of mature biofilms, a substantial reduction in biofilm metabolic activity was observed when FLU (32 mg/L) was combined with low concentrations of FFA (≥8 mg/L).
“Notably, for the treatment of mature biofilms, a substantial reduction in biofilm metabolic activity was observed when FLU (32 mg/L) was combined with low concentrations of FFA (≥8 mg/L).”
Quote in the paperSubstantial claim - Findingown data
CAS alone at 0.125 mg/L caused a high inhibitory effect (95% reduction in prevention and 80% reduction in treatment).
“When CAS was used alone at a concentration of 0.125 mg/L for prevention and treatment of biofilms, as expected a high inhibitory effect was observed (95% reduction in prevention and 80% reduction in treatment).”
Quote in the paperSubstantial claim - Findingown data
CAS combined with FFA for prevention of biofilm formation achieved a maximum inhibitory effect (99.5% reduction in biofilm metabolic activity) at FFA concentrations of ≥16 mg/L.
“When used in combination with FFA for prevention of biofilm formation, a maximum inhibitory effect (99.5% reduction in biofilm metabolic activity) was achieved at concentrations of FFA of ≥16 mg/L (Fig. 2c).”
Quote in the paperSubstantial claim - Findingown data
Treatment of biofilms with AmB alone caused a 50% decrease in biofilm metabolic activity.
“Prevention of biofilm formation with AmB alone caused a 75% decrease in biofilm metabolic activity, and the combination with low concentrations of FFA (≥8 mg/L) caused a maximum inhibitory effect (99% reduction in metabolic activity); similarly, treatment of biofilms with AmB alone caused a 50% decrease in biofilm metabolic activity and the combination with FFA (≥32 mg/mL) caused a 75–90%”
Quote in the paperSubstantial claim - Findingown data
The combination of AmB with low concentrations of FFA (≥8 mg/L) caused a maximum inhibitory effect (99% reduction in metabolic activity) for prevention of biofilm formation.
“Prevention of biofilm formation with AmB alone caused a 75% decrease in biofilm metabolic activity, and the combination with low concentrations of FFA (≥8 mg/L) caused a maximum inhibitory effect (99% reduction in metabolic activity); similarly, treatment of biofilms with AmB alone caused a 50% decrease in biofilm metabolic activity and the combination with FFA (≥32 mg/mL) caused a 75–90%”
Quote in the paperSubstantial claim - Findingown data
Prevention of biofilm formation with AmB alone caused a 75% decrease in biofilm metabolic activity.
“Prevention of biofilm formation with AmB alone caused a 75% decrease in biofilm metabolic activity, and the combination with low concentrations of FFA (≥8 mg/L) caused a maximum inhibitory effect (99% reduction in metabolic activity); similarly, treatment of biofilms with AmB alone caused a 50% decrease in biofilm metabolic activity and the combination with FFA (≥32 mg/mL) caused a 75–90%”
Quote in the paperSubstantial claim - Findingown data
The combination of AmB with FFA (≥32 mg/mL) caused a 75-90% reduction in biofilm metabolic activity for treatment of biofilms.
“Prevention of biofilm formation with AmB alone caused a 75% decrease in biofilm metabolic activity, and the combination with low concentrations of FFA (≥8 mg/L) caused a maximum inhibitory effect (99% reduction in metabolic activity); similarly, treatment of biofilms with AmB alone caused a 50% decrease in biofilm metabolic activity and the combination with FFA (≥32 mg/mL) caused a 75–90%”
Quote in the paperSubstantial claim - Findingown data
When used in combination with FLU (both for prevention and treatment of biofilms), a dose-dependent enhanced effect was observed compared with the effect of either agent alone.
“When used in combination with FLU (both for prevention and treatment of biofilms), a dose-dependent enhanced effect was observed compared with the effect of either agent alone.”
Quote in the paperSubstantial claim - Premisecitation
In a recent high-throughput screen, flufenamic acid (FFA) was identified as having antifungal activity against Candida albicans.
“In a recent high-throughput screen against specific Candida albicans drug targets, several compounds that exhibited non-specific antifungal activity were identified, including the non-steroidal anti-inflammatory drug flufenamic acid (FFA).”
- Also stated in: § 1. Introduction · plain
“In a recent high-throughput screening study performed at the Centre for Molecular Discovery at the University of New Mexico (Albuquerque, NM) (unpublished data), the NSAID flufenamic acid (FFA) was identified as having antifungal activity against C. albicans.”
Quote in the paperSubstantial claim - Also stated in: § 1. Introduction · plain
- Interpretationown data
The activity of combinations of FFA with commonly used antifungals varied according to the specific antifungal type and the concentration of FFA used.
“The activity of combinations of FFA with commonly used antifungals varied according to the specific antifungal type and the concentration of FFA used.”
Quote in the paperSubstantial claim - Interpretationreasoning
Systemic administration of FFA alone would not achieve levels that would be effective against Candida biofilms.
“Although systemic administration of FFA alone would not achieve levels that would be effective against Candida biofilms, high-dose FFA could also serve as a potential antifungal lock strategy for both prevention and treatment of catheter-related Candida biofilm infections.”
Quote in the paperSubstantial claim - Interpretationcitation
FFA might be an effective adjunctive antifungal agent when combined with FLU, AmB and CAS if administered at standard doses.
“According to these pharmacokinetic parameters, FFA might be an effective adjunctive antifungal agent when combined with FLU, AmB and CAS if administered at standard doses (normal dosage varies from 200 mg to 600 mg per day) [6].”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [6] Rajan KT Hill AGS Barr A Whitwell E Flufenamic acid in rheumatoid arthritis Ann Rheum Dis 1967 26 43 6 5334569 PMC1030997 source
Not checked in the free run.
- Premisecitation
Previous findings have indicated little or no effect for treatment of mature biofilms when FLU is used in combination with other antifungals, including AmB and CAS.
“Previous findings have indicated little or no effect for treatment of mature biofilms when FLU is used in combination with other antifungals, including AmB and CAS [10], whereas synergistic effects (similar to the enhanced effect reported in this study) have been observed when FLU is combined with calcium channel inhibitors [11] and doxycycline [12], among others.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [10] Bachmann SP Ramage G VandeWalle K Patterson TF Wickes BL López-Ribot JL Antifungal combinations against Candida albicans biofilms in vitro Antimicrob Agents Chemother 2003 47 3657 9 14576141 PMC253764 source
Not checked in the free run.
- Premisecitation
Synergistic effects have been observed when FLU is combined with calcium channel inhibitors and doxycycline.
“Previous findings have indicated little or no effect for treatment of mature biofilms when FLU is used in combination with other antifungals, including AmB and CAS [10], whereas synergistic effects (similar to the enhanced effect reported in this study) have been observed when FLU is combined with calcium channel inhibitors [11] and doxycycline [12], among others.”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites [11] Uppuluri P Nett J Heitman J Andes D Synergistic effect of calcineurin inhibitors and fluconazole against Candida albicans biofilms Antimicrob Agents Chemother 2008 52 1127 32 18180354 PMC2258509 source
Not checked in the free run.
- cites [12] Kaur R Castano I Cormack BP Functional genomic analysis of fluconazole susceptibility in the pathogenic yeast Candida glabrata: roles of calcium signaling and mitochondria Antimicrob Agents Chemother 2004 48 1600 13 15105111 PMC400560 source
Not checked in the free run.
- Interpretationreasoning
High-dose FFA could serve as a potential antifungal lock strategy for both prevention and treatment of catheter-related Candida biofilm infections.
“Although systemic administration of FFA alone would not achieve levels that would be effective against Candida biofilms, high-dose FFA could also serve as a potential antifungal lock strategy for both prevention and treatment of catheter-related Candida biofilm infections.”
Quote in the paperSubstantial claim
- May combine two claims. 10
- Classified as probably not a substantial claim. 7
May combine two claims.
Findingown dataProliferation of cells within the biofilm was substantially reduced for prevention of biofilm formation, and the multiple layers of a typical biofilm were not apparent.
“Proliferation of cells within the biofilm was substantially reduced for prevention of biofilm formation, and the multiple layers of a typical biofilm were not apparent (Fig. 2d).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataFor treatment of mature biofilms, only AmB and CAS caused a visible reduction in biofilm mass; a gross increase in visual biofilm density was observed, consistent with the paradoxical effect noted by XTT assay for the combination of FFA with CAS.
“For treatment of mature biofilms, only AmB and CAS caused a visible reduction in biofilm mass (Fig. 3d); and a gross increase in the visual biofilm density was observed, consistent with the paradoxical effect noted by XTT assay for the combination of FFA with CAS.”
Quote in the paperSubstantial claimMay combine two claims.
PremisecitationCandida albicans forms biofilms, a characteristic that enhances resistance to antifungal agents and permits colonisation of mucosal surfaces with potential for subsequent dissemination.
“The opportunistic fungal pathogen Candida albicans has the ability to form biofilms, a characteristic that enhances resistance to antifungal agents and permits colonisation of mucosal surfaces with the potential for subsequent dissemination [1].”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [1] Ramage G Saville SP Thomas DP López-Ribot JL Candida biofilms: an update Eukaryot Cell 2005 4 633 8 15821123 PMC1087806 source
Not checked in the free run.
Classified as probably not a substantial claim.
PremisecitationNon-steroidal anti-inflammatory drugs (NSAIDs) have been discovered to possess potent antibacterial activities.
“Furthermore, there has been increasing attention to drugs belonging to different pharmacological classes for possible antimicrobial activity, including a number of non-steroidal anti-inflammatory drugs (NSAIDs), which have been discovered to possess potent antibacterial activities [4,5].”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [4] Alem MAS Douglas LJ Effects of aspirin and other nonsteroidal anti-inflammatory drugs on biofilms and planktonic cells of Candida albicans Antimicrob Agents Chemother 2004 48 41 7 14693516 PMC310207 source
Not checked in the free run.
- cites [5] Alem MAS Douglas LJ Prostaglandin production during growth of Candida albicans biofilms J Med Microbiol 2005 54 1001 5 16192429 source
Not checked in the free run.
May combine two claims.
Interpretationown dataThe combination of FLU with FFA had superior antifungal activity compared with FFA or FLU alone, meeting the criteria for enhanced activity.
“Thus, the combination of FLU with FFA had superior antifungal activity compared with FFA or FLU alone, meeting the criteria for enhanced activity.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
PremisecitationThe antimicrobial activity of NSAIDs is thought to be due to inhibition of prostaglandin biosynthesis.
“The antimicrobial activity of NSAIDs is thought to be due to inhibition of prostaglandin biosynthesis [5]; interestingly, prostaglandin biosynthesis contributes to fungal hyphal formation and biofilm development [5].”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Alem MAS Douglas LJ Prostaglandin production during growth of Candida albicans biofilms J Med Microbiol 2005 54 1001 5 16192429 source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationAntimicrobial lock therapy has been proposed as an alternative strategy for the prevention and treatment of microbial biofilms, particularly for catheter-related bloodstream infections.
“Antimicrobial lock therapy has been proposed as an alternative strategy for the prevention and treatment of microbial biofilms, particularly for catheter-related bloodstream infections [2,3].”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites [2] Mermel LA Farr BM Sherertz RJ Raad II O’Grady N Harris JS Guidelines for the management of intravascular catheter-related infections Clin Infect Dis 2001 32 1249 72 11303260 source
Not checked in the free run.
- cites [3] Walraven CJ Lee SA Antifungal lock therapy Antimicrob Agents Chemother 2013 57 1 8 23070153 PMC3535896 source
Not checked in the free run.
May combine two claims.
PremisecitationProstaglandin biosynthesis contributes to fungal hyphal formation and biofilm development.
“The antimicrobial activity of NSAIDs is thought to be due to inhibition of prostaglandin biosynthesis [5]; interestingly, prostaglandin biosynthesis contributes to fungal hyphal formation and biofilm development [5].”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Alem MAS Douglas LJ Prostaglandin production during growth of Candida albicans biofilms J Med Microbiol 2005 54 1001 5 16192429 source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationFlufenamic acid (FFA) is a NSAID that has been specifically used to treat rheumatoid arthritis.
“FFA is a NSAID that has been specifically used to treat rheumatoid arthritis [6], although its potential as an antifungal agent has not been previously defined.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [6] Rajan KT Hill AGS Barr A Whitwell E Flufenamic acid in rheumatoid arthritis Ann Rheum Dis 1967 26 43 6 5334569 PMC1030997 source
Not checked in the free run.
May combine two claims.
PremisecitationCandida albicans forms biofilms on catheters and medical devices, which may be impossible to cure unless the device is removed.
“Candida albicans also forms biofilms on catheters and medical devices, which may be impossible to cure unless the device is removed [1,2].”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [1] Ramage G Saville SP Thomas DP López-Ribot JL Candida biofilms: an update Eukaryot Cell 2005 4 633 8 15821123 PMC1087806 source
Not checked in the free run.
- cites [2] Mermel LA Farr BM Sherertz RJ Raad II O’Grady N Harris JS Guidelines for the management of intravascular catheter-related infections Clin Infect Dis 2001 32 1249 72 11303260 source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationPrevious studies have linked the paradoxical effect with cell wall stress responses that permit growth in the presence of high concentrations of CAS, such as upregulation of chitin biosynthesis, high-osmolarity glycerol responses and calcium/calcineurin signalling pathways.
“Previous studies have linked the paradoxical effect with cell wall stress responses that permit growth in the presence of high concentration of CAS, such as upregulation of chitin biosynthesis, high-osmolarity glycerol responses and calcium/calcineurin signalling pathways [14].”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [14] Stevens DA Ichinomiya M Koshi Y Horiuchi H Escape of Candida from caspofungin inhibition at concentrations above the MIC (paradoxical effect) accomplished by increased cell wall chitin; evidence for β-1,6-glucan synthesis inhibition by caspofungin Antimicrob Agents Chemother 2006 50 3160 1 16940118 PMC1563524 source
Not checked in the free run.
May combine two claims.
Interpretationown dataHigh doses of FFA exhibited excellent activity for the prevention and treatment of biofilms from six C. albicans isolates, including two echinocandin-resistant strains.
“The results demonstrated that high doses of FFA exhibited excellent activity for the prevention and treatment of biofilms from six C. albicans isolates, including two echinocandin-resistant strains.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim. May combine two claims.
InterpretationreasoningThese findings demonstrate that FFA may be useful in combination with AmB, FLU or CAS as an adjunctive systemic agent, or in high doses as an antifungal lock therapy.
“These findings demonstrate that FFA may be useful in combination with AmB, FLU or CAS as an adjunctive systemic agent, or in high doses as an antifungal lock therapy.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationPrevious studies have suggested that AmB may facilitate the entry of tetracycline derivatives into the fungal cell.
“Previous studies have suggested that AmB may facilitate the entry of tetracycline derivatives into the fungal cell [13]; however, the molecular mechanisms associated with AmB combined with NSAIDs against C. albicans planktonic cells or biofilms remain undefined.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [13] Lew MA Beckett KM Levin MJ Antifungal activity of four tetracycline analogues against Candida albicans in vitro: potentiation by amphotericin B J Infect Dis 1977 136 263 70 408426 source
Not checked in the free run.
May combine two claims.
PremisecitationThe concentration of FFA in plasma varies from 6 mg/L to 20 mg/L and the terminal elimination is long, remaining in plasma for ca. 13.3 h and up to 22 h.
“The concentration of FFA in plasma varies from 6 mg/L to 20 mg/L and the terminal elimination is long, remaining in plasma for ca. 13.3 h and up to 22 h [15].”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [15] Lentjes EG van Ginneken CA Pharmacokinetics of flufenamic acid in man Int J Clin Pharmacol Ther Toxicol 1987 25 185 7 3583467
Not checked in the free run.
The paper relies on this without support.
The inhibitory effect of FFA on C. albicans biofilms has not been reported previously.
“The inhibitory effect of FFA on C. albicans biofilms has not been reported previously, therefore these findings suggest that FFA in combination with traditional antifungals might be useful for the treatment and prevention of C. albicans biofilms.”
“FFA is a NSAID that has been specifically used to treat rheumatoid arthritis [6], although its potential as an antifungal agent has not been previously defined.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Screens of chemical libraries have identified compounds that disrupt formation and/or maintenance of biofilms.
“Screens of chemical libraries have identified compounds that disrupt formation and/or maintenance of biofilms (25, 101, 185, 210).”
From <i>Candida albicans</i> Biofilms and Human Disease (2015) · cites this paper as [25]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Caspofungin concentration differs by 1,000-fold
MajorThe methods specify 0.125 mg/mL caspofungin, but the results specify 0.125 mg/L. These concentrations differ by a factor of 1,000.
“CAS (0.125 mg/mL)”
“CAS was used alone at a concentration of 0.125 mg/L for prevention and treatment of biofilms”
Amphotericin B concentration differs by 1,000-fold
MajorThe methods give the amphotericin B concentration as 0.25 mg/mL, while the abstract gives 0.25 mg/L for the combination experiment. These concentrations differ by a factor of 1,000.
“AmB (0.25 mg/mL)”
“AmB (0.25 mg/L)”
Flufenamic acid treatment threshold exceeds the stated test range
MinorThe reported threshold of ≥16 mg/mL with fluconazole is at least 16,000 mg/L, whereas the stated flufenamic acid test range ends at 1024 mg/L.
“For treatment of mature biofilms, a statistically significant reduction was observed when FLU was combined with FFA at concentrations of ≥16 mg/mL (Fig. 3a).”
“increasing concentrations of FFA (2, 4, 8, 16, 32, 64, 128, 256, 512 and 1024 mg/L in RPMI 1640 medium)”
Amphotericin B combination dose exceeds the stated test range
MinorThe discussion attributes a treatment effect to ≥32 mg/mL flufenamic acid, but the results describe the amphotericin B combination at 32-1024 mg/L. Even 32 mg/mL exceeds the highest stated dose, 1024 mg/L.
“the combination with FFA (≥32 mg/mL) caused a 75–90% reduction in biofilm metabolic activity”
“For treatment of mature biofilms, AmB combined with FFA at concentrations of 32 mg/L and 64 mg/L resulted in a 75% decrease, and a maximum effect (up to 99% reduction) was observed at FFA concentrations of 128–1024 mg/L (Fig. 3b)”
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References 15 · Cited by 26
- Authors
- Chavez-Dozal, A. A., Jahng, M., Rane, H. S., Asare, K., Kulkarny, V. V., Bernardo, S. M., & Lee, S. A.
- Affiliations
- Raymond G. Murphy VA Medical Center (US)
- University of New Mexico (US)
- ORCID
- Lee, Samuel A.
- Venue
- International Journal of Antimicrobial Agents
- Publisher
- Elsevier BV
- Published
- 2014-01
- Volume
- 43
- Issue
- 1
- Pages
- 86-91
- ISSN
- 0924-8579 (ISSN-L)
- PMC
- PMC3902125
- OpenAlex
- W2030859704
- Semantic Scholar
- 3b3de1bce749817a52a3da964bfe4dc696514968
- Cited by
- 26
- References
- 15
- Citations by year
- 2026: 3 · 2025: 3 · 2023: 3 · 2021: 2 · 2020: 2 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 4 · 2014: 2
- FWCI
- 1.95
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- International Journal of Antimicrobial Agents (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (K12GM088021)
- National Institute of Allergy and Infectious Diseases (T32AI007538)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- flufenamic acid, Candida albicans, biofilm, antifungal activity, drug repurposing, nonsteroidal anti-inflammatory drugs, combination therapy, fluconazole, amphotericin B, caspofungin, biofilm inhibition, antifungal resistance
- MeSH terms
- Antifungal Agents, Candida albicans, Flufenamic Acid, Microbial Sensitivity Tests, Biofilms, Microbial Viability
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