Fluconazole-resistant Candida Albicans Research Articles

Synthesis of bio-stabilized silver nanoparticles using Roccella montagnei, their anticandidal capacities & potential to inhibit the virulence factors in fluconazole-resistant Candida albicans.

Candida species is the causative agent in approximately 80% of invasive mycoses and drug-resistant Candida albicans is among the four strains of 'critical priority group' framed by WHO. Lichens are endowed with some rare phytochemicals and a plethora of therapeutics viz. antifungal capacities of Roccella montagnei. Biosynthesis of silver nanoparticles (AgNPs) using lichen could offer an eco-friendly, and cost-effective alternative against emerging 'microbial resistance.' Therefore, the objective was to biosynthesize silver nanoparticles (Rm-AgNPs) using a Hydro-alcoholic (1:1) extract of R. montagnei to develop a potent anticandidal agent against Fluconazole-resistant C. albicans NBC099. UV-Spectroscopy identified AgNPs specific-peak of Rm-AgNPs at 420-440nm and FTIR revealed the presence of amines, alcohol, aromatic compounds, and acids. SEM and TEM analysis indicated that Rm-AgNPs are spherical shaped with a size range of 10-50nm. Zetasizer analysis indicated that particles are highly stable and have a mean hydrodynamic diameter of 116nm with a zeta potential charge of - 41mV. XRD analysis suggested face centered cubic crystal lattice structure. Results indicated that Rm-AgNPs strongly inhibited the growth of NBC099 at a minimum inhibitory concentration (IC50) of ≤ 15µg. C. albicans culture treated with Rm-AgNPs at concentrations below IC50, down-regulates the production of different virulence factors in NBC099, viz. hyphal formation (> 85%), biofilms production (> 80%), phospholipase, esterase, proteinase activity. The apoptosis assay demonstrated the Rm-AgNPs induced apoptosis in NBC099 cells via oxidative stress. Interestingly, Rm-AgNPs showed negligible cytotoxicity (< 6%) in murine RAW 246.7 macrophage cells at a concentration above 15µg/mL. Therefore, Rm-AgNPs have been offered as an anti-candida alternative that can be utilized to improve the efficacy of already available medications.

Unveiling the emerging of fluconazole-resistant Candida albicans in preterm with acute kidney injury: a growing challenge at Vajira hospital

This case presents a challenging scenario in a preterm female newborn born at 25 weeks gestation,who developed respiratory distress syndrome, followed by pneumonia, sepsis, subsequent renal complications, and candida albicans infection. Despite initial susceptibility to fluconazole, the emergence of fluconazole-resistant candida albicans led to a shift in antifungal treatment to amphotericin B in an acute kidney injury neonate. This case underscores the growing concern of drug-resistant candida albicans in neonates, highlighting the importance of judicious selection of empirical antifungal therapy and comprehensive management strategies, especially in the context of acute kidney injury.

Synthesis, structure–activity relationship and biological evaluation of indole derivatives as anti-Candida albicans agents

In this work, we synthesized a series of indole derivatives to cope with the current increasing fungal infections caused by drug-resistant Candida albicans. All compounds were evaluated for antifungal activities against Candida albicans in vitro, and the structure–activity relationships (SARs) were analyzed. The results indicated that indole derivatives used either alone or in combination with fluconazole showed good activities against fluconazole-resistant Candida albicans. Further mechanisms studies demonstrated that compound 1 could inhibit yeast-to-hypha transition and biofilm formation of Candida albicans, increase the activity of the efflux pump, the damage of mitochondrial function, and the decrease of intracellular ATP content. In vivo studies, further proved the anti-Candida albicans activity of compound 1 by histological observation. Therefore, compound 1 could be considered as a novel antifungal agent.

Unleashing the potential of vanillic acid: A new twist on nature's recipe to fight inflammation and circumvent azole-resistant fungal infections

Vanillic acid (VA) — a naturally occurring phenolic compound in plants — is not only used as a flavoring agent but also a prominent metabolite post tea consumption. VA and its associated compounds are believed to play a significant role in preventing diseases, underscoring the need for a systematic investigation. Herein, we report a 4-step synthesis employing the classical organic reactions, such as Willamson’s alkylation, Fischer-Spier reaction, and Steglich esterification, complemented with a protection-deprotection strategy to prepare 46 VA derivatives across the five series (1a–1i, 2a–2i, 3, 3a–3i, 4a–4i, 5a–5i) in high yields. The synthesized compounds were investigated for their antifungal, anti-inflammatory, and toxic effects. Notably, compound 1a demonstrated remarkable ROS inhibition with an IC50 value of 5.1 ± 0.7 µg/mL, which is more than twice as effective as the standard ibuprofen drug. A subset of the synthesized derivatives (2b, 2c, 2e, 3b–3d, 4a–4c, 5a, and 5e) manifested their antifungal effect against drug-resistant Candida strains. Compound 5g, in particular, revealed synergism with the established antifungal drugs amphotericin B (AMB) and fluconazole (FLZ), doubling FLZ's potency against azole resistant Candida albican ATCC 36082. Furthermore, 5g improved the potency of these antifungals against FLZ-sensitive strains, including C. glabrata ATCC 2001 and C. parapsilosis ATCC 22019, as well as various multidrug-resistant (MDR) Candida strains, namely C. albicans ATCC 14053, C. albicans CL1, and C. krusei SH2L OM341600. Additionally, pharmacodynamics of compound 5g was examined using time-kill assay, and a benign safety profile was observed with no hemolytic activity in whole blood, and no cytotoxicity towards the normal BJ human cell line. The synergistic potential of 5g was further investigated through both experimental methods and docking simulations.These findings highlight the therapeutic potential of VA derivatives, particularly in addressing inflammation and circumventing FLZ resistance in Candida albicans.

A mechanism study on the synergistic effects of rifapentine and fluconazole against fluconazole-resistant Candida albicans in vitro

Candida albicans (C. albicans) is one of the most common clinical isolates of systemic fungal infection. Long-term and inappropriate use of antifungal drugs can cause fungal resistance, which poses a great challenge to the clinical treatment of fungal infections. The combination of antifungal drugs and non-antifungal drugs to overcome the problem of fungal resistance has become a research hotspot in recent years. Our previous study found that the combination of rifapentine (RFT) and fluconazole (FLC) has a significant synergistic against FLC-resistant C. albicans. The present study aimed to further verify the synergistic effect between FLC and RFT against the FLC-resistant C. albicans 100, and explore the underlying mechanism. The growth curve and spot assay test not only showed the synergistic effect of FLC and RFT on FLC-resistant C. albicans in vitro but exhibited a dose-dependent effect on RFT, indicating that RFT may play a principal role in the synergic effect of the two drugs. Flow cytometry showed that the combined use of RFT and FLC arrested cells in the G2/M phase, inhibiting the normal division and proliferation of FLC-resistant C. albicans. Transmission electron microscopy (TEM) demonstrated that FLC at a low concentration could still cause a certain degree of damage to the cell membrane in the FLC-resistant C. albicans, as represented by irregular morphologic changes and some defects observed in the cell membrane. When FLC was used in combination with RFT, the nuclear membrane was dissolved and the nucleus was condensed into a mass. Detection of the intracellular drug concentration of fungi revealed that the intracellular concentration of RFT was 31–195 fold that of RFT alone when it was concomitantly used with FLC. This indicated that FLC could significantly increase the concentration of RFT in cells, which may be due to the damage caused to the fungal cell membrane by FLC. In short, the present study revealed a synergistic mechanism in the combined use of RFT and FLC, which may provide a novel strategy for the clinical treatment of FLC-resistant C. albicans.

Promising activity of etomidate against mixed biofilms of fluconazole-resistant Candida albicans and methicillin-resistant Staphylococcus aureus.

Introduction. Candida albicans and Staphylococcus aureus are recognized for their development of resistance and biofilm formation. New therapeutic alternatives are necessary in this context.Hypothesis. Etomidate shows potential application in catheters against mixed biofilms of fluconazole-resistant C. albicans and methicillin-resistant S. aureus (MRSA).Aim. The present study aimed to evaluate the activity of etomidate against mixed biofilms of fluconazole-resistant C. albicans and MRSA.Methodology. The action of etomidate against mature biofilms was verified through the evaluation of biomass and cell viability, and its ability to prevent biofilm formation in peripheral venous catheters was determined based on counts of colony forming units (c.f.u.) and confirmed by morphological analysis through scanning electron microscopy (SEM).Results. Etomidate generated a reduction (P<0.05) in biomass and cell viability starting from a concentration of 250 µg ml-1. In addition, it showed significant ability to prevent the formation of mixed biofilms in a peripheral venous catheter, as shown by a reduction in c.f.u. SEM revealed that treatment with etomidate caused substantial damage to the fungal cells.Conclusion. The results showed the potential of etomidate against polymicrobial biofilms of fluconazole-resistant C. albicans and MRSA.

From Synergy to Monotherapy: Discovery of Novel 2,4,6-Trisubstituted Triazine Hydrazone Derivatives with Potent Antifungal Potency In Vitro and In Vivo.

Invasive fungal infections pose a serious threat to public health and are associated with high mortality and incidence rates. The development of novel antifungal agents is urgently needed. Based on hit-to-lead optimization, a series of 2,4,6-trisubstituted triazine hydrazone compounds were designed, synthesized, and biological evaluation was performed, leading to the identification of compound 28 with excellent in vitro synergy (FICI range: 0.094-0.38) and improved monotherapy potency against fluconazole-resistant Candida albicans and Candida auris (MIC range: 1.0-16.0 μg/mL). Moreover, 28 exhibited broad-spectrum antifungal activity against multiple pathogenic strains. Furthermore, 28 could inhibit hyphal and biofilm formation, which may be related to its ability to disrupt the fungal cell wall. Additionally, 28 significantly reduced the CFU in a mouse model of disseminated infection with candidiasis at a dose of 10 mg/kg. Overall, the triazine-based hydrazone compound 28 with low cytotoxicity, hemolysis, and favorable ADME/T characteristics represents a promising lead to further investigation.

Phenolic compounds and antimicrobial activity of leaves of the genus Sorbus species and natural hybrids

The identification of new sources of biologically active substances is an actual direction of research in recent years, in particular, the demand for antimicrobial compounds and natural preservatives of plant origin is growing. Plants of the genus Sorbus L. are known in folk medicine, but for the rational use of plant material, there is a lack of systematic study of the features of the secondary metabolites’ accumulation and the spectrum of biological activity of plants during vegetation in different environmental conditions. In the present work, the seasonal dynamics of the total content of polyphenols, flavonoids and free phenolic acids in the leaves of rowan plants natural to the flora of Ukraine and introduced in the steppe region were investigated. The highest accumulation of phenolic compounds was found in the leaves of S. domestica, S. latifolia, S. hybrida, S. torminalis and S. aucuparia in the first half of June, and in S. aria and S. intermedia in May, which will serve as a criterion for selecting the best plant material to obtain biologically active compounds. The seasonal dynamics of the content of flavonoids and phenolic acids in the leaves of plants had significant species differences, which require more in-depth studies of the phytocompounds component composition. Antimicrobial activity of leaf ethanol extracts of rowan plant was found to be high against gram-negative bacterial strain Klebsiella pneumoniae B920 (especially the activity of S. aria, S. intermedia and S. aucuparia extracts) and Pseudomonas aeruginosa B907 (especially the activity S. torminalis and S. domestica), clinical gram-positive strains of Staphylococcus epidermidis (the highest activity of S. aria, S. domestica and S. intermedia extracts), as well as against clinical strains of fluconazole-resistant fungus Candida albicans (especially the activity of extracts of S. latifolia S. torminalis and S. intermedia). A conclusion was made about the prospects of using leaf extracts of the genus Sorbus plants as an economical and affordable natural source of biologically active polyphenols with a wide spectrum of antimicrobial activity. Further research may be aimed at elucidating the component composition of rowan leaf extracts and establishing the relationship between individual phytocompounds and the antimicrobial effects of plant extracts.

Fungicidal and antibiofilm activities of 2-Bromo-N-Phenylacetamide Against Fluconazole-Resistant Candida spp.: a promising antifungal agent

Purpose: Amides are proven to possess antimicrobial properties against both fungi and bacteria. The synthetic amide 2-bromo-N-phenylacetamide exhibits antifungal activity according to scientific literature, however, data remains scarce. Given the clinical and epidemiological importance as well as the increasing antifungal resistance in Candida spp., here we aimed to better explore the antifungal potential of 2-bromo-N-phenylacetamide against fluconazole-resistant Candida albicans, Candida glabrata, Candida parapsilosis and for the first time, against Candida tropicalis as well. Methods: Minimum Inhibitory and Minimum Fungicidal Concentrations (MIC/MFC) were established by a Broth Microdilution Technique. Mode of action was explored by Ergosterol and Sorbitol assays. For investigating the combined effect between 2-bromo-N-phenylacetamide and the standard antifungals Amphotericin B and Fluconazole a Checkerboard Technique was performed. Finally, the antibiofilm properties of 2-bromo-N-phenylacetamide were assessed by a Crystal Violet Staining Assay. Results: 2-bromo-N-phenylacetamide exhibited a fungicidal effect, MIC of 32 µg.mL-1 (87.5% of the strains) and MFC of 64 µg.mL-1 (81.25% of the strains). It also demonstrated an activity against Candida albicans mature biofilms as effective as that for Amphotericin B at all tested concentrations. When associated with Fluconazole and Amphotericin B it exerted indifferent effects. Mode of action remained unclear. Conclusion: The synthetic amide 2-bromo-N-phenylacetamide possesses a strong antifungal activity against Candida albicans, Candida tropicalis, Candida glabrata and Candida parapsilosis and an antibiofilm property against Candida albicans as effective as that exhibited for Amphotericin B.

Spectrum and functional properties of &lt;i&gt;ERG11&lt;/i&gt; gene mutations in fluconazole-resistant &lt;i&gt;Candida albicans&lt;/i&gt; strains isolated from HIV-infected patients

Rationale. The low efficacy of azole antimycotics in treatment of Candida infections, especially in HIV-infected patients, is often associated with overexpression of the ERG11 gene in Candida spp., which results in increased production of ergosterol the target of the above antimycotic drugs. Researchers have found ERG11 gene mutations that can modify its overexpression effects by increasing or decreasing it. However, the findings reported by different laboratories and countries are highly contradictory.&#x0D; The purpose of the study is to explore the spectrum and functional properties of ERG11 gene mutations in fluconazole-resistant Candida albicans strains isolated from HIV-infected patients.&#x0D; Materials and methods. The study was performed using 10 C. albicans strains inherently resistant to fluconazole and voriconazole and isolated from the oropharynx of HIV-infected patients; the strains were provided from the collection of the Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology. The strains were assessed by their sensitivity to antimycotic agents: anidulafungin, micafungin, caspofungin, posaconazole, voriconazole, itraconazole, fluconazole, amphotericin B, 5-flucytosine. Expression levels of the ERG11 gene were measured by quantitative PCR. ERG11 gene mutations were identified by Sanger sequencing.&#x0D; Results. Five mutations (E266D, G464S, I471L, D116E, and V488I) were detected in the ERG11 gene in seven C. albicans strains; six strains carried non-associated co-occurring mutations. Increased expression of the ERG11 gene was found in six C. albicans strains. The V488I mutation demonstrated a strong negative association with the increased expression of the ERG11 gene (r = 0.845; p 0.05). The minimum inhibitory concentration (MIC) in strains carrying mutations was a hundred times as low (p 0.05) as MIC in strains without mutations. In mutation carriers, posaconazole and itraconazole MICs were on average 16.5 times as low as MICs of voriconazole and fluconazole (p 0.001). The presence of mutations in the ERG11 gene had almost no effect on MICs of the tested antimycotics of the echinocandin, polyene, and pyrimidine groups.&#x0D; Conclusion. Multiple mutations were detected in the ERG11 gene in most of the C. albicans strains isolated from HIV-infected patients and resistant to fluconazole and voriconazole. Except for the V488I mutation, the detected mutations were not associated with the overexpression of the ERG11 gene and decreased the effects of overexpression of the ERG11 gene by up to 100 times, though they did not eliminate the inherent resistance to triazole antimycotics.

Chelerythrine, a major ingredient isolated from Macleaya cordata (Willd.) R. Br. (Papaveraceae), inhibits fluconazole-resistant Candida albicans biofilms

IntroductionIt has a long history to employ Macleaya cordata (Willd.) R. Br. (Papaveraceae) for the treatment of a variety of diseases in China. Chelerythrine, one of the major of M. cordata with detoxifying and anti-inflammatory effect, is reported to combat bacteria and fungus. This work aims to explore the effect of chelerythrine against fluconazole-resistant Candida albicans and also illustrate its potential mechanism. MethodsThe microdilution method, following CLSI-M27-A3 was used to evaluate the effect of chelerythrine against fluconazole-resistant C. albicans. Meanwhile, biofilm viability was assessed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay and cell morphology was observed by inverted microscope. Real Time Quantitative PCR (RT-qPCR) and confocal laser scanning microscope were employed to detect the mRNA expression and 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) staining, respectively. Fluorescence intensity was quantified by flow cytometry. ResultsThe total alkaloids extract of M. cordata significantly inhibited the viability of C. albicans, concentration of which in 64 μg/ml presented to 40% inhibitory effect. In addition, the minimum inhibitory concentration (MIC)80 for chelerythrine was 8 μM, whereas chelerythrine in 4 μM inhibited biofilm formation by 47.7%. Further studies revealed that chelerythrine reduced the hydrophobicity of cellular surface, inhibited hyphal growth, downregulated the expression of genes related to cyclic adenosine monophosphate (cAMP) pathway, increased cellular permeability and promoted the production of reactive oxygen species (ROS). ConclusionChelerythrine inhibited biofilm formation by decreasing cellular surface hydrophobicity (CSH) and inhibiting the cAMP pathway. Chelerythrine increased cellular permeability and promoted the production of reactive oxygen species, which leads to the cell death of fluconazole-resistant C. albicans.

Design, synthesis and in vitro biological studies of novel triazoles with potent and broad-spectrum antifungal activity

A series of novel triazole derivatives containing aryl-propanamide side chains was designed and synthesised. In vitro antifungal activity studies demonstrated that most of the compounds inhibited the growth of six human pathogenic fungi. In particular, parts of phenyl-propionamide-containing compounds had excellent, broad-spectrum antifungal activity against Candida albicans SC5314, Cryptococcus neoformans 22-21, Candida glabrata 537 and Candida parapsilosis 22-20 with MIC values in the range of ≤0.125 µg/mL–4.0 µg/mL. In addition, compounds A1, A2, A6, A12 and A15 showed inhibitory activities against fluconazole-resistant Candida albicans and Candida auris. Preliminary structure-activity relationships (SARs) are also summarised. Moreover, GC-MS analysis demonstrated that A1, A3, and A9 interfered with the C. albicans ergosterol biosynthesis pathway by inhibiting Cyp51. Molecular docking studies elucidated the binding modes of A3 and A9 with Cyp51. These compounds with low haemolytic activity and favourable ADME/T properties are promising for the development of novel antifungal agents.

Persephacin Is a Broad-Spectrum Antifungal Aureobasidin Metabolite That Overcomes Intrinsic Resistance in Aspergillus fumigatus.

Fungi pose a persistent threat to humankind with worrying indications that emerging and re-emerging pathogens (e.g., Candida auris, Coccidioides spp., drug-resistant Aspergilli, and more) exhibit resistance to the limited number of approved antifungals. To address this problem, our team is exploring endophytic fungi as a resource for the discovery of new antifungal natural products. The rationale behind this decision is based on evidence that endophytes engage with plants in mutualistic relationships wherein some fungi actively participate by producing chemical defense measures that suppress pathogenic microorganisms. To improve the odds of bioactive metabolite discovery, we developed a new hands-free laser-cutting system capable of generating >50 plant samples per minute that, in turn, enabled our team to prepare and screen large numbers of endophytic fungi. One of the fungal isolates obtained in this way was identified as an Elsinoë sp. that produced a unique aureobasidin analogue, persephacin (1). Some distinctive features of 1 are the absence of both phenylalanine residues combined with the incorporation of a novel amino acid residue, persephanine (9). Compound 1 exhibits potent antifungal effects against a large number of pathogenic yeast (including several clinical C. auris strains), as well as phylogenetically diverse filamentous fungi (e.g., Aspergillus fumigatus). In an ex vivo eye infection model, compound 1 outperformed standard-of-care treatments demonstrating the ability to suppress fluconazole-resistant Candida albicans and A. fumigatus at a concentration (0.1% solution) well below the clinically recommended levels used for fluconazole and natamycin (2% and 5% solutions, respectively). In 3D tissue models for acute dermal and ocular safety, 1 was found to be nontoxic and nonirritating at concentrations required to elicit antifungal activity. Natural product 1 appears to be a promising candidate for further investigation as a broad-spectrum antifungal capable of controlling a range of pathogens that negatively impact human, animal, and plant health.

Overcoming Drug Resistance in a Clinical C. albicans Strain Using Photoactivated Curcumin as an Adjuvant.

The limited antifungal drugs available and the rise of multidrug-resistant Candida species have made the efforts to improve antifungal therapies paramount. To this end, our research focused on the effect of a combined treatment between chemical and photodynamic therapy (PDT) towards a fluconazole-resistant clinical Candida albicans strain. The co-treatment of PDT and curcumin in various doses with fluconazole (FLC) had an inhibitory effect on the growth of the FLC-resistant hospital strain of C. albicans in both difusimetric and broth microdilution methods. The proliferation of the cells was inhibited in the presence of curcumin at 3.125 µM and FLC at 41 µM concentrations. The possible involvement of oxidative stress was analyzed by adding menadione and glutathione as a prooxidant and antioxidant, respectively. In addition, we examined the photoactivated curcumin effect on efflux pumps, a mechanism often linked to drug resistance. Nile Red accumulation assays were used to evaluate efflux pumps activity through fluorescence microscopy and spectrofluorometry. The results showed that photoactivated curcumin at 3.125 µM inhibited the transport of the fluorescent substrate that cells usually expel, indicating its potential in combating drug resistance. Overall, the findings suggest that curcumin, particularly when combined with PDT, can effectively inhibit the growth of FLC-resistant C. albicans, addressing the challenge of yeast resistance to azole antifungals through upregulating multidrug transporters.

Atorvastatin liposomes in a 3D-printed polymer film: a repurposing approach for local treatment of oral candidiasis

Oral candidiasis (OC) is an opportunistic fungal infection, common amongst the elderly and the immunocompromised. Unfortunately, the therapeutic efficacy of common antifungals is imperiled by the rise of antifungal drug resistance. An alternative promising therapeutic option possibly contributing to antifungal therapy is drug repurposing. Herein, we aimed to employ novel pharmaceutical drug delivery for enhancing the emerging antifungal potential of the hypocholesterolemic drug atorvastatin (ATV). ATV-propylene-glycol-liposomes (ATV/PG-Lip) were prepared then integrated in 3D-printed (3DP) mucoadhesive films comprising chitosan, polyvinyl-alcohol and hydroxypropyl methylcellulose, as an innovative blend, for the management of OC. ATV/PG-Lip demonstrated good colloidal properties of particle size (223.3 ± 2.1 nm), PDI (0.12 ± 0.001) and zeta potential (-18.2 ± 0.3 mV) with high entrapment efficiency (81.15 ± 1.88%) and sustained drug release. Also, ATV/PG-Lip showed acceptable three-month colloidal stability and in vitro cytocompatibility on human gingival fibroblasts. The developed 3DP-films exhibited controlled ATV release (79.4 ± 1.4% over 24 h), reasonable swelling and mucoadhesion (2388.4 ± 18.4 dyne/cm2). In vitro antifungal activity of ATV/PG-Lip was confirmed against fluconazole-resistant Candida albicans via minimum inhibitory concentration determination, time-dependent antifungal activity, agar diffusion and scanning electron microscopy. Further, ATV/PG-Lip@3DP-film exceeded ATV@3DP-film in amelioration of infection and associated inflammation in an in vivo oral candidiasis rabbit model. Accordingly, the results confirm the superiority of the fabricated ATV/PG-Lip@3DP-film for the management of oral candidiasis and tackling antifungal resistance.Graphical abstract

Isolation and Identification of Flavanone Derivative Eriodictyol from the Methanol Extract of Afzelia africana Bark and Its Antimicrobial and Antioxidant Activities.

Afzelia africana is a plant species with well-documented ethnobotanical and medicinal properties. The plant is reported to have various secondary metabolites and had been applied for the treatment of various diseased conditions. The study objectives include fractionation, isolation, purification, and characterization of eriodictyol from the bark of A. africana, and the determination of its antimicrobial and antioxidant activities. Methodology. The series of methodologies that were employed include fractionations and purification (column chromatography), characterization (HPLC, LC-MS, IR, 1H, 13C, DEPT-135, HSQC, and HMBC), antimicrobial assays (microbroth dilution and checkerboard assay), and antioxidant activities assays (ABTS and DPPH scavenging capacity). The study reports the identification and characterization of eriodictyol from the bark of A. africana which exhibited potent antioxidant activities against ABTS and DPPH radicals with scavenging capacities (SC50) of 2.14 ± 0.05 and 2.51 ± 0.06 µg/mL, respectively. The compound exhibited its antimicrobial activity by reporting good bacteriostatic activities (MBC/MIC > 4) against Staphylococcus aureus (SA), methicillin-resistant Staphylococcus aureus (MRSA), and fluconazole-resistant Candida albicans (CA2). Moreover, a broad spectrum of bactericidal effects (MBC/MIC ≤ 4) was reported against Streptococcus mutans (SM), Escherichia coli (EC), Bacillus subtilis (BS), Klebsiella pneumonia (KP), Pseudomonas aeruginosa (PA), Salmonella typhi (ST), and standard Candida albicans (CA1). The compound further exhibited synergistic effects against EC, KP, ST, and MRSA; ST; and CA2 when combined with ciprofloxacin, tetracycline, and nystatin, respectively. However, antagonistic effects were observed against PA and CA1 when combined with ciprofloxacin and ketoconazole, respectively. The study reports for the first time the identification of eriodictyol from the bark of A. africana which exhibited significant antioxidant and antimicrobial properties.

Discovery of BRD4-HDAC Dual Inhibitors with Improved Fungal Selectivity and Potent Synergistic Antifungal Activity against Fluconazole-Resistant Candida albicans.

Over the past several decades, invasive fungal infections, especially candidiasis, have caused dramatic morbidity and mortality due to ineffective antifungal drugs and severe drug resistance. Herein, new BRD4-histone deacetylase (HDAC) inhibitors were designed to restore the susceptibility of Candida albicans (C. albicans) to fluconazole (FLC). Interestingly, several compounds showed excellent selectivity against fungal HDACs. In particular, compound B2 showed excellent synergistic effect with FLC against resistant C. albicans (FICI = 0.063) with high selectivity against fungal HDACs (SI = 1653) and low cytotoxicity. Compound B2 effectively synergized with FLC and prevented biofilm formation and morphological transition in resistant C. albicans, potentiating the antifungal activity of FLC in vivo and significantly reducing kidney fungal loads. Thus, this drug combination is promising in the treatment of resistant C. albicans infections.

Fluconazole-Resistant Candida albicans Vaginal Infections at a Referral Center and Treated With Boric Acid.

The authors investigate the incidence of clinical and mycological resistance of Candida albicans vulvovaginitis (VVC) at the Jefferson Vulvovaginal Health Center. They also review their experience with boric acid in the treatment of fluconazole-resistant VVC. The authors conducted a retrospective chart review of all patients with C. albicans VVC diagnosed at the Jefferson Vulvovaginal Health Center between November 2019 and December 2021. Patients with clinically defined fluconazole resistance were identified. Information about demographics, in vitro susceptibility testing, and treatment outcomes with boric acid was obtained. Of 970 patients with vaginal C. albicans isolates, 71 (7.3%) with clinically defined fluconazole-resistant C. albicans infections were identified. Relevant demographics included 45.1% African American, 43.7% aged younger than 30 years, and 43.7% with body mass index less than 25. Of the 71 patients, 58 (81.7%) received vaginal boric acid treatment. The mycological and clinical cure rates were 85.7% and 73.7%, respectively. After successful boric acid treatment and negative yeast cultures, 14.3% of patients had a mycological recurrence within 3 months. Of 31 isolates with antifungal susceptibility testing, 83.9% (26/31) were found to have minimal inhibitory concentration results consistent with fluconazole resistance. In a tertiary care vulvovaginal health center, fluconazole-resistant Candida albicans VVC is by no means uncommon and usually responds in the short term to treatment with boric acid. However, in the absence of maintenance boric acid, recurrence of culture-positive VVC is likely.

Anti-Biofilm Activity of Phenyllactic Acid against Clinical Isolates of Fluconazole-Resistant Candida albicans

Commonly found colonizing the human microbiota, Candida albicans is a microorganism known for its ability to cause infections, mainly in the vulvovaginal region, and is responsible for 85% to 90% of vulvovaginal candidiasis (VVC) cases. The development of drug resistance in C. albicans isolates after long-term therapy with fluconazole is an important complication to solve and new therapeutic strategies are required to target this organism and its pathogenicity. In the present study, phenyllactic acid (PLA) an important broad-spectrum antimicrobial compound was investigated for its antifungal and antivirulence activities against clinical isolates of C. albicans. Previously characterized strains of C. albicans isolates from women with VVC and C. albicans ATCC90028 were used to evaluate the antimicrobial and time dependent killing assay activity of PLA showing a MIC 7.5 mg mL-1 and a complete reduction of viable Candida cells detected by killing kinetics after 4 h of treatment with PLA. Additionally, PLA significantly reduced the biomass and the metabolic activity of C. albicans biofilms and impaired biofilm formation also with changes in ERG11, ALS3, and HWP1 genes expression as detected by qPCR. PLA eradicated pre-formed biofilms as showed also with confocal laser scanning microscopy (CLSM) observations. Furthermore, the compound prolonged the survival rate of Galleria mellonella infected by C. albicans isolates. These results indicate that PLA is a promising candidate as novel and safe antifungal agents for the treatment of vulvovaginal candidiasis.

Arthrocolins Synergizing with Fluconazole Inhibit Fluconazole-Resistant Candida albicans by Increasing Riboflavin Metabolism and Causing Mitochondrial Dysfunction and Autophagy.

Our previous study reported that seminaturally occurring arthrocolins A to C with unprecedented carbon skeletons could restore the antifungal activity of fluconazole against fluconazole-resistant Candida albicans. Here, we showed that arthrocolins synergized with fluconazole, reducing the fluconazole minimum and dramatically augmenting the survivals of 293T human cells and nematode Caenorhabditis elegans infected with fluconazole-resistant C. albicans. Mechanistically, fluconazole can induce fungal membrane permeability to arthrocolins, leading to the intracellular arthrocolins that were critical to the antifungal activity of the combination therapy by inducing abnormal cell membranes and mitochondrial dysfunctions in the fungus. Transcriptomics and reverse transcription-quantitative PCR (qRT-PCR) analysis indicated that the intracellular arthrocolins induced the strongest upregulated genes that were involved in membrane transports while the downregulated genes were responsible for fungal pathogenesis. Moreover, riboflavin metabolism and proteasomes were the most upregulated pathways, which were accompanied by inhibition of protein biosynthesis and increased levels of reactive oxygen species (ROS), lipids, and autophagy. Our results suggested that arthrocolins should be a novel class of synergistic antifungal compounds by inducing mitochondrial dysfunctions in combination with fluconazole and provided a new perspective for the design of new bioactive antifungal compounds with potential pharmacological properties. IMPORTANCE The prevalence of antifungal-resistant Candida albicans, which is a common human fungal pathogen causing life-threatening systemic infections, has become a challenge in the treatment of fungal infections. Arthrocolins are a new type of xanthene obtained from Escherichia coli fed with a key fungal precursor toluquinol. Different from those artificially synthesized xanthenes used as important medications, arthrocolins can synergize with fluconazole against fluconazole-resistant Candida albicans. Fluconazole can induce the fungal permeability of arthrocolins into fungal cells, and then the intracellular arthrocolins exerted detrimental effects on the fungus by inducing fungal mitochondrial dysfunctions, leading to dramatically reduced fungal pathogenicity. Importantly, the combination of arthrocolins and fluconazole are effective against C. albicans in two models, including human cell line 293T and nematode Caenorhabditis elegans. Arthrocolins should be a novel class of antifungal compounds with potential pharmacological properties.