Bioluminescence Imaging to Study Mature Biofilm Formation by Candida spp. and Antifungal Activity In Vitro and In Vivo

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125 μg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivo toxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

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Effect of ozonized olive oil on oral levels of Candida spp. in patients with denture stomatitis

Brazilian Dental Science ◽

10.14295/bds.2018.v21i1.1489 ◽

2018 ◽

Vol 21 (1) ◽

pp. 111 ◽

Cited By ~ 1

Author(s):

Erica Crastechini ◽

Cristiane Yumi Koga-Ito ◽

Suzan De Fátima Machado ◽

Guilherme Rodrigues Teodoro ◽

Graziella Nuernberg Back-Brito ◽

...

Keyword(s):

Antifungal Activity ◽

Olive Oil ◽

In Vitro Tests ◽

Candida Spp ◽

Isolation And Identification ◽

Denture Stomatitis ◽

Viable Cells ◽

In Vitro Antifungal Activity

Objective: The aim of this study was to evaluate the effect of ozonized oil (OZ) on the oral levels of Candida spp. in patients with denture stomatitis. Material and Methods: In vitro tests were performed to validate antifungal activity and to standardize OZ conditions. Antifungal activity was screened against C. albicans and five non-albicans species (C. tropicalis, C. dubliniensis, C. krusei, C. guilliermondii, and C. parapsilosis). Also, the effects on C. albicans planktonic and biofilm were evaluated. After validation, OZ was included in a therapeutic protocol of denture stomatitis in vivo. Thirty patients used OZ and 20 used sodium bicarbonate (SB) for 14 days. After 7 and 14 days, clinical evaluation, isolation and identification of yeasts were performed. Isolates were identified by phenotypic and genotypic tests. Ozonized oil showed in vitro antifungal activity against all species of Candida. Ozonized oil reduced the number of viable cells in C. albicans biofilms. Oral candidal levels were lower in relation to baseline both after after 14 days of treatment with SB and OZ. Results: A total of 493 Candida spp. isolates was obtained and 80% were identified as C. albicans. Remission of denture stomatitis was observed in all patients after 7 days of treatment in both groups. Conclusion: Within the limits of the study we can conclude that ozonized olive oil can be a new alternative for the control of biofilm in patients with denture stomatitis. KeywordsOzone; Candida; Antifungal Agents; Stomatitis; Denture.

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Synthesis, Biological Evaluation, and Structure–Activity Relationships of 4-Aminopiperidines as Novel Antifungal Agents Targeting Ergosterol Biosynthesis

Molecules ◽

10.3390/molecules26237208 ◽

2021 ◽

Vol 26 (23) ◽

pp. 7208

Author(s):

Jürgen Krauß ◽

Christoph Müller ◽

Monika Klimt ◽

Leandro Jorquera Valero ◽

José Francisco Martínez ◽

...

Keyword(s):

Antifungal Activity ◽

Galleria Mellonella ◽

Cholesterol Biosynthesis ◽

Biological Evaluation ◽

Ergosterol Biosynthesis ◽

In Vivo Model ◽

Candida Spp ◽

Mcf10a Cells

The aliphatic heterocycles piperidine and morpholine are core structures of well-known antifungals such as fenpropidin and fenpropimorph, commonly used as agrofungicides, and the related morpholine amorolfine is approved for the treatment of dermal mycoses in humans. Inspired by these lead structures, we describe here the synthesis and biological evaluation of 4-aminopiperidines as a novel chemotype of antifungals with remarkable antifungal activity. A library of more than 30 4-aminopiperidines was synthesized, starting from N-substituted 4-piperidone derivatives by reductive amination with appropriate amines using sodium triacetoxyborohydride. Antifungal activity was determined on the model strain Yarrowia lipolytica, and some compounds showed interesting growth-inhibiting activity. These compounds were tested on 20 clinically relevant fungal isolates (Aspergillus spp., Candida spp., Mucormycetes) by standardized microbroth dilution assays. Two of the six compounds, 1-benzyl-N-dodecylpiperidin-4-amine and N-dodecyl-1-phenethylpiperidin-4-amine, were identified as promising candidates for further development based on their in vitro antifungal activity against Candida spp. and Aspergillus spp. Antifungal activity was determined for 18 Aspergillus spp. and 19 Candida spp., and their impact on ergosterol and cholesterol biosynthesis was determined. Toxicity was determined on HL-60, HUVEC, and MCF10A cells, and in the alternative in vivo model Galleria mellonella. Analysis of sterol patterns after incubation gave valuable insights into the putative molecular mechanism of action, indicating inhibition of the enzymes sterol C14-reductase and sterol C8-isomerase in fungal ergosterol biosynthesis.

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Antifungal activity of green and red Brazilian propolis extracts

The Natural Products Journal ◽

10.2174/2210315511666210302161514 ◽

2021 ◽

Vol 11 ◽

Author(s):

Márcia Christina Dornelas de Freitas ◽

Nívea Pereira de Sá ◽

Blenda Fernandes ◽

Anny Caroline Messias ◽

Gabriela Fonseca Lopes ◽

...

Keyword(s):

Antifungal Activity ◽

Amphotericin B ◽

Synergistic Activity ◽

Candida Spp ◽

Physiochemical Parameters ◽

Brazilian Legislation ◽

Buccal Epithelial Cells ◽

Brazilian Propolis

Background: Diseases associated to Candida spp. are recurrent and can be difficult to treat, mainly due to the new strains resistant to the limited number of available antifungals. Objective: Evaluate the in vitro and in vivo activity of Brazilian green propolis (GrProp) and red propolis (RdProp) ethanolic extracts against standard strains of Candida albicans, C. tropicalis, C. krusei, C. glabrata, C. parapsilosis and 10 clinical isolates of C. albicans. Methods: Antifungal activity in vitro was tested using the M-27-A3/CLSI protocol. The in vivo antifungal activity was evaluated using Tenebrio molitor model. And, the effect of extracts on adhesion of C. albicans in human buccal epithelial cells (BECs) was also studied. Results: GrProp and RdProp exhibited antifungal activity against at least one of the Candida strains tested. The adhesion inhibition of C. albicans in BECs was of 45% (GrProp28), 60% (GrProp50) and of 82% (RdProp), in comparison to amphotericin B (82%). All propolis extracts showed synergistic activity with fluconazole and amphotericin B. GrProp50 (10 mg/kg) showed the better protection of T. molitor, blocking the progression of C. albicans infection, increasing survival and delayed the larvae death. Conclusion: Brazilian GrProp and RdProp extracts inhibit the in vitro C. albicans growth and protect T. molitor against infection by this yeast. The physiochemical parameters found for the analyzed samples were in accordance to the standards established by the Brazilian Legislation for propolis and derivatives. GrProp and RdProp have potential to be used against Candida spp. infections, mainly in association with fluconazole or amphotericin B.

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REFRACTORY FUNGAL VAGINITIS TREATED BY TOPICAL AMPHOTERICIN B. Review

Medical Science of Ukraine (MSU) ◽

10.32345/2664-4738.2.2020.10 ◽

2020 ◽

Vol 16 (2) ◽

pp. 55-58

Author(s):

Falah Hasan Obayes AL-Khikani

Keyword(s):

Antifungal Activity ◽

Side Effects ◽

Amphotericin B ◽

Intravenous Injection ◽

Fungal Infections ◽

Wide Spectrum ◽

Candida Spp

Vaginitis is a common problem for women regarding a worldwide health challenge with many side effects. Vaginitis is among the most visiting to gynecology clinics. About 75% of all reproductive women had at least one fungal vaginitis infection in their life, and more than 40% will have two or more than two. Candida spp is the most prevalent in fungal vaginitis, while reports for unusual fungi were observed as mucor spp. Amphotericin B (AmB) belongs to the polyene group has a wide spectrum in vitro and in vivo antifungal activity. All of the known available formulas of AmB are administrated via intravenous injection to treat severe systemic fungal infections, while the development of the topical formula of AmB is still under preliminary development including topical vaginal AmB. Due to the revealing of antimicrobial-resistant fungi in recent years, this study explains the role of topical AmB in treating refractory fungi vaginitis that may not a response to other drugs reported in many cases that may help researchers to develop new effective formula of AmB regarding fungal vaginitis.

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Novel Sulfones with Antifungal Properties: Antifungal Activities and Interactions with Candida spp. Virulence Factors

Mini-Reviews in Medicinal Chemistry ◽

10.2174/1389557518666180924121209 ◽

2018 ◽

Vol 19 (1) ◽

pp. 12-21 ◽

Cited By ~ 4

Author(s):

Małgorzata Gizińska ◽

Monika Staniszewska ◽

Zbigniew Ochal

Keyword(s):

Antifungal Activity ◽

Virulence Factors ◽

Hydrolytic Enzymes ◽

Candida Spp ◽

Pathogenic Potential ◽

Robust Response ◽

New Strategies ◽

Sulfone Derivatives

Since candidiasis is so difficult to eradicate with an antifungal treatment and the existing antimycotics display many limitations, hopefully new sulfone derivatives may overcome these deficiencies. It is pertinent to study new strategies such as sulfone derivatives targeting the virulence attributes of C. albicans that differentiate them from the host. During infections, the pathogenic potential of C. albicans relies on the virulence factors as follows: hydrolytic enzymes, transcriptional factors, adhesion, and development of biofilms. In the article we explored how the above-presented C. albicans fitness and virulence attributes provided a robust response to the environmental stress exerted by sulfones upon C. albicans; C. albicans fitness and virulence attributes are fungal properties whose inactivation attenuates virulence. Our understanding of how these mechanisms and factors are inhibited by sulfones has increased over the last years. As lack of toxicity is a prerequisite for medical approaches, sulfones (non-toxic as assessed in vitro and in vivo) may prove to be useful for reducing C. albicans pathogenesis in humans. The antifungal activity of sulfones dealing with these multiple virulence factors and fitness attributes is discussed.

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Antifungal activity of β-lapachone against azole-resistant Candida spp. and its aspects upon biofilm formation

Future Microbiology ◽

10.2217/fmb-2020-0011 ◽

2020 ◽

Vol 15 (16) ◽

pp. 1543-1554

Author(s):

Cecília R da Silva ◽

Rosana de S Campos ◽

João B de A Neto ◽

Letícia S Sampaio ◽

Francisca BSA do Nascimento ◽

...

Keyword(s):

Flow Cytometry ◽

Antifungal Activity ◽

Comet Assay ◽

Biofilm Formation ◽

Broth Microdilution ◽

Candida Spp ◽

Antifungal Effect ◽

Planktonic Form ◽

Resistant Strains

Aim: The purpose of this study was to assess the antifungal effect of β-lapachone (β-lap) on azole-resistant strains of Candida spp. in both planktonic and biofilm form. Materials & methods: The antifungal activity of β-lap was evaluated by broth microdilution, flow cytometry and the comet assay. The cell viability of the biofilms was assessed using the MTT assay. Results: β-lap showed antifungal activity against resistant strains of Candida spp. in planktonic form. In addition, β-lap decreased the viability of mature biofilms and inhibited the formation of biofilms in vitro. Conclusion: β-lap showed antifungal activity against Candida spp., suggesting that the compound can be utilized as an adjunct agent in the treatment of candidiasis.

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Antifungal Activity of Oleylphosphocholine on In Vitro and In Vivo Candida albicans Biofilms

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01767-17 ◽

2017 ◽

Vol 62 (1) ◽

Cited By ~ 4

Author(s):

Michelle Holtappels ◽

Erwin Swinnen ◽

Lies De Groef ◽

Jurgen Wuyts ◽

Lieve Moons ◽

...

Keyword(s):

Candida Albicans ◽

Antifungal Activity ◽

Biofilm Formation ◽

Ex Vivo ◽

Oral Candidiasis ◽

Sprague Dawley ◽

Content Type ◽

Polyurethane Catheters

ABSTRACT In this study, we investigated the potential antifungal activity of the alkylphospholipid oleylphosphocholine (OlPC), a structural analogue of miltefosine, on in vitro and in vivo Candida albicans biofilm formation. The effect of OlPC on in vitro and in vivo C. albicans biofilms inside triple-lumen polyurethane catheters was studied. In vivo biofilms were developed subcutaneously after catheter implantation on the lower back of Sprague-Dawley rats. Animals were treated orally with OlPC (20 mg/kg of body weight/day) for 7 days. The effect of OlPC on biofilms that developed on the mucosal surface was studied in an ex vivo model of oral candidiasis. The role of OlPC in C. albicans morphogenesis was investigated by using hypha-inducing media, namely, Lee, Spider, and RPMI 1640 media. OlPC displayed activity against both planktonic cells and in vitro C. albicans biofilms. To completely abolish preformed, 24-h-old biofilms, higher concentrations (8, 10, and 13 mg/liter) were needed. Moreover, OlPC was able to reduce C. albicans biofilms formed by caspofungin-resistant clinical isolates and acted synergistically when combined with caspofungin. The daily oral administration of OlPC significantly reduced in vivo C. albicans biofilms that developed subcutaneously. In addition, OlPC decreased biofilm formation on mucosal surfaces. Interestingly, the application of subinhibitory concentrations of OlPC already inhibited the yeast-to-hypha transition, a crucial virulence factor of C. albicans. We document, for the first time, the effects of OlPC on C. albicans cells and suggest the potential use of OlPC for the treatment of C. albicans biofilm-associated infections.

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A Peptide Found in Human Serum, Derived from the C-Terminus of Albumin, Shows Antifungal Activity In Vitro and In Vivo

Microorganisms ◽

10.3390/microorganisms8101627 ◽

2020 ◽

Vol 8 (10) ◽

pp. 1627

Author(s):

Tecla Ciociola ◽

Pier Paolo Zanello ◽

Tiziana D’Adda ◽

Serena Galati ◽

Stefania Conti ◽

...

Keyword(s):

Antifungal Activity ◽

Human Serum ◽

Fungicidal Activity ◽

Galleria Mellonella ◽

Serum Proteins ◽

Morphological Alterations ◽

C Terminus ◽

Different Sources

The growing problem of antimicrobial resistance highlights the need for alternative strategies to combat infections. From this perspective, there is a considerable interest in natural molecules obtained from different sources, which are shown to be active against microorganisms, either alone or in association with conventional drugs. In this paper, peptides with the same sequence of fragments, found in human serum, derived from physiological proteins, were evaluated for their antifungal activity. A 13-residue peptide, representing the 597–609 fragment within the albumin C-terminus, was proved to exert a fungicidal activity in vitro against pathogenic yeasts and a therapeutic effect in vivo in the experimental model of candidal infection in Galleria mellonella. Studies by confocal microscopy and transmission and scanning electron microscopy demonstrated that the peptide penetrates and accumulates in Candida albicans cells, causing gross morphological alterations in cellular structure. These findings add albumin to the group of proteins, which already includes hemoglobin and antibodies, that could give rise to cryptic antimicrobial fragments, and could suggest their role in anti-infective homeostasis. The study of bioactive fragments from serum proteins could open interesting perspectives for the development of new antimicrobial molecules derived by natural sources.

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Copper(II) and Zinc(II) Complexes with the Clinically Used Fluconazole: Comparison of Antifungal Activity and Therapeutic Potential

Pharmaceuticals ◽

10.3390/ph14010024 ◽

2020 ◽

Vol 14 (1) ◽

pp. 24

Author(s):

Nevena Lj. Stevanović ◽

Ivana Aleksic ◽

Jakob Kljun ◽

Sanja Skaro Bogojevic ◽

Aleksandar Veselinovic ◽

...

Keyword(s):

Antifungal Activity ◽

Biofilm Formation ◽

Candida Parapsilosis ◽

Therapeutic Potential ◽

A549 Cells ◽

Candida Krusei ◽

Strong Inhibition ◽

Subinhibitory Concentrations ◽

Pyocyanin Production

Copper(II) and zinc(II) complexes with clinically used antifungal drug fluconazole (fcz), {[CuCl2(fcz)2].5H2O}n, 1, and {[ZnCl2(fcz)2]·2C2H5OH}n, 2, were prepared and characterized by spectroscopic and crystallographic methods. The polymeric structure of the complexes comprises four fluconazole molecules monodentately coordinated via the triazole nitrogen and two chlorido ligands. With respect to fluconazole, complex 2 showed significantly higher antifungal activity against Candida krusei and Candida parapsilosis. All tested compounds reduced the total amount of ergosterol at subinhibitory concentrations, indicating that the mode of activity of fluconazole was retained within the complexes, which was corroborated via molecular docking with cytochrome P450 sterol 14α-demethylase (CYP51) as a target. Electrostatic, steric and internal energy interactions between the complexes and enzyme showed that 2 has higher binding potency to this target. Both complexes showed strong inhibition of C. albicans filamentation and biofilm formation at subinhibitory concentrations, with 2 being able to reduce the adherence of C. albicans to A549 cells in vitro. Complex 2 was able to reduce pyocyanin production in Pseudomonas aeruginosa between 10% and 25% and to inhibit its biofilm formation by 20% in comparison to the untreated control. These results suggest that complex 2 may be further examined in the mixed Candida-P. aeruginosa infections.

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