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1.
Sci Rep ; 14(1): 16760, 2024 07 21.
Article in English | MEDLINE | ID: mdl-39033245

ABSTRACT

Gut fungal imbalances, particularly increased Candida spp., are linked to obesity. This study explored the potential of Lactiplantibacillus plantarum cell-free extracts (postbiotics) to modulate the growth of Candida albicans and Candida kefyr, key members of the gut mycobiota. A minimal synthetic gut model was employed to evaluate the effects of Lactiplantibacillus plantarum postbiotics on fungal growth in mono- and mixed cultures. Microreactors were employed for culturing, fungal growth was quantified using CFU counting, and regression analysis was used to evaluate the effects of postbiotics on fungal growth. Postbiotics at a concentration of 12.5% significantly reduced the growth of both Candida species. At 24 h, both C. albicans and C. kefyr in monocultures exhibited a decrease in growth of 0.11 log CFU/mL. In contrast, mixed cultures showed a more pronounced antifungal effect, with C. albicans and C. kefyr reductions of 0.62 log CFU/mL and 0.64 log CFU/mL, respectively. Regression analysis using the Gompertz model supported the antifungal activity of postbiotics and revealed species-specific differences in growth parameters. These findings suggest that L. plantarum postbiotics have the potential to modulate the gut mycobiota by reducing Candida growth, potentially offering a therapeutic approach for combating fungal overgrowth associated with obesity.


Subject(s)
Candida , Gastrointestinal Microbiome , Obesity , Obesity/microbiology , Candida/drug effects , Candida/growth & development , Gastrointestinal Microbiome/drug effects , Humans , Probiotics/pharmacology , Candida albicans/drug effects , Models, Biological , Antifungal Agents/pharmacology
2.
J Med Microbiol ; 73(7)2024 Jul.
Article in English | MEDLINE | ID: mdl-38979984

ABSTRACT

Introduction. The development of new antifungal drugs has become a global priority, given the increasing cases of fungal diseases together with the rising resistance to available antifungal drugs. In this scenario, drug repositioning has emerged as an alternative for such development, with advantages such as reduced research time and costs.Gap statement. Propafenone is an antiarrhythmic drug whose antifungal activity is poorly described, being a good candidate for further study.Aim. This study aims to evaluate propafenone activity against different species of Candida spp. to evaluate its combination with standard antifungals, as well as its possible action mechanism.Methodology. To this end, we carried out tests against strains of Candida albicans, Candida auris, Candida parapsilosis, Candida tropicalis, Candida glabrata and Candida krusei based on the evaluation of the MIC, minimum fungicidal concentration and tolerance level, along with checkerboard and flow cytometry tests with clinical strains and cell structure analysis by scanning electron microscopy (SEM).Results. The results showed that propafenone has a 50% MIC ranging from 32 to 256 µg ml-1, with fungicidal activity and positive interactions with itraconazole in 83.3% of the strains evaluated. The effects of the treatments observed by SEM were extensive damage to the cell structure, while flow cytometry revealed the apoptotic potential of propafenone against Candida spp.Conclusion. Taken together, these results indicate that propafenone has the potential for repositioning as an antifungal drug.


Subject(s)
Antifungal Agents , Candida , Microbial Sensitivity Tests , Propafenone , Antifungal Agents/pharmacology , Candida/drug effects , Candida/growth & development , Propafenone/pharmacology , Humans , Itraconazole/pharmacology , Drug Synergism , Drug Resistance, Fungal/drug effects , Candidiasis/microbiology , Candidiasis/drug therapy , Drug Repositioning
3.
PLoS One ; 19(6): e0305405, 2024.
Article in English | MEDLINE | ID: mdl-38889118

ABSTRACT

Syzigium aromaticum essential oil (EO), eugenol, and ß-caryophyllene were evaluated regarding antifungal, antibiofilm, and in vitro toxicity. Additionally, in vivo toxicity of EO was observed. Anti-Candida activity was assessed through broth microdilution assay for all compounds. Time-kill assay (0, 1, 10, 30 min, 1, 2, and 4 h) was used to determine the influence of EO and eugenol on Candida Growth kinetics. Thereafter, both compounds were evaluated regarding their capacity to act on a biofilm formation and on mature biofilm, based on CFU/ml/g of dry weight. Cell Titer Blue Viability Assay was used for in vitro cytotoxicity, using oral epithelial cells (TR146) and human monocytes (THP-1). Lastly, Galleria mellonella model defined the EO in vivo acute toxicity. All compounds, except ß-cariofilene (MIC > 8000 µg/ml), presented antifungal activity against Candida strains (MIC 500-1000 µg/ml). The growth kinetics of Candida was affected by the EO (5xMIC 30 min onward; 10xMIC 10 min onward) and eugenol (5xMIC 10 min onward; 10xMIC 1 min onward). Fungal viability was also affected by 5xMIC and 10xMIC of both compounds during biofilm formation and upon mature biofilms. LD50 was defined for TR146 and THP1 cells at, respectively, 59.37 and 79.54 µg/ml for the EO and 55.35 and 84.16 µg/ml for eugenol. No sign of toxicity was seen in vivo up to 10mg/ml (20 x MIC) for the EO. S. aromaticum and eugenol presented antifungal and antibiofilm activity, with action on cell growth kinetics. In vivo acute toxicity showed a safe parameter for the EO up to 10 mg/ml.


Subject(s)
Antifungal Agents , Biofilms , Candida , Eugenol , Microbial Sensitivity Tests , Oils, Volatile , Syzygium , Oils, Volatile/pharmacology , Oils, Volatile/toxicity , Humans , Biofilms/drug effects , Biofilms/growth & development , Candida/drug effects , Candida/growth & development , Syzygium/chemistry , Antifungal Agents/pharmacology , Antifungal Agents/toxicity , Animals , Eugenol/pharmacology , Eugenol/toxicity , Cell Line
4.
Arch Microbiol ; 206(6): 257, 2024 May 11.
Article in English | MEDLINE | ID: mdl-38734773

ABSTRACT

There is a growing imperative for research into alternative compounds for the treatment of the fungal infections. Thus, many studies have focused on the analysis of antifungal proteins and peptides from different plant sources. Among these molecules are protease inhibitors (PIs). Previously, PIs present in the peptide-rich fractions called PEF1, PEF2 and PEF3 were identified from Capsicum chinense seeds, which have strong activity against phytopathogenic fungi. The aim of this study was to evaluate the mechanism of action and antimicrobial activity of PIs from PEF2 and PEF3 on the growth of yeasts of the genus Candida. In this work, analyses of their antimicrobial activity and cell viability were carried out. Subsequently, the mechanism of action by which the PIs cause the death of the yeasts was evaluated. Cytotoxicity was assessed in vitro by erythrocytes lysis and in vivo in Galleria mellonella larvae. PEF2 and PEF3 caused 100% of the growth inhibition of C. tropicalis and C. buinensis. For C. albicans inhibition was approximately 60% for both fractions. The PEF2 and PEF3 caused a reduction in mitochondrial functionality of 54% and 46% for C. albicans, 26% and 30% for C. tropicalis, and 71% and 68% for C. buinensis, respectively. These fractions induced morphological alterations, led to membrane permeabilization, elevated ROS levels, and resulted in necrotic cell death in C. tropicalis, whilst demonstrating low toxicity toward host cells. From the results obtained here, we intend to contribute to the understanding of the action of PIs in the control of fungal diseases of medical importance.


Subject(s)
Antifungal Agents , Candida , Protease Inhibitors , Antifungal Agents/pharmacology , Candida/drug effects , Candida/growth & development , Protease Inhibitors/pharmacology , Microbial Sensitivity Tests , Animals , Capsicum/microbiology , Reactive Oxygen Species/metabolism , Seeds/growth & development , Plant Extracts/pharmacology , Plant Extracts/chemistry , Erythrocytes/drug effects , Larva/microbiology , Larva/growth & development , Larva/drug effects
5.
Drug Des Devel Ther ; 15: 2091-2098, 2021.
Article in English | MEDLINE | ID: mdl-34040347

ABSTRACT

PURPOSE: The incidence of fungal infection after corneal transplant has increased significantly in recent years, especially Candida spp. This study aimed to evaluate the efficacy and safety of the addition of cycloheximide in Optisol-GS media in decreasing the growth of Candida spp. strains. METHODS: This in vitro laboratory efficacy study measured fungal colony growth in 24 vials of Optisol-GS that were divided into 6 groups of 4 vials each, as follows: (1) MIC/2 cycloheximide, (2) MIC cycloheximide, (3) MICx5 cycloheximide, (4) MICx10 cycloheximide, from MIC values obtained for each strain, (5) unsupplemented optisol-GS as a positive control (added inoculum), and (6) unsupplemented optisol-GS as a negative control (no inoculum). In each group was added Candida albicans, C. glabrata and C. parapsilosis, except in the negative control. The evaluated variables were fungal colony growth from the Optisol-GS vials, corneal endothelial cell density and endothelial cell viability at different concentrations of cycloheximide. RESULTS: In the efficacy study, all strains showed a reduction in fungal cell growth from the second day at all evaluated concentrations of optisol-GS supplemented with cycloheximide, even at subinhibitory concentrations (MIC/2). For C. glabrata, the colony count was reduced to 99%. No evidence of corneal endothelial toxicity was found at any concentration, in the safety study, compared with the paired control. CONCLUSION: The addition of cycloheximide to optisol-GS decreased the fungal growth, demonstrating fungicide action against C. glabrata and fungistatic action against C. albicans and C. parapsilosis. This drug did not demonstrate toxicity to the corneal endothelium at different concentrations.


Subject(s)
Antifungal Agents/pharmacology , Candida/drug effects , Chondroitin Sulfates/pharmacology , Cycloheximide/pharmacology , Dextrans/pharmacology , Gentamicins/pharmacology , Candida/growth & development , Complex Mixtures/pharmacology , Microbial Sensitivity Tests
6.
J Microbiol Methods ; 184: 106200, 2021 05.
Article in English | MEDLINE | ID: mdl-33713728

ABSTRACT

Isolating high quality RNA is a limiting factor in molecular analysis, since it is the base for transcriptional studies. The RNA extraction method can directly affect the RNA quality and quantity, as well as, its overall cost. The industrial importance of the yeast genus Candida in several sectors comes from their capacity to produce Lipases. These enzymes are one of the main metabolites produced by some Candida species, and it has been shown that Candida yeast can biodegrade petroleum hydrocarbons and diesel oil from biosurfactants that they can produce, a feature that turns these organisms into potential combatants for bioremediation techniques. Thus, this study aimed to determine an efficient method for isolating high quality RNA from Candida viswanathii biomass. To achieve this aim, three different RNA extraction methods, TRIzol, Hot Acid Phenol, and CTAB (Cetyltrimethylammonium Bromide), were tested. The three tested methods allowed the isolation of high-quality RNA from C. viswanathii biomass and yielded suitable RNA quantity for carrying out RT-qPCR studies. In addition, all methods displayed high sensitivity for the expression analysis of the CvGPH1 gene through RT-qPCR, with TRIzol and CTAB showing the best results and the CTAB method displaying the best cost-benefit ratio (US$0.35/sample).


Subject(s)
Candida/genetics , Chemical Fractionation/methods , RNA, Fungal/isolation & purification , Candida/growth & development , Candida/isolation & purification , Cetrimonium/chemistry , Chemical Fractionation/instrumentation , Phenol/chemistry , Polymerase Chain Reaction , RNA, Fungal/genetics
7.
Med Mycol ; 59(7): 744-747, 2021 Jul 06.
Article in English | MEDLINE | ID: mdl-33594432

ABSTRACT

Previous studies showed that the crude extract obtained from Streptococcus mutans inhibited the growth of Candida albicans reference strains. In this study, we evaluated whether the antifungal effects of S. mutans extract can be extended to clinical Candida isolates, including C. albicans and non-abicans strains with different susceptibilities to fluconazole. We verified that S. mutans extract increased the survival of Galleria mellonella larvae infected with C. albicans and C. glabrata and inhibited the fungal cells in hemolymph. These antifungal effects occurred for both fluconazole-susceptible and fluconazole-resistant strains. However, larvae infected by C. krusei were not affected by S. mutans extract. LAY SUMMARY: Streptococcus mutans crude extract shows antifungal effects on clinical Candida strains susceptible and resistant to fluconazole in Galleria mellonella model.


Subject(s)
Antifungal Agents/pharmacology , Candida albicans/drug effects , Candida/drug effects , Candida/growth & development , Streptococcus mutans/chemistry , Animals , Candida/classification , Candida albicans/growth & development , Complex Mixtures/pharmacology , Drug Resistance, Fungal , Larva/microbiology , Microbial Sensitivity Tests , Moths/microbiology
8.
Future Microbiol ; 16(2): 71-81, 2021 01.
Article in English | MEDLINE | ID: mdl-33459560

ABSTRACT

Aim: The purpose of this study was to evaluate the antifungal activity of midazolam, alone and in association with azoles, against isolates of clinical Candida spp. in planktonic and biofilm form. Materials & methods: The antifungal activity was observed using the broth microdilution technique. Flow cytometry tests were performed to investigate the probable mechanism of action and the comet test and cytotoxicity test were applied to evaluate DNA damage. Results: Midazolam (MIDAZ) showed antifungal activity against planktonic cells (125-250 µg/ml) and reduced the viability of Candida spp. biofilms (125 a 2500 µg/ml). The interaction of MIDAZ against Candida spp. biofilms was observed through scanning electron microscopy, causing alteration of their appearance. Therefore, MIDAZ has antifungal potential against Candida spp.


Subject(s)
Antifungal Agents/pharmacology , Candida/drug effects , Candidiasis/microbiology , Midazolam/pharmacology , Biofilms/drug effects , Candida/genetics , Candida/growth & development , Candida/physiology , Drug Evaluation, Preclinical , Drug Resistance, Fungal , Fluconazole/pharmacology , Humans , Microbial Sensitivity Tests
9.
Future Microbiol ; 15: 1439-1452, 2020 10.
Article in English | MEDLINE | ID: mdl-33156698

ABSTRACT

Aim: This work aimed to develop a membrane based on voriconazole (VCZ)-loaded natural rubber latex (NRL) for treating infected ulcers with Candida spp. and study their interaction, drug release, antifungal activity against Candida parapsilosis and biological characterization. Materials & methods: VCZ-loaded NRL membrane was produced by casting method. Results: Infrared spectrum showed that the incorporation of VCZ into the NRL membrane maintained its characteristics. Its mechanical properties were considered suitable for dermal application. The VCZ was able to release from NRL membrane, maintaining its antifungal activity against C. parapsilosis, besides did not present hemolytic effects. Conclusion: The VCZ-NRL membrane showed good results in mechanical, antifungal and biological assays, representing an interesting alternative to treatment of infected wound with Candida spp.


Subject(s)
Antifungal Agents/pharmacology , Bandages/microbiology , Candida/drug effects , Latex/chemistry , Skin Ulcer/microbiology , Voriconazole/pharmacology , Antifungal Agents/chemistry , Biomechanical Phenomena , Candida/growth & development , Humans , Microbial Sensitivity Tests , Skin Ulcer/drug therapy , Voriconazole/chemistry
10.
Toxins (Basel) ; 12(9)2020 08 20.
Article in English | MEDLINE | ID: mdl-32825220

ABSTRACT

The growing number of oral infections caused by the Candida species are becoming harder to treat as the commonly used antibiotics become less effective. This drawback has led to the search for alternative strategies of treatment, which include the use of antifungal molecules derived from natural products. Herein, crotoxin (CTX), the main toxin of Crotalus durissus terrificus venom, was challenged against Candida tropicalis (CBS94) and Candida dubliniensis (CBS7987) strains by in vitro antimicrobial susceptibility tests. Minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and inhibition of biofilm formation were evaluated after CTX treatment. In addition, CTX-induced cytotoxicity in HaCaT cells was assessed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) colorimetric assay. Native CTX showed a higher antimicrobial activity (MIC = 47 µg/mL) when compared to CTX-containing mouthwash (MIC = 750 µg/mL) and nystatin (MIC = 375 µg/mL). Candida spp biofilm formation was more sensitive to both CTX and CTX-containing mouthwash (IC100 = 12 µg/mL) when compared to nystatin (IC100 > 47 µg/mL). Moreover, significant membrane permeabilization at concentrations of 1.5 and 47 µg/mL was observed. Native CTX was less cytotoxic to HaCaT cells than CTX-containing mouthwash or nystatin between 24 and 48 h. These preliminary findings highlight the potential use of CTX in the treatment of oral candidiasis caused by resistant strains.


Subject(s)
Anti-Infective Agents, Local/pharmacology , Biofilms/drug effects , Candida tropicalis/drug effects , Candida/drug effects , Crotoxin/pharmacology , Mouthwashes/pharmacology , Anti-Infective Agents, Local/chemistry , Anti-Infective Agents, Local/isolation & purification , Biofilms/growth & development , Candida/growth & development , Candida tropicalis/growth & development , Cell Line, Transformed , Cell Membrane/drug effects , Cell Membrane/physiology , Cell Survival/drug effects , Cell Survival/physiology , Crotoxin/chemistry , Crotoxin/isolation & purification , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical/methods , Humans , Male , Middle Aged , Mouthwashes/chemistry , Treatment Outcome
11.
Biomolecules ; 10(8)2020 07 24.
Article in English | MEDLINE | ID: mdl-32722158

ABSTRACT

Candida species cause an opportunistic yeast infection called Candidiasis, which is responsible for more than 50,000 deaths every year around the world. Effective treatments against candidiasis caused by non-albicans Candida species such as C. glabrata, C. parapsilosis, C. aureus, and C.krusei are limited due to severe resistance to conventional antifungal drugs. Natural drimane sesquiterpenoids have shown promising antifungal properties against Candida yeast and have emerged as valuable candidates for developing new candidiasis therapies. In this work, we isolated isodrimeninol (C1) from barks of Drimys winteri and used it as starting material for the hemi-synthesis of four sesquiterpenoids by oxidation with pyridinium chlorochromate (PCC). The structure of the products (C2, C3, C4, and C5) was elucidated by 1D and 2D NMR spectroscopy resulting in C4 being a novel compound. Antifungal activity assays against C. albicans, C. glabrata, and C. krusei revealed that C4 exhibited an increased activity (IC50 of 75 µg/mL) compared to C1 (IC50 of 125 µg/mL) in all yeast strains. The antifungal activity of C1 and C4 was rationalized in terms of their capability to inhibit lanosterol 14-alpha demethylase using molecular docking, molecular dynamics simulations, and MM/GBSA binding free energy calculations. In silico analysis revealed that C1 and C4 bind to the outermost region of the catalytic site of 14-alpha demethylase and block the entrance of lanosterol (LAN) to the catalytic pocket. Binding free energy estimates suggested that C4 forms a more stable complex with the enzyme than C1, in agreement with the experimental evidence. Based on this new approach it is possible to design new drimane-type sesquiterpenoids for the control of Candida species as inhibitors of 14-alpha demethylase.


Subject(s)
14-alpha Demethylase Inhibitors/chemistry , Candida/growth & development , Polycyclic Sesquiterpenes/chemistry , Pyridinium Compounds/chemistry , Sesquiterpenes/chemistry , Sterol 14-Demethylase/chemistry , 14-alpha Demethylase Inhibitors/chemical synthesis , 14-alpha Demethylase Inhibitors/pharmacology , Antifungal Agents/chemical synthesis , Antifungal Agents/chemistry , Antifungal Agents/pharmacology , Candida/classification , Candida/drug effects , Catalytic Domain , Humans , Magnetic Resonance Spectroscopy , Molecular Docking Simulation , Molecular Structure , Oxidation-Reduction , Polycyclic Sesquiterpenes/chemical synthesis , Polycyclic Sesquiterpenes/pharmacology , Protein Domains , Pyridinium Compounds/metabolism , Sesquiterpenes/chemical synthesis , Sesquiterpenes/pharmacology , Sterol 14-Demethylase/metabolism
12.
Molecules ; 25(12)2020 Jun 19.
Article in English | MEDLINE | ID: mdl-32575630

ABSTRACT

Heparin-based silver nanoparticles (AgHep-NPs) and gold nanoparticles (AuHep-NPs) were produced by a photochemical method using silver nitrate and chloroauric acid as metal precursors and UV light at 254 nm. UV-Vis spectroscopy graphs showed absorption for AgHep-NPs and AuHep-NPs at 420 nm and 530 nm, respectively. TEM revealed a pseudospherical morphology and a small size, corresponding to 10-25 nm for AgHep-NPs and 1.5-7.5 nm for AuHep-NPs. Their antifungal activity against Candida albicans, Issatchenkia orientalis (Candida krusei), and Candida parapsilosis was assessed by the microdilution method. We show that AgHep-NPs were effective in decreasing fungus density, whereas AuHep-NPs were not. Additionally, the viability of human gingival fibroblasts was preserved by both nanoparticle types at a level above 80%, indicating a slight cytotoxicity. These results are potentially useful for applications of the described NPs mainly in dentistry and, to a lesser extent, in other biomedical areas.


Subject(s)
Antifungal Agents , Candida/growth & development , Cytotoxins , Fibroblasts/metabolism , Gingiva/metabolism , Gold , Metal Nanoparticles/chemistry , Photochemical Processes , Silver/chemistry , Antifungal Agents/chemical synthesis , Antifungal Agents/chemistry , Antifungal Agents/pharmacology , Cell Line , Cytotoxins/chemical synthesis , Cytotoxins/chemistry , Cytotoxins/pharmacology , Gold/chemistry , Gold/pharmacology , Humans
13.
Folia Microbiol (Praha) ; 65(2): 393-405, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31401762

ABSTRACT

Nosocomial infections are an important cause of morbi-mortality worldwide. The increase in the rate of resistance to conventional drugs in these microorganisms has stimulated the search for new therapeutic options. The nitro moiety (NO2) is an important pharmacophore of molecules with high anti-infective activity. We aimed to synthesize new nitro-derivates and to evaluate their antibacterial and anti-Candida potential in vitro. Five compounds [3-nitro-2-phenylchroman-4-ol (3); 3-nitro-2-phenyl-2H-chromene (4a); 3-nitro-2-(4-chlorophenyl)-2H-chromene (4b); 3-nitro-2-(4-fluorophenyl)-2H-chromene (4c), and 3-Nitro-2-(2,3-dichlorophenyl)-2H-chromene (4d)] were efficiently synthesized by Michael-aldol reaction of 2-hydroxybenzaldehyde with nitrostyrene, resulting in one ß-nitro-alcohol (3) and four nitro-olefins (4a-4d). The antibacterial and anti-Candida potentials were evaluated by assaying minimal inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and minimum bactericidal concentration (MBC). Mono-halogenated nitro-compounds (4b and 4c) showed anti-staphylococcal activity with MIC values of 15.6-62.5 µg/mL and MBC of 62.5 µg/mL. However, the activity against Gram-negative strains was showed to be considerably lower and our data suggests that this effect was associated with the outer membrane. Furthermore, nitro-compounds 4c and 4d presented activity against Candida spp. with MIC values ranging from 7.8-31.25 µg/mL and MFC of 15.6-500 µg/mL. In addition, these compounds were able to induce damage in fungal cells increasing the release of intracellular material, which was associated with actions on the cell wall independent of quantitative changes in chitin and ß-glucan. Together, these findings show that nitro-compounds can be exploited as anti-staphylococcal and anti-Candida prototypes.


Subject(s)
Anti-Infective Agents/pharmacology , Nitro Compounds/pharmacology , Anti-Infective Agents/chemical synthesis , Anti-Infective Agents/chemistry , Bacteria/drug effects , Bacteria/growth & development , Candida/drug effects , Candida/growth & development , Cross Infection/drug therapy , Cross Infection/microbiology , Drug Design , Humans , Microbial Sensitivity Tests , Nitro Compounds/chemical synthesis , Nitro Compounds/chemistry
14.
Microbiol Res ; 230: 126330, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31541842

ABSTRACT

Glycoconjugates found on cell walls of Candida species are fundamental for their pathogenicity. Laborious techniques have been employed to investigate the sugar composition of these microorganisms. Herein, we prepared a nanotool, based on the fluorescence of quantum dots (QDs) combined with the specificity of Cramoll lectin, to evaluate glucose/mannose profiles on three Candida species. The QDs-Cramoll conjugates presented specificity and bright fluorescence emission. The lectin preserved its biological activity after the conjugation process mediated by adsorption interactions. The labeling of Candida species was analyzed by fluorescence microscopy and quantified by flow cytometry. Morphological analyses of yeasts labeled with QDs-Cramoll conjugates indicated that C. glabrata (2.7 µm) was smaller when compared to C. albicans (4.0 µm) and C. parapsilosis sensu stricto (3.8 µm). Also, C. parapsilosis population was heterogeneous, presenting rod-shaped blastoconidia. More than 90% of cells of the three species were labeled by conjugates. Inhibition and saturation assays indicated that C. parapsilosis had a higher content of exposed glucose/mannose than the other two species. Therefore, QDs-Cramoll conjugates demonstrated to be effective fluorescent nanoprobes for evaluation of glucose/mannose constitution on the cell walls of fungal species frequently involved in candidiasis.


Subject(s)
Candida/chemistry , Fluorescent Dyes/chemistry , Glucose/analysis , Lectins/chemistry , Mannose/analysis , Microscopy, Fluorescence/methods , Candida/growth & development , Candida/isolation & purification , Candida/metabolism , Candidiasis/diagnosis , Candidiasis/microbiology , Cell Wall/chemistry , Cell Wall/metabolism , Glucose/metabolism , Humans , Mannose/metabolism , Microscopy, Fluorescence/instrumentation , Nanoparticles/chemistry , Quantum Dots/chemistry
15.
Biofouling ; 35(9): 997-1006, 2019 10.
Article in English | MEDLINE | ID: mdl-31710252

ABSTRACT

Surface pre-reacted glass-ionomer (S-PRG) is a bioactive filler produced by PRG technology, which is applied to various dental materials. The inhibitory effects of S-PRG eluate against Candida, the most common fungal oral pathogen, were investigated. Minimum inhibitory concentrations (MIC) and anti-biofilm activities were tested against Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis. For the in vivo study, Galleria mellonella was used as a model to evaluate the effects of S-PRG on toxicity, hemocyte counts and candidiasis. The MIC of S-PRG ranged from 5 to 40% (v/v). S-PRG eluate exhibited anti-biofilm activity for all the Candida species tested. Furthermore, injection of S-PRG eluate into G. mellonella was not toxic to the larvae and protected G. mellonella against experimental candidiasis. In addition, S-PRG eluate inhibited biofilm formation by C. albicans, C. glabrata, C. krusei, and C. tropicalis and exerted protective effects on G. mellonella against experimental candidiasis in vivo.


Subject(s)
Antifungal Agents/pharmacology , Biofilms/drug effects , Candida/drug effects , Candidiasis, Oral/prevention & control , Glass Ionomer Cements/pharmacology , Moths/drug effects , Acrylic Resins/pharmacology , Animals , Antifungal Agents/toxicity , Biofilms/growth & development , Candida/growth & development , Glass Ionomer Cements/toxicity , Larva/drug effects , Microbial Sensitivity Tests , Microbial Viability/drug effects , Moths/microbiology , Silicon Dioxide/pharmacology
16.
J Appl Microbiol ; 127(5): 1362-1372, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31297951

ABSTRACT

AIM: The increase in the number of fungal infections worldwide, coupled with the limitations of current antifungal chemotherapy, demand the development of safe and effective new antifungals. Here, we presented the synthesis of a novel acridone (M14) and its antifungal properties against Candida and dermatophytes species. METHODS AND RESULTS: A series of 17 acridones was designed, synthesized and tested for its antifungal activity. The minimum inhibitory concentration (MIC) was determined by the broth microdilution method. Only the acridone M14 showed growth-inhibitory activity against reference strains and clinical isolates of Candida and dermatophytes, with MIC range of 7·81-31·25 µg ml-1 . Moreover, M14 exhibited fungicidal activity and prevented biofilm formation by C. albicans as well as reduced the viability of preformed biofilms, even at sub-MICs. The confocal laser scanning microscopy analysis revealed that C. albicans hyphal growth was completely inhibited in the presence of M14. Similarly, there was a severe inhibition on hyphal growth of Trichophyton rubrum. We also found that M14 has relatively low toxicity to human fibroblasts. CONCLUSIONS: The new acridone M14 has antifungal properties against Candida spp. and dermatophytes, and antibiofilm activity against C. albicans. In addition, M14 is relatively selective to fungal cells compared to human normal cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its in vitro antifungal activity, anti-Candida biofilm effect and moderate cytotoxicity towards normal human cell, M14 may serve as a valuable lead compound to develop a new antifungal agent.


Subject(s)
Acridones/pharmacology , Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Biofilms/drug effects , Candida/drug effects , Acridones/chemical synthesis , Antifungal Agents/chemical synthesis , Biofilms/growth & development , Candida/growth & development , Candida albicans/drug effects , Candida albicans/growth & development , Cell Survival , Humans , Hyphae/drug effects , Hyphae/growth & development , Microbial Sensitivity Tests , Trichophyton/drug effects , Trichophyton/growth & development
17.
Int J Mol Sci ; 20(14)2019 Jul 23.
Article in English | MEDLINE | ID: mdl-31340425

ABSTRACT

The aim of this study was to perform a systematic review of the literature followed by a meta-analysis about the efficacy of photodynamic therapy (PDT) on the microorganisms responsible for dental caries. The research question and the keywords were constructed according to the PICO strategy. The article search was done in Embase, Lilacs, Scielo, Medline, Scopus, Cochrane Library, Web of Science, Science Direct, and Pubmed databases. Randomized clinical trials and in vitro studies were selected in the review. The study was conducted according the PRISMA guideline for systematic review. A total of 34 articles were included in the qualitative analysis and four articles were divided into two subgroups to perform the meta-analysis. Few studies have achieved an effective microbial reduction in microorganisms associated with the pathogenesis of dental caries. The results highlight that there is no consensus about the study protocols for PDT against cariogenic microorganisms, although the results showed the PDT could be a good alternative for the treatment of dental caries.


Subject(s)
Bacteroidaceae Infections/drug therapy , Candidiasis/drug therapy , Dental Caries/drug therapy , Photochemotherapy/methods , Photosensitizing Agents/pharmacology , Streptococcal Infections/drug therapy , Bacteroidaceae Infections/microbiology , Biofilms/drug effects , Biofilms/growth & development , Candida/drug effects , Candida/growth & development , Candida/pathogenicity , Candidiasis/microbiology , Curcumin/pharmacology , Dental Caries/microbiology , Humans , Methylene Blue/pharmacology , Porphyromonas gingivalis/drug effects , Porphyromonas gingivalis/growth & development , Porphyromonas gingivalis/pathogenicity , Rosaniline Dyes/pharmacology , Streptococcal Infections/microbiology , Streptococcus/drug effects , Streptococcus/growth & development , Streptococcus/pathogenicity , Tolonium Chloride/pharmacology , Treatment Outcome
18.
Biomed Res Int ; 2019: 3941242, 2019.
Article in English | MEDLINE | ID: mdl-31179320

ABSTRACT

The microbial resistance of fungi and bacteria is currently considered a major public health problem. Esters derived from cinnamic acid have a broad spectrum of pharmacological properties that include antimicrobial activity. In this study, a collection of structurally related 4-chlorocinnamic acid esters was prepared using Fischer esterification reactions, alkyl or aryl halide esterification, and Mitsunobu and Steglich reactions. All of the esters were submitted to antimicrobial tests against strains of the species Candida albicans, Candida glabrata, Candida krusei, Candida guilliermondii, Pseudomonas aeruginosa, and Staphylococcus aureus. The compounds also were subjected to molecular docking study with the enzyme 14α-demethylase. Twelve esters derived from 4-chlorocinnamic acid were obtained, with yields varying from 26.3% to 97.6%, three of which were unpublished. The ester methyl 4-chlorocinnamate (1) presented activity against S. aureus at the highest concentration tested. In the antifungal evaluation, all of the esters were bioactive, but methoxyethyl 4-chlorocinnamate (4) and perillyl 4-chlorocinnamate (11) were the most potent (MIC = 0.13 and 0.024 µmol/mL, respectively). The data of molecular docking suggested that all the compounds present good affinity towards the active site related to antifungal activity. Therefore, the esters tested may be inhibitors of the enzyme 14α-demethylase. In addition, the results demonstrate that substituents of short alkyl chains with presence of heteroatom, such as oxygen, or those with a perillyl type terpenic substructure promote better antifungal profiles.


Subject(s)
Anti-Infective Agents , Candida/growth & development , Cinnamates , Pseudomonas aeruginosa/growth & development , Staphylococcus aureus/growth & development , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Cinnamates/chemistry , Cinnamates/pharmacology
19.
Braz J Microbiol ; 50(2): 379-388, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30826999

ABSTRACT

The Asparagaceae family is endemic from America, being the Agave genus the most important. The Agave species possess economic relevance and are use as raw material to produce several distilled alcoholic beverages, as bacanora, tequila, and mezcal. The fermentation process has been carry out either spontaneously or by adding a selected yeast strain. The latter is generally responsible for the production of ethanol and volatile compounds. This study comprised five Agave species (A. angustifolia, A. cupreata, A. durangensis, A. salmiana, and A. tequilana) and eight endogenous yeast strains: five of them were non-Saccharomyces (Torulaspora delbrueckii, Zygosaccharomyces bisporus, Candida ethanolica, and two Kluyveromyces marxianus) and three Saccharomyces cerevisiae strains. The results showed that the S. cerevisiae strains were not able to grow on A. durangensis and A. salmiana juices. The Kluyveromyces marxianus strains grew and fermented all the agave juices and displayed high ethanol production (48-52 g L-1) and volatile compounds. The ethanol production was higher on A. angustifolia juice (1.1-2.8-fold), whereas the volatile compound was dependent on both yeast strain and the Agave species. The use of endogenous non-Saccharomyces yeast strains is feasible, as they may outperform S. cerevisiae regarding the production of fermented beverages from agave plants with a high content of ethanol and aromatic compounds. Graphical abstract.


Subject(s)
Agave/microbiology , Alcoholic Beverages/microbiology , Candida/metabolism , Kluyveromyces/metabolism , Saccharomyces cerevisiae/metabolism , Torulaspora/metabolism , Zygosaccharomyces/metabolism , Candida/growth & development , Ethanol/metabolism , Fermentation/physiology , Kluyveromyces/growth & development , Saccharomyces cerevisiae/growth & development , Torulaspora/growth & development , Zygosaccharomyces/growth & development
20.
J Biotechnol ; 294: 30-37, 2019 Mar 20.
Article in English | MEDLINE | ID: mdl-30769000

ABSTRACT

The aim of this work was the production of bioactive metabolites by submerged fermentation from the fungus Diaporthe schini, followed by their extraction, separation and characterization. Different solvents (methanol, dichloromethane and hexane) were used for the extraction of metabolites from the fermentation broth and the extracts obtained were evaluated by in vitro antibacterial and antifungal activity. The separation and characterization of the extract from the hexane extraction was performed by column chromatography and GC-MS, respectively. The extracts had a great inhibitory action on the Gram-positive bacteria Staphylococcus epidermidis and Staphylococcus aureus, on the Gram-negative bacteria Enterobacter aerogenes and Klebsiella pneumoniae and on the fungus Candida krusei. The main metabolites produced were: 13-docosenamide, (Z)-; 2-hexadecene, 3,7,11,15-tetramethyl; 9-octadecenamide and 11-octadecenoic acid. Studies related to the antibacterial and antifungal activities of metabolites extracted from microorganisms are found in the literature. However, works about the identification of metabolites produced by submerged fermentation from Diaporthe schini were not found until the present moment. This work is an initial study where the conditions of the process can be optimized by looking for the production of a specific compound and can be a promising source for obtaining new drugs.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Ascomycota/metabolism , Anti-Bacterial Agents/isolation & purification , Antifungal Agents/isolation & purification , Ascomycota/genetics , Bacteria/drug effects , Bacteria/growth & development , Candida/drug effects , Candida/growth & development , DNA, Fungal/analysis , Fermentation , Microbial Sensitivity Tests , Solanum/microbiology
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