Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Oligonucleotide primers were used to evaluate the relative gene expression of T. rubrum ATCC 28189, treated with nonyl protocatechuate, and compared to the untreated fungus.

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques: Expressing

Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Scanning electron microscopy of T. rubrum and T. mentagrophytes mature biofilms, untreated ( A , F , K , P ) and treated with fluconazole ( B , G , L , Q ); griseofulvin ( C , H , M , R ); terbinafine ( D , I , N , S ) and nonyl 3,4-dihydroxybenzoate ( E , J , O , T ). White arrows denote small structures, similar to atrophied microconidia or extracellular vesicles ( E , J ).

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques: Electron Microscopy

Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Orthogonal sections obtained by confocal laser scanning microscopy of mature T. rubrum and T. mentagrophytes biofilms, untreated ( A , D , G , J ); treated with terbinafine 32 mg/L ( B , E , H , K ) and treated with nonyl 3, 4-dihydroxybenzoate 250 mg/L ( C , F , I , L ), showed that both treatments reduced the thickness and/or mass of biofilms. Pictures were taken at 63× magnification, 20 µm bars. Metabolically active cells are stained red (FUN 1), while cells surrounded by polysaccharide material are stained green (ConA). The image has been adjusted for brightness.

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques: Confocal Laser Scanning Microscopy, Metabolic Labelling, Staining

Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Confocal laser scanning microscopy images of untreated T. rubrum ATCC 28189 ( A ), treated with 7.8 mg/L nonyl ( B ), 128 mg/L fluconazole ( C ), and 1 mg/L amphotericin B ( D ) at a magnification of 100×; bars, 10 µm. The panels show cell walls stained with calcofluor white, even in those in which more extensive damage and inhibition of hyphal development occurred. Lower panels represent enlargements of the areas demarcated by rectangles for better visualization of the damage caused by the treatments to the fungi cell walls at the magnification of 255×; bars, 5 µm.

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques: Confocal Laser Scanning Microscopy, Staining, Inhibition

Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Mechanism of cell death by necrosis ( A ) and apoptosis ( B ) induced after treatment of T. rubrum ATCC 28189 with nonyl, fluconazole, and amphotericin B. Both significantly induced necrotic death ( p < 0.05). * p < 0.05; ** p <0.01; *** p <0.001.

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques:

Journal: Pharmaceutics

Article Title: Membranolytic Activity Profile of Nonyl 3,4-Dihydroxybenzoate: A New Anti-Biofilm Compound for the Treatment of Dermatophytosis

doi: 10.3390/pharmaceutics15051402

Figure Lengend Snippet: Relative expression levels of Erg 1 and Erg 11 genes represented in Log2Fold change in T. rubrum ATCC 28189 strain treated with nonyl protocatechuate, compared to the untreated control. Results from three independent experiments are expressed as the mean ± standard deviation. Statistical significance was determined by the Student’s t -test and indicated by asterisks: ** p < 0.01.

Article Snippet: For the T. rubrum ATCC strain, treatment with terbinafine or nonyl significantly reduced the thickness of the biofilms on average to 33.23 µm ( B) and 39.28 µm ( C), respectively.

Techniques: Expressing, Standard Deviation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Success of Topical Treatment of Onychomycosis Seems to Be Influenced by Fungal Features

doi: 10.1155/2021/5553634

Figure Lengend Snippet: Illustration of the invasiveness of Trichophyton rubrum on a healthy human nail in ten days. CMRP2912: isolated from patient 1; CMRP2918: isolated from patient 2. T. rubrum ATCC 40051. Topline: macroscopic aspects of fungal growth in nail fragments. Bottom line: features of fungal invasion as seen under a light microscope. Histological sections of nails stained by GMS, at 400 ×.

Article Snippet: Some laboratory tests were performed on the clinical isolates obtained from the two patients, T. rubrum CMRP2912 and T. rubrum CMRP2918, using the reference T. rubrum ATCC 40051 strain.

Techniques: Isolation, Light Microscopy, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Nitric Oxide-Releasing Nanoparticles Are Similar to Efinaconazole in Their Capacity to Eradicate Trichophyton rubrum Biofilms

doi: 10.3389/fcimb.2021.684150

Figure Lengend Snippet: T. rubrum ATCC MYA-4438 strain forms stronger biofilms than the clinical isolate BR1A strain. (A) Kinetics of T. rubrum biofilm formation in 96-well plates determined by XTT reduction assay. (B) Quantification of the biomass of mature T. rubrum biofilms after 72 h incubation was performed using crystal violet staining. For panels a and b, each time point denotes the average of 3 independent wells. Error bars indicate standard deviations. p values (**p < 0.01; ***p < 0.001) were calculated by student ’ s t test analysis.

Article Snippet: The kinetics of biofilm formation by the T. rubrum strains ATCC MYA 4438 and BR1A was quantified by the XTT reduction assay ( ).

Techniques: Incubation, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Nitric Oxide-Releasing Nanoparticles Are Similar to Efinaconazole in Their Capacity to Eradicate Trichophyton rubrum Biofilms

doi: 10.3389/fcimb.2021.684150

Figure Lengend Snippet: Susceptibility of T. rubrum ATCC MYA 4438 or BR1A planktonic cells or biofilms to nitric oxide nanoparticles (NO-np), control nanoparticles (np), fluconazole (FCZ), terbinafine (TRB), or efinaconazole (EFCZ).

Article Snippet: The kinetics of biofilm formation by the T. rubrum strains ATCC MYA 4438 and BR1A was quantified by the XTT reduction assay ( ).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Nitric Oxide-Releasing Nanoparticles Are Similar to Efinaconazole in Their Capacity to Eradicate Trichophyton rubrum Biofilms

doi: 10.3389/fcimb.2021.684150

Figure Lengend Snippet: Scanning electron microscopy showing the architecture of biofilms for (A) T. rubrum ATCC MYA-4438 and (B) BR1A strains. Different magnifications (upper panel; 1000 and lower panel; 3000X) are shown for untreated biofilms and those treated with NO-np 40 mg/mL, fluconazole (FCZ) 0.512 mg/mL, terbinafine (TRB) 0.032 mg/mL, or efinaconazole (EFCZ) 0.320 mg/mL. Red arrows denote collapse of the hyphal walls, while white arrows denote reduced hyphal density.

Article Snippet: The kinetics of biofilm formation by the T. rubrum strains ATCC MYA 4438 and BR1A was quantified by the XTT reduction assay ( ).

Techniques: Electron Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Nitric Oxide-Releasing Nanoparticles Are Similar to Efinaconazole in Their Capacity to Eradicate Trichophyton rubrum Biofilms

doi: 10.3389/fcimb.2021.684150

Figure Lengend Snippet: Confocal microscopy images of mature T. rubrum biofilms formed on glass-bottom plates for 72 h at 37°C (A, D) and treated with NO-np (B, E) or EFCZ (C, F) . Orthogonal images of mature T. rubrum biofilms showed metabolically active (red, FUN-1-stained) cells embedded in the polysaccharide extracellular material (green, ConA), while the yellow-brownish areas represent metabolically inactive or nonviable cells. Images were obtained after 72 h of exposure of the fungal cells to 40 mg/mL of NO-np or 0.32 mg/mL of EFCZ, and the images were compared with those of biofilms incubated in presence of RPMI. The pictures were taken at a magnification of ×63. Bars, 50 μm. The thickness of the fungal biofilms grown under these conditions was measured by z -stack reconstruction. The results are representative of those of two experiments.

Article Snippet: The kinetics of biofilm formation by the T. rubrum strains ATCC MYA 4438 and BR1A was quantified by the XTT reduction assay ( ).

Techniques: Confocal Microscopy, Metabolic Labelling, Staining, Incubation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on early-stage biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine inhibited the metabolic activity of early-stage biofilms from the concentrations of 15.6 and 32 mg/L in all strains tested. Biofilms formed by both the T. rubrum strains conferred resistance to fluconazole at all concentrations tested. Biofilms of the T. mentagrophytes strain were inhibited from the concentration of 64 mg/L (**p < 0.01; ***p < 0.001). Tr, T. rubrum ; Tm, T. mentagrophytes .

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on mature biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured by the XTT reduction assay. Only 2-chalcone showed action against mature biofilms from a concentration of 31.2 mg/L. On the contrary, biofilms formed by all tested strains were resistant to the drugs terbinafine and fluconazole even at the highest concentrations (*p < 0.05; **p < 0.001). Tr- T. rubrum , Tm- T. mentagrophytes .

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on planktonic cells of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine were more potent against planktonic cells than fluconazole. The compounds 2-chalcone, terbinafine, and fluconazole inhibited the cellular metabolic activity in the planktonic form at the concentration used for biofilm formation (10 6 cells/mL) of all strains tested, from the concentration of 15.6, 0.06, and 512 mg/L respectively. (*p < 0.05; **p < 0.01; ***p < 0.001). Tr, T. rubrum , Tm- T. mentagrophytes .

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone-mediated PDT using LED irradiation at a dose of 150 J/cm 2 in planktonic cells (10 6 cell/mL) (A) , early-stage (B) , and mature biofilms (C) of T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438, measured by the XTT reduction assay. Potentiation of 2-chalcone was shown when it was mediated PDT resulting in the inhibition of planktonic forms and biofilms (early-stage and mature) from 2, 4, and 7.8 mg/L. (*p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Irradiation, Inhibition

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy (SEM) images of early stage (A) and mature biofilms (B) of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 untreated (a, c, e) and treated with 2-chalcone in the dark (b, d, f). The images of untreated biofilms show a robust biofilm, formed with the entanglement of integral hyphae and covered with a polymeric extracellular matrix (red arrows). Biofilms treated in the early-stage with 2-chalcone present a low density showing the action of 2-chalcone in inhibiting their maturation. In mature biofilms, as in the early-stage biofilms, 2-chalcone promoted total hyphal collapse (blue arrows).

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Electron Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy images of mature T. rubrum ATCC 28189 biofilms treated with 2-chalcone combined with blue LED at a dose of 150 J/cm 2 (C, D) and irradiated only at a dose of 150 J/cm 2 without the photosensitizer (A, B) . The blue arrows indicate empty spaces within the biofilm showing that the biofilm had become less dense. The red arrows show collapsed hyphae.

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Electron Microscopy, Irradiation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Values of IC 50 and selectivity index (SI) in HaCat cell monolayers treated with 2-chalcone in the dark and with photosensitization.

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: The amount of extracted ergosterol (A) and confocal laser scanning microscopy images (CLSM) (B) of T. rubrum ATCC 28189 treated with 2-chalcone, fluconazole, and amphotericin B. The ergosterol quantification graph shows that treatments with 2-chalcone in the dark and 2-chalcone-mediated PDT reduced the amount of sterols extracted, suggesting that this molecule may have an action on ergosterol or its synthesis chain. Amphotericin B and fluconazole also reduced the amount of steroids extracted, proving their direct action and on the synthesis chain, respectively. In CLSM images, the cell wall is stained with calcofluor white. Cells treated with 2-chalcone showed changes in the cell wall with compromised structure. However, cells treated with fluconazole showed cell wall integrity. **p < 0.01; ***p < 0.001.

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques: Confocal Laser Scanning Microscopy, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Measurement of ROS production after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and mediated PDT (2-chalcone + LED). The compounds amphotericin B, hydrogen peroxide, and 2-chalcone in the dark induced ROS formation when compared to the control without treatment in the dark. Treatment with 2-chalcone-mediated PDT did not induce ROS formation when compared to the control + LED. ***p <0.001.

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Mechanism of death due to apoptosis (A) and necrosis (B) induced after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and with 2-chalcone mediated PDT (2-chalcone +LED), compared with the untreated control. Further, 2-chalcone in the dark and amphotericin B induced cell death by both apoptosis and necrosis, whereas 2-chalcone +LED only induced death by necrosis. *p < 0.05; ***p < 0.001.

Article Snippet: In contrast, most FLZ-treated cells remained alive because the T. rubrum ATCC 28189 strain was resistant to FLZ as shown in the susceptibility assay.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on early-stage biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine inhibited the metabolic activity of early-stage biofilms from the concentrations of 15.6 and 32 mg/L in all strains tested. Biofilms formed by both the T. rubrum strains conferred resistance to fluconazole at all concentrations tested. Biofilms of the T. mentagrophytes strain were inhibited from the concentration of 64 mg/L (**p < 0.01; ***p < 0.001). Tr, T. rubrum ; Tm, T. mentagrophytes .

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on mature biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured by the XTT reduction assay. Only 2-chalcone showed action against mature biofilms from a concentration of 31.2 mg/L. On the contrary, biofilms formed by all tested strains were resistant to the drugs terbinafine and fluconazole even at the highest concentrations (*p < 0.05; **p < 0.001). Tr- T. rubrum , Tm- T. mentagrophytes .

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on planktonic cells of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine were more potent against planktonic cells than fluconazole. The compounds 2-chalcone, terbinafine, and fluconazole inhibited the cellular metabolic activity in the planktonic form at the concentration used for biofilm formation (10 6 cells/mL) of all strains tested, from the concentration of 15.6, 0.06, and 512 mg/L respectively. (*p < 0.05; **p < 0.01; ***p < 0.001). Tr, T. rubrum , Tm- T. mentagrophytes .

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone-mediated PDT using LED irradiation at a dose of 150 J/cm 2 in planktonic cells (10 6 cell/mL) (A) , early-stage (B) , and mature biofilms (C) of T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438, measured by the XTT reduction assay. Potentiation of 2-chalcone was shown when it was mediated PDT resulting in the inhibition of planktonic forms and biofilms (early-stage and mature) from 2, 4, and 7.8 mg/L. (*p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Irradiation, Inhibition

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy (SEM) images of early stage (A) and mature biofilms (B) of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 untreated (a, c, e) and treated with 2-chalcone in the dark (b, d, f). The images of untreated biofilms show a robust biofilm, formed with the entanglement of integral hyphae and covered with a polymeric extracellular matrix (red arrows). Biofilms treated in the early-stage with 2-chalcone present a low density showing the action of 2-chalcone in inhibiting their maturation. In mature biofilms, as in the early-stage biofilms, 2-chalcone promoted total hyphal collapse (blue arrows).

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Electron Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy images of mature T. rubrum ATCC 28189 biofilms treated with 2-chalcone combined with blue LED at a dose of 150 J/cm 2 (C, D) and irradiated only at a dose of 150 J/cm 2 without the photosensitizer (A, B) . The blue arrows indicate empty spaces within the biofilm showing that the biofilm had become less dense. The red arrows show collapsed hyphae.

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Electron Microscopy, Irradiation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Values of IC 50 and selectivity index (SI) in HaCat cell monolayers treated with 2-chalcone in the dark and with photosensitization.

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: The amount of extracted ergosterol (A) and confocal laser scanning microscopy images (CLSM) (B) of T. rubrum ATCC 28189 treated with 2-chalcone, fluconazole, and amphotericin B. The ergosterol quantification graph shows that treatments with 2-chalcone in the dark and 2-chalcone-mediated PDT reduced the amount of sterols extracted, suggesting that this molecule may have an action on ergosterol or its synthesis chain. Amphotericin B and fluconazole also reduced the amount of steroids extracted, proving their direct action and on the synthesis chain, respectively. In CLSM images, the cell wall is stained with calcofluor white. Cells treated with 2-chalcone showed changes in the cell wall with compromised structure. However, cells treated with fluconazole showed cell wall integrity. **p < 0.01; ***p < 0.001.

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques: Confocal Laser Scanning Microscopy, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Measurement of ROS production after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and mediated PDT (2-chalcone + LED). The compounds amphotericin B, hydrogen peroxide, and 2-chalcone in the dark induced ROS formation when compared to the control without treatment in the dark. Treatment with 2-chalcone-mediated PDT did not induce ROS formation when compared to the control + LED. ***p <0.001.

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Mechanism of death due to apoptosis (A) and necrosis (B) induced after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and with 2-chalcone mediated PDT (2-chalcone +LED), compared with the untreated control. Further, 2-chalcone in the dark and amphotericin B induced cell death by both apoptosis and necrosis, whereas 2-chalcone +LED only induced death by necrosis. *p < 0.05; ***p < 0.001.

Article Snippet: The photodynamic therapy assay was applied using 2-chalcone as a photosensitizer against planktonic cells (10 6 cells/mL) and against early-stage and mature biofilms of the strains T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Antifungal activity (expressed in mg/L) of 2-chalcone, terbinafine, and fluconazole against dermatophyte species.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Activity Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on early-stage biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine inhibited the metabolic activity of early-stage biofilms from the concentrations of 15.6 and 32 mg/L in all strains tested. Biofilms formed by both the T. rubrum strains conferred resistance to fluconazole at all concentrations tested. Biofilms of the T. mentagrophytes strain were inhibited from the concentration of 64 mg/L (**p < 0.01; ***p < 0.001). Tr, T. rubrum ; Tm, T. mentagrophytes .

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on mature biofilms of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured by the XTT reduction assay. Only 2-chalcone showed action against mature biofilms from a concentration of 31.2 mg/L. On the contrary, biofilms formed by all tested strains were resistant to the drugs terbinafine and fluconazole even at the highest concentrations (*p < 0.05; **p < 0.001). Tr- T. rubrum , Tm- T. mentagrophytes .

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone (A) , terbinafine (B) , and fluconazole (C) on planktonic cells of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 measured using the XTT reduction assay. The compounds 2-chalcone and terbinafine were more potent against planktonic cells than fluconazole. The compounds 2-chalcone, terbinafine, and fluconazole inhibited the cellular metabolic activity in the planktonic form at the concentration used for biofilm formation (10 6 cells/mL) of all strains tested, from the concentration of 15.6, 0.06, and 512 mg/L respectively. (*p < 0.05; **p < 0.01; ***p < 0.001). Tr, T. rubrum , Tm- T. mentagrophytes .

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Activity Assay, Concentration Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Effect of 2-chalcone-mediated PDT using LED irradiation at a dose of 150 J/cm 2 in planktonic cells (10 6 cell/mL) (A) , early-stage (B) , and mature biofilms (C) of T. rubrum ATCC 28189, T. mentagrophytes ATCC 11481, and T. rubrum ATCC MYA-4438, measured by the XTT reduction assay. Potentiation of 2-chalcone was shown when it was mediated PDT resulting in the inhibition of planktonic forms and biofilms (early-stage and mature) from 2, 4, and 7.8 mg/L. (*p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Irradiation, Inhibition

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy (SEM) images of early stage (A) and mature biofilms (B) of T. rubrum ATCC 28189, T. rubrum ATCC MYA-4438, and T. mentagrophytes ATCC 11481 untreated (a, c, e) and treated with 2-chalcone in the dark (b, d, f). The images of untreated biofilms show a robust biofilm, formed with the entanglement of integral hyphae and covered with a polymeric extracellular matrix (red arrows). Biofilms treated in the early-stage with 2-chalcone present a low density showing the action of 2-chalcone in inhibiting their maturation. In mature biofilms, as in the early-stage biofilms, 2-chalcone promoted total hyphal collapse (blue arrows).

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Electron Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Scanning electron microscopy images of mature T. rubrum ATCC 28189 biofilms treated with 2-chalcone combined with blue LED at a dose of 150 J/cm 2 (C, D) and irradiated only at a dose of 150 J/cm 2 without the photosensitizer (A, B) . The blue arrows indicate empty spaces within the biofilm showing that the biofilm had become less dense. The red arrows show collapsed hyphae.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Electron Microscopy, Irradiation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Values of IC 50 and selectivity index (SI) in HaCat cell monolayers treated with 2-chalcone in the dark and with photosensitization.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: The amount of extracted ergosterol (A) and confocal laser scanning microscopy images (CLSM) (B) of T. rubrum ATCC 28189 treated with 2-chalcone, fluconazole, and amphotericin B. The ergosterol quantification graph shows that treatments with 2-chalcone in the dark and 2-chalcone-mediated PDT reduced the amount of sterols extracted, suggesting that this molecule may have an action on ergosterol or its synthesis chain. Amphotericin B and fluconazole also reduced the amount of steroids extracted, proving their direct action and on the synthesis chain, respectively. In CLSM images, the cell wall is stained with calcofluor white. Cells treated with 2-chalcone showed changes in the cell wall with compromised structure. However, cells treated with fluconazole showed cell wall integrity. **p < 0.01; ***p < 0.001.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: Confocal Laser Scanning Microscopy, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Measurement of ROS production after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and mediated PDT (2-chalcone + LED). The compounds amphotericin B, hydrogen peroxide, and 2-chalcone in the dark induced ROS formation when compared to the control without treatment in the dark. Treatment with 2-chalcone-mediated PDT did not induce ROS formation when compared to the control + LED. ***p <0.001.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: 2-Hydroxychalcone as a Potent Compound and Photosensitizer Against Dermatophyte Biofilms

doi: 10.3389/fcimb.2021.679470

Figure Lengend Snippet: Mechanism of death due to apoptosis (A) and necrosis (B) induced after treatment of T. rubrum ATCC 28189 with 2-chalcone in the dark and with 2-chalcone mediated PDT (2-chalcone +LED), compared with the untreated control. Further, 2-chalcone in the dark and amphotericin B induced cell death by both apoptosis and necrosis, whereas 2-chalcone +LED only induced death by necrosis. *p < 0.05; ***p < 0.001.

Article Snippet: The strain T. rubrum ATCC 28189 demonstrated resistance to FLZ even in the planktonic form as described previously ( ).

Techniques: