RESUMEN
The primary aim of this study was to investigate the impact of treatment with low-temperature plasma (LTP) for varying exposure durations on a multispecies cariogenic biofilm comprising C. albicans, L. casei, and S. mutans, as well as on single-species biofilms of L. casei and C. albicans, cultured on hydroxyapatite discs. Biofilms were treated with LTP-argon at a 10 mm distance for 30 s, 60 s, and 120 s. Chlorhexidine solution (0.12%) and NaCl (0.89%) were used as positive (PC) and negative controls (NC), respectively. Argon flow only was also used as gas flow control (F). Colony-forming units (CFU) recovery and confocal laser scanning microscopy (CLSM) were used to analyze biofilm viability. LTP starting at 30 s of application significantly reduced the viability of multispecies biofilms by more than 2 log10 in all treated samples (p < 0.0001). For single-species biofilms, L. casei showed a significant reduction compared to PC and NC of over 1 log10 at all exposure times (p < 0.0001). In the case of C. albicans biofilms, LTP treatment compared to PC and NC resulted in a significant decrease in bacterial counts when applied for 60 and 120 s (1.55 and 1.90 log10 CFU/mL, respectively) (p < 0.0001). A significant effect (p ≤ 0.05) of LTP in single-species biofilms was observed to start at 60 s of LTP application compared to F, suggesting a time-dependent effect of LTP for the single-species biofilms of C. albicans and L. casei. LTP is a potential mechanism in treating dental caries by being an effective anti-biofilm therapy of both single and multispecies cariogenic biofilms.
Asunto(s)
Biopelículas , Candida albicans , Gases em Plasma , Streptococcus mutans , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Gases em Plasma/farmacología , Candida albicans/fisiología , Candida albicans/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/fisiología , Caries Dental/microbiología , Caries Dental/terapia , Lacticaseibacillus casei/fisiología , Humanos , Viabilidad Microbiana/efectos de los fármacos , Microscopía Confocal , FríoRESUMEN
Candida auris has emerged as a significant healthcare-associated pathogen due to its multidrug-resistant nature. Ongoing constraints in the discovery and provision of new antifungals create an urgent imperative to design effective remedies to this pressing global blight. Herein, we screened a chemical library and identified aryl-carbohydrazide analogs with potent activity against both C. auris and the most prevalent human fungal pathogen, C. albicans. SPB00525 [N'-(2,6-dichlorophenyl)-5-nitro-furan-2-carbohydrazide] exhibited potent activity against different strains that were resistant to standard antifungals. Using drug-induced haploinsufficient profiling, transcriptomics and metabolomic analysis, we uncovered that Ole1, a Δ(9) fatty acid desaturase, is the likely target of SPB00525. An analog of the latter, HTS06170 [N'-(2,6-dichlorophenyl)-4-methyl-1,2,3-thiadiazole-5-carbohydrazide], had a superior antifungal activity against both C. auris and C. albicans. Both SPB00525 and HTS06170 act as antivirulence agents and inhibited the invasive hyphal growth and biofilm formation of C. albicans. SPB00525 and HTS06170 attenuated fungal damage to human enterocytes and ameliorate the survival of Galleria mellonella larvae used as systemic candidiasis model. These data suggest that inhibiting fungal Δ(9) fatty acid desaturase activity represents a potential therapeutic approach for treating fungal infection caused by the superbug C. auris and the most prevalent human fungal pathogen, C. albicans.
Asunto(s)
Antifúngicos , Candida auris , Candidiasis , Pruebas de Sensibilidad Microbiana , Antifúngicos/farmacología , Animales , Candidiasis/tratamiento farmacológico , Candidiasis/microbiología , Candida auris/efectos de los fármacos , Candida auris/genética , Ácido Graso Desaturasas/metabolismo , Ácido Graso Desaturasas/genética , Ácido Graso Desaturasas/antagonistas & inhibidores , Candida albicans/efectos de los fármacos , Candida albicans/enzimología , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Humanos , Inhibidores Enzimáticos/farmacología , Mariposas Nocturnas/microbiología , Mariposas Nocturnas/efectos de los fármacos , Metabolómica , Larva/microbiología , Larva/efectos de los fármacos , Modelos Animales de Enfermedad , Hidrazinas/farmacología , Bibliotecas de Moléculas Pequeñas/farmacología , Perfilación de la Expresión GénicaRESUMEN
Background: Polymeric denture materials can be susceptible to colonization by oral microorganisms. Zein-coated magnesium oxide nanoparticles (zMgO NPs) demonstrate antimicrobial activity. The aim of this study was to investigate the antimicrobial effect and adherence of different oral microorganisms on hybrid polymeric denture materials incorporated with zMgO NPs. Methods: Five types of polymeric denture materials were used. A total of 480 disc-shaped specimens were divided by material type (n=96/grp), then subdivided by zMgO NPs concentration: control with no nanoparticles and other groups with zMgO NPs concentrations of 0.3%, 0.5% and 1% by weight. Characterization of the polymeric denture materials incorporating zMgO NPs was done, and the antimicrobial activity of all groups was tested against four types of microorganisms: 1) Streptococcus mutans, 2) Staphylococcus aureus, 3) Enterococcus faecalis and 4) Candida albicans. The samples underwent an adherence test and an agar diffusion test. Experiments were done in triplicates. Results: The characterization of the hybrid samples revealed variation in the molecular composition, as well as a uniform distribution of the zMgO NPs in the polymeric denture materials. All hybrid polymeric denture materials groups induced a statistically significant antimicrobial activity, while the control groups showed the least antimicrobial activity. The agar diffusion test revealed no release of the zMgO NPs from the hybrid samples, indicating the NPs did not seep out of the matrix. Conclusion: The zMgO NPs were effective in reducing the adherence of the tested microorganisms and enhancing the antimicrobial activity of the polymeric denture materials. This antimicrobial effect with the polymeric dentures could aid in resisting microbial issues such as denture stomatitis.
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Antiinfecciosos , Candida albicans , Staphylococcus aureus , Streptococcus mutans , Zeína , Zeína/química , Zeína/farmacología , Candida albicans/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Antiinfecciosos/farmacología , Antiinfecciosos/química , Óxido de Magnesio/química , Óxido de Magnesio/farmacología , Nanopartículas/química , Enterococcus faecalis/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Humanos , Materiales Dentales/farmacología , Materiales Dentales/química , Dentaduras/microbiología , Polímeros/química , Polímeros/farmacologíaRESUMEN
Background: In the face of increasing antifungal resistance among Candida albicans biofilms, this study explores the efficacy of a combined treatment using Kangbainian lotion (KBN) and miconazole nitrate (MN) to address this challenge. Methods: Using UPLC-Q-TOF/MS Analysis for Identification of Active Compounds in KBN Lotion; FICI for synergy evaluation, XTT and ROS assays for biofilm viability and oxidative stress, fluorescence and confocal laser scanning microscopy (CLSM) for structural and viability analysis, and real-time fluorescence for gene expression. Conclusion: Our study indicates that the combined application of KBN and MN somewhat impacts the structural integrity of Candida albicans biofilms and affects the expression of several key genes involved in biofilm formation, including ALS1, ALS3, HWP1, HSP90, and CSH1. These preliminary findings suggest that there may be a synergistic effect between KBN and MN, potentially influencing not only the structural aspects of fungal biofilms but also involving the modulation of genetic pathways during their formation.
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Antifúngicos , Biopelículas , Candida albicans , Farmacorresistencia Fúngica , Sinergismo Farmacológico , Miconazol , Pruebas de Sensibilidad Microbiana , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Antifúngicos/farmacología , Miconazol/farmacología , Especies Reactivas de Oxígeno/metabolismo , Viabilidad Microbiana/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , HumanosRESUMEN
Candida albicans is an opportunistic fungal pathogen that can cause systemic infections in immunocompromised individuals. Morphological transition and biofilm formation are major virulence factors of C. albicans. Moreover, biofilm enhances resistance to antifungal agents. Therefore, it is urgent to identify new and effective compounds to target the biofilm of C. albicans. In the present study, the antifungal activities of equol against C. albicans were investigated. In vitro, the microdilution analysis and spot assay result showed that equol exhibited potent inhibitory activities against C. albicans. Further investigations confirmed that the antifungal effects of equol involved interference with the transition from yeast to hypha and biofilm formation of C. albicans. In addition, transcriptome sequencing and reverse transcription-quantitative PCR (qRT-PCR) analysis showed that equol significantly downregulated the expression of several genes in the Ras1-cAMP-PKA pathway related to hyphae and biofilm formation and significantly upregulated the expression of the negative transcriptional repressors RFG1 and TUP1. Moreover, equol effectively reduced the production of cAMP, a key messenger in the Ras1-cAMP-PKA pathway, while supplementation with cAMP partly rescued the equol-induced defects in hyphal development. Furthermore, in a mouse model of systemic candidiasis (SC), equol treatment significantly decreased the fungal burden (liver, kidneys, and lung) in mice and local tissue damage, while enhancing the production of interleukin-10 (IL-10). Together, these findings confirm that equol is a potentially effective agent for treatment of SC.
Asunto(s)
Antifúngicos , Biopelículas , Candida albicans , Candidiasis , Equol , Candida albicans/efectos de los fármacos , Candida albicans/genética , Animales , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Ratones , Candidiasis/microbiología , Candidiasis/tratamiento farmacológico , Equol/farmacología , Femenino , Modelos Animales de Enfermedad , Pruebas de Sensibilidad Microbiana , Hifa/efectos de los fármacos , Hifa/crecimiento & desarrollo , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Ratones Endogámicos BALB C , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismoRESUMEN
Systemic infection with Candida albicans poses a significant risk for people with weakened immune systems and carries a mortality rate of up to 60%. However, current therapeutic options have several limitations, including increasing drug tolerance, notable off-target effects, and severe adverse reactions. Over the past four decades, the progress in developing drugs to treat Candida albicans infections has been sluggish. This comprehensive review addresses the limitations of existing drugs and summarizes the efforts made toward redesigning and innovating existing or novel drugs through nanotechnology. The discussion explores the potential applications of nanomedicine in Candida albicans infections from four perspectives: nano-preparations for anti-biofilm therapy, innovative formulations of "old drugs" targeting the cell membrane and cell wall, reverse drug resistance therapy targeting subcellular organelles, and virulence deprivation therapy leveraging the unique polymorphism of Candida albicans. These therapeutic approaches are promising to address the above challenges and enhance the efficiency of drug development for Candida albicans infections. By harnessing nano-preparation technology to transform existing and preclinical drugs, novel therapeutic targets will be uncovered, providing effective solutions and broader horizons to improve patient survival rates.
Asunto(s)
Antifúngicos , Candida albicans , Candidiasis , Nanotecnología , Humanos , Candida albicans/efectos de los fármacos , Antifúngicos/uso terapéutico , Antifúngicos/farmacología , Candidiasis/tratamiento farmacológico , Nanotecnología/métodos , Animales , Farmacorresistencia Fúngica/efectos de los fármacos , Biopelículas/efectos de los fármacos , Nanomedicina/métodos , Nanopartículas/química , Nanopartículas/uso terapéutico , Sistemas de Liberación de Medicamentos/métodosRESUMEN
Human lactoferrin (hLf) is an innate host defense protein that inhibits microbial H+-ATPases. This protein includes an ancestral structural motif (i.e., γ-core motif) intimately associated with the antimicrobial activity of many natural Cys-rich peptides. Peptides containing a complete γ-core motif from hLf or other phylogenetically diverse antimicrobial peptides (i.e., afnA, SolyC, PA1b, PvD1, thanatin) showed microbicidal activity with similar features to those previously reported for hLf and defensins. Common mechanistic characteristics included (1) cell death independent of plasma membrane (PM) lysis, (2) loss of intracellular K+ (mediated by Tok1p K+ channels in yeast), (3) inhibition of microbicidal activity by high extracellular K+, (4) influence of cellular respiration on microbicidal activity, (5) involvement of mitochondrial ATP synthase in yeast cell death processes, and (6) increment of intracellular ATP. Similar features were also observed with the BM2 peptide, a fungal PM H+-ATPase inhibitor. Collectively, these findings suggest host defense peptides containing a homologous γ-core motif inhibit PM H+-ATPases. Based on this discovery, we propose that the γ-core motif is an archetypal effector involved in the inhibition of PM H+-ATPases across kingdoms of life and contributes to the in vitro microbicidal activity of Cys-rich antimicrobial peptides.
Asunto(s)
Secuencias de Aminoácidos , ATPasas de Translocación de Protón , Humanos , ATPasas de Translocación de Protón/metabolismo , ATPasas de Translocación de Protón/antagonistas & inhibidores , Péptidos Antimicrobianos/farmacología , Péptidos Antimicrobianos/química , Lactoferrina/farmacología , Lactoferrina/química , Antiinfecciosos/farmacología , Antiinfecciosos/química , Cisteína/metabolismo , Cisteína/química , Candida albicans/efectos de los fármacos , Membrana Celular/metabolismo , Membrana Celular/efectos de los fármacosRESUMEN
As an important small organic molecule, cyclopropane is widely used in drug design. In this paper, fifty-three amide derivatives containing cyclopropane were designed and synthesized by introducing amide groups and aryl groups into cyclopropane through the active splicing method, and their antibacterial and antifungal activities were evaluated in vitro. Among them, thirty-five compounds were new compounds, and eighteen compounds were known compounds (F14, F15, F18, F20-F26, F36, and F38-F44). Bioassay results disclosed that four, three, and nine of the compounds showed moderate activity against Staphylococcus aureus, Escherichia coli, and Candida albicans, respectively. Three compounds were sensitive to Candida albicans, with excellent antifungal activity (MIC80 = 16 µg/mL). The molecular docking results show that compounds F8, F24, and F42 have good affinity with the potential antifungal drug target CYP51 protein.
Asunto(s)
Amidas , Antifúngicos , Candida albicans , Ciclopropanos , Diseño de Fármacos , Pruebas de Sensibilidad Microbiana , Simulación del Acoplamiento Molecular , Staphylococcus aureus , Ciclopropanos/farmacología , Ciclopropanos/química , Ciclopropanos/síntesis química , Amidas/química , Amidas/farmacología , Amidas/síntesis química , Candida albicans/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/síntesis química , Antifúngicos/química , Escherichia coli/efectos de los fármacos , Relación Estructura-Actividad , Antiinfecciosos/farmacología , Antiinfecciosos/síntesis química , Antiinfecciosos/química , Antibacterianos/farmacología , Antibacterianos/síntesis química , Antibacterianos/química , Estructura MolecularRESUMEN
In Candida albicans, Cdr1 pumps azole drugs out of the cells to reduce intracellular accumulation at detrimental concentrations, leading to azole-drug resistance. Milbemycin oxime, a veterinary anti-parasitic drug, strongly and specifically inhibits Cdr1. However, how Cdr1 recognizes and exports azole drugs, and how milbemycin oxime inhibits Cdr1 remain unclear. Here, we report three cryo-EM structures of Cdr1 in distinct states: the apo state (Cdr1Apo), fluconazole-bound state (Cdr1Flu), and milbemycin oxime-inhibited state (Cdr1Mil). Both the fluconazole substrate and the milbemycin oxime inhibitor are primarily recognized within the central cavity of Cdr1 through hydrophobic interactions. The fluconazole is suggested to be exported from the binding site into the environment through a lateral pathway driven by TM2, TM5, TM8 and TM11. Our findings uncover the inhibitory mechanism of milbemycin oxime, which inhibits Cdr1 through competition, hindering export, and obstructing substrate entry. These discoveries advance our understanding of Cdr1-mediated azole resistance in C. albicans and provide the foundation for the development of innovative antifungal drugs targeting Cdr1 to combat azole-drug resistance.
Asunto(s)
Antifúngicos , Azoles , Candida albicans , Microscopía por Crioelectrón , Proteínas Fúngicas , Proteínas de Transporte de Membrana , Candida albicans/efectos de los fármacos , Candida albicans/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/antagonistas & inhibidores , Antifúngicos/farmacología , Antifúngicos/química , Azoles/farmacología , Azoles/química , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/ultraestructura , Farmacorresistencia Fúngica , Fluconazol/farmacología , Sitios de UniónRESUMEN
Microalgae are of great interest due to their ability to produce valuable compounds, such as pigments, omega-3 fatty acids, antioxidants, and antimicrobials. The dinoflagellate genus Amphidinium is particularly notable for its amphidinol-like compounds, which exhibit antibacterial and antifungal properties. This study utilized a two-stage cultivation method to grow Amphidinium carterae CCAP 1102/8 under varying conditions, such as blue LED light, increased salinity, and the addition of sodium carbonate or hydrogen peroxide. After cultivation, the biomass was extracted and fractionated using solid-phase extraction, yielding six fractions per treatment. These fractions were analyzed using Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS/MS) to identify their chemical components. Key amphidinol compounds (AM-B, AM-C, AM-22, and AM-A) were identified, with AM-B being the most abundant in Fraction 4, followed by AM-C. Fraction 5 also contained a significant amount of AM-C along with an unknown compound. Fraction 4 returned the highest antimicrobial activity against the pathogens Staphylococcus aureus, Enterococcus faecalis, and Candida albicans, with Minimal Biocidal Concentrations (MBCs) ranging from 1 to 512 µg/mL. Results indicate that the modulation of both amphidinol profile and fraction bioactivity can be induced by adjusting the cultivation parameters used to grow two-stage batch cultures of A. carterae.
Asunto(s)
Candida albicans , Dinoflagelados , Pruebas de Sensibilidad Microbiana , Dinoflagelados/química , Dinoflagelados/crecimiento & desarrollo , Dinoflagelados/efectos de los fármacos , Candida albicans/efectos de los fármacos , Candida albicans/crecimiento & desarrollo , Antibacterianos/farmacología , Antibacterianos/química , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/crecimiento & desarrollo , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Cromatografía Liquida , Antifúngicos/farmacología , Antifúngicos/química , Antiinfecciosos/farmacología , Antiinfecciosos/química , AnfidinolesRESUMEN
BACKGROUND AND OBJECTIVES: Mineral Trioxide Aggregate (MTA) is one of the main retrograde filling materials that is used today as a root end filling material and perforation repair material. This study was conducted with the aim of investigating the antibacterial and antifungal properties of four types of bio-ceramic materials, AGM MTA, Ortho MTA, Pro root MTA and Cem cement for oral and dental health. METHODS: In this study, the antibacterial activity of four types of bio-ceramic materials against two bacterial strains of Enterococcus faecalis (ATTC 29212), Escherichia coli (ATTC 35318) and antifungal activity against Candida albicans (ATTC 10231) were investigated using the well diffusion method. RESULTS: In the context of the relationship between the type of microorganism and the diameter of the growth inhibitory zone for each type of bio-ceramic material, there was no significant difference for Enterococcus faecalis, and a significant difference was observed for Escherichia coli and Candida albicans (p < 0.05). CONCLUSION: The results show that each of the bio-ceramic materials AGM, Pro root, Cem cement and Ortho have antibacterial and antifungal properties. AGM MTA bio-ceramic material on Candida albicans fungus and Ortho MTA bio-ceramic material had the most effect on Escherichia coli bacteria. Therefore, the mentioned bio-ceramic materials can play a significant role in oral and dental health by providing a suitable material for restoration.
Asunto(s)
Compuestos de Aluminio , Compuestos de Calcio , Candida albicans , Cerámica , Combinación de Medicamentos , Enterococcus faecalis , Escherichia coli , Óxidos , Materiales de Obturación del Conducto Radicular , Silicatos , Compuestos de Calcio/farmacología , Silicatos/farmacología , Candida albicans/efectos de los fármacos , Enterococcus faecalis/efectos de los fármacos , Óxidos/farmacología , Compuestos de Aluminio/farmacología , Escherichia coli/efectos de los fármacos , Materiales de Obturación del Conducto Radicular/farmacología , Humanos , Cementos Dentales/farmacología , Antibacterianos/farmacología , Ensayo de Materiales , Antifúngicos/farmacologíaRESUMEN
OBJECTIVE: To develop a novel calcium silver zeolite (Ca-Ag-Zeo) and assess its biocompatibility, physiochemical properties and antimicrobial effects. METHODS: Ca-Ag-Zeo was synthesized using ion-exchange method with calcium chloride, silver nitrate and Zeolite X (Zeo). Silver zeolite X (Ag-Zeo) and Zeo were set as control. The chemical structure, morphology, crystal structure and elemental composition of Ca-Ag-Zeo was characterized by X-ray diffraction spectrum, scanning electron microscopy, transmission electron microscopy and energy dispersive spectroscopy, respectively. Its biocompatibility on the human gingival fibroblasts was assessed by cell counting kit-8 assay. Its physiochemical properties were determined by the released calcium and silver ion using Inductive Coupled Plasma Emission Spectrometry for up to 12 weeks. The antimicrobial properties on Streptococcus mutans, Lactobacillus acidophilus, Lactobacillus casei, and Candida albicans were assessed by minimum bactericidal concentration (MBC) or minimum fungicidal concentration (MFC) assay. RESULTS: Ca-Ag-Zeo with a hexagonal cage structure was synthesized. As for biocompatibility, the half-maximal inhibitory concentration (± SD in mg/mL) of Ca-Ag-Zeo, Ag-Zeo and Zeo in human gingival fibroblasts were 0.52 ± 0.05, 0.15 ± 0.01 and 3.35 ± 0.58, respectively (Zeo > Ca-Ag-Zeo > Ag-Zeo; p < 0.05). As for physiochemical properties, the accumulated ion release (± SD in mg) of Ca-Ag-Zeo, Ag-Zeo and Zeo were 0.011 ± 0.003, 0 and 0 for calcium ion, respectively (Ca-Ag-Zeo > Ag-Zeo, Zeo; p < 0.001), and 0.213 ± 0.032, 0.209 ± 0.019 and 0 for silver ion, respectively (Ca-Ag-Zeo, Ag-Zeo > Zeo; p < 0.001). As for anti-microbial ability, the MBC/MFC (mg/mL) of Ca-Ag-Zeo, Ag-Zeo and Zeo were 32, 16 and > 256 against Streptococcus mutans; 32, 16, > 256 against Lactobacillus acidophilus; 16, 16, and 256 against Lactobacillus casei; 0.25, 0.125; and 2, 1, > 256 against Candida albicans, respectively. CONCLUSION: A novel Ca-Ag-Zeo was developed. It presented better biocompatibility compared to Ag-Zeo. It released calcium and silver ions sustainably, and it could inhibit the growth of common cariogenic microorganisms.
Asunto(s)
Calcio , Candida albicans , Caries Dental , Fibroblastos , Pruebas de Sensibilidad Microbiana , Plata , Streptococcus mutans , Zeolitas , Humanos , Zeolitas/farmacología , Zeolitas/química , Streptococcus mutans/efectos de los fármacos , Candida albicans/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Caries Dental/prevención & control , Caries Dental/microbiología , Plata/farmacología , Plata/química , Lactobacillus acidophilus/efectos de los fármacos , Difracción de Rayos X , Encía/efectos de los fármacos , Encía/citología , Lacticaseibacillus casei/efectos de los fármacos , Microscopía Electrónica de Rastreo , Materiales Biocompatibles/farmacología , Microscopía Electrónica de Transmisión , Ensayo de Materiales , Nitrato de Plata/farmacología , Antibacterianos/farmacología , Antiinfecciosos/farmacologíaRESUMEN
This study proposes an affordable plasma device that utilizes a parallel-plate dielectric barrier discharge geometry with a metallic mesh electrode, featuring a straightforward 3D-printed design. Powered by a high-voltage supply adapted from a cosmetic plasma device, it operates on atmospheric air, eliminating the need for gas flux. Surface modification of polyethylene treated with this device was characterized and showed that the elemental composition after 15 min of plasma treatment decreased the amount of C to ~80 at% due to the insertion of O (~15 at%). Tested against Candida albicans and Staphylococcus aureus, the device achieved a reduction of over 99% in microbial load with exposure times ranging from 1 to 10 min. Simultaneously, the Vero cell viability remained consistently high, namely between 91% and 96% across exposure times. These results highlight this device's potential for the surface modification of materials and various infection-related applications, boasting affordability and facilitating effective antimicrobial interventions.
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Candida albicans , Gases em Plasma , Staphylococcus aureus , Propiedades de Superficie , Candida albicans/efectos de los fármacos , Gases em Plasma/química , Gases em Plasma/farmacología , Staphylococcus aureus/efectos de los fármacos , Animales , Células Vero , Chlorocebus aethiops , Viabilidad Microbiana/efectos de los fármacos , Polímeros/químicaRESUMEN
Candida albicans, the most common pathogenic fungus, can form biofilms on the surface of medical devices and often causes bloodstream infections. Biofilms have a complex structure composed of microorganisms and a surrounding extracellular matrix. Biofilms are difficult to treat because they are resistant to antifungal drugs and the host environment. Nearly one in four patients with candidemia have a polymicrobial infection. These polymicrobial biofilms, especially those comprising cross-kingdom species of fungi and bacteria, can lead to long hospital stays and high mortality rates. This review outlines the unique interactions of dual-species biofilms with Candida albicans and the clinically important bacteria Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli.
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Biopelículas , Candida albicans , Pseudomonas aeruginosa , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Humanos , Candida albicans/efectos de los fármacos , Candida albicans/fisiología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/fisiología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/fisiología , Staphylococcus aureus/crecimiento & desarrollo , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Antifúngicos/farmacología , Coinfección/microbiología , Bacterias/efectos de los fármacos , Bacterias/crecimiento & desarrolloRESUMEN
This study investigated crotamine (CTA), a peptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, known for its exceptional cell penetration potential. The objective was to explore the antibacterial and antifungal activity of CTA, its ability to inhibit efflux pumps and evaluate the effectiveness of its pharmacological combination with antibiotics and antifungals. In microbiological assays, CTA in combination with antibiotics was tested against strains of S. aureus and the inhibition of NorA, Tet(K) and MepA efflux pumps was also evaluated. CTA alone did not present clinically relevant direct antibacterial action, presenting MIC > 209.7 µM against strains S. aureus 1199B, IS-58, K2068. The standard efflux pump inhibitor CCCP showed significant effects in all negative relationships to assay reproducibility. Against the S. aureus 1199B strain, CTA (20.5 µM) associated with norfloxacin diluted 10 × (320.67 µM) showed a potentiating effect, in relation to the control. Against the S. aureus IS-58 strain, the CTA associated with tetracycline did not show a significant combinatorial effect, either with 2304 or 230.4 µM tetracycline. CTA at a concentration of 2.05 µM associated with ciprofloxacin at a concentration of 309.4 µM showed a significant potentiating effect. In association with EtBr, CTA at concentrations of 2.05 and 20.5 µM potentiated the effect in all strains tested, reducing the prevention of NorA, Tet(K) and MepA efflux pumps. In the C. albicans strain, a potentiating effect of fluconazole (334.3 µM) was observed when combined with CTA (2.05 µM). Against the C. tropicalis strain, a significant effect was also observed in the association of fluconazole 334.3 µM, where CTA 2.05 µM considerably reduced fungal growth and decreased the potentiation of fluconazole. Against the C. krusei strain, no significant potentiating effect of fluconazole was obtained by CTA. Our results indicate that CTA in pharmacological combination potentiates the effects of antibiotics and antifungal. This represents a new and promising antimicrobial strategy for treating a wide variety of infections.
Asunto(s)
Antibacterianos , Antifúngicos , Venenos de Crotálidos , Crotalus , Pruebas de Sensibilidad Microbiana , Antifúngicos/farmacología , Antifúngicos/química , Antibacterianos/farmacología , Venenos de Crotálidos/farmacología , Animales , Staphylococcus aureus/efectos de los fármacos , Sinergismo Farmacológico , Candida albicans/efectos de los fármacos , Serpientes VenenosasRESUMEN
Bacterial growing resistance to antibiotics poses a critical threat to global health. This study investigates, for the first time, the antibiofilm properties of Vicia ervilia agglutinin (VEA) from six different V. ervilia accessions against pathogenic bacteria, and the yeast Candida albicans. In the absence of antimicrobial properties, purified VEA significantly inhibited biofilm formation, both in Gram-positive and Gram-negative bacteria, but not in C. albicans. With an inhibitory concentration ranging from 100 to 500 µg/ml, the VEA antibiofilm activity was more relevant against the Gram-positive bacteria Streptococcus aureus and Staphylococcus epidermidis, whose biofilm was reduced up to 50% by VEA purified from accessions #5 and #36. VEA antibiofilm variability between accessions was observed, likely due to co-purified small molecules rather than differences in VEA protein sequences. In conclusion, VEA seed extracts from the accessions with the highest antibiofilm activity could represent a valid approach for the development of an effective antibiofilm agent.
Asunto(s)
Antibacterianos , Biopelículas , Candida albicans , Bacterias Gramnegativas , Bacterias Grampositivas , Pruebas de Sensibilidad Microbiana , Biopelículas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Bacterias Gramnegativas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Candida albicans/fisiología , Antibacterianos/farmacología , Lectinas de Plantas/farmacología , Extractos Vegetales/farmacología , Extractos Vegetales/química , Semillas/químicaRESUMEN
OBJECTIVE: To investigate caries preventive effects of 38 % silver diamine fluoride (SDF) pretreatment on neighboring tooth proximal to glass ionomer cement (GIC), including conventional GIC (CGIC) and resin-modified GIC (RMGIC) restorations in an in vitro model. METHODS: HUMAN TOOTH BLOCKS WERE RESTORED WITH: SDF+CGIC (Group 1), CGIC (Group 2), SDF+RMGIC (Group 3) or RMGIC (Group 4). Enamel specimen simulating proximal surface of neighboring tooth was placed in proximity to the restorations. The specimen underwent cariogenic challenge with cross-kingdom biofilm of Streptococcus mutans, Lacticaseibacillus casei and Candida albicans. After cariogenic challenge, the biofilm's growth kinetics, viability, and morphology were evaluated by propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), respectively. The enamel lesion depth, surface morphology and crystal characteristics were determined by micro-computed tomography (micro-CT), SEM and X-ray diffraction (XRD), respectively. RESULTS: PMA-qPCR demonstrated lower microbial growth in Group 1 and 3 compared with Group 2 and 4 (p < 0.05). CLSM showed the dead-to-live ratio in Groups 1-4 were 1.15±0.12, 0.53±0.13, 1.10±0.24 and 0.63±0.10, respectively (Group 1,3 > 2,4, p < 0.05). SEM revealed Groups 1 and 3 had scattered biofilm whereas Group 2 and 4 had confluent biofilm. Micro-CT showed the enamel lesion depths (µm) were 98±9, 126±7, 103±6 and 128±7 for Group 1 to 4, respectively (Group 1,3 < 2,4, p < 0.05). SEM revealed oriented and ordered enamel prismatic patterns in Group 1 and 3, not in Group 2 and 4. XRD showed the reflections of hydroxyapatite in Groups 1 and 3 were sharper than Groups 2 and 4. CONCLUSION: SDF pretreatment enhances the preventive effect of GIC on proximal enamel surface on neighboring tooth through inhibiting cariogenic biofilm, reducing enamel demineralization and promoting enamel remineralization. CLINICAL SIGNIFICANCE: SDF pretreatment of GIC restorations can help prevent caries on neighboring teeth, particular for patients with high caries risk.
Asunto(s)
Biopelículas , Cariostáticos , Caries Dental , Esmalte Dental , Fluoruros Tópicos , Cementos de Ionómero Vítreo , Microscopía Electrónica de Rastreo , Compuestos de Amonio Cuaternario , Compuestos de Plata , Streptococcus mutans , Compuestos de Plata/uso terapéutico , Compuestos de Plata/farmacología , Humanos , Cementos de Ionómero Vítreo/uso terapéutico , Cementos de Ionómero Vítreo/farmacología , Caries Dental/prevención & control , Caries Dental/microbiología , Compuestos de Amonio Cuaternario/farmacología , Compuestos de Amonio Cuaternario/uso terapéutico , Biopelículas/efectos de los fármacos , Fluoruros Tópicos/uso terapéutico , Fluoruros Tópicos/farmacología , Cariostáticos/uso terapéutico , Cariostáticos/farmacología , Esmalte Dental/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Restauración Dental Permanente/métodos , Microtomografía por Rayos X , Candida albicans/efectos de los fármacos , Difracción de Rayos X , Microscopía ConfocalRESUMEN
Candida albicans has been listed in the critical priority group by the WHO in 2022 depending upon its contribution in invasive candidiasis and increased resistance to conventional drugs. Drug repurposing offers an efficient, rapid, and cost-effective solution to develop alternative therapeutics against pathogenic microbes. Alexidine dihydrochloride (AXD) and hexachlorophene (HCP) are FDA approved anti-cancer and anti-septic drugs, respectively. In this study, we have shown antifungal properties of AXD and HCP against the wild type (reference strain) and clinical isolates of C. albicans. The minimum inhibitory concentrations (MIC50) of AXD and HCP against C. albicans ranged between 0.34 and 0.69 µM and 19.66-24.58 µM, respectively. The biofilm inhibitory and eradication concentration of AXD was reported comparatively lower than that of HCP for the strains used in the study. Further investigations were performed to understand the antifungal mode of action of AXD and HCP by studying virulence features like cell surface hydrophobicity, adhesion, and yeast to hyphae transition, were also reduced upon exposure to both the drugs. Ergosterol content in cell membrane of the wild type strain was upregulated on exposure to AXD and HCP both. Biochemical analyses of the exposed biofilm indicated reduced contents of carbohydrate, protein, and e-DNA in the extracellular matrix of the biofilm when compared to the untreated control biofilm. AXD exposure downregulated activity of tissue invading enzyme, phospholipase in the reference strain. In wild type strain, ROS level, and activities of antioxidant enzymes were found elevated upon exposure to both drugs. FESEM analysis of the drug treated biofilms revealed degraded biofilm. This study has indicated mode of action of antifungal potential of alexidine dihydrochloride and hexachlorophene in C. albicans.
Asunto(s)
Antifúngicos , Biopelículas , Candida albicans , Reposicionamiento de Medicamentos , Pruebas de Sensibilidad Microbiana , Candida albicans/efectos de los fármacos , Candida albicans/genética , Antifúngicos/farmacología , Biopelículas/efectos de los fármacos , Humanos , Amidinas/farmacología , Hifa/efectos de los fármacos , Hifa/crecimiento & desarrollo , Ergosterol/metabolismo , Candidiasis/tratamiento farmacológico , Candidiasis/microbiología , Virulencia/efectos de los fármacos , BiguanidasRESUMEN
Previous reports have demonstrated that the peptide derived from LfcinB, R-1-R, exhibits anti-Candida activity, which is enhanced when combined with an extract from the Bidens pilosa plant. However, the mechanism of action remains unexplored. In this research, a proteomic study was carried out, followed by a bioinformatic analysis and biological assays in both the SC5314 strain and a fluconazole-resistant isolate of Candida albicans after incubation with R-1-R. The proteomic data revealed that treatment with R-1-R led to the up-regulation of most differentially expressed proteins compared to the controls in both strains. These proteins are primarily involved in membrane and cell wall biosynthesis, membrane transport, oxidative stress response, the mitochondrial respiratory chain, and DNA damage response. Additionally, proteomic analysis of the C. albicans parental strain SC5314 treated with R-1-R combined with an ethanolic extract of B. pilosa was performed. The differentially expressed proteins following this combined treatment were involved in similar functional processes as those treated with the R-1-R peptide alone but were mostly down-regulated (data are available through ProteomeXchange with identifier PXD053558). Biological assays validated the proteomic results, evidencing cell surface damage, reactive oxygen species generation, and decreased mitochondrial membrane potential. These findings provide insights into the complex antifungal mechanisms of the R-1-R peptide and its combination with the B. pilosa extract, potentially informing future studies on natural product derivatives.
Asunto(s)
Antifúngicos , Bidens , Candida albicans , Extractos Vegetales , Proteómica , Antifúngicos/farmacología , Proteómica/métodos , Bidens/química , Extractos Vegetales/farmacología , Extractos Vegetales/química , Candida albicans/efectos de los fármacos , Sinergismo Farmacológico , Proteínas Fúngicas/metabolismo , Péptidos/farmacología , Péptidos/química , Pruebas de Sensibilidad Microbiana , Farmacorresistencia Fúngica/efectos de los fármacos , Fluconazol/farmacologíaRESUMEN
BACKGROUND: The incidence of invasive fungal diseases (IFDs), represented by Candida albicans infection, is increasing year by year. However, clinically available antifungal drugs are very limited and encounter challenges such as limited efficacy, drug resistance, high toxicity, and exorbitant cost. Therefore, there is an urgent need for new antifungal drugs. PURPOSE: This study aims to find new antifungal compounds from plants, preferably those with good activity and low toxicity, and reveal their antifungal targets. METHODS: In vitro antifungal activities of compounds were investigated using broth microdilution method, spot assay, hyphal growth assay and biofilm formation assay. Synergistic effects were assessed using broth microdilution checkerboard technique. In vivo antifungal activities were evaluated using Galleria mellonella and murine candidiasis models. Cytotoxicity of compounds was investigated using Cell Counting Kit-8 (CCK-8). Discovery and validation of antifungal targets of compounds were conducted by using monoallelic knockout library of C. albicans, haploinsufficiency profiling (HIP), thermal shift assay (TSA), enzyme inhibitory effect assay, molecular docking, and in vitro and in vivo antifungal studies. RESULTS: 814 plant products were screened, among which petroselinic acid (PeAc) was found as an antifungal molecule. As a rare fatty acid isolated from coriander (Coriandrum sativum), carrot (Daucus carota) and other plants of the Apiaceae family, PeAc had not previously been found to have antifungal effects. In this study, PeAc was revealed to inhibit the growth of various pathogenic fungi, exhibited synergistic effects with fluconazole (FLC), inhibited the formation of C. albicans hyphae and biofilms, and showed antifungal effects in vivo. PeAc was less toxic to mammalian cells. Fructose-1,6-bisphosphate aldolase (Fba1p) was identified as a target of PeAc by using HIP, TSA, enzyme inhibitory effect assay and molecular docking methods. PeAc exerted antifungal effects more effectively on fba1Δ/FBA1 than wild-type (WT) strain both in vitro and in vivo. CONCLUSIONS: PeAc is an effective and low toxic antifungal compound. The target of PeAc is Fba1p. Fba1p is a promising target for antifungal drug development.