RESUMO
Available treatments against human fungal pathogens present high levels of resistance, motivating the development of new antifungal therapies. In this context, the present work aimed to analyze direct electric current (DC) antifungal action, using an in vitro apparatus equipped with platinum electrodes. Candida albicans yeast cells were submitted to three distinct conditions of DC treatment (anodic flow-AF; electroionic flow-EIF; and cathodic flow-CF), as well as different charges, ranging from 0.03 to 2.40 C. Our results indicated C. albicans presented distinct sensibility depending on the DC intensity and polarity applied. Both the colony-forming unit assay and the cytometry flow with propidium iodide indicated a drastic reduction on cellular viability after AF treatment with 0.15 C, while CF- and EIF-treated cells stayed alive when DC doses were increased up to 2.40 C. Additionally, transmission electron microscopy revealed important ultrastructural alterations in AF-treated yeasts, including cell structure disorganization, ruptures in plasmatic membrane, and cytoplasmic rarefaction. This work emphasizes the importance of physical parameters (polarity and doses) in cellular damage, and brings new evidence for using electrotherapy to treat C. albicans pathology process. Bioelectromagnetics. 38:95-108, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Candida albicans/citologia , Eletricidade , Candida albicans/metabolismo , Candida albicans/fisiologia , Candida albicans/ultraestrutura , Adesão Celular , Linhagem Celular , Eletrodos , Células Epiteliais/citologia , Humanos , Masculino , Viabilidade Microbiana , Pessoa de Meia-Idade , Platina/química , Propídio/metabolismoRESUMO
The commensal fungal pathogen Candida albicans is a leading cause of lethal systemic infections in immunocompromised patients. One of the main mechanisms of host immune evasion and virulence by this pathogen is the switch from yeast form to hyphal growth morphologies. Micro RNAs (miRNAs), a small regulatory non-coding RNA, has been identified as an important part of the immune response to a wide variety of pathogens. In general, miRNAs act by modulating the intensity of inflammatory responses. miRNAs act by base-paring binding to specific sequences of target mRNAs, generally causing their silencing through mRNA degradation or translational repression. To study the impact of C. albicans cell morphology upon host miRNA expression, we investigated the differential modulation of 9 different immune response-related miRNAs in primary murine bone marrow-derived macrophages (BMDMs) exposed to either yeasts or hyphal forms of Candida albicans. Here, we show that the different growth morphologies induce distinct miRNA expression patterns in BMDMs. Interestingly, our data suggest that the C-Type lectin receptor Dectin-1 is a major PRR that orchestrates miR155 upregulation in a Syk-dependent manner. Our results suggest that PRR-mediating signaling events are key drivers of miRNA-mediated gene regulation during fungal pathogenesis.
Assuntos
Candida albicans/citologia , Candida albicans/patogenicidade , Lectinas Tipo C/metabolismo , Macrófagos/microbiologia , MicroRNAs/genética , Animais , Candida albicans/crescimento & desenvolvimento , Candida albicans/imunologia , Regulação Fúngica da Expressão Gênica , Hifas/imunologia , Hifas/patogenicidade , Hifas/fisiologia , Evasão da Resposta Imune , Lectinas Tipo C/genética , Macrófagos/imunologia , Camundongos , Receptores de Reconhecimento de Padrão/metabolismo , Transdução de Sinais , Quinase Syk/metabolismo , Ativação Transcricional , Regulação para CimaRESUMO
Candida albicans is a major fungal pathogen of humans, causing approximately 400,000 life-threatening systemic infections world-wide each year in severely immunocompromised patients. An important fungicidal mechanism employed by innate immune cells involves the generation of toxic reactive oxygen species (ROS), such as superoxide and hydrogen peroxide. Consequently, there is much interest in the strategies employed by C. albicans to evade the oxidative killing by macrophages and neutrophils. Our understanding of how C. albicans senses and responds to ROS has significantly increased in recent years. Key findings include the observations that hydrogen peroxide triggers the filamentation of this polymorphic fungus and that a superoxide dismutase enzyme with a novel mode of action is expressed at the cell surface of C. albicans. Furthermore, recent studies have indicated that combinations of the chemical stresses generated by phagocytes can actively prevent C. albicans oxidative stress responses through a mechanism termed the stress pathway interference. In this review, we present an up-date of our current understanding of the role and regulation of oxidative stress responses in this important human fungal pathogen.
Assuntos
Candida albicans/metabolismo , Interações Hospedeiro-Patógeno , Estresse Oxidativo , Candida albicans/citologia , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Humanos , Imunidade Inata , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/farmacologia , Transdução de Sinais/efeitos dos fármacosRESUMO
AIM: This study aimed to evaluate the influence of surface free energy (SFE) of denture base and liner materials on Candida albicans biofilm development. METHODS: Discs were fabricated using poly(methyl methacrylate) acrylic resin and poly(ethyl methacrylate) denture liner, according to the manufacturers' instructions. For SFE test, discs were pellicle-coated with saliva alone, saliva + blood plasma, or blood plasma alone. Candida albicans biofilms were allowed to form on pellicle-coated discs for 48 h. Biofilms were evaluated for cell counts, metabolic activity, and structural characteristics at adhesion phase (after 1.5 h of development) and at biofilm maturity (after 48 h of development). Data were analyzed by anova and Tukey tests using a significance level of 5%. RESULTS: Saliva + blood plasma pellicles had a higher SFE compared to pellicles of saliva or blood plasma alone (P < 0.001). Differences in SFE by pellicle-coating did not affect the cell counts, metabolic activity, or structure at the adhesion phase (P > 0.05). In contrast, the presence of blood plasma resulted in higher cell counts, biovolume, and thickness of mature biofilms on both materials (P < 0.001). CONCLUSIONS: Increases in SFE from pellicle-coating leads to robust mature C. albicans biofilms on both denture materials.
Assuntos
Biofilmes , Candida albicans/fisiologia , Materiais Dentários/química , Bases de Dentadura/microbiologia , Reembasadores de Dentadura/microbiologia , Candida albicans/citologia , Contagem de Colônia Microbiana , Película Dentária/microbiologia , Humanos , Teste de Materiais , Metilmetacrilatos/química , Microscopia Confocal , Plasma , Polimetil Metacrilato/química , Distribuição Aleatória , Propriedades de Superfície , Tensão SuperficialRESUMO
Candida albicans can be a yeast that is a commensal on the human body but can cause opportunistic or pathogenic infections. Candida infections may create serious health problems and as a result has initiated a search for new drugs with an antifungal action. Geraniol is an acyclic monoterpene alcohol with known pharmacological properties, including antimicrobial activity. The aim of this work was to evaluate the antifungal activity and mechanism(s) of geraniol against C. albicans strains. The minimum inhibitory concentration (MIC) was determined through broth microdilution techniques. We investigated possible geraniol activity on the fungal cell wall (sorbitol protect effect), cell membrane (geraniol to ergosterol binding), the time-kill curve, and its biological activity on the yeast's morphology. Amphotericin B was used as control, and all tests were performed in duplicate. The MIC of geraniol was 16 µg/ml (for 90% of isolates) but its probable mechanism of action did not involve the cell wall and ergosterol binding. In the morphological interference assay, we observed that the product inhibited pseudohyphae and chlamydoconidia formation. Time-dependent kill curve assay demonstrated that the fungicidal activity for MIC × 2 started at 2 h for the ATCC 76485 strain, and at 4 h for the LM-70 strain. Geraniol showed in vitro antifungal potential against strains of C. albicans but did not involve action on the cell wall or ergosterol. This study contributes to the development of new antifungal drugs, especially against Candida spp.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Terpenos/farmacologia , Monoterpenos Acíclicos , Candida albicans/citologia , Candida albicans/fisiologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacosRESUMO
The release of extracellular vesicles (EV) by fungal organisms is considered an alternative transport mechanism to trans-cell wall passage of macromolecules. Previous studies have revealed the presence of EV in culture supernatants from fungal pathogens, such as Cryptococcus neoformans, Histoplasma capsulatum, Paracoccidioides brasiliensis, Sporothrix schenckii, Malassezia sympodialis and Candida albicans. Here we investigated the size, composition, kinetics of internalization by bone marrow-derived murine macrophages (MO) and dendritic cells (DC), and the immunomodulatory activity of C. albicans EV. We also evaluated the impact of EV on fungal virulence using the Galleria mellonella larvae model. By transmission electron microscopy and dynamic light scattering, we identified two populations ranging from 50 to 100 nm and 350 to 850 nm. Two predominant seroreactive proteins (27 kDa and 37 kDa) and a group of polydispersed mannoproteins were observed in EV by immunoblotting analysis. Proteomic analysis of C. albicans EV revealed proteins related to pathogenesis, cell organization, carbohydrate and lipid metabolism, response to stress, and several other functions. The major lipids detected by thin-layer chromatography were ergosterol, lanosterol and glucosylceramide. Short exposure of MO to EV resulted in internalization of these vesicles and production of nitric oxide, interleukin (IL)-12, transforming growth factor-beta (TGF-ß) and IL-10. Similarly, EV-treated DC produced IL-12p40, IL-10 and tumour necrosis factor-alpha. In addition, EV treatment induced the up-regulation of CD86 and major histocompatibility complex class-II (MHC-II). Inoculation of G. mellonella larvae with EV followed by challenge with C. albicans reduced the number of recovered viable yeasts in comparison with infected larvae control. Taken together, our results demonstrate that C. albicans EV were immunologically active and could potentially interfere with the host responses in the setting of invasive candidiasis.
Assuntos
Candida albicans/química , Candida albicans/imunologia , Fatores Imunológicos/química , Fatores Imunológicos/imunologia , Vesículas Secretórias/química , Vesículas Secretórias/imunologia , Animais , Antígenos de Fungos/análise , Antígenos de Fungos/química , Antígenos de Fungos/imunologia , Candida albicans/citologia , Células Cultivadas , Cromatografia em Camada Fina , Células Dendríticas/metabolismo , Endocitose , Proteínas Fúngicas/análise , Proteínas Fúngicas/química , Proteínas Fúngicas/imunologia , Interleucina-12/metabolismo , Lipídeos/análise , Macrófagos/metabolismo , Camundongos , Microscopia Eletrônica de Transmissão , Peso Molecular , Óxido Nítrico/metabolismo , Proteoma/análise , Vesículas Secretórias/ultraestrutura , Fator de Crescimento Transformador beta/metabolismoRESUMO
PROJECT: The opportunistic fungal Candida albicans can produce superficial and systemic infections in immunocompromised patients. An essential stage to both colonization and virulence by C. albicans is the transition from budding yeast form to filamentous form, producing biofilms. PROCEDURE: In this work, we studied the effect of the organochalcogenide compound (PhSe)2 on both cell growth and biofilm formation by C. albicans. RESULTS: (PhSe)2 inhibited both growth and biofilm formation by C. albicans. The inhibitory effects of (PhSe)2 depended on the cell density and (PhSe)2 concentration. We have also observed that (PhSe)2 stimulated ROS production (67%) and increased cell membrane permeability (2.94-fold) in C. albicans. In addition, (PhSe)2 caused a marked decrease in proteinase activity (6.8-fold) in relation to non-treated group. CONCLUSIONS: (PhSe)2 decreased both cell growth and biofilm development, decreasing the release of extracellular proteinases, which is an important facet of C. albicans pathogenicity. The toxicity of (PhSe)2 towards C. albicans can be associated with an increase in ROS production, which can increase cell permeability. The permanent damage to the cell membranes can culminate in cell death.
Assuntos
Derivados de Benzeno/farmacologia , Biofilmes/crescimento & desenvolvimento , Candida albicans/citologia , Candida albicans/fisiologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Compostos Organosselênicos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Biofilmes/efeitos dos fármacos , Candida albicans/enzimologia , Candida albicans/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Peptídeo Hidrolases/metabolismoRESUMO
Candida albicans is an opportunistic dimorphic fungus that inhabits various host mucosal sites. It can cause both superficial and serious systemic disease. Conversion from the yeast to the hyphal form has been associated with increased virulence and mucosal invasiveness. The aim of this study was to investigate the effect of sodium diclofenac and aspirin on germs tube formation of different Candida albicans strains. Prostaglandins may play an important role in fungal colonization. Nonsteroidal anti-inflammatory drugs are inhibitors of the cyclooxygenase isoenzymes. These drugs specifically block the biosynthesis of mammalian prostaglandins by inhibiting one or both of cyclooxygenase isoenzymes. In tests for germ tube formation sodium diclofenac reduced the filamentation to the 12.5%- 5.1%. In the presence of aspirin the filamentation was reduced up to 85-45% depending on the tested strain. Our results suggest that cyclooxygenase-depending synthesis of fungal prostaglandins is important for morphogenesis and fungal virulence. Inhibitors of cyclooxygenase isoensymes (aspirin and diclofenac) are effective in decreasing germ tube formation of Candida albicans.
Assuntos
Anti-Inflamatórios/farmacologia , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Aspirina/farmacologia , Candida albicans/citologia , Diclofenaco/farmacologia , Hifas/citologiaRESUMO
Candida albicans is an opportunistic dimorphic fungus that inhabits various host mucosal sites. It can cause both superficial and serious systemic disease. Conversion from the yeast to the hyphal form has been associated with increased virulence and mucosal invasiveness. The aim of this study was to investigate the effect of sodium diclofenac and aspirin on germs tube formation of different Candida albicans strains. Prostaglandins may play an important role in fungal colonization. Nonsteroidal anti-inflammatory drugs are inhibitors of the cyclooxygenase isoenzymes. These drugs specifically block the biosynthesis of mammalian prostaglandins by inhibiting one or both of cyclooxygenase isoenzymes. In tests for germ tube formation sodium diclofenac reduced the filamentation to the 12.5%- 5.1%. In the presence of aspirin the filamentation was reduced up to 85-45% depending on the tested strain. Our results suggest that cyclooxygenase-depending synthesis of fungal prostaglandins is important for morphogenesis and fungal virulence. Inhibitors of cyclooxygenase isoensymes (aspirin and diclofenac) are effective in decreasing germ tube formation of Candida albicans.
Assuntos
Anti-Inflamatórios/farmacologia , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Aspirina/farmacologia , Candida albicans/citologia , Diclofenaco/farmacologia , Hifas/citologiaRESUMO
Candida albicans is a commensal fungus, but circumstantially it may cause superficial infections of the mucous membranes, such as denture stomatitis, when a biofilm is formed on the surface of dental prostheses. This study evaluated the cell viability of C. albicans biofilms against the antifungal activity of thymol when compared with miconazole, by the fluorescence imaging using SYTO 9 and propidium iodide dyes, and counting of colony forming units. C. albicans standard strains (ATCC 11006) were used. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of drugs were determined by broth microdilution tests and the inoculum was standardized to match 0.5 on the McFarland scale (106 cfu/mL). Biofilms were grown on the surface of acrylic resin disks in parallel flow chambers from Sabouraud broth supplemented with 10% dextrose. For counting of colony forming units, the fungal solution was sequentially diluted and plated in Sabouraud dextrose agar. Data were analyzed using two-way ANOVA and Tukey's test (a=5%). Biofilms treated with thymol and miconazole presented low numbers of viable cells at the evaluated exposure times. There was statistically significant difference (p<0.05) when compared with control, and the mean value of the exposure times between miconazole and thymol did not differ significantly (p>0.05). In conclusion, both drugs have similar efficiency as antifungal agents against biofilms of C. albicans formed on acrylic surfaces.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Timol/farmacologia , Candida albicans/citologiaRESUMO
Candida albicans is an opportunistic dimorphic fungus that inhabits various host mucosal sites. It can cause both superficial and serious systemic disease. Conversion from the yeast to the hyphal form has been associated with increased virulence and mucosal invasiveness. The aim of this study was to investigate the effect of sodium diclofenac and aspirin on germs tube formation of different Candida albicans strains. Prostaglandins may play an important role in fungal colonization. Nonsteroidal anti-inflammatory drugs are inhibitors of the cyclooxygenase isoenzymes. These drugs specifically block the biosynthesis of mammalian prostaglandins by inhibiting one or both of cyclooxygenase isoenzymes. In tests for germ tube formation sodium diclofenac reduced the filamentation to the 12.5%- 5.1%. In the presence of aspirin the filamentation was reduced up to 85-45% depending on the tested strain. Our results suggest that cyclooxygenase-depending synthesis of fungal prostaglandins is important for morphogenesis and fungal virulence. Inhibitors of cyclooxygenase isoensymes (aspirin and diclofenac) are effective in decreasing germ tube formation of Candida albicans.
Assuntos
Anti-Inflamatórios/farmacologia , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Aspirina/farmacologia , Candida albicans/citologia , Diclofenaco/farmacologia , Hifas/citologiaRESUMO
Candida albicans is a commensal fungus, but circumstantially it may cause superficial infections of the mucous membranes, such as denture stomatitis, when a biofilm is formed on the surface of dental prostheses. This study evaluated the cell viability of C. albicans biofilms against the antifungal activity of thymol when compared with miconazole, by the fluorescence imaging using SYTO 9 and propidium iodide dyes, and counting of colony forming units. C. albicans standard strains (ATCC 11006) were used. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of drugs were determined by broth microdilution tests and the inoculum was standardized to match 0.5 on the McFarland scale (106 cfu/mL). Biofilms were grown on the surface of acrylic resin disks in parallel flow chambers from Sabouraud broth supplemented with 10% dextrose. For counting of colony forming units, the fungal solution was sequentially diluted and plated in Sabouraud dextrose agar. Data were analyzed using two-way ANOVA and Tukey's test (a=5%). Biofilms treated with thymol and miconazole presented low numbers of viable cells at the evaluated exposure times. There was statistically significant difference (p<0.05) when compared with control, and the mean value of the exposure times between miconazole and thymol did not differ significantly (p>0.05). In conclusion, both drugs have similar efficiency as antifungal agents against biofilms of C. albicans formed on acrylic surfaces.
Candida albicans é um fungo comensal que circunstancialmente pode causar infecções superficiais das mucosas, como a estomatite protética, na qual o biofilme se forma sobre a superfície das próteses dentárias. Este estudo avaliou a viabilidade celular de biofilmes de C. albicans frente à ação antifúngica do timol em comparação com o miconazol por meio da técnica de fluorescência, empregando os corantes SYTO 9 e iodeto de propídio, e contagem das unidades formadoras de colônia. Utilizaram-se cepas de C. albicans (ATCC 11006). As concentrações inibitórias mínimas (CIM) e fungicidas mínimas (CFM) das drogas foram determinadas com testes de microdiluição em caldo, sendo o inóculo padronizado para corresponder a 0,5 da escala de McFarland (106 UFC/mL). Os biofilmes foram cultivados sobre a superfície de discos de resina acrílica, em células paralelas de fluxo, a partir de caldo Sabouraud suplementado com dextrose 10%. Para a contagem das unidades formadoras de colônia, as soluções fúngicas foram sequencialmente diluídas e semeadas em agar Sabouraud dextrose Os dados foram analisados por meio de estatística ANOVA A dois fatores e teste de Tukey (α = 5%). Biofilmes tratados com o timol e miconazol apresentaram baixos números percentuais de células viáveis nos tempos de exposição avaliados. Houve diferença estatística significante (p<0.05) em comparação com o controle, e o valor médio dos tempos de exposição entre miconazol e timol não diferiu estatisticamente (p>0.05). Concluindo, ambas as drogas possuem similar eficiência como agentes antifúngicos contra a viabilidade de biofilmes de C. albicans formados em superfícies de resinas acrílicas.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Timol/farmacologia , Candida albicans/citologiaRESUMO
Candida albicans es una levadura comensal, que bajo ciertas circunstancia puede convertirse en patógeno. La capacidad de cambiar se forma constituye un factor de virulencia, permitiéndole a la levadura invadir y diseminarse. El sistema inmune innato es capaz de reconocer las diferentes morfologías de C. albicans activando receptores (PRRs) que reconocen PAMPs o patrones moleculares conservados. Las células inmunes más importantes son los macrófagos y los neutrófilos que desencadenan una respuesta efectora a través de la fagocitosis y la activación de estrés oxidativo contra C. albicans. Las células dendríticas, por su parte expresan la mayoría de los PRRs involucrados en el reconocimiento de C. albicans activando la secreción de citoquinas hacia una respuesta tipo TH1 (inducida por INF tipo1, IL-12, INFγ, ), Treg (inducida por TGFβ, IL-10) y TH17 (inducida por IL-23, IL6). Las células epiteliales no sólo constituyen una barrera física frente a la infección por C. albicans, sino son fundamentales en el reconocimiento primario de este microorganismo. Mediante una respuesta bifásica estas células activan diferencialmente, vías de transducción de señales que determinan que no se active una respuesta de citoquinas frente a la presencia de blastoconidios (forma comensal) y que se active frente a presencia de hifas (forma invasora). Por su parte, C. albicans es capaz de desarrollar mecanismos evasivos de la respuesta inmune. Esta compleja interacción y hongo-hospedero determina si el hospedero será capaz de eliminar a este microorganismo o si éste finalmente invadirá expresando su virulencia.
Candida albicans is a comensal microorganism that under certain circumstances is able to transform into a pathogen. This ability to switch constitute a virulence factor that C. albicans uses to invade and spread. The innate immune system recognize the different forms of C. albicans activating receptors (PRRs) that recognize PAMPs or conserved molecular patterns. The most important immune cells are macrophages and neutrophils that generate an effector response through phagocytosis and oxidative burst against C. albicans. Dendritic cells express the most of PRRs involved in the recognition of C. albicans activating the cytokines synthesis forward to a TH1 (induced by INF tipo1, IL-12, INFγ), Treg (induced by TGFβ, IL-10) y TH17 (induced by IL- 23, IL6) immune response. Epithelial cells not only constitute a physical barrier against C. albicans, but they are crucial in the first recognition of this microorganism. Through a biphasic response these cells differentially activate pathways that determine a no cytokine response when blastoconidia (commensal form) is present and an inflammatory response in presence of hyphae (pathogenic form). C. albicans develops immune evasive mechanism. This complex host-fungi interaction determines if the host will eradicate the infection or if this microorganism will invade the host, expressing it virulence.
Assuntos
Humanos , Candida albicans/citologia , Candida albicans/patogenicidade , Interações Hospedeiro-Patógeno , Imunidade Inata , Receptores de Reconhecimento de PadrãoRESUMO
In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.
Assuntos
Álcoois/metabolismo , Biofilmes/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Candida albicans/citologia , Candida albicans/crescimento & desenvolvimento , Viabilidade Microbiana/efeitos dos fármacosRESUMO
The photodynamic mechanism of action induced by N,N-dimethyl-2-(4'-N,N,N-trimethylaminophenyl)fulleropyrrolidinium iodide (DTC60(2+)) was investigated on Candida albicans and Escherichia coli cells. First, photogeneration of superoxide anion radical by DTC60(2+) in the presence of NADH was detected using nitro blue tetrazolium method in reverse micelles. In C. albicans suspensions, 10 µM DTC60(2+) was an effective photosensitizer, producing a â¼5log decrease of cell survival when the cultures were irradiated for 30 min with visible light. Also, C. albicans cells growth was not detected in the presence of 10 µM DTC60(2+) and irradiation. Photodynamic mechanism investigations were compared in both C. albicans and E. coli cells. Studies under anoxic conditions indicated that oxygen was required for the photodynamic inactivation of these microorganisms. The photocytotoxicity induced by DTC60(2+) was similar in D2O than in water cell suspensions. Furthermore, photoinactivation of microbial cells was negligible in the presence of azide ion, while the addition of mannitol produced a photoprotective effect on the cellular survival. These results indicate that DTC60(2+) has potential as agent to the photodynamic inactivation of microbial cells. Also, the photocytotoxicity activity induced by this cationic fullerene derivative can involve the intermediacy of both superoxide anion radical and singlet molecular oxygen.
Assuntos
Candida albicans/citologia , Candida albicans/efeitos dos fármacos , Ácidos Carboxílicos/farmacologia , Escherichia coli/citologia , Escherichia coli/efeitos dos fármacos , Fulerenos/farmacologia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Carga Bacteriana/efeitos dos fármacos , Carga Bacteriana/efeitos da radiação , Candida albicans/efeitos da radiação , Ácidos Carboxílicos/química , Cátions Bivalentes , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Relação Dose-Resposta a Droga , Escherichia coli/efeitos da radiação , Fulerenos/química , Doses de RadiaçãoRESUMO
Candida infections are very common in cancer patients and it is a common practice to prescribe antifungal antibiotics along with anticancer drugs. Yeast to hyphal form switching is considered to be important in invasive candidiasis. Targeting morphogenetic switching may be useful against invasive candidiasis. In this study, we report the antimorphogenetic properties of thirty cancer drugs.
Assuntos
Humanos , Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Candida albicans/efeitos dos fármacos , Hifas/efeitos dos fármacos , Candida albicans/citologia , Candida albicans/crescimento & desenvolvimento , Reposicionamento de Medicamentos , Hifas/citologia , Hifas/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , MicroscopiaRESUMO
Candida infections are very common in cancer patients and it is a common practice to prescribe antifungal antibiotics along with anticancer drugs. Yeast to hyphal form switching is considered to be important in invasive candidiasis. Targeting morphogenetic switching may be useful against invasive candidiasis. In this study, we report the antimorphogenetic properties of thirty cancer drugs.
Assuntos
Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Candida albicans/efeitos dos fármacos , Hifas/efeitos dos fármacos , Candida albicans/citologia , Candida albicans/crescimento & desenvolvimento , Reposicionamento de Medicamentos , Humanos , Hifas/citologia , Hifas/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , MicroscopiaRESUMO
In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.
Assuntos
Álcoois/metabolismo , Biofilmes/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Candida albicans/citologia , Candida albicans/crescimento & desenvolvimento , Viabilidade Microbiana/efeitos dos fármacosRESUMO
Photodynamic inactivation of Candida albicans produced by 5-(4-trifluorophenyl)-10,15,20-tris(4-N,N,N-trimethylammoniumphenyl)porphyrin (TFAP(3+)), 5,10,15,20-tetrakis(4-N,N,N-trimethylammoniumphenyl)porphyrin (TMAP(4+)) and 5,10,15,20-tetrakis(4-N-methylpyridyl)porphyrin (TMPyP(4+)) was investigated to obtain insight about the mechanism of cellular damage. In solution, absorption spectroscopic studies showed that these cationic porphyrins interact strongly with calf thymus DNA. The electrophoretic analysis indicated that photocleavage of DNA induced by TFAP(3+) took place after long irradiation periods (>5 h). In contrast, TMAP(4+) produced a marked reduction in DNA band after 1 h irradiation. In C. albicans, these cationic porphyrins produced a â¼3.5 log decrease in survival when the cell suspensions (10(7) cells/mL) were incubated with 5 µM photosensitizer and irradiated for 30 min with visible light (fluence 162 J/cm(2)). After this treatment, modifications of genomic DNA isolated from C. albicans cells were not found by electrophoresis. Furthermore, transmission electron microscopy showed structural changes with appearance of low density areas into the cells and irregularities in cell barriers. However, the photodamage to the cell envelope was insufficient to cause the release of intracellular biopolymers. Therefore, modifications in the cytoplasmic biomolecules and alteration in the cell barriers could be mainly involved in C. albicans photoinactivation.
Assuntos
Candida albicans/efeitos dos fármacos , Luz , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Animais , Candida albicans/citologia , Candida albicans/genética , Cátions/síntese química , Cátions/química , Cátions/farmacologia , Bovinos , DNA/química , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/química , Porfirinas/síntese química , Porfirinas/química , Relação Estrutura-AtividadeRESUMO
Candida albicans y Candida dubliniensis presentan una estrecha relación filogenética. La similitud de estas especies puede hacer que en un laboratorio microbiológico se identifique en forma errónea C. dubliniensis como C. albicans. El objetivo de esta investigación fue evaluar diversos métodos fenotípicos para la diferenciación entre Candida dubliniensis y Candida albicans. Se utilizaron 6 cepas control de C. dubliniensis y una de C. albicans, provenientes de colecciones reconocidas y sometidas a genotipificación. También se utilizaron 70 aislados identificados como posibles C. albicans utilizando el medio CHROMagar Candida y el medio de bilis agar Feo. Los métodos evaluados fueron: agar Sabouraud dextrosa a 45°C, agar Sabouraud con NaCl al 6,5%, agar Tween 80, agar tabaco, agar Pals, agar tomate-zanahoria y aglutinación con partículas de látex (Bichro-Dubli Fumouze®). Encontramos que las técnicas más confiables para realizar la diferenciación fenotípica entre estas dos especies fueron: el agar tomate-zanahoria, el agar Pals, el agar tabaco y la aglutinación con partículas de látex (Bichro-Dubli Fumouze®). Además en este estudio, de los 70 aislados considerados como C. albicans, encontramos 1 (1.4%) posible Candida dubliniensis. Sin embargo, las pruebas de biología molecular son las más adecuadas para el diagnóstico certero de estas dos especies.
Candida albicans and Candida dubliniensis have a close phylogenetic relationship. The similarity between these species can cause a microbiology laboratory to identify C. dubliniensis erroneously as C. albicans. The objective of this research was to evaluate diverse phenotypic methods for differentiating between Candida dubliniensis and Candida albicans. Six control strains of C. dubliniensis and one of C. albicans from recognized collections were used and submitted to genotypification. Also, 70 isolates were used, identified as possible C. albicans utilizing CHROMagar Candida and bilis agar Feo mediums. The methods evaluated were: Sabouraud dextrosa agar at 45°C, Sabouraud agar with NaCl at 6.5%, Tween 80 agar, tobacco agar, Pals agar, tomato-carrot agar and agglutination with latex particles (Bichro-Dubli Fumouze®). It was found that the most reliable techniques for performing phenotype differentiation between these two species were tomato-carrot agar, Pals agar, tobacco agar and agglutination with latex particles (Bichro-Dubli Fumouze®). Of the 70 isolates considered to be C. albicans, one (1.4%) possible Candida dubliniensis was found. Nevertheless, molecular biology tests are the most appropriate means for achieving an accurate diagnosis of these two species.