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1.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 40(7): 1049-1055, 2020 May 25.
Artigo em Chinês | MEDLINE | ID: mdl-32701244

RESUMO

OBJECTIVE: To investigate the characteristics of growth and metabolism and the in vivo toxicity of Candida auris under different conditions. METHODS: We observed the growth of Candida auris and Candida albicans under routine culture conditions and in different pH and salt concentrations, and compared their activities of sugar fermentation using microbiochemical reaction tubes. Four-week-old nude mice were randomized into Candida auris infection group (n=5), Candida albicans infection group (n=5) and control group (n=5) for intragastric administration of 0.3 mL suspension the two Candida species (5×109 cfu/mL) or 0.3 mL normal saline. Samples of the liver, kidney, intestine, feces and blood were taken for analysis of the in vivo distribution and toxicity of Candida albicans by fungal culture and histopathological examination. RESULTS: Candida auris exhibited logarithmic growth at 8-24 h after inoculation and showed stable growth after 24 h. Candida auris showed optimal growth within the pH value range of 5-7 with a growth pattern identical to that of Candida albicans. Candida auris grew better than Candida albicans in media containing 5% and 10% NaCl, and could ferment glucose, sucrose, trehalose and sorbitol. Candida auris could be isolated from the feces, blood, liver and kidney of infected nude mice, and the liver had the highest fungal load (5.7 log10 cfu/g). Candida auris could cause pathological changes in the liver and intestine of the mice, but with a lesser severity as compared with Candida albicans. CONCLUSIONS: Candida auris exhibits optimal growth in mildly acidic or neutral conditions with a high salt tolerance, and can potentially penetrate the intestinal barrier into blood and lead to tissue injuries in hosts with immunosuppression.


Assuntos
Candida , Candidíase , Animais , Candida/crescimento & desenvolvimento , Candida/isolamento & purificação , Candida albicans/crescimento & desenvolvimento , Candidíase/microbiologia , Meios de Cultura , Camundongos , Camundongos Nus , Distribuição Aleatória
2.
J Vis Exp ; (161)2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32716381

RESUMO

Candida albicans hyphal morphogenesis in the gastrointestinal (GI) tract is tightly controlled by various environmental signals, and plays an important role in the dissemination and pathogenesis of this opportunistic fungal pathogen. However, methods to visualize fungal hyphae in the GI tract in vivo are challenging which limits the understanding of environmental signals in controlling this morphogenesis process. The protocol described here demonstrates a novel ex vivo method for visualization of hyphal morphogenesis in gut homogenate extracts. Using an ex vivo assay, this study demonstrates that cecal contents from antibiotic treated mice, but not from untreated control mice, promote C. albicans hyphal morphogenesis in the gut content. Further, adding back specific groups of gut metabolites to the cecal contents from antibiotic-treated mice differentially regulates hyphal morphogenesis ex vivo. Taken together, this protocol represents a novel method to identify and investigate the environmental signals that control C. albicans hyphal morphogenesis in the GI tract.


Assuntos
Bioensaio/métodos , Candida albicans/crescimento & desenvolvimento , Trato Gastrointestinal/microbiologia , Hifas/crescimento & desenvolvimento , Morfogênese , Animais , Candida albicans/efeitos dos fármacos , Ceco/microbiologia , Cefoperazona/farmacologia , Feminino , Proteínas Fúngicas/metabolismo , Trato Gastrointestinal/efeitos dos fármacos , Hifas/efeitos dos fármacos , Masculino , Metaboloma/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Morfogênese/efeitos dos fármacos
3.
Braz Oral Res ; 34: e050, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32578760

RESUMO

Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.


Assuntos
Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Óleo de Melaleuca/farmacologia , Terpenos/farmacologia , Resinas Acrílicas , Análise de Variância , Antifúngicos/farmacologia , Biofilmes/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Bases de Dentadura/microbiologia , Testes de Sensibilidade Microbiana , Microscopia Confocal , Valores de Referência , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Óleo de Melaleuca/química , Terpenos/química
4.
Int J Nanomedicine ; 15: 3681-3693, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32547022

RESUMO

Background: Fungal infections are becoming more prevalent and threatening because of the continuous emergence of azole-resistant fungal infections. The present study was aimed to assess the activity of free Methylglyoxal (MG) or MG-conjugated chitosan nanoparticles (MGCN) against fluconazole-resistant Candida albicans. Materials and Methods: A novel formulation of MGCN was prepared and characterized to determine their size, shape and polydispersity index. Moreover, the efficacy of fluconazole or MG or MGCN was determined against intracellular C. albicans in macrophages and the systematic candidiasis in a murine model. The safety of MG or MGCN was tested in mice by analyzing the levels of hepatic and renal toxicity parameters. Results: Candida albicans did not respond to fluconazole, even at the highest dose of 20 mg/kg, whereas MG and MGCN effectively eliminated C. albicans from the macrophages and infected mice. Mice in the group treated with MGCN at a dose of 10 mg/kg exhibited a 90% survival rate and showed the lowest fungal load in the kidney, whereas the mice treated with free MG at the same dose exhibited 50% survival rate. Moreover, the administration of MG or MGCN did not induce any liver and kidney toxicity in the treated mice. Conclusion: The findings of the present work suggest that MGCN may be proved a promising therapeutic formulation to treat azole-resistant C. albicans infections.


Assuntos
Candidíase/tratamento farmacológico , Quitosana/química , Farmacorresistência Fúngica , Fluconazol/uso terapêutico , Nanopartículas/química , Aldeído Pirúvico/uso terapêutico , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candidíase/microbiologia , Modelos Animais de Doenças , Farmacorresistência Fúngica/efeitos dos fármacos , Feminino , Fluconazol/farmacologia , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Nanopartículas/ultraestrutura , Tamanho da Partícula , Aldeído Pirúvico/farmacologia
5.
J Appl Oral Sci ; 28: e20190578, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32348446

RESUMO

Objective This study sought to analyze the gene expression of Candida albicans in sound root surface and root caries lesions, exploring its role in root caries pathogenesis. Methodology The differential gene expression of C. albicans and the specific genes related to cariogenic traits were studied in association with samples of biofilm collected from exposed sound root surface (SRS, n=10) and from biofilm and carious dentin of active root carious lesions (RC, n=9). The total microbial RNA was extracted, and the cDNA libraries were prepared and sequenced on the Illumina Hi-Seq2500. Unique reads were mapped to 163 oral microbial reference genomes including two chromosomes of C. albicans SC5314 (14,217 genes). The putative presence of C. albicans was estimated (sum of reads/total number of genes≥1) in each sample. Count data were normalized (using the DESeq method package) to analyze differential gene expression (using the DESeq2R package) applying the Benjamini-Hochberg correction (FDR<0.05). Results Two genes (CaO19.610, FDR=0.009; CaO19.2506, FDR=0.018) were up-regulated on SRS, and their functions are related to biofilm formation. Seven genes ( UTP20 , FDR=0.018; ITR1 , FDR=0.036; DHN6 , FDR=0.046; CaO19.7197 , FDR=0.046; CaO19.7838 , FDR=0.046; STT4 , FDR=0.046; GUT1 , FDR=0.046) were up-regulated on RC and their functions are related to metabolic activity, sugar transport, stress tolerance, invasion and pH regulation. The use of alternative carbon sources, including lactate, and the ability to form hypha may be a unique trait of C. albicans influencing biofilm virulence. Conclusions C. albicans is metabolically active in SRS and RC biofilm, with different roles in health and disease.


Assuntos
Biofilmes/crescimento & desenvolvimento , Candida albicans/genética , RNA Fúngico/genética , Cárie Radicular/microbiologia , Candida albicans/crescimento & desenvolvimento , Candida albicans/isolamento & purificação , Expressão Gênica , Regulação Fúngica da Expressão Gênica , Humanos , Morfogênese , RNA-Seq/métodos , Valores de Referência , Raiz Dentária/microbiologia , Regulação para Cima , Fatores de Virulência
6.
Ecotoxicol Environ Saf ; 197: 110625, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32302863

RESUMO

Due to the potential of release and accumulation in the environment, nanoplastics have attracted an increasing attention. In this study, we investigated the effect of exposure to nanopolystyrene (30 nm) in nematode Caenorhabditis elegans after the fungal infection. After Candida albicans infection, exposure to nanopolystyrene (10 and 100 µg/L) for 24-h could cause the more severe toxicity on lifespan and locomotion behavior compared with fungal infection alone. The more severe activation of oxidative stress and suppression of SOD-3:GFP expression and mitochondrial unfolded protein response (mt UPR) were associated with this observed toxicity enhancement induced by nanopolystyrene exposure. Moreover, the more severe C. albicans colony formation and suppression of innate immune response as indicated by the alteration in expression of anti-microbial genes (abf-2, cnc-4, cnc-7, and fipr-22/23) further contributed to the formation of this toxicity enhancement induced by nanopolystyrene exposure. Our results demonstrated that short-term exposure to nanopolystyrene in the range of µg/L potentially enhances the adverse effects of fungal infection on organisms.


Assuntos
Caenorhabditis elegans , Candidíase/induzido quimicamente , Locomoção/efeitos dos fármacos , Longevidade/efeitos dos fármacos , Poliestirenos/toxicidade , Animais , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/microbiologia , Proteínas de Caenorhabditis elegans/metabolismo , Candida albicans/crescimento & desenvolvimento , Candidíase/microbiologia , Estresse Oxidativo/efeitos dos fármacos
7.
Biofouling ; 36(2): 210-221, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32292058

RESUMO

Surfactin is a type of cyclic lipopeptide biosurfactant implicated in a wide range of applications. Although its antimicrobial activity has been characterized, its effect on Candida albicans physiology remains to be elucidated. The present study evaluated the influence of surfactin-C15 (SF) and its complexes with divalent counterions on C. albicans biofilm formation and preformed biofilms. The SF and metal(II)-SF complexes inhibited biofilm formation and reduced the metabolic activity of mature biofilms in a concentration-dependent manner. The same concentrations of the compounds studied dislodged preexisting biofilms grown on polystyrene plates. Moreover, SF and its metal(II) complexes reduced the mRNA expression of hypha-specific genes HWP1, ALS1, ALS3, ECE1 and SAP4 without exhibiting significant growth inhibition. Further research showed that the compounds tested reduced cellular surface hydrophobicity (CSH). These results suggest that SF and metal(II)-SF complexes could be used as anti-biofilm agents against C. albicans hypha-related infections in clinical practice.


Assuntos
Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Complexos de Coordenação/farmacologia , Hifas/efeitos dos fármacos , Tensoativos/metabolismo , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Candida albicans/metabolismo , Proteínas Fúngicas/genética , Proteínas de Fluorescência Verde/genética , Hifas/crescimento & desenvolvimento
8.
Biofouling ; 36(2): 234-244, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32321306

RESUMO

This study evaluated adhesion and biofilm formation by Candida albicans, Pseudomonas aeruginosa and Staphylococcus epidermidis on surfaces of titanium (Ti) and titanium coated with F18 Bioactive Glass (BGF18). Biofilms were grown and the areas coated with biofilm were determined after 2, 4 and 8 h. Microscopy techniques were applied in order to visualize the structure of the mature biofilm and the extracellular matrix. On the BGF18 specimens, there was less biofilm formation by C. albicans and S. epidermidis after incubation for 8 h. For P. aeruginosa biofilm, a reduction was observed after incubation for 4 h, and it remained reduced after 8 h on BGF18 specimens. All biofilm matrices seemed to be thicker on BGF18 surface than on titanium surfaces. BGF18 showed significant anti-biofilm activity in comparison with Ti in the initial periods of biofilm formation; however, there was extensive biofilm after incubation for 48 h.


Assuntos
Materiais Biocompatíveis/química , Biofilmes/crescimento & desenvolvimento , Vidro/química , Próteses e Implantes/microbiologia , Titânio/química , Candida albicans/crescimento & desenvolvimento , Pseudomonas aeruginosa/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Propriedades de Superfície
9.
Biofouling ; 36(3): 245-255, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32326753

RESUMO

Dental plaque is a biofilm composed of a complex oral microbial community. The accumulation of plaque in the pit and fissures of dental elements often leads to the development of tooth decay (dental caries). Here, potent anti-biofilm materials were developed by incorporating zinc methacrylates or di-n-butyl-dimethacrylate-tin into the light-curable sealant and their physical, mechanical, and biological properties were evaluated. The data revealed that 5% di-n-butyl-dimethacrylate-tin (SnM 5%) incorporated sealant showed strong anti-biofilm efficacy against various single-species (Streptococcus mutans or Streptococcus oralis or Candida albicans) and S. mutans-C. albicans cross-kingdom dual-species biofilms without either impairing the mechanical properties of the sealant or causing cytotoxicities against mouse fibroblasts. The findings indicate that the incorporation of SnM 5% in the experimental pit and fissure self-adhesive sealant may have the potential to be part of current chemotherapeutic strategies to prevent the formation of cariogenic oral biofilms that cause dental caries.


Assuntos
Adesivos/farmacologia , Biofilmes/efeitos dos fármacos , Cárie Dentária/prevenção & controle , Selantes de Fossas e Fissuras/farmacologia , Zinco/química , Adesivos/química , Animais , Biofilmes/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Cárie Dentária/microbiologia , Humanos , Metacrilatos/química , Camundongos , Microbiota/efeitos dos fármacos , Selantes de Fossas e Fissuras/química , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus oralis/efeitos dos fármacos , Streptococcus oralis/crescimento & desenvolvimento
10.
Nat Commun ; 11(1): 2031, 2020 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-32341348

RESUMO

Neutrophils employ several mechanisms to restrict fungi, including the action of enzymes such as myeloperoxidase (MPO) or NADPH oxidase, and the release of neutrophil extracellular traps (NETs). Moreover, they cooperate, forming "swarms" to attack fungi that are larger than individual neutrophils. Here, we designed an assay for studying how these mechanisms work together and contribute to neutrophil's ability to contain clusters of live Candida. We find that neutrophil swarming over Candida clusters delays germination through the action of MPO and NADPH oxidase, and restricts fungal growth through NET release within the swarm. In comparison with neutrophils from healthy subjects, those from patients with chronic granulomatous disease produce larger swarms against Candida, but their release of NETs is delayed, resulting in impaired control of fungal growth. We also show that granulocyte colony-stimulating factors (GCSF and GM-CSF) enhance swarming and neutrophil ability to restrict fungal growth, even during treatment with chemical inhibitors that disrupt neutrophil function.


Assuntos
Candida albicans/crescimento & desenvolvimento , Neutrófilos/citologia , Neutrófilos/microbiologia , Sistemas CRISPR-Cas , Candidíase/microbiologia , Linhagem Celular , Armadilhas Extracelulares/metabolismo , Fator Estimulador de Colônias de Granulócitos/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Doença Granulomatosa Crônica/microbiologia , Humanos , Processamento de Imagem Assistida por Computador , Análise em Microsséries , NADPH Oxidases/metabolismo , Peroxidase/metabolismo , Espécies Reativas de Oxigênio/metabolismo
11.
J Mycol Med ; 30(2): 100939, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32111506

RESUMO

Nosocomial infections by fungi are important causes of morbidity and mortality, and the adhesion capacity of yeast on abiotic and biotic surfaces has been considered an important step in this process. Als3 proteins are widely studied for their ability to allow Candida albicans to bind to various surfaces. The objective of the present study was to verify, with more details, the action of F2768-0318 in relation to its antifungal activity as well as its ability to act on C. albicans virulence factors related to adhesion and biofilm formation in vitro and in vivo by inhibiting the Als3 protein. F2768-0318 was assessed in tests of biofilm formation and adhesion on abiotic surfaces (polystyrene plates) and adherence on biotic surfaces, including human endocervical (HeLa) cells, human umbilical vein endothelial cells (HUVECs), and fresh buccal epithelial cells (BEC). Our results showed F2768-0318 was useful in reducing the adhesion and biofilm formation of C. albicans on abiotic surfaces, indicating the possibility of treating hospital materials and preventing biofilm formation on these types of equipment. Further studies are still needed, including optimization of the molecule to allow this molecule to be effective on other types of surfaces, such as human cells.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Animais , Antifúngicos/uso terapêutico , Candida albicans/crescimento & desenvolvimento , Candida albicans/fisiologia , Candidemia/tratamento farmacológico , Candidemia/microbiologia , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/microbiologia , Células Epiteliais/fisiologia , Feminino , Células HeLa , Células Endoteliais da Veia Umbilical Humana , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Testes de Toxicidade
12.
BMC Biotechnol ; 20(1): 15, 2020 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-32164664

RESUMO

BACKGROUND: Infections caused by fungi are often refractory to conventional therapies and urgently require the development of novel options, such as immunotherapy. To produce therapeutic antibodies, a plant-based expression platform is an attractive biotechnological strategy compared to mammalian cell cultures. In addition to whole plants, hairy roots (HR) cultures can be used, representing an expression system easy to build up, with indefinite growth while handled under containment conditions. RESULTS: In this study the production in HR of a recombinant antibody, proved to be a good candidate for human immunotherapy against fungal infections, is reported. Expression and secretion of this antibody, in an engineered single chain (scFvFc) format, by HR from Nicotiana benthamiana and Solanum lycopersicum have been evaluated with the aim of directly using the deriving extract or culture medium against pathogenic fungi. Although both Solanaceae HR showed good expression levels (up to 68 mg/kg), an optimization of rhizosecretion was only obtained for N. benthamiana HR. A preliminary assessment to explain this result highlighted the fact that not only the presence of proteases, but also the chemical characteristics of the growth medium, can influence antibody yield, with implications on recombinant protein production in HR. Finally, the antifungal activity of scFvFc 2G8 antibody produced in N. benthamiana HR was evaluated in Candida albicans growth inhibition assays, evidencing encouraging results. CONCLUSIONS: Production of this anti-fungal antibody in HR of N. benthamiana and S. lycopersicum elucidated factors affecting pharming in this system and allowed to obtain promising ready-to-use immunotherapeutics against C. albicans.


Assuntos
Antifúngicos/farmacologia , Candida albicans/crescimento & desenvolvimento , Anticorpos de Cadeia Única/farmacologia , Solanaceae/citologia , Candida albicans/efeitos dos fármacos , Recombinação Homóloga , Lycopersicon esculentum/citologia , Lycopersicon esculentum/genética , Raízes de Plantas/citologia , Raízes de Plantas/genética , Plantas Geneticamente Modificadas , Engenharia de Proteínas , Proteínas Recombinantes/farmacologia , Anticorpos de Cadeia Única/genética , Solanaceae/genética , Tabaco/citologia , Tabaco/genética
13.
Biofouling ; 36(2): 126-137, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32093497

RESUMO

Microbial biofilms are associated with persistent infections because of their high tolerance to antimicrobial agents and host defenses. The effects of centipede oil from Scolopendra subspinipes mutilans and its main components were investigated to identify non-toxic biofilm inhibitors. Centipede oil and linoleic acid at 20 µg ml-1 markedly inhibited biofilm formation by two fluconazole-resistant Candida albicans strains and three Staphylococcus aureus strains without affecting their planktonic cell growth. Also, both centipede oil and linoleic acid inhibited hyphal growth and cell aggregation by C. albicans. In addition, centipede oil and linoleic acid showed anti-biofilm activities against mixed C. albicans and S. aureus biofilms. Transcriptomic analysis showed that centipede oil and linoleic acid downregulated the expressions of several hypha/biofilm-related genes in C. albicans and α-hemolysin in S. aureus. Furthermore, both compounds effectively reduced C. albicans virulence in a nematode infection model with minimal toxicity.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Ácido Linoleico/farmacologia , Óleos Voláteis/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Animais , Anti-Infecciosos/toxicidade , Artrópodes/química , Biofilmes/crescimento & desenvolvimento , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/microbiologia , Candida albicans/crescimento & desenvolvimento , Candida albicans/patogenicidade , Hifas/crescimento & desenvolvimento , Ácido Linoleico/toxicidade , Testes de Sensibilidade Microbiana , Óleos Voláteis/toxicidade , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/patogenicidade , Virulência/efeitos dos fármacos
14.
Molecules ; 25(4)2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-32093422

RESUMO

Edible mushrooms are an important source of nutraceuticals and for the discovery of bioactive metabolites as pharmaceuticals. In this work, the OSMAC (One Strain, Many Active Compounds) approach was used to isolate two new compounds (1 and 2) along with seven known compounds (3-9) from a mycelial culture of a unique North American edible mushroom Hericium sp. The fruiting body was collected in Marine on St. Croix, Minnesota (USA), and mycelial cultures were grown on four different solid and liquid media. Extracts from the mycelial cultures were screened for antimicrobial activity and only the extract from the Cheerios substrate culture exhibited antifungal activity. Bioassay guided fractionation and HPLC analysis were used to isolate nine pure compounds and the structures of the known compounds were established by analysis of the NMR and mass spectrometry data and comparison to published reports. Compound 1 is a new erinacerin alkaloid and 2 is an aldehyde derivative of 4-hydroxy chroman. Four chlorinated orcinol derivatives (3-6), a pyran (7), erinaceolactone (8), and erinacine (9) were identified. Compound 4 showed antifungal activity against C. albicans and C. neoformans (MIC = 31.3-62.5 µg/mL, respectively). Compound 4 also inhibited biofilm formation of C. albicans and C. neoformans at 7.8 µg/mL. These results suggest that mycelial cultures of edible fungi may provide useful, bioactive compounds.


Assuntos
Agaricales/química , Antifúngicos , Candida albicans/crescimento & desenvolvimento , Micélio/química , Agaricales/crescimento & desenvolvimento , Antifúngicos/química , Antifúngicos/farmacologia , Biofilmes , Micélio/crescimento & desenvolvimento
15.
Int J Mol Sci ; 21(3)2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-32013007

RESUMO

Pathogenic yeasts Candida albicans and Candida parapsilosis possess a ß-type carbonic anhydrase Nce103p, which is involved in CO2 hydration and signaling. C. albicans lacking Nce103p cannot survive in low CO2 concentrations, e.g., in atmospheric growth conditions. Candida carbonic anhydrases are orthologous to the Saccharomyces cerevisiae enzyme, which had originally been detected as a substrate of a non-classical export pathway. However, experimental evidence on localization of C. albicans and C. parapsilosis carbonic anhydrases has not been reported to date. Immunogold labeling and electron microscopy used in the present study showed that carbonic anhydrases are localized in the cell wall and plasmatic membrane of both Candida species. This localization was confirmed by Western blot and mass spectrometry analyses of isolated cell wall and plasma membrane fractions. Further analysis of C. albicans and C. parapsilosis subcellular fractions revealed presence of carbonic anhydrases also in the cytosolic and mitochondrial fractions of Candida cells cultivated in shaken liquid cultures, under the atmospheric conditions.


Assuntos
Candida albicans/crescimento & desenvolvimento , Candida parapsilosis/crescimento & desenvolvimento , Anidrases Carbônicas/metabolismo , Técnicas de Cultura Celular por Lotes , Candida albicans/enzimologia , Candida parapsilosis/enzimologia , Membrana Celular/enzimologia , Parede Celular/enzimologia , Citosol/enzimologia , Proteínas Fúngicas/metabolismo , Espectrometria de Massas , Microscopia Eletrônica , Mitocôndrias/enzimologia
16.
Biofouling ; 36(1): 56-72, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31997658

RESUMO

The effects of two prominent copper oxide nanoparticles (CuO-NP and Cu2O-NP), with the oxidation state of Cu++ (cupric) and Cu+ (cuprous), on Candida albicans were evaluated. CuO-NP and Cu2O-NP were synthesized and characterized by XRD, FESEM, HR-TEM and Zeta potential. At sub-MIC (50 µg ml-1), both cupric and cuprous oxide NPs prevented yeast-to-hyphae switching and wrinkling behaviour in C. albicans. The mechanism for the antifungal action of the two NPs differed; CuO-NP significantly elicited reactive oxygen species, whereas membrane damage was more pronounced with Cu2O-NP. Real time PCR analysis revealed that CuO-NP suppressed the morphological switching of yeast-to-hyphae by down-regulating cph1, hst7 and ras1 and by up-regulation of the negative regulator tup1. In comparison, Cu2O-NP resulted in down-regulation of ras1 and up-regulation of the negative regulators nrg1 and tup1. Between the two NPs, CuO exhibited increased antifungal activity due to its stable oxidation state (Cu++) and its smaller dimensions compared with Cu2O-NP.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Cobre/farmacologia , Hifas/efeitos dos fármacos , Nanopartículas Metálicas/química , Candida albicans/crescimento & desenvolvimento , Candida albicans/metabolismo , Membrana Celular/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Testes de Sensibilidade Microbiana , Espécies Reativas de Oxigênio/metabolismo
17.
Biochim Biophys Acta Proteins Proteom ; 1868(3): 140135, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31964485

RESUMO

Heat shock proteins are best known for their role as chaperonins involved in general proteostasis, but they can also participate in specific cellular regulatory pathways, e.g. via their post-translational modification. Hsp70/Ssa1 is a central cytoplasmic chaperonin in eukaryotes, which also participates in cell cycle regulation via its phosphorylation at a specific residue. Here we analyze the role of Ssa1 phosphorylation in the morphogenesis of the fungus Candida albicans, a common human opportunistic pathogen. C. albicans can assume alternative yeast and hyphal (mold) morphologies, an ability that contributes to its virulence. We identified 11 phosphorylation sites on C. albicans Ssa1, of which 8 were only detected in the hyphal cells. Genetic analysis of these sites revealed allele-specific effects on growth or hyphae formation at 42 °C. Colony morphology, which is normally wrinkled or crenellated at 37 °C, reverted to smooth in several mutants, but this colony morphology phenotype was unrelated to cellular morphology. Two mutants exhibited a mild increase in sensitivity to the cell wall-active compounds caspofungin and calcofluor white. We suggest that this analysis could help direct screens for Ssa1-specific drugs to combat C. albicans virulence. The pleiotropic effects of many Ssa1 mutations are consistent with the large number of Ssa1 client proteins, whereas the lack of concordance between the phenotypes of the different alleles suggests that different sites on Ssa1 can affect interaction with specific classes of client proteins, and that modification of these sites can play cellular regulatory roles, consistent with the "chaperone code" hypothesis.


Assuntos
Candida albicans/citologia , Candida albicans/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Candida albicans/genética , Candida albicans/crescimento & desenvolvimento , Parede Celular/efeitos dos fármacos , Proteínas Fúngicas/química , Proteínas de Choque Térmico HSP70/química , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Morfogênese , Fosforilação
18.
Molecules ; 25(2)2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31941142

RESUMO

Reaction of 4-(1-adamantyl)-3-thiosemicarbazide (1) with numerous substituted acetophenones and benzaldehydes yielded the corresponding thiosemicarbazones containing adamantane skeletons. The synthesized compounds were evaluated for their in vitro activities against some Gram-positive and Gram-negative bacteria, and the fungus Candida albicans, and cytotoxicity against four cancer cell lines (Hep3B, HeLa, A549, and MCF-7). All of them showed good antifungal activity against Candida albicans. Compounds 2c, 2d, 2g, 2j and 3a, 3e, 3g displayed significant inhibitory activity against Enterococcus faecalis. Compounds 2a, 2e, 2h, 2k and 3j had moderate inhibitory potency against Staphylococcus aureus. Compounds 2a, 2e and 2g found so good inhibitory effect on Bacillus cereus. Compounds 2d and 2h, which contain (ortho) hydroxyl groups on the phenyl ring, were shown to be good candidates as potential agents for killing the tested cancer cell lines, i.e., Hep3B, A549, and MCF-7. Compounds 2a-c, 2f, 2g, 2j, 2k, 3g, and 3i were moderate inhibitors against MCF-7.


Assuntos
Adamantano/química , Antibacterianos , Antifúngicos , Antineoplásicos , Bactérias/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Neoplasias , Tiossemicarbazonas , Células A549 , Antibacterianos/síntese química , Antibacterianos/química , Antibacterianos/farmacologia , Antifúngicos/síntese química , Antifúngicos/química , Antifúngicos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Células HeLa , Humanos , Células MCF-7 , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Neoplasias/patologia , Tiossemicarbazonas/síntese química , Tiossemicarbazonas/química , Tiossemicarbazonas/farmacologia
19.
Sci Rep ; 10(1): 498, 2020 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-31949170

RESUMO

The incidence of resistant Candida isolates, especially Candida albicans, has increased continuously. To overcome the resistance, research on antifungal agent sensitizers has attracted considerable attention. Omeprazole and lansoprazole were found to inhibit the growth of sensitive C. albicans and hyphae formation in a high dose, respectively. This study aimed to determine the interactions of common clinically proton pump inhibitors (PPIs) and fluconazole both in vitro and in vivo and to further explore the possible mechanisms. In vitro, the tested PPIs all acted synergistically with fluconazole against both resistant C. albicans planktonic cells and biofilms preformed for ≤12 h with the minimum inhibitory concentration of fluconazole decreased from >512 µg/mL to 1-4 µg/mL. In vivo, PPIs plus fluconazole prolonged the survival rate of infected Galleria mellonella larvae by two-fold compared with that for the fluconazole monotherapy group and significantly reduced the tissue damage of infected larvae. Mechanism studies showed that PPIs significantly suppressed efflux pump activity, which is the common resistance mechanism of C. albicans, and significantly inhibited the virulence factors: phospholipase activity and morphology switching. These findings will provide new insights into antifungal agent discovery and potential approaches for the treatment of candidiasis caused by resistant C. albicans.


Assuntos
Candida albicans/efeitos dos fármacos , Farmacorresistência Fúngica/efeitos dos fármacos , Fluconazol/farmacologia , Inibidores da Bomba de Prótons/farmacologia , Animais , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Sinergismo Farmacológico , Holometábolos/crescimento & desenvolvimento , Holometábolos/parasitologia , Hifas/efeitos dos fármacos , Testes de Sensibilidade Microbiana
20.
Chem Biodivers ; 17(1): e1900462, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31788939

RESUMO

A series of coumarin-tagged ß-lactam triazole hybrids (10a-10o) were synthesized and tested for their cytotoxic activity against MDA-MB-231 (triple negative breast cancer), MCF-7 (estrogen receptor positive breast cancer (ER+)) and A549 (human lung carcinoma) cancer cell lines including one normal cell line, HEK-293 (human embryonic kidney). Two compounds 10b and 10d exhibited substantial cytotoxic effect against MCF-7 cancer cell lines with IC50 values of 53.55 and 58.62 µm, respectively. More importantly, compounds 10b and 10d were non-cytotoxic against HEK-293 cell lines. Structure-activity relationship (SAR) studies suggested that the nitro and chloro group at the C-3 position of phenyl ring are favorable for anticancer activity, particularly against MCF-7 cell lines. Furthermore, antimicrobial evaluation of these compounds revealed modest inhibition of examined pathogenic strains with compounds 10c and 10i being the most promising antimicrobial agents against Pseudomonas aeruginosa and Candida albicans, respectively.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Cumarínicos/farmacologia , Triazóis/farmacologia , beta-Lactamas/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Antifúngicos/síntese química , Antifúngicos/química , Antineoplásicos/síntese química , Antineoplásicos/química , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Proliferação de Células/efeitos dos fármacos , Cumarínicos/síntese química , Cumarínicos/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Células HEK293 , Humanos , Células MCF-7 , Testes de Sensibilidade Microbiana , Estrutura Molecular , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Relação Estrutura-Atividade , Triazóis/química , beta-Lactamas/química
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