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1.
Insect Biochem Mol Biol ; 116: 103258, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31678582

RESUMO

The protease inhibitors found in silkworm cocoons can be divided into several families, a majority of which contain serpin, TIL, or Kunitz domains. Previously, it has been reported that TIL-type protease inhibitors have antimicrobial activity. To date, however, it has not been determined whether the Kunitz-type protease inhibitor BmSPI51, the most abundant of cocoon protease inhibitors, plays an antimicrobial role. Thus, in this study, we sought to determine the biological role of BmSPI51 in silkworm cocoons. Our results obtained from real-time quantitative reverse transcription PCR and immunofluorescence analyses indicate that BmSPI51 is expressed exclusively in the silk glands during the larval fifth instar stage and is subsequently secreted into cocoon silk. Moreover, at a molar ratio of 1:1, BmSPI51 produced via prokaryotic expression exhibited inhibitory activity against trypsin and also proved to be highly stable over wide ranges of temperature and pH values. The expression of BmSPI51 was also found to be significantly upregulated in the larval fat body after infection with three species of fungi, namely, Candida albicans, Beauveria bassiana, and Saccharomyces cerevisiae. In vitro inhibition tests revealed that BmSPI51 significantly inhibited the sporular growth of all three of these fungal species. Further, results obtained from a binding assay showed that BmSPI51 binds to ß-d-glucan and mannan on the surface of fungal cells. In this study, we, thus, revealed the antimicrobial activity of BmSPI51 and its underlying mechanism in silkworm, thereby contributing to our present understanding of defense mechanisms in silkworm cocoons.


Assuntos
Bombyx/metabolismo , Bombyx/microbiologia , Proteínas de Insetos/genética , Inibidores de Serino Proteinase/genética , Animais , Antifúngicos/metabolismo , Beauveria/fisiologia , Bombyx/crescimento & desenvolvimento , Candida albicans/fisiologia , Corpo Adiposo/química , Proteínas de Insetos/metabolismo , Larva/crescimento & desenvolvimento , Larva/metabolismo , Larva/microbiologia , Saccharomyces cerevisiae/fisiologia , Inibidores de Serino Proteinase/metabolismo
2.
Microbiol Res ; 230: 126346, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31563763

RESUMO

In this study, we aimed to determine the interspecies interactions between Proteus mirabilis and Candida albicans. Mono and dual-species biofilms were grown in a microtiter plate and metabolomic analysis of the biofilms was performed. The effects of togetherness of two species on the expression levels of candidal virulence genes and urease and swarming activities of P.mirabilis were investigated. The growth of C.albicans was inhibited by P.mirabilis whereas the growth and swarming activity of P.mirabilis were increased by C.albicans. The inhibition of Candida cell growth was found to be biofilm specific. The alteration was not detected in urease activity. The expressions of EFG1, HWP1 and SAP2 genes were significantly down-regulated, however, LIP1 was upregulated by P.mirabilis. In the presence of P.mirabilis carbonhydrates, amino acids, polyamine and lipid metabolisms were altered in C.albicans. Interestingly, the putrescine level was increased up to 230 fold in dual-species biofilm compared to monospecies C.albicans biofilm. To our knowledge, this is the first study to investigate the impact of each microbial pathogen on the dual microbial environment by integration of metabolomic data.


Assuntos
Proteínas de Bactérias/metabolismo , Candida albicans/fisiologia , Proteus mirabilis/fisiologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Candida albicans/química , Candida albicans/genética , Candida albicans/crescimento & desenvolvimento , Metabolômica , Proteus mirabilis/química , Proteus mirabilis/genética , Proteus mirabilis/crescimento & desenvolvimento
3.
Carbohydr Polym ; 228: 115391, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31635733

RESUMO

Fungal biofilm formation is an emerging problem in a wide range of health-related applications. This study aims to design and synthesize amphiphilic quaternary ammonium chitosans (AQACs) that could bind onto fungal biofilms to kill adherent fungal cells, and establish their structural/fungicidal activity relationships. AQACs with different hydrophobic alkyl chain length (C4, C8, and C12) were synthesized by quaternization of 3-bromopropionic acid with the corresponding tertiary amines, followed by reacting with chitosan using the EDC/NHS chemistry. The new AQACs were soluble in water, yet formed self-aggregates in the solution with different sizes. In antifungal tests against free-floating Candida albicans, shorter alkyl chains (C4) in the AQACs resulted in the most potent fungicidal effect. However, in the treatment of Candida biofilms formed on solid surfaces, AQACs with longer alkyl chains (C8 and C12) were much more effective than their shorter chain counterpart (C4). The effects of alkyl chain self-aggregation on the opposite trend in fungicidal and anti-biofilm activities were discussed. All the AQACs showed excellent cytocompatibility with mammalian cells.


Assuntos
Antifúngicos/química , Biofilmes/efeitos dos fármacos , Quitosana/química , Compostos de Amônio Quaternário/química , Animais , Materiais Biocompatíveis/química , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Linhagem Celular , Conformação Molecular
4.
J Med Microbiol ; 68(12): 1802-1812, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31702539

RESUMO

Introduction. Candida albicans is responsible for several types of oral and systemic infections. In light of emerging resistance to antifungals, studies have demonstrated the antifungal effect of lactoferrin (LF), which is part of the innate immune system, has anticandidal activities.Methodology. C. albicans (2×106 c.f.u. ml-1) were incubated either with PBS or human LF (hLF) (100 µg ml-1) at 37 °C for 24 h and then RNA was isolated and virulence factors analysed. C. albicans (1×105 c.f.u.) was injected into the tail vein of immunocompromised wild-type and Ltf -/-. Then, 24 h later, the Ltf -/-I mice received hLF intravenously (100 µg g-1 body weight), while the control group received PBS. Then, 48 h later, the organs were collected, homogenized and C. albicans c.f.u.s were counted. In addition, the inflammatory mediators of kidneys and the virulence factors of C. albicans were analysed.Results. hLF-treated Ltf -/-I mice showed significant clearance of C. albicans in different organ tissues when compared to untreated Ltf -/-I mice. The inflammatory cytokines, such as IL-1ß, IL-6 , TNF-α and MPO and iNOS were downregulated in hLF-treated Ltf -/-I mice when compared to untreated Ltf -/-I mice. Whereas, IL-10 and IL-17A were upregulated at 72 h post infection when compared to Ltf -/-C mice. Histological analysis also revealed a significant decrease in the size and number of infectious foci in the hLF-treated groups. hLF treatment significantly downregulated several virulence factors of C. albicans both in vitro and in vivo.Conclusion. We concluded that hLF-treated Ltf -/- mice can reduce the severity of C. albicans-induced systemic infection.


Assuntos
Candidíase/tratamento farmacológico , Lactoferrina/uso terapêutico , Animais , Aderência Bacteriana , Candida albicans/fisiologia , Candidíase/imunologia , Candidíase/patologia , Citocinas/análise , Humanos , Rim/microbiologia , Rim/patologia , Lactoferrina/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
5.
BMC Complement Altern Med ; 19(1): 303, 2019 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-31703673

RESUMO

BACKGROUND: Candida albicans is an opportunistic pathogen that causes oral candidiasis and denture stomatitis. It has also been reported to infect oral mucositis lesions in patients who suffer from cancer affecting the head and neck and who receive chemotherapy and radiotherapy treatments. This study aimed to investigate the effects of two cinnamon bark fractions, i.e., an essential oil and an aqueous extract enriched in proanthocyanidins (Cinnulin PF®) on growth, biofilm formation, and adherence properties of C. albicans as well as on oral epithelial cells (barrier integrity, inflammatory response). METHODS: A microplate dilution assay was used to determine antifungal and anti-biofilm properties. A fluorescent assay was used to determine C. albicans adherence to oral epithelial cells. Cytotoxicity toward oral epithelial cells was assessed by determination of cell metabolic activity. Tight junction integrity of gingival keratinocytes was assessed by determination of transepithelial electrical resistance. IL-6 and IL-8 secretion by TNFα-stimulated oral epithelial cells was quantified by ELISA. RESULTS: While Cinnulin PF® did not reduce C. albicans growth, the cinnamon bark oil exhibited high antifungal activity with minimum inhibitory concentrations and minimum fungicidal concentrations in the range of 0.039 to 0.078%. The cinnamon oil was also active against a pre-formed C. albicans biofilm. Interestingly, Cinnulin PF® prevented biofilm formation by C. albicans and attenuated its adherence to oral epithelial cells. At their effective concentrations, the cinnamon oil and the Cinnulin PF® displayed no significant cytotoxicity against oral epithelial cells. In an in vitro model, both cinnamon fractions reinforced the integrity of the oral epithelial barrier. Lastly, Cinnulin PF® inhibited the secretion of interleukin-6 and interleukin-8 by oral epithelial cells stimulated with TNF-α. CONCLUSION: By their ability to attenuate growth, biofilm formation and adherence property of C. albicans, to reinforce the epithelial barrier function, and to exert anti-inflammatory properties the two cinnamon fractions (essential oil, Cinnulin PF®) investigated in the present study may be promising agents for treating oral infections involving C. albicans.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candidíase Bucal/microbiologia , Cinnamomum zeylanicum/química , Células Epiteliais/microbiologia , Boca/microbiologia , Óleos Voláteis/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candida albicans/fisiologia , Candidíase Bucal/tratamento farmacológico , Linhagem Celular , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Boca/metabolismo , Casca de Planta/química
6.
BMC Complement Altern Med ; 19(1): 308, 2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31718633

RESUMO

BACKGROUND: Dental caries is a biofilm-diet-dependent worldwide public health problem, and approaches against microorganisms in cariogenic biofilms are necessary. METHODS: The antimicrobial and antibiofilm activities of 12 Casearia sylvestris extracts (0.50 mg/mL) from different Brazilian biomes (Atlantic Forest, Cerrado, Caatinga, Pampa, and Pantanal) and varieties (sylvestris, lingua, and intermediate) were tested against two species found in cariogenic biofilms (Streptococcus mutans and Candida albicans). The extracts effective against S. mutans were used to evaluate the "adhesion strength" of this bacterium to the salivary pellicle and initial glucan matrix and the S. mutans-GtfB activity. Also, the antimicrobial activity against S. mutans of three fractions (methanol, ethyl acetate, and hexane; 0.25 mg/mL) from the extracts was evaluated. RESULTS: Three extracts from the Atlantic Forest variety sylvestris (FLO/SC, GUA/CE, PRE/SP) reduced ≥50% (> 3 logs) S. mutans viable population (p < 0.0001 vs. vehicle), while two extracts from the same biome and variety (PAC/CE, PRE/SP) decreased ≥50% of the viable counts of C. albicans (p < 0.0001 vs. vehicle). For S. mutans biofilms, three extracts (GUA/CE, PAC/CE, PRE/SP) reduced the biomass by ≥91% (p > 0.0001 vs. vehicle) and 100% of the microbial population (p < 0.0001 vs. vehicle). However, for the fungal biofilm, two extracts (PAC/CE, PRE/SP) reduced the viable counts by ≥52% (p < 0.0001 vs. vehicle), but none reduced biomass. The extracts with higher antimicrobial and antibiofilm activities presented higher content of clerodane-type diterpenes and lower content of glycosylated flavonoids than the less active extracts. The extracts had no effect on the removal of cells adhered to the pellicle (p > 0.05 vs. vehicle) while promoted the detachment of a larger number of S. mutans cells from GtfB-glucan matrix (p < 0.0031 vs. vehicle), and FLO/SC, GUA/CE and PRE/SP reduced the quantity of glucans (p < 0.0136 vs. vehicle). Only the ethyl acetate fractions reduced the microbial population of S. mutans (p < 0.0001 vs. vehicle), except for one (PAC/CE). Among the ethyl acetate fractions, three from var. lingua (two from Cerrado, and one from Cerrado/Caatinga) reduced ≥83% of the microbial population. CONCLUSIONS: C. sylvestris extracts from Atlantic Forest var. sylvestris and ethyl acetate fractions from Cerrado and Cerrado/Caatinga var. lingua may be used as a strategy against cariogenic microorganisms.


Assuntos
Anti-Infecciosos/farmacologia , Candida albicans/efeitos dos fármacos , Casearia/química , Cárie Dentária/microbiologia , Extratos Vegetais/farmacologia , Streptococcus mutans/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Brasil , Candida albicans/fisiologia , Ecossistema , Humanos , Testes de Sensibilidade Microbiana , Streptococcus mutans/fisiologia
7.
Adv Exp Med Biol ; 1197: 69-78, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31732935

RESUMO

Candida albicans is an opportunistic pathogen colonizing the oropharyngeal, esophageal, and gastrointestinal mucosa in most healthy humans. In immunocompromised hosts, this fungal organism can cause mucosal candidiasis in these sites. C. albicans also causes fungemia, a serious consequence of cancer cytotoxic chemotherapy, which is thought to develop from fungal translocation through compromised mucosal barriers. Changes in endogenous bacterial population size or composition as well as changes in the host environment can transform fungal commensals into opportunistic pathogens in the upper and lower GI tract. Pioneering studies from our group have shown that a ubiquitous oral commensal of the mitis streptococcal group (Streptococcus oralis) has a mutualistic relationship with C. albicans, with C. albicans enabling streptococcal biofilm growth at mucosal sites, and S. oralis facilitating invasion of the oral and esophageal mucosa by C. albicans. In these studies, we used a cortisone-induced immunosuppression mouse model. More recently, the development of a novel mouse chemotherapy model has allowed us to examine the interactions of C. albicans with the endogenous bacterial microbiota in the oral and small intestinal mucosa, two sites adversely affected by cytotoxic chemotherapy. In this model, oral inoculation with C. albicans causes severe dysbiosis in the mucosal bacterial composition in both sites. We also found that antibiotic treatment ameliorates invasion of the oral mucosa but aggravates dissemination through the intestinal mucosa. In this chapter, we discuss work from our laboratory and others examining the relationships of C. albicans with oral bacteria and their role in mucosal homeostasis or disease.


Assuntos
Candida albicans , Microbiota , Mucosa Bucal , Animais , Candida albicans/fisiologia , Candidíase/microbiologia , Modelos Animais de Doenças , Homeostase , Humanos , Camundongos , Microbiota/fisiologia , Mucosa Bucal/microbiologia , Streptococcus oralis
8.
Adv Exp Med Biol ; 1197: 119-141, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31732939

RESUMO

Oral cavity harbors a complex and highly diverse microbial community. Cross-kingdom interactions between Candida and oral bacteria are critical for their co-existence, which may also affect the course and the severity of biofilm-mediated bacterial-mediated diseases. C. albicans has been found in polymicrobial biofilms associated with denture stomatitis, oral mucositis, dental caries, periodontal diseases, peri-implantitis, and root canal infection. Thus, it is of utmost importance to unravel the mechanisms of Candida-bacterial interactions and their impact on the onset and severity of cross-kingdom biofilm-related diseases. Here, we highlight the potential role of Candida-bacterial biofilm interactions in the pathogenesis of oral diseases, especially mucosal infections and dental caries. The influence of Candida-bacterial biofilms on the mucosal host immune response is also discussed. Finally, we present some of the current and prospective therapeutic strategies for controlling these cross-kingdom interactions and their virulence properties associated with oral diseases.


Assuntos
Fenômenos Fisiológicos Bacterianos , Biofilmes , Candida , Interações Hospedeiro-Patógeno , Doenças da Boca , Candida/fisiologia , Candida albicans/fisiologia , Humanos , Doenças da Boca/microbiologia , Estudos Prospectivos
9.
Pak J Pharm Sci ; 32(4): 1519-1528, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31608870

RESUMO

In order to enhance essential oil's stability and water insolubility, Massoia aromatica oil nanoemulsion was formulated and tested on the planktonic growth and biofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans; macrophage phagocytosis and on Vero cells viability. Oil in water nanoemulsion formula was optimized by using several solvents and co-solvents composition. The stability test of the formula was conducted by using a six cycle's freeze-thaw technique. Particle size and morphology were analyzed using a particle size analyzer and transmission electron microscopy. Microbial growth, biofilm formation inhibition, and cytotoxicity assays were performed on the optimized formula by using micro dilution methods. Mice macrophage phagocytosis activities against latex and C. albicans in the presence of samples were evaluated. Massoia nanoemulsion was obtained as a transparent yellowish emulsion having 99.6-99.9% of transmittance; physically and chemically stable; showed stronger antibacterial and antibiofilm on P. aeruginosa and S. aureus, moderate to C. albicans; no significant different on phagocytic activities. The IC50 of massoia oil nanoemulsion and massoia oil towards Vero cells were 35.9µg/mL and 107.5µg/mL respectively. Massoia oil nanoemulsion can protect the stability and decreases the hydrophobicity of the oil, conserve the antimicrobial and immunomodulatory activities, but increases its cytotoxicity.


Assuntos
Anti-Infecciosos/farmacologia , Cryptocarya/química , Emulsões/toxicidade , Óleos Vegetais/farmacologia , Animais , Anti-Infecciosos/química , Anti-Infecciosos/toxicidade , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Emulsões/química , Macrófagos Peritoneais/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Nanoestruturas/química , Nanoestruturas/toxicidade , Tamanho da Partícula , Fagocitose/efeitos dos fármacos , Óleos Vegetais/química , Óleos Vegetais/toxicidade , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Testes de Toxicidade , Células Vero
10.
J Appl Oral Sci ; 27: e20180593, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31508792

RESUMO

There is growing evidence that C. albicans is associated with dental caries, but its role on caries development needs to be better clarified. Label="OBJECTIVE">To evaluate at the hard tissue level the effect of C. albicans on the cariogenic potential of S. mutans biofilms focusing on the mineral profile of induced carious lesions. This study also aimed to evaluate the effect of C. albicans on the acidogenic potential of S. mutans biofilms. METHODOLOGY Dual-species (CA+SM) and single-species biofilms (CA or SM) were grown on the surface of enamel slabs in the presence of glucose/sucrose supplemented culture medium for 24, 48 and 72 hours. Demineralization was evaluated through percentage of surface microhardness change (%SMC) and transversal microradiography analysis (ILM and LD) and pH of the spent medium was recorded daily. Data were analyzed by two-way ANOVA followed by Bonferroni correction. RESULTS%SMC was statistically different among the biofilms at each time point being the highest for SM biofilms and the lowest for CA biofilms which also differed from CA+SM biofilms [SM (24 h: 47.0±7.3; 48 h: 66.3±8.3; 72 h: 75.4±3.9); CA (24 h: 7.3±3.3; 48 h: 7.1±6.4; 72 h: 6.6±3.6); CA+SM (24 h: 35.9±7.39.1; 48 h: 47.2±9.5; 72 h: 47.6±9.5)]. pH of spent medium was statistically lower for SM biofilms compared to the other biofilms at each time point and remained constant over time while pH values increased from 24 to 72 h for both CA and CA+SM biofilms [SM (24 h: 4.4±0.1; 48 h: 4.4±0.1; 72 h: 4.5±0.1); CA (24 h: 6.9±0.3; 48 h: 7.2±0.2; 72 h: 7.5±0.2); CA+MS (24 h: 4.7±0.2; 48 h: 5.1±0.1; 72 h: 6.1±0.6)]. IML and LD for SM biofilms increased over time while no difference was observed from 24 to 72 h for the other biofilms. CONCLUSIONS The present data suggest that C. albicans has low enamel demineralization potential and the presence of C. albicans can reduce both the cariogenic and acidogenic potentials of S. mutans biofilms.


Assuntos
Biofilmes/crescimento & desenvolvimento , Candida albicans/fisiologia , Esmalte Dentário/microbiologia , Streptococcus mutans/metabolismo , Desmineralização do Dente/microbiologia , Ácidos/metabolismo , Animais , Bovinos , Contagem de Colônia Microbiana , Esmalte Dentário/química , Testes de Dureza , Concentração de Íons de Hidrogênio , Microrradiografia/métodos , Valores de Referência , Propriedades de Superfície , Fatores de Tempo
11.
J Mater Sci Mater Med ; 30(9): 108, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31535222

RESUMO

Biological effects of titanium (Ti) alloys were analyzed on biofilms of Candida albicans, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans, and Streptococcus sanguinis, as well as on osteoblast-like cells (MG63) and murine macrophages (RAW 264.7). Standard samples composed of aluminum and vanadium (Ti-6Al-4V), and sample containing niobium (Ti-35Nb) and zirconium (Ti-13Nb-13Zr) were analyzed. Monomicrobial biofilms were formed on the Ti alloys. MG63 cells were grown with the alloys and the biocompatibility (MTT), total protein (TP) level, alkaline phosphatase (ALP) activity, and mineralization nodules (MN) formation were verified. Levels of interleukins (IL-1ß and IL-17), tumor necrosis factor alpha (TNF-α), and oxide nitric (NO) were checked, from RAW 264.7 cells supernatants. Data were statically analyzed by one-way analysis of variance (ANOVA) and Tukey's test, or T-test (P ≤ 0.05). Concerning the biofilm formation, Ti-13Nb-13Zr alloy showed the best inhibitory effect on E. faecalis, P. aeruginosa, and S. aureus. And, it also acted similarly to the Ti-6Al-4V alloy on C. albicans and Streptococcus spp. Both alloys were biocompatible and similar to the Ti-6Al-4V alloy. Additionally, Ti-13Nb-13Zr alloy was more effective for cell differentiation, as observed in the assays of ALP and MN. Regarding the stimulation for release of IL-1ß and TNF-α, Ti-35Nb and Ti-13Nb-13Zr alloys inhibited similarly the synthesis of these molecules. However, both alloys stimulated the production of IL-17. Additionally, all Ti alloys showed the same effect for NO generation. Thus, Ti-13Nb-13Zr alloy was the most effective for inhibition of biofilm formation, cell differentiation, and stimulation for release of immune mediators.


Assuntos
Ligas/farmacologia , Materiais Biocompatíveis/farmacologia , Biofilmes/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Titânio/farmacologia , Ligas/química , Animais , Materiais Biocompatíveis/química , Biofilmes/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Células Cultivadas , Teste de Materiais , Camundongos , Testes de Sensibilidade Microbiana , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Pseudomonas/efeitos dos fármacos , Pseudomonas/fisiologia , Células RAW 264.7 , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Streptococcus/efeitos dos fármacos , Streptococcus/fisiologia , Propriedades de Superfície , Titânio/química
12.
J Microbiol Biotechnol ; 29(11): 1806-1816, 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31546294

RESUMO

Candida albicans is an opportunistic fungus possessing multiple virulence factors controlling pathogenicity. Cell wall proteins are the most important among these factors, being the first elements contacting the host. Ddr48 is a cell wall protein consisting of 212 amino acids. A DDR48 haploinsufficient mutant strain was previously found necessary for proper oxidative stress response and drug resistance. In this study, we aimed to further elucidate the role of Ddr48 by performing additional phenotypic characterization assays. A combinatory proteomic and bioinformatics approach was also undertaken to determine differentially expressed cell wall proteins. Results showed that the mutant strain exhibited a 10% decrease in adhesion mirrored by a 20% decrease in biofilm formation, and slight sensitivity to menadione, diamide, and SDS. Both strains showed similar hyphae formation, virulence, temperature tolerance, and calcofluor white and Congo red sensitivities. Furthermore, a total of 8 and 10 proteins were identified exclusively in the wild-type strain grown under filamentous and nonfilamentous conditions respectively. Results included proteins responsible for superoxide stress resistance (Sod4 and Sod6), adhesion (Als3, Hyr4, Pmt1, and Utr2), biofilm formation (Hsp90, Ece1, Rim9, Ipp1, and Pra1) and cell wall integrity (Utr2 and Pga4). The lack of detection of these proteins in the mutant strain correlates with the observed phenotypes.


Assuntos
Candida albicans/fisiologia , Parede Celular/metabolismo , Proteínas Fúngicas/genética , Estresse Oxidativo/genética , Fatores de Virulência/genética , Aderência Bacteriana/genética , Biofilmes/crescimento & desenvolvimento , Candida albicans/genética , Candida albicans/metabolismo , Parede Celular/genética , Farmacorresistência Fúngica/genética , Proteínas Fúngicas/metabolismo , Hifas/genética , Hifas/metabolismo , Mutação , Fenótipo , Proteômica , Fatores de Virulência/metabolismo
13.
Chin J Nat Med ; 17(8): 616-623, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31472899

RESUMO

Loureirin A is a major active component of Draconis sanguis, a traditional Chinese medicine. This work aimed to investigate the activity of loureirin A against Candida albicans biofilms. 2, 3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT)reduction assay and scanning electron microscopy were used to investigate the anti-biofilm effect. Minimal inhibitory concentration testing and time-kill curve assay were used to evaluate fungicidal activity. Cell surface hydrophobicity (CSH) assay and hyphal formation experiment were respectively carried out to investigate adhesion and morphological transition, two virulence traits of C. albicans. Real-time RT-PCR was used to investigate gene expression. Galleria mellonella-C. albicans and Caenorhabditis elegans-C. albicans infection models were used to evaluate the in-vivo antifungal effect. Human umbilical vein endothelial cells and C. elegans nematodes were used to evaluate the toxicity ofloureirin A. Our data indicated that loureirin A had a significant effect on inhibiting C. albicans biofilms, decreasing CSH, and suppressing hyphal formation. Consistently, loureirin A down-regulated the expression of some adhesion-related genes and hypha/biofilm-related genes. Moreover, loureirin A prolonged the survival of Galleria mellonella and Caenorhabditis elegans in C. albicans infection models and exhibited low toxicity. Collectively, loureirin A inhibits fungal biofilms, and this effect may be associated with the suppression of pathogenic traits, adhesion and hyphal formation.


Assuntos
Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Chalconas/farmacologia , Animais , Biofilmes/crescimento & desenvolvimento , Caenorhabditis elegans , Candida albicans/genética , Candidíase/microbiologia , Adesão Celular/efeitos dos fármacos , Adesão Celular/genética , Células Cultivadas , Modelos Animais de Doenças , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Humanos , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Hifas/patogenicidade , Medicina Tradicional Chinesa , Testes de Sensibilidade Microbiana , Mariposas
14.
J Med Microbiol ; 68(10): 1479-1488, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31380734

RESUMO

Purpose. Fungal infections have increased in recent decades, with Candida albicans being the fourth most common aetiological agent of nosocomial infections. Disaccharide trehalose has been proposed as a target for the development of new antifungals. In C. albicans we have examined the susceptibility shown by two mutants deficient in trehalose biosynthesis, namely tps1Δ and tps2Δ, to amphotericin B (AmB) and micafungin (MF).Methodology. Minimum inhibitory concentrations (MICs) were calculated according to the Clinical and Laboratory Standards Institute (CLSI) criteria. Cell viability was assessed by cell counting. Intracellular reactive oxygen species (ROS) and the mitochondrial membrane potential were measured by flow cytometry, while the trehalose content and biofilm formation were determined by enzymatic assays.Results. While the tps1Δ mutant was highly sensitive to AmB exposure, its resistance to MF was similar to that of the wild-type. Notably, the opposite phenotype was recorded in the tps2Δ mutant. In turn, MF induced a significant level of endogenous ROS production in the parental SC5314 and tps2Δ cells, whereas the ROS formation in tps1Δ cells was virtually undetectable. The level of endogenous ROS correlated positively with the rise in mitochondrial activity. Only AmB was able to promote intracellular synthesis of trehalose in the parental strain; it was absent from tps1Δ cells and showed low levels in tps2Δ, confirming the unspecific dephosphorylation of trehalose-6P in C. albicans. Furthermore, the capacity of both tps1Δ and tps2Δ mutants to form biofilms was drastically reduced after AmB exposure, whereas it increased in tps1Δ cells treated with MF.Conclusion. Our data lend weight to the idea of using trehalose biosynthesis as a potential target for antifungal therapy.


Assuntos
Anfotericina B/farmacologia , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/enzimologia , Proteínas Fúngicas/genética , Glucosiltransferases/genética , Micafungina/farmacologia , Trealose/biossíntese , Biofilmes/efeitos dos fármacos , Candida albicans/genética , Candida albicans/fisiologia , Candidíase/microbiologia , Proteínas Fúngicas/metabolismo , Glucosiltransferases/metabolismo , Humanos , Testes de Sensibilidade Microbiana , Espécies Reativas de Oxigênio/metabolismo , Deleção de Sequência
15.
EMBO J ; 38(19): e101597, 2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31448850

RESUMO

Understanding how cellular activities impact genome stability is critical to multiple biological processes including tumorigenesis and reproductive biology. The fungal pathogen Candida albicans displays striking genome dynamics during its parasexual cycle as tetraploid cells, but not diploid cells, exhibit genome instability and reduce their ploidy when grown on a glucose-rich "pre-sporulation" medium. Here, we reveal that C. albicans tetraploid cells are metabolically hyperactive on this medium with higher rates of fermentation and oxidative respiration relative to diploid cells. This heightened metabolism results in elevated levels of reactive oxygen species (ROS), activation of the ROS-responsive transcription factor Cap1, and the formation of DNA double-strand breaks. Genetic or chemical suppression of ROS levels suppresses each of these phenotypes and also protects against genome instability. These studies reveal how endogenous metabolic processes can generate sufficient ROS to trigger genome instability in polyploid C. albicans cells. We also discuss potential parallels with metabolism-induced instability in cancer cells and speculate that ROS-induced DNA damage could have facilitated ploidy cycling prior to a conventional meiosis in eukaryotes.


Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Candida albicans/fisiologia , Proteínas de Ciclo Celular/genética , Dano ao DNA , Proteínas Fúngicas/genética , Instabilidade Genômica , Fermentação , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Genoma Fúngico , Metabolômica , Estresse Oxidativo , Poliploidia , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima
16.
Biomed Res Int ; 2019: 9130806, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31309119

RESUMO

In the article has been presented an analysis of susceptibility of selected dental materials, made in the CAD/CAM technology. The morphology and structural properties of selected dental materials and their composites were determined by using XRPD (X-ray powder diffraction) techniques, as well as the IR (infrared) spectroscopy. Moreover, an adhesion as well as development of biofilm by oral microorganisms has been studied. It has been shown that a degree of the biofilm development on the tested dental materials depended on microorganism genus and species. Streptococcus mutans has demonstrated the best adhesion to the tested materials in comparison with Candida albicans and Lactobacillus rhamnosus. However, the sintered materials such as IPS e.max® and the polished IPS e.max® have showed the best "anti-adhesive properties" in relation to S. mutans and L. rhamnosus that have not formed the biofilm on the polished IPS e.max® sample. Furthermore, S. mutans have not formed the biofilm on both surfaces. On the contrary to S. mutans and L. rhamnosus, C. albicans has demonstrated the adhesive properties in relation to the above-mentioned surfaces. Moreover, in contrast to S. mutans and C. albicans, L. rhamnosus has not formed the biofilm on the polished IPS Empress material.


Assuntos
Biofilmes/crescimento & desenvolvimento , Candida albicans/fisiologia , Projeto Auxiliado por Computador , Porcelana Dentária/química , Lactobacillus rhamnosus/fisiologia , Teste de Materiais , Streptococcus mutans/fisiologia , Propriedades de Superfície
17.
Cells ; 8(7)2019 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336838

RESUMO

The presence of Candida albicans in the biofilm underlying the dental prosthesis is related to denture stomatitis (DS), an inflammatory reaction of the oral mucosa. The oral epithelium, a component of the innate immune response, has the ability to react to fungal invasion. In this study, we evaluated the in vitro effect of viable C. albicans on the apoptosis, nitric oxide (NO) production, and ß-defensin 2 (hBD-2) expression and production of human palate epithelial cells (HPECs). We further determined whether or not these effects were correlated with fungal invasion of epithelial cells. Interaction between HPEC primary culture and C. albicans was obtained through either direct or indirect cell-cell contact with a supernatant from a hyphal fungus. We found that the hyphae supernatants were sufficient to induce slight HPEC apoptosis, which occurred prior to the activation of the specific mechanisms of epithelial defense. The epithelial defense responses were found to occur via NO and antimicrobial peptide hBD-2 production only during direct contact between C. albicans and HPECs and coincided with the fungus's intraepithelial invasion. However, although the hBD-2 levels remained constant in the HPEC supernatants over time, the NO release and hBD-2 gene expression were reduced at a later time (10 h), indicating that the epithelial defense capacity against the fungal invasion was not maintained in later phases. This aspect of the immune response was associated with increased epithelial invasion and apoptosis maintenance.


Assuntos
Fibroblastos , Queratinócitos , Mucosa Bucal , Óxido Nítrico/metabolismo , Palato , beta-Defensinas/metabolismo , Biofilmes , Candida albicans/fisiologia , Candidíase/imunologia , Candidíase/microbiologia , Linhagem Celular , Fibroblastos/citologia , Fibroblastos/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Queratinócitos/citologia , Queratinócitos/metabolismo , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Palato/citologia , Palato/metabolismo
18.
Phytomedicine ; 63: 153033, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31352284

RESUMO

BACKGROUND: The emergence of antibiotic resistant microorganisms presents a worldwide problem that requires novel antibiotic and non-antibiotic strategies, and biofilm formation is a mechanism of drug resistance utilized by diverse microorganisms. The majority of microorganisms live in biofilms that help their survival against starvation, antimicrobial agents, and immunological defense systems. Therefore, it is important novel compounds be identified that inhibit biofilm formation and cell survival without drug resistance. STUDY DESIGN: In this study, the antimicrobial and antibiofilm activities of five prenylated flavanones (Okinawan propolins) isolated from fruits of Macaranga tanarius (L.) were investigated against 14 microorganisms including 10 pathogens. RESULTS: Of these five propolins, propolin D at 5-10 µg/ml significantly inhibited biofilm formation by three Staphylococcus aureus strains, a Staphylococcus epidermidis strain, and a Candida albicans with MICs from 10 to 50 µg/ml, and in C. albicans, propolin D was found to inhibit biofilm formation by reducing cell aggregation and downregulated the expressions of hypha/biofilm-related genes including ECE1 and HWP1. Interestingly, at sub-MIC concentrations (10-50 µg/ml), propolin D significantly inhibited biofilm formation by enterohemorrhagic E. coli O157:H7, uropathogenic E. coli O6:H1, and Acinetobacter baumannii without affecting planktonic cell growth, but did not inhibit biofilm formation by a commensal E. coli K-12 strain, three probiotic Lactobacillus plantarum strains, or two Pseudomonas aeruginosa strains. And, propolin D reduced fimbriae production by E. coli O157:H7 and repressed gene expression of curli fimbriae genes (csgA and csgB). Also, propolin D was minimally toxic in a Caenorhabditis elegans nematode model. CONCLUSION: These findings show that prenylated flavanones, especially propolin D from Macaranga tanarius (Okinawan propolis), should be considered potential candidates for the development of non-toxic antibacterial and antifungal agents against persistent microorganisms.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Euphorbiaceae/química , Flavanonas/farmacologia , Flavonoides/farmacologia , Animais , Antibacterianos/química , Antibacterianos/toxicidade , Antifúngicos/química , Antifúngicos/toxicidade , Biofilmes/efeitos dos fármacos , Caenorhabditis elegans/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Avaliação Pré-Clínica de Medicamentos , Escherichia coli O157/efeitos dos fármacos , Flavanonas/química , Flavanonas/toxicidade , Flavonoides/química , Flavonoides/toxicidade , Testes de Sensibilidade Microbiana , Prenilação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Testes de Toxicidade
19.
BMC Res Notes ; 12(1): 383, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31287001

RESUMO

OBJECTIVE: Chitosan nanoparticle (nanochitosan) has a broad antimicrobial spectrum against diverse pathogenic microorganisms. However, its effect on dental caries-associated microorganisms, such as Streptococcus mutans and Candida albicans is yet to be explored. These microorganisms are known for causing early childhood caries. Therefore, this study was aimed at investigating nanochitosan inhibition capacity against dual-species biofilms of S. mutans and C. albicans. In this study, nanochitosan antimicrobial activity is reported against mono and dual biofilm species of S. mutans and/or C. albicans at 3 and 18 h incubation time. Nanochitosan inhibition capacity was observed through biofilm mass quantity and cell viability. RESULTS: The present study successfully synthesized nanochitosan with average diameter of approximately 20-30 nm, and also established dual-species biofilms of S. mutans and C. albicans in vitro. With nanochitosan treatment, the cell viability of both microorganisms significantly decreased with the increasing concentration of nanochitosan. There was no significant decrease in biofilm mass both in the dual and single-species biofilms after 3 h of incubation. However, greater inhibition of biofilm was observed at 18 h incubation.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Quitosana/química , Nanopartículas/química , Streptococcus mutans/efeitos dos fármacos , Anti-Infecciosos/química , Biofilmes/crescimento & desenvolvimento , Candida albicans/fisiologia , Criança , Cárie Dentária/tratamento farmacológico , Cárie Dentária/microbiologia , Placa Dentária/tratamento farmacológico , Placa Dentária/microbiologia , Humanos , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Transmissão , Nanopartículas/ultraestrutura , Tamanho da Partícula , Streptococcus mutans/fisiologia , Fatores de Tempo
20.
Biomed Res Int ; 2019: 1851740, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31275963

RESUMO

The human opportunistic fungal pathogen Candida albicans causes a severe health burden while the biofilms formed by C. albicans present a kind of infections that are hard to cure, highlighting the pressing need for new antifungal drugs against C. albicans. This study was to explore the antifungal activities of lycorine hydrochloride (LH) against C. albicans. The minimal inhibitory concentration (MIC) of LH against C. albicans SC5314 was 64 µM. Below its MIC, LH demonstrated antivirulence property by suppressing adhesion, filamentation, biofilm formation, and development, as well as the production of extracellular phospholipase and exopolymeric substances (EPS). The cytotoxicity of LH against mammalian cells was low, with half maximal inhibitory concentrations (IC50) above 256 µM. Moreover, LH showed a synergistic effect with AmB, although its interaction with fluconazole, as well as caspofungin, was indifferent. Thus, our study reports the potential use of LH, alone or in combination with current antifungal drugs, to fight C. albicans infections.


Assuntos
Alcaloides de Amaryllidaceae/farmacologia , Candida albicans/patogenicidade , Fenantridinas/farmacologia , Adesividade/efeitos dos fármacos , Alcaloides de Amaryllidaceae/química , Alcaloides de Amaryllidaceae/toxicidade , Antifúngicos/química , Antifúngicos/farmacologia , Antifúngicos/toxicidade , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Morte Celular/efeitos dos fármacos , AMP Cíclico/metabolismo , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Hifas/efeitos dos fármacos , Fenantridinas/química , Fenantridinas/toxicidade , Fosfolipases/metabolismo , Virulência/efeitos dos fármacos
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