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1.
Int J Nanomedicine ; 18: 3231-3246, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37337577

RESUMO

Purpose: Magnetic separation of microbes can be an effective tool for pathogen identification and diagnostic applications to reduce the time needed for sample preparation. After peptide functionalization of superparamagnetic iron oxide nanoparticles (SPIONs) with an appropriate interface, they can be used for the separation of sepsis-associated yeasts like Candida albicans. Due to their magnetic properties, the magnetic extraction of the particles in the presence of an external magnetic field ensures the accumulation of the targeted yeast. Materials and Methods: In this study, we used SPIONs coated with 3-aminopropyltriethoxysilane (APTES) and functionalized with a peptide originating from GP340 (SPION-APTES-Pep). For the first time, we investigate whether this system is suitable for the separation and enrichment of Candida albicans, we investigated its physicochemical properties and by thermogravimetric analysis we determined the amount of peptide on the SPIONs. Further, the toxicological profile was evaluated by recording cell cycle and DNA degradation. The separation efficiency was investigated using Candida albicans in different experimental settings, and regrowth experiments were carried out to show the use of SPION-APTES-Pep as a sample preparation method for the identification of fungal infections. Conclusion: SPION-APTES-Pep can magnetically remove more than 80% of the microorganism and with a high selective host-pathogen distinction Candida albicans from water-based media and about 55% in blood after 8 minutes processing without compromising effects on the cell cycle of human blood cells. Moreover, the separated fungal cells could be regrown without any restrictions.


Assuntos
Candida albicans , Nanopartículas Magnéticas de Óxido de Ferro , Proteínas e Peptídeos Salivares , Humanos , Candida albicans/isolamento & purificação , Fenômenos Magnéticos
2.
Proc Natl Acad Sci U S A ; 119(40): e2209607119, 2022 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-36161889

RESUMO

Blood stream infections (BSIs) cause high mortality, and their rapid detection remains a significant diagnostic challenge. Timely and informed administration of antibiotics can significantly improve patient outcomes. However, blood culture, which takes up to 5 d for a negative result, followed by PCR remains the gold standard in diagnosing BSI. Here, we introduce a new approach to blood-based diagnostics where large blood volumes can be rapidly dried, resulting in inactivation of the inhibitory components in blood. Further thermal treatments then generate a physical microscale and nanoscale fluidic network inside the dried matrix to allow access to target nucleic acid. The amplification enzymes and primers initiate the reaction within the dried blood matrix through these networks, precluding any need for conventional nucleic acid purification. High heme background is confined to the solid phase, while amplicons are enriched in the clear supernatant (liquid phase), giving fluorescence change comparable to purified DNA reactions. We demonstrate single-molecule sensitivity using a loop-mediated isothermal amplification reaction in our platform and detect a broad spectrum of pathogens, including gram-positive methicillin-resistant and methicillin-susceptible Staphylococcus aureus bacteria, gram-negative Escherichia coli bacteria, and Candida albicans (fungus) from whole blood with a limit of detection (LOD) of 1.2 colony-forming units (CFU)/mL from 0.8 to 1 mL of starting blood volume. We validated our assay using 63 clinical samples (100% sensitivity and specificity) and significantly reduced sample-to-result time from over 20 h to <2.5 h. The reduction in instrumentation complexity and costs compared to blood culture and alternate molecular diagnostic platforms can have broad applications in healthcare systems in developed world and resource-limited settings.


Assuntos
DNA Bacteriano , DNA Fúngico , Teste em Amostras de Sangue Seco , Reação em Cadeia da Polimerase , Sepse , Antibacterianos/farmacologia , Candida albicans/genética , Candida albicans/isolamento & purificação , DNA Bacteriano/sangue , DNA Fúngico/sangue , Teste em Amostras de Sangue Seco/métodos , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Heme/química , Humanos , Limite de Detecção , Meticilina/farmacologia , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Sepse/sangue , Sepse/diagnóstico , Sepse/microbiologia , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Células-Tronco
3.
Bol. micol. (Valparaiso En linea) ; 37(1): 2-8, jun. 2022. ilus, tab
Artigo em Espanhol | LILACS | ID: biblio-1396937

RESUMO

Introducción: C. albicans es reconocida como la especie más virulenta del género y representa la causa más frecuente de candidiasis en humanos. A nivel taxonómico, C.albicans se clasifica como un complejo de especies estrechamente relacionadas que incluye a C. albicans sensu stricto (s.s), C. dubliniensis y C. africana. Objetivo: identificar las especies del complejo C. albicans aisladas desde distintas muestras de pacientes de la quinta región de Valparaíso. Materiales y método: Se identificaron 103 cepas del complejo C. albicans, aisladas desde muestras superficiales y profundas durante el año 2020. La identificación se realizó en base a morfofisiología y la amplificación del gen HWP1. Resultados: Se identificaron 100 cepas como C. albicans s.s, 2 como C. dubliniensis y 1 como C. africana. Dentro de las cepas identificadas como C. albicans s.s se observaron cuatro patrones de tamaños de fragmentos genéticos. Conclusiones: C. albicans s.s fue la especie más frecuente y en base al genotipo de HPW1 se describen cuatro patrones ( H1 a H4). (AU)


Introduction: C. albicans is recognized as the most virulent species of the genus and represents the major cause of candidiasis in humans. At the taxonomic level, C. albicansis classified as a complex of closely related species that includes C. albicans sensu stricto (s.s), C. dubliniensis, and C. africana. Objective: to identify the species of the C. albicans complex isolated from different samples of patients from the fifth region of Valparaíso. Materials and method: 103 strains of the C. albicans complex were identified, isolated from superficial and deep samples during the year 2020. The identification was carried out based on morphophysiology and the amplification of the HWP1 gene. Results: 100 strains were identified as C. albicans s.s, 2 as C. dubliniensis and 1 as C. africana. Within the strains identified as C. albicans s.s, 4 patterns of fragment sizes were observed. Conclusions: C. albicans s.s was the most frequent species and based on the HPW1 genotype, four patterns are described (H1 to H4).(AU)


Assuntos
Humanos , Candida albicans/isolamento & purificação , Candida albicans/genética , Candida albicans/classificação , Chile , Estudos Prospectivos , Genótipo
4.
Stomatologiia (Mosk) ; 101(3): 18-21, 2022.
Artigo em Russo | MEDLINE | ID: mdl-35640174

RESUMO

THE AIM OF THE STUDY: The study by the method of tissue polymerase chain reaction of the species composition of the microbiota of lesions of the oral mucosa in patients with bullous lesions. MATERIAL AND METHODS: Biopsy specimens of the oral mucosa of 51 patients were studied by the polymerase chain reaction method, of which 14 patients with pemphigus vulgaris, 17 patients with pemphigoid bullosa, and 20 patients with the bullous form of ruber lichen planus. 4 types of microorganisms have been identified - Fusobacterium, Streptococcus pneumoniae, Candida albicans, Ureaplasma spp. and viruses - Human Papillomavirus 16, Epstein-Barr virus and Citomegalovirus. RESULTS: In the study of the microbiota of bullous lesions, associations of microorganisms and viruses were established in a significant number of cases. Associations of Str.pneumoniae and C. albicans were quite common in patients with pemphigus vulgaris in 26.3%, pemphigoid bullosa in 20.0%, and in patients with the bullous form of ruber lichen planus in 14.3% of cases. In patients with pemphigus vulgaris, the association of Str.pneumoniae, C. albicans and EBV was noted in 31.6% of cases. In patients with the bullous form of ruber lichen planus in a high percentage of cases (28.6%), the associations of Str. pneumoniae, EBV and CMV. CONCLUSION: Identification at earlier stages of management of patients with bullous lesions Str. pneumoniae, Candida albicans, and Fusobacterium associated with herpes viruses should be regarded as one of the triggering mechanisms of an autoimmune conflict, which subsequently causes a specific clinical picture of these diseases.


Assuntos
Microbiota , Mucosa Bucal/patologia , Vesícula/microbiologia , Vesícula/virologia , Candida albicans/isolamento & purificação , Infecções por Vírus Epstein-Barr/complicações , Infecções por Vírus Epstein-Barr/patologia , Fusobacterium/isolamento & purificação , Herpesvirus Humano 4/isolamento & purificação , Humanos , Líquen Plano/complicações , Líquen Plano/patologia , Mucosa Bucal/microbiologia , Mucosa Bucal/virologia , Penfigoide Bolhoso/patologia , Pênfigo/complicações , Pênfigo/patologia , Streptococcus pneumoniae/isolamento & purificação
5.
J Clin Lab Anal ; 36(2): e24208, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34997991

RESUMO

INTRODUCTION: Resistance to azole drugs has been observed in candidiasis due to their long-term use and poor response to treatment. Resistance to azole drugs in Candida albicans isolates is controlled by several genes including ERG11, CDR1, CDR2, and MDR1. In this study, the expression of the mentioned genes was evaluated in C. albicans isolates susceptible and resistant to fluconazole. METHODS: After identifying the Candida isolates using morphological and molecular methods, the minimum inhibitory concentration (MIC) and drug susceptibility were determined using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) method. RNA was then extracted and cDNA was synthesized from 24 C. albicans isolates from patients with cancer. Then, the mean expressions of these genes were compared in two groups using real-time polymerase chain reaction (RT-PCR). RESULTS: A total of 74 Candida isolates were obtained from the oral cavity of 61 cancer patients with oral candidiasis. After 24 h, 21.6% of the isolates were fluconazole-resistant, 10.8% were identified as dose-dependent, and the rest of the isolates (67.6%) were fluconazole-sensitive. The mean expressions of the CDR1 and MDR1 genes were significantly higher in the resistant isolates than in the sensitive ones. However, the ERG11 and CDR2 genes were not significantly increased in the resistant isolates. CONCLUSION: The increased mean expressions of the CDR1 and MDR1 genes had a greater effect on fluconazole resistance among the drug-resistant strains of C. albicans in chemotherapy patients. It seemed that the accumulation of chemotherapeutic drugs in this organism stimulated some regulatory factors and increased the expression of these two genes and ultimately helped to further increase their expression and resistance to fluconazole.


Assuntos
Candida albicans/genética , Candidíase Bucal/metabolismo , Farmacorresistência Fúngica/genética , Fluconazol/farmacologia , Proteínas Fúngicas/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candidíase Bucal/etiologia , Proteínas Fúngicas/metabolismo , Expressão Gênica , Humanos , Irã (Geográfico) , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Neoplasias/complicações , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Esterol 14-Desmetilase/genética , Esterol 14-Desmetilase/metabolismo
6.
Med Mycol ; 60(1)2021 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-34718724

RESUMO

Whole genome sequencing has been extensively used to describe infection outbreaks, although with limited application on Candida albicans and Candida parapsilosis.We retrospectively studied all patients admitted to the neonatal care unit diagnosed with candidemia caused by C. albicans (n = 46) or C. parapsilosis (n = 31) between 2007 and 2010 (Period 1) and 2011 and 2014 (Period 2). All isolates were genotyped by microsatellite markers. A cluster was defined as a group of ≥ 2 patients infected by strains with identical genotypes. For the validation of microsatellite markers and outbreak investigation, phylogenetic analyses and whole genome pairwise strain comparisons were performed.The number of episodes was significantly higher in Period 1 than in Period 2 (51 vs 32; P = 0.003); the reduction in the number of cases coincided with the educational campaign for catheter care implementation in 2011. Overall, eight genotypes were clusters involving 29 patients. All C. albicans (n = 5) and C. parapsilosis (n = 3) clusters were found during Period 1 before the educational campaign. No statistically significant differences were found between the percentage of C. albicans and C. parapsilosis clusters, but the percentage of patients associated to the clusters was significantly higher for C. parapsilosis clusters in comparison to C. albicans clusters (52 vs 28.2%; P = 0.03). Whole genome sequencing confirmed microsatellite-defined clusters with high bootstrap values.Whole genome sequences confirmed microsatellite-defined clusters, corroborating the presence of outbreaks. Persistent or sporadic Candida clusters causing candidemia in neonates disappeared after the implementation of catheter care educational campaigns. LAY SUMMARY: We retrospectively studied all patients admitted to the neonatal care unit diagnosed with candidemia caused by C. albicans or C. parapsilosis. Reliable whole genome sequences confirmed microsatellite-defined clusters, corroborating the presence of outbreaks before educational campaigns for catheter care.


Assuntos
Candida albicans/isolamento & purificação , Candida parapsilosis/isolamento & purificação , Candidemia/diagnóstico , Sequenciamento Completo do Genoma/métodos , Candida albicans/genética , Candida parapsilosis/genética , Candidemia/epidemiologia , Células Clonais , Surtos de Doenças , Feminino , Genótipo , Humanos , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Masculino , Repetições de Microssatélites , Estudos Retrospectivos
7.
Anal Biochem ; 633: 114428, 2021 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-34678249

RESUMO

Owing to modern lifestyles and increasing amounts of medical intervention, clinical infections caused by conditionally pathogenic fungi are becoming increasingly serious. Among these, Candida albicans is the most common. Therefore, the rapid and accurate detection of this pathogenic fungus is important to guiding the selection of clinical therapeutic agents. Recombinase polymerase amplification (RPA) combined with lateral flow strips (LFS) is a promising molecular detection method with the advantages of rapidity, simplicity of operation and high sensitivity. However, this simplicity brings with it the inherent and non-negligible risk of false-positive signals from primer-dimers. In this study, primer-dependent artifacts were eliminated by using probes in the RPA reaction, introducing specific base substitutions to the primer and probe sequences and analyzing and screening the formation of primer-probe complexes. These measures were rigorously tested for efficacy, leading to the creation of an improved RPA-LFS system. The standardized method enabled the specific detection of C. albicans within 25 min at 37 °C without interference. The system had a detection limit of 1 CFU per reaction without DNA purification or 102 fg genomic DNA/50 µL. The detection sensitivity was not affected by the presence of other fungal DNA. The RPA-LFS method can therefore be used to detect clinical samples, and the results are accurate and consistent in comparison with those obtained using quantitative PCR. This study provides a paradigm for eliminating the risk of false-positive primer dimers in isothermal amplification assays and establishes a simple and easy method for the detection of C. albicans.


Assuntos
Candida albicans/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico , Fitas Reagentes/química , Recombinases/genética , Reação em Cadeia da Polimerase em Tempo Real
8.
Exp Eye Res ; 212: 108788, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34637791

RESUMO

This study aimed to investigate the efficacy of intravitreal povidone-iodine (PI) administration for the treatment of Candida albicans endophthalmitis. Forty New Zealand white rabbits were divided into four groups (n = 10 per group). After the induction of endophthalmitis using Candida albicans, groups A, B, and C received single intravitreal injections of 0.035 mg voriconazole, 0.3 mg PI, and their combination, respectively. Rabbits that were administered sham injections were in group D as controls. Fundus photography, vitreous culture, electroretinography (ERG), and histologic examinations of the retina were conducted on day 7. The anterior chamber flare (grade 0 to 4), severity of iritis (grade 0 to 4), and vitreous opacity (grade 0 to 3) were scored. Candida albicans was cultured in the vitreous sample. On day 7, the vitreous opacities were found in all groups. Compared to that in group D, groups A, B, and C showed a lower score for flare (p < 0.001) and iritis (p < 0.001) and less fungal growth in the vitreous culture (n = 2, 1, 1, and 10 in groups A, B, C, and D, respectively; p < 0.001). Furthermore, ERG and histologic findings demonstrated less affected a- and b-waves and damaged retinal tissues in groups A, B, and C. However, these findings were not different among groups A, B, and C. PI significantly improved Candida albicans endophthalmitis, and the effect was comparable that of the voriconazole, although some vitreous opacities remained. No synergistic effect of the combination of PI and voriconazole was observed. Intravitreal PI may be useful to treat Candida albicans endophthalmitis.


Assuntos
Candida albicans/isolamento & purificação , Candidíase/tratamento farmacológico , Endoftalmite/tratamento farmacológico , Infecções Oculares Fúngicas/tratamento farmacológico , Povidona-Iodo/administração & dosagem , Animais , Anti-Infecciosos Locais/administração & dosagem , Candidíase/microbiologia , Modelos Animais de Doenças , Endoftalmite/microbiologia , Infecções Oculares Fúngicas/microbiologia , Feminino , Injeções Intravítreas , Coelhos
9.
Diagn Microbiol Infect Dis ; 101(4): 115533, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34587571

RESUMO

In the present report, we describe an unusual case of mixed infection of Candida albicans and Talaromyces marneffei in the oral cavity and oropharynx with cutaneous involvement. On the CHROMagar Candida plate, green colonies (identified as C. albicans) and tiny violet colonies (identified as T. marneffei) grew from the throat swab after incubation for 96 hours. 10 clinical isolates of T. marneffei were used to verify their color production on CHROMagar Candida. All colonies were violet on the fourth, seventh and ninth day incubated at 37 °C. T. marneffei appears violet on the CHROMagar Candida plate, but it may be easily ignored because of its slow growth and small colony size, especially after incubation for 48 hours. Therefore, when using CHROMagar Candida plate to detect specimens in AIDS patients, special attention must be paid to detect non-yeasts such as T. marneffei for up to 96 hours.


Assuntos
Candida albicans/isolamento & purificação , Coinfecção/diagnóstico , Micoses/diagnóstico , Talaromyces/isolamento & purificação , Infecções Oportunistas Relacionadas com a AIDS/diagnóstico , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Ágar , Candida albicans/crescimento & desenvolvimento , Coinfecção/microbiologia , Meios de Cultura , Humanos , Masculino , Pessoa de Meia-Idade , Boca/microbiologia , Técnicas de Tipagem Micológica , Micoses/microbiologia , Orofaringe/microbiologia , Talaromyces/crescimento & desenvolvimento , Fatores de Tempo
10.
Int J Antimicrob Agents ; 58(5): 106431, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34520832

RESUMO

Extracorporeal membrane oxygenation (ECMO) can affect antimicrobial pharmacokinetics. This case report describes a 33-year-old male with newly diagnosed acquired immunodeficiency syndrome presenting in acute severe type 1 respiratory failure. On investigation, the patient had positive cultures for Candida albicans from respiratory specimens and high blood cytomegalovirus titres, and required venovenous ECMO therapy for refractory respiratory failure. Intravenous fluconazole (6 mg/kg, 24-h) and ganciclovir (5 mg/kg, 12-h) was commenced. Pre-oxygenator, post-oxygenator and arterial blood samples were collected after antibiotic administration, and were analysed for total fluconazole and ganciclovir concentrations. Although there was a 40% increase in the volume of distribution for fluconazole relative to healthy volunteers, the pharmacodynamic targets for prophylaxis were still met. The area under the curve exposure of ganciclovir (50.78 mg•h/L) achieved target thresholds. The ECMO circuit had no appreciable effect on achievement of therapeutic exposures of fluconazole and ganciclovir.


Assuntos
Candidíase/tratamento farmacológico , Infecções por Citomegalovirus/tratamento farmacológico , Oxigenação por Membrana Extracorpórea/efeitos adversos , Fluconazol/farmacocinética , Ganciclovir/farmacocinética , Insuficiência Respiratória/terapia , Síndrome de Imunodeficiência Adquirida/imunologia , Síndrome de Imunodeficiência Adquirida/microbiologia , Adulto , Antifúngicos/uso terapêutico , Antivirais/uso terapêutico , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Citomegalovirus/efeitos dos fármacos , Citomegalovirus/isolamento & purificação , Quimioterapia Combinada , Fluconazol/uso terapêutico , Ganciclovir/uso terapêutico , Humanos , Masculino
11.
Biochem Biophys Res Commun ; 578: 136-141, 2021 11 12.
Artigo em Inglês | MEDLINE | ID: mdl-34562653

RESUMO

Vps75 is a histone chaperone that interacts with the fungal-specific histone acetyltransferase Rtt109 and stimulates its acetylation activity on histone H3. Here we report the crystal structure of Vps75 of Candida albicans, one of the most common fungal pathogens. CaVps75 exists as a headphone-like dimer that forms a large negatively charged region on its concave side, showing the potential to bind positively charged regions of histones. The distal ends of the concave side of the CaVps75 dimer are positively charged and each has one more α helix than yeast Vps75. CaVps75 exhibits ionic strength- and concentration-dependent higher oligomerization in solution. In the crystal, two dimers are bound through electrostatic interactions between charged regions on the concave side of their earmuff domains, and this inter-dimer interaction differs from the currently known inter-dimer interactions of Vps75s. Our results will help to understand the role of Vps75 in C. albicans.


Assuntos
Candida albicans/química , Candidíase/microbiologia , Proteínas Fúngicas/química , Chaperonas de Histonas/química , Candida albicans/isolamento & purificação , Candidíase/metabolismo , Candidíase/patologia , Cristalografia por Raios X , Dimerização , Proteínas Fúngicas/metabolismo , Histona Acetiltransferases/genética , Histona Acetiltransferases/metabolismo , Chaperonas de Histonas/metabolismo , Histonas/química , Histonas/metabolismo , Concentração Osmolar , Eletricidade Estática
13.
Braz J Microbiol ; 52(4): 2351-2356, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34235705

RESUMO

In the past decade, researchers have focused on the emergence of drug resistance in fungal pathogens such as Candida albicans, also considered as pathobionts that occur harmlessly in the human body but could potentially be triggered to cause diseases. The increasing rate of antifungal resistance in commensal gut fungi is alarming and should be further investigated. Here, we report seven novel MLST (Multi Locus Sequence Typing) genotypes of multi-drug resistant C. albicans isolates obtained from participants of a community study in Segamat, a district in the state of Johor, Malaysia. A total of eight C. albicans were isolated from four individuals, which were found to express high resistance against fluconazole, itraconazole, voriconazole and 5-fluorocytosine antifungals. MLST was performed to assess the clonal relatedness of these drug resistant isolates among themselves and against other strains isolated from other geographical regions. The novel MLST C. albicans sequence types suggest significant genetic changes compared to previous genotypes.


Assuntos
Candida albicans , Farmacorresistência Fúngica , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/genética , Candida albicans/isolamento & purificação , Candidíase/microbiologia , Farmacorresistência Fúngica/efeitos dos fármacos , Farmacorresistência Fúngica/genética , Resistência a Múltiplos Medicamentos/genética , Humanos , Malásia , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus
14.
Future Microbiol ; 16: 619-621, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34082566

RESUMO

Here, we present a case of off-label successful use of the T2 MR (T2Candida® test) for the diagnosis of invasive candidiasis (Candida albicans endolphthalmitis). This case demonstrates that T2Candida could be performed in sterile body fluids to improve microbiological diagnosis of invasive candidiasis.


Assuntos
Candidíase Invasiva/diagnóstico , Técnicas Microbiológicas , Uso Off-Label , Corpo Vítreo/microbiologia , Antifúngicos/uso terapêutico , Candida albicans/isolamento & purificação , Candidíase Invasiva/tratamento farmacológico , Endoftalmite/diagnóstico , Endoftalmite/tratamento farmacológico , Feminino , Humanos , Lactente , Fatores de Tempo
15.
J Cell Mol Med ; 25(16): 7948-7960, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34155778

RESUMO

Smoking and Candida albicans (C. albicans) infection are risk factors for many oral diseases. Several studies have reported a close relationship between smoking and the occurrence of C. albicans infection. However, the exact underlying mechanism of this relationship remains unclear. We established a rat infection model and a C. albicans-Leuk1 epithelial cell co-culture model with and without smoke exposure to investigate the mechanism by which smoking contributes to C. albicans infection. Oral mucosa samples from healthy individuals and patients with oral leucoplakia were also analysed according to their smoking status. Our results indicated that smoking induced oxidative stress and redox dysfunction in the oral mucosa. Smoking-induced Nrf2 negatively regulated the NLRP3 inflammasome, impaired the oral mucosal defence response and increased the oral mucosa susceptibility to C. albicans. The results suggest that the Nrf2 pathway could be involved in the pathogenesis of oral diseases by mediating an antioxidative response to cigarette smoke exposure and suppressing host immunity against C. albicans.


Assuntos
Candida albicans/patogenicidade , Candidíase/microbiologia , Fumar Cigarros/efeitos adversos , Inflamassomos/metabolismo , Mucosa Bucal/microbiologia , Fator 2 Relacionado a NF-E2/metabolismo , Animais , Candida albicans/isolamento & purificação , Candidíase/metabolismo , Candidíase/patologia , Linhagem Celular , Modelos Animais de Doenças , Feminino , Humanos , Técnicas In Vitro , Masculino , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia , Fator 2 Relacionado a NF-E2/genética , Ratos , Ratos Wistar
16.
BMC Infect Dis ; 21(1): 523, 2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-34082699

RESUMO

BACKGROUND: Vaginal candidiasis is frequent in women of reproductive age. Accurate identification Candida provides helpful information for successful therapy and epidemiology study; however, there are very limited data from the Vietnam have been reported. This study was performed to determine the prevalence, species distribution of yeast causing vaginal discharge and antifungal susceptibility patterns of Candida albicans among symptomatic non-pregnant women of reproductive age. METHODS: Vaginal discharge samples were collected from 462 women of reproductive age in Hanoi, Vietnam between Sep 2019 and Oct 2020. Vaginal swabs from these patients were examined by direct microscopic examination (10% KOH). CHROMagar™ Candida medium and Sabouraud dextrose agar supplemented with chloramphenicol (0.5 g/l) were used to isolate yeast, and species identification was performed using morphological tests and molecular tools (PCR and sequencing). Antifungal susceptibility testing was determined according to the Clinical and Laboratory Standards Institute guidelines (M27-A3 and M27-S4). RESULTS: The prevalence of vaginal yeast colonization in non-pregnant women was 51.3% of 462 participants. Nine different yeast species were identified. Among these isolates, C. albicans (51.37%) was the most frequent, followed by C. parapsilosis (25.88%), C. glabrata (11.37%), C. tropicalis (4.31%), C. krusei (3.92%), C. africana (1.57%), Saccharomyces cerevisiae (0.78%), C. nivariensis (1 isolates, 0.39%), and C. lusitaniae (1 isolates, 0.39%), respectively. Among C. albicans, all 46 isolates were 100% susceptible to micafungin, caspofungin, and miconazole. The susceptibility rates to amphotericine B, 5-flucytosine, fluconazole, itraconazole and voriconazole were 95.65, 91.30, 91.30, 82.61 and 86.95%, respectively. CONCLUSIONS: The prevalence of VVC among symptomatic non-pregnant women of reproductive age in Vietnam was higher than many parts of the world. The high frequency of non-albicans Candida species, which were often more resistant to antifungal agents, was a notable feature. Resistance rates of vaginal C. albicans isolates to antifungal agents was low. Our findings suggest that continued surveillance of changes in species distribution and susceptibility to antifungals should be routinely screened and treated.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candidíase Vulvovaginal/microbiologia , Descarga Vaginal/microbiologia , Adolescente , Adulto , Candida/classificação , Candida/efeitos dos fármacos , Candida/isolamento & purificação , Candida albicans/classificação , Candida albicans/isolamento & purificação , Candidíase Vulvovaginal/epidemiologia , Farmacorresistência Fúngica/efeitos dos fármacos , Feminino , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Prevalência , Centros de Atenção Terciária , Descarga Vaginal/epidemiologia , Vietnã/epidemiologia , Adulto Jovem
17.
J Biol Chem ; 297(2): 100917, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34181946

RESUMO

Across eukaryotes, Rho GTPases such as Rac and Cdc42 play important roles in establishing cell polarity, which is a key feature of cell growth. In mammals and filamentous fungi, Rac targets large protein complexes containing NADPH oxidases (NOX) that produce reactive oxygen species (ROS). In comparison, Rho GTPases of unicellular eukaryotes were believed to signal cell polarity without ROS, and it was unclear whether Rho GTPases were required for ROS production in these organisms. We document here the first example of Rho GTPase-mediated post-transcriptional control of ROS in a unicellular microbe. Specifically, Cdc42 is required for ROS production by the NOX Fre8 of the opportunistic fungal pathogen Candida albicans. During morphogenesis to a hyphal form, a filamentous growth state, C. albicans FRE8 mRNA is induced, which leads to a burst in ROS. Fre8-ROS is also induced during morphogenesis when FRE8 is driven by an ectopic promoter; hence, Fre8 ROS production is in addition controlled at the post-transcriptional level. Using fluorescently tagged Fre8, we observe that the majority of the protein is associated with the vacuolar system. Interestingly, much of Fre8 in the vacuolar system appears inactive, and Fre8-induced ROS is only produced at sites near the hyphal tip, where Cdc42 is also localized during morphogenesis. We observe that Cdc42 is necessary to activate Fre8-mediated ROS production during morphogenesis. Cdc42 regulation of Fre8 occurs without the large NOX protein complexes typical of higher eukaryotes and therefore represents a novel form of ROS control by Rho GTPases.


Assuntos
Candida albicans/patogenicidade , Candidíase/patologia , Hifas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína cdc42 de Ligação ao GTP/metabolismo , Candida albicans/isolamento & purificação , Candidíase/metabolismo , Candidíase/microbiologia , Polaridade Celular , Proteínas Fúngicas/metabolismo , Morfogênese
18.
PLoS One ; 16(6): e0252555, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34061886

RESUMO

C. albicans is a commensal organism present in the human microbiome of more than 60% of the healthy population. Transition from commensalism to invasive candidiasis may occur after a local or a general failure of host's immune system. This transition to a more virulent phenotype may reside either on the capacity to form hyphae or on an acquired resistance to antifungal drugs. Indeed, overexpression of genes coding drug efflux pumps or adhesins, cell wall proteins facilitating the contact between the fungus and the host, usually marks the virulence profile of invasive Candida spp. In this paper, we compare virulence of two clinical isolates of C. albicans with that of laboratory-induced resistant strains by challenging G. mellonella larvae with these pathogens along with monitoring transcriptional profiles of drug efflux pumps genes CDR1, CDR2, MDR1 and the adhesin genes ALS1 and HWP1. Although both clinical isolates were found resistant to both fluconazole and micafungin they were found less virulent than laboratory-induced resistant strains. An unexpected behavior emerged for the former clinical isolate in which three genes, CDR1, CDR2 and HWP1, usually correlated with virulence, although hyperexpressed, conferred a less aggressive phenotype. On the contrary, in the other isolate, we observed a decreased expression of CDR1, CDR2 and HWP1as well as of MDR1 and ALS1 that may be consistent with the less aggressive performance observed in this strain. These altered gene expressions might directly influence Candida virulence or they might be an epiphenomenon of a vaster rearrangement occurred in these strains during the challenge with the host's environment. An in-deepth comprehension of this scenario could be crucial for developing interventions able to counteract C. albicans invasiveness and lethality.


Assuntos
Candida albicans/genética , Candida albicans/patogenicidade , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Expressão Gênica , Glicoproteínas de Membrana/genética , Proteínas de Membrana Transportadoras/genética , Animais , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candidíase/microbiologia , Farmacorresistência Fúngica/efeitos dos fármacos , Farmacorresistência Fúngica/genética , Feminino , Fluconazol/farmacologia , Humanos , Hifas/genética , Larva/microbiologia , Lepidópteros/microbiologia , Micafungina/farmacologia , Testes de Sensibilidade Microbiana , Fenótipo , Virulência/genética
19.
J Clin Lab Anal ; 35(7): e23826, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33988259

RESUMO

BACKGROUND: Esophageal candidiasis is the most frequent form of esophagitis. The pathogenicity of Candida spp. is related to a combination of microbial factors, hydrolytic enzyme secretion and phenotypic switching. This study was designed to investigate esophageal candidiasis, antifungal activity, enzymatic activity patterns, phenotyping, and genotyping profiles of Candida albicans species. METHODS: Nine hundred thirty-three visited patients were evaluated, and esophageal biopsies from patients were included in this study during 2019-2020. Direct smear, Gram staining, and culture on CHROMagar were performed for each sample. Isolated species were identified with conventional procedures and PCR-RFLP. Susceptibility to antifungals was determined according to CLSI guidelines. ABC typing, phenotype switching, hemolysin, proteinase, phospholipase, and esterase activity were also determined with the appropriate protocols. RESULTS: Twenty-three (2.5%) patients (mean age 55.2 years) were diagnosed with esophageal candidiasis. The species isolated were 19(82.6%) C. albicans, 3(13.1%) C. glabrata, and 1(4.3%) C. tropicalis. Genotype A (57.9%) was the predominant type in C. albicans isolates. 50% of C. albicans isolates exhibited a white phenotype. A high level of phospholipase (47.4%), hemolysin (68.4%), and proteinase activity (36.8%) was observed in the C. albicans isolates. Only three C. glabrata isolates displayed non-wild type susceptibility to voriconazole and itraconazole. CONCLUSION: This study shows that C. albicans are still the most frequent isolates from patients with esophageal candidiasis. The predominance of genotype A, the white phenotype, and strong hemolysin activity may indicate a high prevalence of pathogenicity in these isolates. Sensitivity to antifungal drugs was greatest for amphotericin and fluconazole.


Assuntos
Antifúngicos/farmacologia , Candida albicans/enzimologia , Candida albicans/genética , Candidíase/microbiologia , Esofagite/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candidíase/complicações , Esofagite/complicações , Feminino , Fluconazol/farmacologia , Genótipo , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Fenótipo , Adulto Jovem
20.
Sci Rep ; 11(1): 10802, 2021 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-34031498

RESUMO

Early childhood caries (ECC) recurrence occurs in approximately 40% of treated cases within one year. The association of Streptococcus mutans and Candida albicans with the onset of ECC is well known. Also, S. mutans strains harboring collagen-binding proteins (Cbps) avidly bind to collagen-rich dentin and are linked to increased caries risk. Here, we investigated the presence of Cbp+ S. mutans and C. albicans in saliva and dental plaque of children with varying caries statuses, and their salivary microbiome. In this cross-sectional study, 143 children who were caries-free (n = 73), treated for ECC with no signs of recurrence after 6 months (n = 45), or treated for ECC and experiencing recurrence within 6 months following treatment (n = 25) were enrolled. Co-infection with C. albicans and S. mutans, especially Cbp+ S. mutans, was strongly associated with caries recurrence. Subjects of the recurrence group infected with Cbp+ S. mutans showed a greater burden of Candida spp. and of Mutans streptococci in dentin than those infected with Cbp- strains. Salivary microbiome analysis revealed that Streptococcus parasanguinis was overrepresented in the caries recurrence group. Our findings indicate that Cbp+ S. mutans and C. albicans are intimately associated with caries recurrence, contributing to the establishment of recalcitrant biofilms.


Assuntos
Proteínas de Bactérias/metabolismo , Candida albicans/patogenicidade , Coinfecção/microbiologia , Cárie Dentária/microbiologia , Streptococcus mutans/patogenicidade , Candida albicans/isolamento & purificação , Candida albicans/metabolismo , Pré-Escolar , Estudos Transversais , Cárie Dentária/metabolismo , Suscetibilidade à Cárie Dentária , Dentina/metabolismo , Feminino , Humanos , Masculino , Recidiva , Saliva/microbiologia , Streptococcus/isolamento & purificação , Streptococcus mutans/isolamento & purificação , Streptococcus mutans/metabolismo
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