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Antimicrobial action of photodynamic therapy in gene expression in Candida albicans biofilms

Grant number: 13/22897-2
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2014
Effective date (End): November 30, 2017
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Antonio Olavo Cardoso Jorge
Grantee:Fernanda Freire
Home Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
Associated scholarship(s):14/25772-9 - Monitoring the effect of photodynamic therapy oral infection with Candida albicans in immunosuppressed mice by bioluminescence imaging, BE.EP.DR

Abstract

The micro-organisms are becoming increasingly resistant to antimicrobials and strains of Candida albicans are resistant to antifungal thus becomes important and necessary to conduct research to evaluate the effects of new therapeutic methods, such as photodynamic therapy (PDT) on virulence factors of C. albicans related adhesion and hyphal formation of biofilms. The objective of this study is to verify the effects of photodynamic therapy on Candida albicans biofilms, evaluating its effects on the expression of genes TEC1 (transcription factor), HWP1 (cell wall protein hyphae), EFG1 (transcriptional regulator related morphogenesis), BCR1 (regulator of biofilm formation and cell wall), ACE2 (transcriptional regulator involved in morphogenesis) and ALS1 (adhesin) by yeast. Will be evaluated for quantification of biofilms 30 isolates from HIV patients and 30 samples from patients with denture stomatitis, as the production of biofilm, dry weight and XTT. Of these, 5 samples of each group having better ability of biofilm formation will be selected. Thus, will be used clinical samples of C. albicans were isolated from HIV patients (n=5), of patients with denture stomatitis (n=5) and a standard strain ATCC 18804.The quantification of gene expression will be related to production of these genes in clinical samples and the reference strain assay using real-time PCR. The presence and expression of genes are analyzed before and after photodynamic therapy. We will used for photodynamic therapy the photosensitizers methylene blue at 300 mM and erytrosine at 5 mM, laser sensitized aluminum gallium arsenide low - power (visible red, 660 nm) and green LED (532 ± 10 nm), respectively. Will be assessed four experimental groups for each sample: a) F+L+: sensitization with the dye and light irradiation, b) F+L-: Only treatment with the photosensitizer, c) F-L+ : Only light irradiation and d) F-L-: control group without sensitization with dye and absence of light. In addition from evaluating the expression of TEC1, HWP1, EFG1, BCR1, ALS1 and ACE2 genes will be checked metabolic activity of cells from each sample for the four groups tested by XTT colorimetric assay. The results will be analyzed by Minitab software with a significance level of 5% and by the software of real-time equipment. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
FREIRE, FERNANDA; DE BARROS, PATRICIA PIMENTEL; PEREIRA, CRISTIANE APARECIDA; JUNQUEIRA, JULIANA CAMPOS; CARDOSO JORGE, ANTONIO OLAVO. Photodynamic inactivation in the expression of the Candida albicans genes ALS3, HWP1, BCR1, TEC1, CPH1, and EFG1 in biofilms. Lasers in Medical Science, v. 33, n. 7, p. 1447-1454, SEP 2018. Web of Science Citations: 4.
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FREIRE, Fernanda. Expressão dos genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 de Candida albicans em biofilmes após inativação fotodinâmica. 2017. 64 f. Doctoral Thesis - Universidade Estadual Paulista "Julio de Mesquita Filho" Instituto de Ciência e Tecnologia (Campus de São José dos Campos)..

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