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Deep proteomic analysis of cervicovaginal fluid in bacterial vaginosis: association with bacterial composition and response to metronidazole treatment

Grant number: 14/22463-5
Support Opportunities:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): February 16, 2015
Effective date (End): May 15, 2015
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal Investigator:Márcia Guimarães da Silva
Grantee:Camila Marconi
Supervisor: Valerie C. Wasinger
Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Research place: University of New South Wales (UNSW), Australia  
Associated to the scholarship:12/10403-2 - Characterization of the vaginal microbiome of Brazilian women of reproductive age, BP.PD


Normal vaginal flora is commonly defined as the prevalence of Lactobacillus spp. over the many other bacteria that may colonize this milieu. Recent studies have been providing increasing evidence of the strict relation between the bacterial composition of vaginal flora and host's immune response. Moreover, alterations in the levels of inflammatory mediators caused by abnormal vaginal flora there have been proposed as the mechanism by which women with decreased or absent lactobacilli are at higher risk for many sexually transmitted infections, - including the Human Immunodeficiency Virus (HIV). Bacterial vaginosis is the most common type of abnormal vaginal flora, but its treatment is troublesome. Although the recommended treatment for bacterial vaginosis is based in a 7-days regimen of oral metronidazole, failure rates of 40% are observed within one month-period. Despite the current efforts, literature is still scarce regarding the full cervicovaginal proteomic profile and the exact microbiological composition of bacterial vaginosis. More information regarding these aspects is needed to elucidate how host relates with the local bacterial composition and which mechanisms are implicated in the difficulty to reestablish a lactobacilli-dominated after bacterial vaginosis treatment. This cross-sectional study aims to evaluate the association of cervicovaginal proteome with the types of vaginal bacterial communities and with the cure or persistence of bacterial vaginosis after metronidazole therapy. From 200 non-pregnant reproductive-aged women prospectively enrolled in Botucatu/SP as part of a current largest study "Characterization of the vaginal microbiome of Brazilian women in reproductive age", a total of 40 samples from women with normal flora and 40 from women with bacterial vaginosis will be submitted to proteomic and microbiome analysis. Women with bacterial vaginosis will be treated with metronidazole for 7 days of therapy, and re-evaluated in 6 weeks. Cervicovaginal samples will be taken at each visit. Classification of the vaginal flora in normal, intermediate and bacterial vaginosis will be based on Nugent criteria. Vaginal bacterial communities will be identified using microbiome sequencing analyses of the V3-V4 hypervariable region of 16S rRNA gene at Illumina MiSeq platform. Cervicovaginal proteomic profile will be determined by MS/MS spectrometry (Orbitrap XL mass) after MD10 fractionation system that increases the proteome coverage, particularly by detecting low-mass and low abundance proteins and peptides. All proteome data will be processed using the adequate bioinformatics tools. Cervicovaginal proteomic profiles will be described and compared between among the different bacterial communities types and between the cases of persistent bacterial vaginosis after metronidazole therapy with those successfully treated using the appropriate statistical tests in MetaCoreTM, computational platform (version 6.8, St Joseph, MI). (AU)

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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)
DURSKI, MARIANA; RAVEL, JACQUES; SPAUTZ, ANA C. P. G.; CARVALHO, NEWTON S.; SILVA, MARCIA G.; MARCONI, CAMILA. Comparison of two microscopic interpretations of vaginal microbiota with molecular profiling. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, v. 104, n. 1, p. 5-pg., . (12/10403-2, 12/16800-3, 14/22463-5)
MARCONI, CAMILA; EL-ZEIN, MARIAM; RAVEL, JACQUES; MA, BING; LIMA, MOISES D.; CARVALHO, NEWTON SERGIO; ALVES, ROSANE R. F.; PARADA, CRISTINA M. G. L.; LEITE, SANDRA H. M.; GIRALDO, PAULO C.; et al. Characterization of the Vaginal Microbiome in Women of Reproductive Age From 5 Regions in Brazil. SEXUALLY TRANSMITTED DISEASES, v. 47, n. 8, p. 562-569, . (12/10403-2, 14/22463-5, 12/16800-3)
NOVAK, J.; FERREIRA, C. S. T.; GOLIM, M. A.; SILVA, M. G.; MARCONI, C.. Covariates of vaginal microbiota and pro-inflammatory cytokine levels in women of reproductive age. BENEFICIAL MICROBES, v. 14, n. 2, p. 11-pg., . (14/22463-5, 12/16800-3, 16/02390-9)

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