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Lipidomics analysis of the vaginal discharge in women with vulvovaginal candidiasis and cytolytic vaginosis

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Author(s):
José Marcos Sanches Júnior
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas
Defense date:
Examining board members:
Paulo César Giraldo; Cristina Laguna Benetti Pinto; Iara Moreno Linhares
Advisor: Paulo César Giraldo
Abstract

Background: Vulvovaginal candisiasis (VVC) and cytolytic vaginosis (CV) are gynecological conditions that often include vaginal itching, burning, abnormal discharge, dysuria and dyspareunia. Although the symptoms are similar, the vaginal microenvironment and the local inflammatory response are completely different, a fact that implies in different treatments. The triggering factors and pathophysiology of both diseases are unclear. Advances in mass spectrometry and bioinformatics techniques contribute to a better understanding of the health and disease processes, having the lipidomics as a great tool to characterize potential biomarkers for many diseases. Objective: Characterize the lipids in the vaginal contents in women with VVC and CV, and relate them to these diseases. Materials and methods: Cross-sectional study analyzed the vaginal contents of 24 non-pregnant women, aged between 18 and 44 years, attended at the Genital Infections Outpatient Clinic (CAISM-UNICAMP), 8 with CVV, 8 with CV (vaginal discharge and pruritus/burning) and another 8 women without any vulvovaginal complaint. All the women signed the free and informed consent term, underwent detailed anamnesis and specular examination, when vaginal material was collected for cytological, microbiological and lipid studies. Samples of the lateral vaginal wall were collected with sterile Dacron swab and put on glass slides for Gram staining, in Sabouraud culture for fungal screening and stored in Falcon dry tubes at -80 ° C for the lipidomics analysis by mass spectrometry (MS). The diagnosis of vaginal candidiasis was made considering the presence in the vaginal contents of hyphae or blastocodides in the smears stained by Gram and the positivity of the specific fungal culture. Cytolytic vaginosis was characterized by the absence of mycotic elements and inflammatory process (leukocytes), presence of epithelial cells lysis in the vaginal smear. The lipidomics samples were resuspended in deionized water followed by the addition of chloroform and methanol, shaken and centrifuged and sonicated for lysis of the material, and then dried on SpeedVac. Chromatographic separation was performed on an Agilent1290-Infinity automated ultra-high performance liquid chromatography system plus a C-18 column. The data were obtained in positive and negative mode by MassHunter Qualitative software (Agilent). All metabolites were assessed by mass error (? 5 ppm), isotope standard, fragmentation patterns and plausible retention time, characterized from the descriptions of the compounds by The Human Metabolome Database, XCMS and Lipid Maps. Multivariate data analysis techniques were used to process the data found. Segregation between groups was analyzed by PCA (main component analysis) and PLS-DA (discriminate partial least squares analysis). Results: The PCA analysis showed an important separation that characterized the metabolic difference between the three groups, identifying a total of 38 lipids by the PLS-DA analysis. The major biomarkers that correlate with the pathophysiology of CVV were: 15-methyl-15S-Prostaglandin D2, O-adipoylcarnitine, fosfoserine, undecanoic acid and formyl dodecanoate. In VC there were 1- (11Z-docosenoyl)-glycerol-3-phosphate and 5-aminopentanoic acid, 1-oleoyl cyclic phosphatidic acid and palmitolic acid as biomarkers. In the control group, the main lipids found were eicosadienoic acid, 1-stearoyl cyclic phosphate, myristic acid, 7Z, 10Z-hexadecadienoic acid, and glycerol phosphate acids. Conclusion: Women with CVV and VC present similar clinical complaints (pruritus, burning and vaginal discharge), but the lipid composition is different for these conditions, being also different in women without vaginal complaints. Lipid potential biomarkers can elucidate physiopathological mechanisms of CVV and VC not yet described, such as factors that lead to adhesion of molecules to the vaginal epithelium, cell lysis, lactobacillary growth, oxidative stress and apoptosis, collaborating to better understand these conditions (AU)

FAPESP's process: 16/18850-9 - Lipidomics analysis of the vaginal discharge in women with vulvovaginal candidiasis and cytolytic vaginosis
Grantee:José Marcos Sanches Junior
Support type: Scholarships in Brazil - Master