Salivary proteomic profile of patients with Parkinson´s disease under high intensity body exercises practices

• Grant number: 20/02175-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2020
• End date: November 30, 2022
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Ana Carolina Magalhães
• Grantee: Karina Oliveira Santos
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The aim of this work will be to evaluate the salivary protein profile of adults with Parkinson's disease and control adults, before and after high intensity physical exercise, to identify possible biomarkers for monitoring disease progression. Twenty adults with Parkinson's disease and twenty adults, matched by age and gender, without Parkinson's disease, will participate. These participants will be recruited from "Laboratório de Pesquisa em Movimento Humano" (MOVI-LAB) at UNESP/Bauru. Total unstimulated saliva will be collected from each participant by adding it to 50mL Falcon tubes immersed in ice for 10 to 15 minutes, to obtain at least 10 mL of saliva per participant. Saliva will be collected before and immediately after the aerobic stimulus done during thirty minutes in an ergonomic stool, under supervision (Inbramed ATL, Inbrasport, Porto Alegre, Brazil) at MOVI-LAB. The collected saliva will be centrifuged, and supernatant stored in cryogenic tube at -80 ºC. For protein extraction, 100 µL of each sample will be aliquoted into separate tubes and diluted 1:1 with 6 M urea extraction solution, 2 M tourea and 50 mM NH4CO3 (pH 7.8). Following this, they will be agitated for 10 minutes at 4ºC and placed in an ultrasonic bath for 5 minutes and centrifuged, repeating this sequence of procedures twice. After extraction, 3 pools will be formed per participant and concentrated at 150 µL in Falcon Amicon tubes. Amino acid chains will be cleaved using 2% (w/w) trypsin (Promega®, USA) for 14 h at 37ºC. The samples will be purified and desalted and 1 µL of each sample will be used for protein quantification by the Bradford method (Bio-Rad®, USA). The samples will be resuspended and subjected to Nano Liquid Chromatography Electron Spray Ionization Tandem Mass Spectrometry analysis (LC-ESI-MS, Waters, Wilmslow, United Kingdom). Subsequently, typical peptide analysis will be performed on the ACQUITY UPLC system (Waters, USA) coupled to the Xevo Q-TOF G2 mass spectrometer (Waters, USA). Protein identification will be performed according to Protein Lynx Global Server (PLGS) software version 3.03 (Waters, USA) and results based on the UniProt (Universal ProteinResource) catalog Homo Sapiens database. The difference in expression between groups will be calculated using Monte Carlo algorithm and expressed as p<0,05 for proteins present in lower abundance and 1-p>0,95 for proteins present in higher abundance. At the end, the salivary proteins presented in each group will be compared and classified according to the terms of the Generic Gene Ontologv - GO - TermFinder. The IBI-IMIM research tool will be used to identify candidate to biomarker proteins (disease-related biomarker database).

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)

OLIVEIRA VENTURA, TALITA MENDES; SANTOS, KARINA OLIVEIRA; BRAGA, ALINE SILVA; GRIZZO THOMASSIAN, LARISSA TERCILIA; RABELO BUZALAF, MARILIA AFONSO; BARBIERI, FABIO AUGUSTO; KALVA-FILHO, CARLOS AUGUSTO; FARIA, MURILO HENRIQUE; MAGALHAES, ANA CAROLINA. Salivary proteomic profile of young adults before and after the practice of interval exercise: preliminary results. SPORT SCIENCES FOR HEALTH, v. 18, n. 3, p. 15-pg., 2022-01-10. (19/21797-0, 20/02175-6)

VENTURA, TALITA MENDES OLIVEIRA; SANTOS, KARINA OLIVEIRA; BRAGA, ALINE SILVA; THOMASSIAN, LARISSA TERCILIA GRIZZO; BUZALAF, MARILIA AFONSO RABELO; KALVA-FILHO, CARLOS AUGUSTO; FARIA, MURILO HENRIQUE; BARBIERI, FABIO AUGUSTO; MAGALHAES, ANA CAROLINA. Proteomic profile of saliva in patients with Parkinson's disease after the practice of interval exercise. PARKINSONISM & RELATED DISORDERS, v. 98, p. 2-pg., 2022-05-01. (20/02175-6, 19/21797-0)