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Effect of synbiotic activity on intestinal epithelial integrity in obese and lean microbiota

Grant number: 16/20336-1
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): February 01, 2017
Effective date (End): January 31, 2018
Field of knowledge:Biological Sciences - Microbiology
Principal researcher:Katia Sivieri
Grantee:Fernanda Bianchi
Supervisor abroad: Lene Jespersen
Home Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Research place: University of Copenhagen, Frederiksberg, Denmark  
Associated to the scholarship:15/08228-6 - Selection and assessment of the combination between waste of tropical fruits and probiotic strains in the modulation of intestinal microbiota of obese compared to the normal weight using the simulator of the human intestinal microbial ecosystem, BP.DR


The human intestinal epithelium is a barrier between the outside and the inside preventing harmful substances from entering the body. Studies have shown that increased intestinal permeability is associated with several disorders such as obesity. The gut microbiota target various intracellular pathways and regulate intestinal barrier function as well as the presence of some prebiotics fibers and probiotic strains which can enhance intestinal barrier function and, in some cases, prevents or reverses the adverse effects of pathogens. The aim of this work is to assess the effect of a symbiotic combination between a probiotic strain and a by-product of tropical fruit on the integrity of the intestinal barrier by transepithelial electrical resistance (TEER), tight junction mRNA and to analyze the Caco-2 cytotoxicity during a treatment in a Simulator of Human Intestinal Microbial Ecosystem (SHIME®), using feces of obese and lean volunteers. The samples will be collected from each colon vessels (ascending, transverse and descending colon) of the SHIME® reactor, during the control period, treatment with the combination and washout period using obese and lean microbiota. Transepithelial electrical resistance (TEER) across polarized monolayers of intestinal epithelial cells of human (Caco-2) will be determined during all periods of SHIME® experiments (control, treatment and washout). It will be performed the tight junction mRNA expression by qPCR (Fluidigm qPCR system) of vessels with changes in TEER. Caco-2 cytotoxicity test is going to be performed using samples of SHIME® by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). rRNA 16S gene sequencing of SHIME® samples will be performed using Illumina MiSeq® platform. SPSS will be used for the statistical analyses of the MTT assay results, TEER measurements and mRNA expression. The data will be evaluated using one-way analysis of variance followed by the Tuke test if the data were normally distributed and Kruskal-Walls nonparametric test with Bonferroni corrections if the data were not normally distributed. (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)
BIANCHI, FERNANDA; LARSEN, NADJA; TIEGHI, THATIANA DE MELLO; TALLARICO ADORNO, MARIA ANGELA; KOT, WITOLD; ISAY SAAD, SUSANA MARTA; JESPERSEN, LENE; SIVIERI, KATIA. Modulation of gut microbiota from obese individuals by in vitro fermentation of citrus pectin in combination with Bifidobacterium longum BB-46. Applied Microbiology and Biotechnology, v. 102, n. 20, p. 8827-8840, . (15/08228-6, 13/50506-8, 15/13965-0, 16/20336-1)
BIANCHI, FERNANDA; LARSEN, NADJA; TIEGHI, THATIANA DE MELLO; ADORNO, MARIA ANGELA T.; SAAD, I, SUSANA M.; JESPERSEN, LENE; SIVIERI, KATIA. In vitro modulation of human gut microbiota composition and metabolites by Bifidobacterium longum BB-46 and a citric pectin. Food Research International, v. 120, p. 595-602, . (15/13965-0, 15/08228-6, 13/50506-8, 16/20336-1)
SRIMAHAEAK, THANYAPORN; BIANCHI, FERNANDA; CHLUMSKY, ONDREJ; LARSEN, NADJA; JESPERSEN, LENE. In-vitro study of Limosilactobacillus fermentum PCC adhesion to and integrity of the Caco-2 cell monolayers as affected by pectins. Journal of Functional Foods, v. 79, . (16/20336-1)

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