Validation of quantitative fluorescent polymerase chain reaction (QF-PCR) test to detect fetal aneuploidies

Author(s)

Renata de Castro Gonçalves

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD)
• Defense date: 2016-12-14

Examining board members

Denise de Castro Fernandes; Miriam Helena Fonseca Alaniz; Marilene Demasi

• Advisor: Denise de Castro Fernandes

Abstract

Vascular smooth muscle cell (VSMC) phenotype switch depends on extrinsic/ intrinsic cues including NADPH oxidase (Nox) - dependent redox signalling. Growth factor- triggered Nox1 expression/ activity is regulated by the chaperone oxidoreductase protein disulphide isomerase- A1 (PDI). PDI is required for VSMC migration and cytoskeleton organization, and extracellular PDI counteracts constrictive vascular remodelling via cytoskeleton reshaping. Such pattern of PDI effects led us to hypothesize that PDI may oschestrate VSMC phenotypic alterations, and test it we developed VSMC which overexpress PDI by doxyxycline treatment (VSMC-PDIteton) . We observed that sustained PDI overexpression (72h) increased cell length and induced the expression of differentiation markers calponin, ?-actin and smoothelin, which were not further upregulated upon catalase incubation. Intracelular superoxide production increased alter 48h of PDI overexpression, produced partially from Nox1, based on inhibiton with GKT136901 or NOXA1ds peptide. Acute PDI overexpression (40h) increased VSMC distance during single cell migration assay. Finally, PDI silencing spontaneously decreased differentiation marker calponin expression. These data suggest that PDI overexpression induces transiently NADPH oxidase overexpression and probably the major isoform is Nox1, while sustained PDI overexpression indices VSMC phenotypic changes (AU)

• FAPESP's process: 13/03363-7 - Study of the mechanism by which Protein Disulfide Isomerase overexpression modulates Angiotensin II Receptor (AT1R) activation in vascular smooth muscle cells
• Grantee: Renata de Castro Gonçalves
• Support Opportunities: Scholarships in Brazil - Master