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In vitro evaluation of p-cresol on the oxidative metabolism and apoptosis of neutrophils from dogs

Grant number: 11/16018-0
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): January 01, 2013
Effective date (End): February 28, 2014
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
Principal Investigator:Paulo César Ciarlini
Grantee:Anelise Maria Bosco
Host Institution: Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil


In human immunosuppression that occurs in uremic patients is associated with dysfunction of uremic toxins caused by neutrophils. The effect of uremic toxin p-cresol on the function of human neutrophils have been extensively investigated. This toxin is not properly filtered during hemodialysis and its accumulation in the blood promotes the inhibition of neutrophil oxidative metabolism, thus affecting its bactericidal function. The neutrophil dysfunction in nephropathic dogs has not been adequately investigated. Our group recently obtained evidence that the in vitro neutrophil oxidative metabolism of healthy dogs is affected when incubated as uremic serum, but to date no studies on the specific effect of p-cresol on neutrophil function in this species. In this sense, the present study aims to measure plasma concentrations of p-cresol in dogs with CRF and test the hypothesis that similarly to what happens in humans toxinaa this increase is associated with changes in oxidative metabolism and apoptosis of neutrophils. This will be a standardized methodology for measuring liquid-chromatographic plasma concentrations of this toxin in dogs healthy controls and patients with CRF equivalent to stage IV. It will also be evaluated and compared in vitro the specific effect of p-cresol on superoxide production, lipid peroxidation, and apoptosis rate of neutrophils isolated from dogs healthy controls, whereas plasma concentrations were observed "in vivo" in dogs with CRF . Cells isolated from 20 healthy dogs are incubated in RPMI medium with and without p-cresol, and plasma of dogs with CRF. The oxidative metabolism of neutrophils is evaluated by flow cytometry using the capillary tube hidroetidina. The viability and apoptosis of neutrophils is measured in capillary flow cytometry using Annexin V-PE system. Oxidative stress is assessed by quantification of plasma thiobarbituric reactive species, total antioxidant status, uric acid, total bilirubin and albumin. (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)
BOSCO, A. M.; ALMEIDA, B. F. M.; PEREIRA, P. P.; DOS SANTOS, D. B.; NETO, A. J. S.; FERREIRA, W. L.; CIARLINI, P. C.. The uremic toxin methylguanidine increases the oxidative metabolism and accelerates the apoptosis of canine neutrophils. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, v. 185, p. 14-19, . (11/16018-0, 11/18746-3)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
BOSCO, Anelise Maria. Avaliação in vitro do P-cresol sobre o metabolismo oxidativo e apoptose dos neutrófilos de cães. 2014. Master's Dissertation - Universidade Estadual Paulista (Unesp). Faculdade de Medicina Veterinária. Araçatuba Araçatuba.

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