Background: Urinary proteome is a topic not deeply investigated in dogs. Leishmaniasis (L) and Cushing’s Syndrome (CS) are frequently associated with persistent proteinuria. Evaluation of urine proteome with sensitive methods is considered a non invasive procedure to monitor the therapy and select novel biomarkers. Objectives: Comparison of urinary electrophoretic profiles between healthy (H) and diseased dogs (L, CS) at the time of diagnosis and during the treatment monitoring. Methods: Urine samples were collected from 49 dogs (12 H, 26 L, 11 CS) and Urine Protein and Albumin to Creatinine ratios (UPC, UAC), HRE and SDS-PAGE were performed. Electrophoretic profiles were analyzed by ImageJ©; proteins were identified by ESI-Q-TOF. Results: UPC and UAC (mean±SD) were significantly higher (p<0.01) in L (3.47±4.76 and 1.69±3.19) and CS (2.55±3.69 and 1.79±3.65) compared to H dogs (0.11±0.11 and 0.008±0.008). HRE was useful with urine protein concentration higher than 40 mg/dl, and revealed different electrophoretic patterns between healthy (mainly albumin band is present) and diseased dogs (also other bands can be present). Urinary albumin concentration determined by HRE correlated well with the data obtained by routine immunoturbidimetric method (r=0.91, p<0.01). SDS-PAGE allowed to visualize an “healthy profile” with 20-25 different bands in urine of H dogs; the two most abundant ones have been identified as uromodulin and albumin. Urine samples from L and CS dogs presented a significantly higher number of protein bands (35-40), a decrease of uromodulin and an increase of albumin band intensities and the appearance of other bands, particularly in the range of 55-14 kDa; among them haptoglobin, superoxide dismutase1 and arginine esterase were identified by ESI-Q-TOF in urine of L dogs. Qualitative changes of protein profiles were evidenced during the treatment in CS and L dogs, indicating in some cases an improvement of renal damage. Conclusion: HRE could represent a rapid screening test for diagnosis of renal damage. SDS-PAGE is a sensitive and promising technique to investigate qualitative proteinuria in healthy and diseased dogs. Urine proteome analysis could help clinicians to better characterize the proteinuria even in absence of renal histopathology.
URINE PROTEOME IN DOGS AFFECTED BY LEISHMANIASIS AND CUSHING’S SYNDROME
FERLIZZA, ENEA;FRACASSI, FEDERICO;DONDI, FRANCESCO;CARPENE', EMILIO;ISANI, GLORIA
2013
Abstract
Background: Urinary proteome is a topic not deeply investigated in dogs. Leishmaniasis (L) and Cushing’s Syndrome (CS) are frequently associated with persistent proteinuria. Evaluation of urine proteome with sensitive methods is considered a non invasive procedure to monitor the therapy and select novel biomarkers. Objectives: Comparison of urinary electrophoretic profiles between healthy (H) and diseased dogs (L, CS) at the time of diagnosis and during the treatment monitoring. Methods: Urine samples were collected from 49 dogs (12 H, 26 L, 11 CS) and Urine Protein and Albumin to Creatinine ratios (UPC, UAC), HRE and SDS-PAGE were performed. Electrophoretic profiles were analyzed by ImageJ©; proteins were identified by ESI-Q-TOF. Results: UPC and UAC (mean±SD) were significantly higher (p<0.01) in L (3.47±4.76 and 1.69±3.19) and CS (2.55±3.69 and 1.79±3.65) compared to H dogs (0.11±0.11 and 0.008±0.008). HRE was useful with urine protein concentration higher than 40 mg/dl, and revealed different electrophoretic patterns between healthy (mainly albumin band is present) and diseased dogs (also other bands can be present). Urinary albumin concentration determined by HRE correlated well with the data obtained by routine immunoturbidimetric method (r=0.91, p<0.01). SDS-PAGE allowed to visualize an “healthy profile” with 20-25 different bands in urine of H dogs; the two most abundant ones have been identified as uromodulin and albumin. Urine samples from L and CS dogs presented a significantly higher number of protein bands (35-40), a decrease of uromodulin and an increase of albumin band intensities and the appearance of other bands, particularly in the range of 55-14 kDa; among them haptoglobin, superoxide dismutase1 and arginine esterase were identified by ESI-Q-TOF in urine of L dogs. Qualitative changes of protein profiles were evidenced during the treatment in CS and L dogs, indicating in some cases an improvement of renal damage. Conclusion: HRE could represent a rapid screening test for diagnosis of renal damage. SDS-PAGE is a sensitive and promising technique to investigate qualitative proteinuria in healthy and diseased dogs. Urine proteome analysis could help clinicians to better characterize the proteinuria even in absence of renal histopathology.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
