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Urine markers are advocated to early detect kidney damage in the clinical practice, nevertheless histology remains the gold standard. The aim of this study was to evaluate quali-quantitative proteinuria and possible renal damage using different non-invasive tests in dogs affected by leishmaniasis.Based on clinical signs and serology/cytology, 26 affected dogs (Leish) were included. Fifteen healthy, non-proteinuric dogs were selected as Control. Upon admission, all dogs underwent to physical examination, Systolic Blood Pressure (SBP) measurement, clinicopathological evaluation (CBC, Urea, Creatinine, ALT, ALP, Glucose, Calcium, Phosphorous, Sodium, Potassium, Cholesterol, Triglyceride, Albumin, Total Protein), serology, urinalysis, ultrasound examination and Renal Resistive Index (RRI) determination. Urine Total Protein and Urine Albumin to Creatinine ratios (UPC; UAC), urine High Resolution agarose and Silver Staining Sodium-Dodecyl-Sulphate-PolyAcrylamide gel electrophoresis (HRE; SDS-PAGE) were performed. A cut-off of 66 kDa was selected to classify bands in High or Low Molecular Weight (HMW; LMW). Data were analyzed with non-parametric statistics and ROC curve analysis (ROC). A difference was considered significant for p0.5) was detected in 16/26 dogs (P), 5/26 were borderline proteinuric (BLP; UPC 0.2-0.5) and 5/26 were nonproteinuric (NP, UPC 0.17, 89% Sensitivity and 88% Specificity and for UAC >0.013, 94% Sensitivity and 82% Specificity. The Areas under the Curve for UPC and UAC were 0.89 and 0.92, respectively. Non-invasive methods, particularly urinary SDS-PAGE, could be useful to detect an early renal damage in canine leishmaniasis. Further studies are required to correlate these findings to renal histology

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

Background: Leishmaniasis is a frequent cause of nephropathy in dogs. Evaluate urine proteome with sensitive methods is a useful and non invasive method to monitor the therapy and discover novel biomarkers. Objective: Comparing different electrophoretic methods for urine protein separation and monitoring the therapy in dogs affected by leishmaniasis. Methods: Urine samples were collected from 43 leishmaniotic dogs. UP/C, UA/C and high resolution electrophoresis (HRE) were performed for all samples. Moreover, SDS-PAGE (26 samples) and a semi-automated SDS-AGE (12 samples) were tested. Results: UP/C and UA/C median (range) values were 1.45 (0.06-20.08) and 0.38 (0.01-14.8) respectively. With HRE different electrophoretic patterns were evidenced and urinary albumin concentrations (median 32 mg/dL; range 5-463 mg/dL) and percentage (mean±SD; 39±13.7) were determined. With SDS-AGE different patterns of proteinuria were found: 9 dogs presented mixed proteinuria (complete or incomplete) and 3 presented non selective glomerular proteinuria. With SDS-PAGE in most of the samples 35-40 different bands were visualized; the most abundant ones were at apparent molecular weights (expressed in kDa) of 71, 66, 55, 46, 37, 26, 22, 14. Conclusion: HRE was a useful and rapid method to monitor renal function and the therapy effectiveness, in addition urinary albumin can also be quantified. SDS-AGE, according to literature, allowed the localization of nephronal damage. SDS-PAGE, due to its high sensitivity, permitted the visualization of many urine proteins, that could be studied as novel biomarkers of nephropathy.

Background: Data on the urinary proteome of domestic animals are limited, as a consequence the study of urinary proteome of healthy domestic animals is the first step to discover new biomarkers of nephropathy. Objective: The study of the urine proteome from healthy dogs, cats, horses and cows. Methods: Urine samples were collected from clinically healthy dogs (15), cats (10), horses (37) and cows (10). For all samples, UP/C was calculated and SDS-PAGE with silver staining was performed. Results: All samples had UP/C lower than 0.2. After SDS-PAGE, all samples showed many bands, which were undetectable by routine methods. In dogs were found 15-19 bands, in cats 17-20, in horses 13-15 and in cows 15-18. On the basis of the apparent molecular weight (MW) reported in kDa and comparing with human urine proteome, we tried an identification of the most represented proteins: P90 (Tamm-Horsfall), P76 (transferrin), P64 (albumin) P54 (immunoglobulin heavy chain) P27 (immunoglobulin light chain) and P14 (lysozyme). All samples presented putative albumin, transferrin, IG heavy and light chains; Tamm-Horsfall was clearly evident in dogs, horses and cats. Dogs and cows presented proteins mainly at MW lower than albumin, cats presented proteins mainly at MW higher than albumin, while horses presented proteins at both high and low MW. Conclusion: SDS-PAGE/silver staining is a very sensitive method to detect low abundance proteins in urine of healthy domestic animals, allowing the identification of different species-specific electrophoretic profiles. Work is in progress to confirm the identity of the different proteins.

Diabetes mellitus (DM) is a common disorder in middle-aged to older dogs. Treatment options, similar to those for humans, include insulin injection, dietary changes and exercise. Since some diabetic dogs may develop humoral immune responses to exogenous insulin resulting in treatment failure, the use of alternative medicine could represent an interesting new therapeutic strategy for DM in addition to the traditional insulin therapy. The main objective of this report was to evaluate a new therapeutic strategy for DM, based on the association of insulin injections with an orally administered extract of Boswellia serrata to improve glycemic control in a diabetic dog. A nine year old female neutered mixed breed dog diagnosed with DM was treated with increasing doses of a porcine insulin zinc suspension starting from 0.2 U/kg up to 0.6 U/kg q 12 hours. Twenty weeks after the start of insulin therapy the duration of effect and glucose nadir were ideal and clinical symptoms had disappeared but hyperglycemia, although less severe, persisted. Supplementation with a dry extract from the gum- resin of Boswellia serrata was initiated, at a dosage of 15 mg/kg q 12 hours, to improve insulin sensitivity or possible remaining endogenous insulin secretion. B. serrata supplementation led to good glycemic control. No side-effect or adverse reaction were observed during the study. The present case report provides the first evidence in veterinary medicine of a positive effect of dietary supplementation with boswellic acids associated with traditional insulin therapy on glycemic control in a diabetic dog. A major pitfall of the study is the lack of a control. More extensive clinical trials are required to provide definitive evidence of B. serrata efficacy. Key words: diabetes mellitus; dog; boswellic acids; glycemic control  VPLIV DODAJANJA BOSVELIJE ( Boswellia serrata ) OB ZDRAVLJENJU Z INZULINOM NA UREJANJE RAVNI GLUKOZE V KRVI PRI PSU S SLADKORNO BOLEZNIJO Sladkorna bolezen oz. diabetes mellitus (DM) je pogosta motnja pri psih v srednjih letih in starejAiih psih. MoA¾nosti zdravljenja, ki so podobne kot pri ljudeh, vkljuAujejo dodajanje inzulina, spremembe v prehrani in telesno aktivnost. Ker lahko nekateri psi s sladkorno boleznijo razvijejo imunski odziv na dodajanje inzulina, kar lahko vodi do neuspeAinega zdravljenja, lahko uporaba alternativne medicine predstavlja zanimivo novo metodo zdravljenja DM poleg tradicionalnega zdravljenja z inzulinom. Glavni cilj raziskave je bil ovrednotiti novo metodo zdravljenja DM, ki temelji na hkratnem dodajanju inzulina in peroralnem dodajanju izvleAka bosvelije ( Boswellia serrata ) za izboljAianje urejanja ravni glukoze v krvi pri psu s sladkorno boleznijo. Devet let stara sterilizirana samica meAiane pasme, ki je bila diagnosticirana s sladkorno boleznijo, je bila zdravljena z naraAiAajoAimi odmerki suspenzije praAiiAjega inzulina in cinka (od 0,2 E/kg do 0,6 U/kg), ki jih je dobivala vsakih 12 ur. Dvajset tednov po zaAetku zdravljenja z inzulinom so bili uAinki in najniA¾ja vrednost izmerjene glukoze v normalnih mejah vrednosti, kliniAni simptomi so izginili, le hiperglikemija je bila Aie vedno prisotna, vendar pa je bila manj resna. Dodatek suhega izvleAka iz gumijeve smole bosvelije Boswellia serrata v odmerku 15 mg/kg vsakih 12 ur je poveAal obAutljivost na inzulin oz. morebitne preostanke endogenega izloAanja inzulina. Dodatek B. serrata je pozitivno vplival na nadzorovanje sladkorne bolezni. Med zdravljenjem niso bili opaA¾eni stranski ali nezaA¾eleni uAinki. PoroAilo o opisanem primeru vsebuje prve dokaze o pozitivnem uAinku prehranskega dopolnila z bosveliAnimi kislinami v veterinarski medicini, povezanimi z obiAajnim zdravljenjem z inzulinom, pri nadzoru glukoze v krvi pri psu s sladkorno boleznijo. Najbolj neugoden del te Aitudije je odsotnost kontrole. Za zagotovitev dokonAnega dokaza o uAinkovitosti B. serrata so potrebni obseA¾nejAii kliniAni poskusi. KljuAne besede: diabetes mellitus ; pes; bosveliAna kislina; nadzor sladkorne bolezn