Protein profiling of urine from dogs with renal disease using ProteinChip analysis.

Measurement of total urinary proteins in individuals that tested positive by urinary dipstick is a typical method for assessing the presence of potentially serious renal disorders. In the absence of such overt proteinuria, however, measurement of specific urinary proteins may be useful in the diagnosis of nephropathies and may provide greater insight into the pathogenesis. The urine of 28 dogs (16 with renal disease and 12 healthy) was evaluated to determine whether specific low-molecular-weight proteins or the pattern of protein excretion could also be used as a marker of tubular dysfunction in dogs. Specific proteins were assessed by immunological methods, whereas protein profiles were determined by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (MS). In particular, changes in the excretion of retinol-binding protein (RBP) and Tamm-Horsfall protein (THP) appear to be of clinical relevance in the diagnosis of canine kidney diseases. The pattern of urinary protein and peptides revealed specific changes in abundance in dogs with renal disease at molecular masses (kD) of 11.58, 12.41, 12.60, 14.58, 20.95 (RBP), 27.85, and 65.69 (albumin). In conclusion, comparable proteins as in humans might be used as urinary markers for proximal (RBP) and distal (THP) tubular dysfunction in dogs. Surface-enhanced laser desorption/ionization time-of-flight MS is a promising tool for the study of kidney physiology and pathophysiology and might aid in the discovery of new biomarkers of renal disease.     

Levels of retinol and retinyl esters in plasma and urine of dogs with urolithiasis

Vitamin A (VA) deficiency and Tamm-Horsfall glycoprotein (THP), a protein that binds retinol and retinyl esters in canine urine, might be involved in the pathogenesis of urolithiasis in dogs. In the present study, we assessed levels of retinol, retinyl esters, retinol-binding protein (RBP) and THP in plasma and urine of dogs with a history of urolithiasis (n = 25) compared with clinically healthy controls (n = 18). Plasma retinol concentrations were higher in dogs with uroliths of struvit (P < 0.01), calcium oxalate (P < 0.05), urate (P < 0.01) and cysteine, but there were no differences in the concentrations of plasma RBP and retinyl esters. Excretion of urinary retinol and retinyl esters were tentatively, but not significantly higher in the stone-forming groups, which was accompanied by increased levels of urinary RBP (P < 0.01) and lower excretions in THP (P < 0.01). The results show that VA deficiency may be excluded as a potential cause for canine urolithiasis. However, the occurrence of RBP and a concomitant reduction of THP in urine indicates a disturbed kidney function as cause or consequence of stone formation in dogs.     

Proteinuria in the dog: a clinicopathological study in 51 proteinuric dogs

In 51 dogs with predominantly massive urinary protein loss, the daily loss was quantified and glomerular and tubulointerstitial lesions from renal biopsies were characterised and graded using histology, immune fluorescence and electron microscopy. The highest median daily urinary protein loss occurred in dogs with membranous glomerulonephritis (median 380 mg kg-1 d-1) and renal amyloidosis (median 257 mg kg-1 d-1). Although in nine febrile dogs the urinary protein loss was transient, both glomerular and tubular lesions were diagnosed in five and seven of these dogs, respectively. The pattern of urinary proteins was determined using sodium dodecyl-sulphate polyacrylamide gel electrophoresis. The albumin fractional clearance (FC) was raised in 46 dogs, whereas the FCS of the low molecular weight (MW) protein fraction (MW less than 66,000) and high molecular weight protein fraction (MW more than 66,000) were raised in 42 and 28 dogs, respectively. Both the high molecular weight protein FC and albumin FC significantly correlated to the grade of glomerular lesions, whereas the low molecular weight protein FC only moderately significantly correlated to the grade of tubular lesions. The selectivity index, calculated as (formula; see text) did not differentiate between the various forms of glomerulopathies. The urinary lysozyme concentration was significantly correlated to the grade of tubular lesions. It is concluded that although quantitative and qualitative measurements of urinary proteins can provide additional clinical information, they do not have a reliable predictive value and histopathological examination of renal tissue is still necessary for the final diagnosis.     

Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis of Canine Urinary Proteins for the Analysis and Differentiation of Tubular and Glomerular Diseases

A preliminary investigation was performed to evaluate the use of a new, noninvasive technique for the localization of canine renal lesions by electrophoresis of urinary proteins. Urine specimens from six clinically healthy, nonproteinuric dogs and 12 dogs with persistent proteinuria were examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). Urine electrophoretic patterns of proteinuric dogs were classified as glomerular (n=4), tubular (n=2), or mixed (glomerular and tubular) (n=6), based on the number and molecular weight of the silver-stained protein bands. Renal tissues from biopsies or necropsies were obtained from eight of the dogs with proteinuric disease. Interpretation of seven of eight electrophoretograms agreed with the histologic interpretation of renal lesions. We concluded SDS PAGE is a potentially valuable technique for detection and localization of renal lesions in dogs with proteinuric disease.     

Sodium dodecyl sulfate polyacrylamide gel electrophoresis of canine urinary proteins for the analysis and differentiation of tubular and glomerular diseases

A preliminary investigation was performed to evaluate the use of a new, noninvasive technique for the localization of canine renal lesions by electrophoresis of urinary proteins. Urine specimens from six clinically healthy, nonproteinuric dogs and 12 dogs with persistent proteinuria were examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). Urine electrophoretic patterns of proteinuric dogs were classified as glomerular (n = 4), tubular (n = 2), or mixed (glomerular and tubular) (n = 6), based on the number and molecular weight of the silver-stained protein bands. Renal tissues from biopsies or necropsies were obtained from eight of the dogs with proteinuric disease. Interpretation of seven of eight electrophoretograms agreed with the histologic interpretation of renal lesions. We concluded SDS PAGE is a potentially valuable technique for detection and localization of renal lesions in dogs with proteinuric disease.     

Electrohydraulic Shock-wave Lithotripsy in Experimental Canine Struvite Bladder Stone Disease

Electrohydraulic shock-wave lithotripsy (EHL), the fragmentation of urinary stones with hydraulic shock waves, was performed through a cystoscope in two female dogs with surgically implanted struvite bladder stones. Stone fragmentation was achieved after about an hour, but complete removal of fragments by irrigation through the cystoscope sheath was not successful. After the procedure, small stone fragments and pieces of the shock-wave electrode remained in the bladders of both dogs. All stone fragments and electrode pieces were spontaneously voided by one dog; further cystoscopy was required in the other dog to remove three larger stone fragments with a stone basket, and pieces of the electode with grasping forceps. Bladder-stone removal in the female dog by EHL, irrigation, and using a stone basket is technically feasible, but requires expensive equipment.     

Diagnostic relevance of qualitative proteinuria evaluated by use of sodium dodecyl sulfate-agarose gel electrophoresis and comparison with renal histologic findings in dogs

OBJECTIVE: To evaluate results of SDS-agarose gel electrophoresis (AGE) of urinary proteins for use in defining glomerular and tubulointerstitial derangements, investigate patterns of high-molecular-weight (HMW) proteins for differentiating among glomerular disorders, and assess low-molecular-weight (LMW) proteins as markers of severity of tubulointerstitial disease in dogs. ANIMALS: 49 dogs with increased serum creatinine concentrations or abnormal renal protein loss. PROCEDURE: Urinary proteins were examined by use of SDS-AGE and differentiated on the basis of molecular weight. The HMW proteins (> or = 69 kd) were considered indicative of glomerular origin, whereas LMW proteins (< 69 kd) were of tubular origin. Renal specimens were examined by use of light microscopy. Glomerular and tubulointerstitial lesions were differentiated by use of the classification for the World Health Organization and semiquantitative grading, respectively. RESULTS: Sensitivity of SDS-AGE was 100% for detection of glomerular lesions and 92.6% for tubulointerstitial lesions; specificity was 40% and 62.5%, respectively. Although HMW urinary proteins were not significantly associated with the type of glomerular lesion, LMW urinary proteins were significantly associated with the grade of tubulointerstitial damage. Detection of 12- or 15-kd proteins or both was highly indicative of a severe tubulointerstitial lesion. CONCLUSIONS AND CLINICAL RELEVANCE: SDS-AGE of urinary proteins in dogs represents a noninvasive test with high sensitivity for identifying glomerular and tubulointerstitial damage, but low specificity limits its validity as a stand-alone test to differentiate between glomerular and tubulointerstitial lesions. The test is particularly useful for identifying dogs with advanced tubulointerstitial disease but cannot be used to characterize glomerular disorders.     

Percutaneous Nephrolithotomy in the Dog An Experimental Study

Percutaneous nephrostomies (PN) were created with teflon fascial dilators under fluoroscopic guidance, to implant stones bilaterally into the renal pelves of five dogs weighing 7 to 35 kg. Between one and seven stones of 0.25 to 1.00 cm diameter were implanted into each renal pelvis. Two to 4 weeks later, PN tracts were recreated, and the implanted renal stones were successfully removed under fluoroscopic and endoscopic visualization. Results of gross and histopathologic evaluation of the kidneys 2, 7, 14, 30, and 90 days after stone removal showed minimal, localized renal parenchymal trauma. Complications, although infrequent, included proximal ureteral tears and hemorrhage. Difficulty was encountered in maneuvering equipment around large or numerous stones. Canine percutaneous nephrolithotomy was successful despite great variation in kidney size, as well as variations in the number, size, and shape of stones in the upper urinary tract.     

Multiple meningiomas in three dogs

Three dogs with seizures were diagnosed with multiple intracranial meningiomas. Two of the three dogs were golden retrievers, and ages ranged from 9 to 11 years. Treatment consisted of surgery and radiation (n=2) or chemotherapy (n=1). In all three cases, the masses were two distinct tumors as determined by imaging, surgery, or necropsy. In two dogs, the meningiomas had the same histological pattern, while in one dog the histological subtypes were different.     

Cats and calcium oxalate: strategies for managing lower and upper tract stone disease

PRACTICAL RELEVANCE: Calcium oxalate (CaOx) containing stones are among the most common of the urinary tract stones identified in cats. RISK FACTORS: Risk factors for CaOx stone formation include such things as breed, gender and diet; stress and obesity have also been hypothesized to be risk factors for this disease. MANAGEMENT APPROACH: A tailored, individual management strategy for preventing CaOx stone recurrence is important and should include addressing the diet, environment and any other comorbid conditions present. Increasing the cat's moisture intake is one of the key mechanisms for preventing recurrence. CLINICAL CHALLENGES: CaOx ureterolithiasis has emerged as a difficult and sometimes life-threatening problem for cats. In those cats where stones are found incidentally, periodic monitoring may be required to assess for disease progression. Interventional procedures such as ureteral stent placements are now increasingly being performed for recurrent cases or those with larger stone burdens. Periodic radiographs for more severe cases and frequent client communication can help ensure successful outcomes for cats with lower and upper CaOx stone disease. EVIDENCE BASE: Limited evidence-based studies are published regarding management of feline upper and lower urinary tract CaOx stone disease, making this a difficult condition to manage in some cats. Studies designed to evaluate the relationship to dietary modifications, medical management, stress, obesity and surgical techniques are warranted in cats with upper and lower urinary tract CaOx stones.     

Medical dissolution of struvite nephrolithiasis using amino acid preparation in dogs

A new chemolytic therapy with amino acid preparation was tried for use on struvite renal stones in two dogs, which were admitted to Azabu University Veterinary Teaching Hospital. No other special diet, for the purpose of dissolving the stones, was fed during the treatment. Dissolution of the stones began at week 1, and no stone was detected on radiography at week 4 in both cases. The amino acid preparation thereafter has been administered every 2-3 days to prevent formation of the stones. After 6 months from treatment, complete dissolution was confirmed, and no relapse has been observed. Therefore, it is concluded that chemolytic therapy with amino acid preparation might be useful for the treatment of nephrolithiasis with struvite renal stones in dogs.     

母犬二次膀胱尿道结石症的诊治

犬尿路结石在徐州地区发病较高,并且复发率较高,给家庭养犬业造成了极大的影响。犬尿路结石主要发生于老龄公犬,笔者最近在临床上接诊一例复杂的母犬膀胱尿道结石病例,尤其尿道结石较大,处理起来非常困难,最后采用尿道口切开术,将结石顺利取出。犬尿路结石的手术疗法国内一般采用膀胱切开术和尿道切开术,采用尿道口切开术尚属首例。     

Characterization of the renal response to protein ingestion in dogs with experimentally induced renal failure.

Effects of a protein meal (2.7 g of casein/kg of body weight) on glomerular filtration rate (GFR) and renal plasma flow (RPF) were assessed in dogs after 15/16 nephrectomy (n = 10), and were compared with observations in dogs with intact kidneys (n = 5). Increase in GFR and RPF was observed in both groups of dogs between 1.5 and 8 hours after protein ingestion. A maximal value for GFR was observed between 4 and 5 hours after protein ingestion in dogs of both groups. Enhancement of urinary protein excretion was evident in partially nephrectomized dogs after protein ingestion (P less than 0.05), a result that was confirmed by 24-hour total urine collection from partially nephrectomized dogs fed a balanced ration. A qualitatively similar vasodilatory response was observed in partially nephrectomized dogs and in dogs with intact kidneys, and the mean maximal increase of GFR and RPF expressed as a percentage of baseline values in the latter dogs (47.0 +/- 8.1 and 43.6 +/- 10.3%, respectively) exceeded that observed in partially nephrectomized dogs (20.8 +/- 2.2 and 22.7 +/- 6.3%, respectively; P less than 0.01). The incremental response of the kidneys to protein ingestion was directly related to the degree of renal function, as reflected in the linear regression relationship between the incremental increase in GFR and the baseline value for GFR (P less than 0.01, R2 = 0.721).     

Identification of allergens responsible for canine cutaneous adverse food reactions to lamb, beef and cow's milk

Lamb, beef and cow's milk are common causes of cutaneous adverse food reactions in dogs. The aim of this study was to identify the proteins responsible for cutaneous adverse reactions to these foods. Ten dogs with allergen-specific serum immunoglobulin (Ig)E to lamb, beef and cow's milk were included in the study. These dogs had been diagnosed with cutaneous adverse food reactions by convincing clinical history and food-elimination diet trials followed by challenge exposure. Sera were analysed by enzyme-linked immunosorbent assay with bovine proteins and SDS-PAGE immunoblots with lamb, beef and cow's milk extracts. All the dogs had specific IgE against bovine IgG, and it was the only protein in the cow's milk extract that bound IgE from the sera studied. In the lamb and beef extracts, the major allergens recognized by the specific IgE of most sera had molecular masses between 51 and 58 kDa, which were identified as phosphoglucomutase and the IgG heavy chain. Other IgE-binding proteins with molecular masses of 27, 31, 33, 37 and 42 kDa were also detected with some sera. Our results indicate that bovine IgG is a major allergen in cow's milk and hence it appears to be a source of cross-reactivity with beef and probably with lamb because of the high homology with ovine immunoglobulins. These results are similar to those found for meat allergy in humans. However, this is the first time that phosphoglucomutase has been identified as an important allergen involved in allergic reactions to lamb and beef.     

动物尿结石研究进展

随着现代科学技术的不断发展,尿结石的形成机理成为研究的热点.泌尿系结石,现代医学称其为尿结石或尿石症,是指在泌尿系统中晶体物质和有机物异常积聚形成的石样物质,因结石所在的部位不同分为肾石样物质、输尿管结石、膀胱结石以及尿道结石.近年来动物尿结石给畜牧业造成了很大的损失,尤其是反刍动物尿结石发病率较高,但现今仍无有效的预防和治疗方法.无机金属离子和有机基质在尿结石形成过程中起着重要的作用,此文对尿结石形成学说、成因分析以及相关研究进展进行综述.     

Markers of bone metabolism in dog breeds of different size

Serum and urinary markers of bone turnover may be of value in animals as noninvasive tools for determining the response of the skeleton to disease and injury. Although normal values for bone markers have been reported for the Beagle, concerns remain that breed to breed differences will complicate the interpretation of bone marker data in dogs. To explore this, we examined serum bone markers in two breeds of vastly different size, Pomeranians and Irish Wolfhounds. Our hypothesis was that serum concentrations of bone markers are similar in toy and giant dog breeds and fall within the same range as those reported for Beagles. Bone alkaline phosphatase (BALP) and carboxy-terminal telopeptide of type I collagen (ICTP), respectively markers of bone formation and bone resorption, were measured in age matched Pomeranians (n=14) and Irish Wolfhounds (n=14). No statistically significant differences between the mean BALP and mean ICTP serum concentrations from Pomeranians and Irish Wolfhounds were found. All BALP and ICTP concentrations were within the reference range reported for Beagles. The results of this study suggest that serum BALP and ICTP concentrations in giant and toy breeds are the same as in Beagles and that these assays may be used for dogs of all sizes.     

Cystine-containing urinary calculi in dogs: 102 cases (1981-1989).

One hundred and seven cystine-containing urinary calculi from 1 female and 101 male dogs were analyzed. Cystine-containing calculi accounted for 2% (107 of 5,375) of all canine urinary calculi submitted to the urinary stone analysis laboratory from July 1981 through December 1989. Male dogs that formed cystine calculi were compared with 3 other canine populations to determine whether certain breeds were apparently at increased or decreased risk for cystine calculus formation. In one or more of 3 population comparisons, significantly increased risk of cystine calculus formation was found in Mastiffs, Australian Cattle Dogs, English Bulldogs, Chihuahuas, Bullmastiffs, Newfoundlands, Dachshunds, Basenjis, Australian Shepherd Dogs, Scottish Deerhounds, Staffordshire Terriers, Miniature Pinschers, pitbull terriers, Welsh Corgis, Silky Terriers, and Bichon Frises. Significantly low risk of cystine calculus formation was found in German Shepherd Dogs, Poodles, Schnauzers, and mixed-breed dogs.     

Analysis of serum proteins, using agarose electrophoresis in normal dogs and in dogs naturally infected with Dirofilaria immitis.

The electrophoretic pattern of 130 serum samples from clinically normal dogs was evaluated, using agarose as the supporting matrix. The relative mobility of each globulin fraction in relationship to the mobility of albumin was determined in 70 dogs, and nomenclature based on the mobilities (Rf values) was proposed. Biuret protein determinations were done, and relative and absolute values of each serum protein fraction were determined. Changes in the fractions of serum proteins were evaluated in regard to sex and age of the dogs and the presence of microfilariae of Dirofilaria immitis, as determined by modified Knott tests.     

Assessment of IgE binding to native and hydrolyzed soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity

OBJECTIVE: To assess binding of IgE to native, whole hydrolyzed, and separated hydrolyzed fractions of soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity. ANIMALS: 8 na?ve Beagles (6 experimentally sensitized to native soy protein and 2 control dogs). PROCEDURES: 6 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization protocol was completed, serum concentrations of soy-specific IgE were measured and intradermal skin tests were performed in all 6 dogs to confirm that the dogs were sensitized against soy protein. Serum samples from each sensitized and control dog underwent western blot analysis to assess the molecular mass band pattern of the different allergenic soy fractions and evaluate reactivities to native and hydrolyzed soy protein. RESULTS: In sera from sensitized dogs, a characteristic band pattern with 2 major bands (approx 75 and 50 kd) and 2 minor bands (approx 31 and 20 kd) was detected, whereas only a diffuse band pattern associated with whole hydrolyzed soy protein was detected in the most reactive dog. Reactivity was evident only for the higher molecular mass peptide fraction. In control dogs, no IgE reaction to native or hydrolyzed soy protein was detected. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that the binding of soy-specific IgE to the hydrolyzed soy protein used in the study was significantly reduced, compared with binding of soy-specific IgE to the native soy protein, in dogs with experimentally induced soy hypersensitivity.     

Effects of phosphorus/calcium-restricted and phosphorus/calcium-replete 32% protein diets in dogs with chronic renal failure.

Twenty-four dogs with induced, severe chronic renal failure were allotted to 2 groups of 12 each. Group-A dogs were fed a 0.4% phosphorus (P)/0.6% calcium, 32% protein diet, and group-B dogs were fed a 1.4% P/1.9% calcium, 32% protein diet. Dogs were studied over 24 months to determine clinical status, survival, blood biochemical alterations, glomerular filtration rate (GFR), urinary excretion of P and protein, renal morphologic changes, and renal tissue concentrations of calcium, P, and magnesium. Group-A dogs developed statistically significant differences from group-B dogs in several blood biochemical values (PCV and total solids, calcium, P, potassium, sodium, chloride, total CO2 (TCO2), anion gap, and parathyroid hormone concentrations) and in urinary P excretion. Mean (+/- SEM) GFR values in group-A and group-B dogs were nearly identical when diets were initiated (group A = 0.73 +/- 0.05 ml/min/kg of body weight; group B = 0.72 +/- 0.08 ml/min/kg), but significantly (P = 0.0346) lower GFR developed in group-B than in group-A dogs over time. At 24 months, GFR in survivors was 0.83 +/- 0.08 and 0.63 +/- 0.15 ml/min/kg for dogs of groups A and B, respectively. Other measurements favored the hypothesis that P/calcium restriction was beneficial, but values failed to reach statistical significance. Survival was greater at 24 months in group-A than in group-B (7 vs 5) dogs, and renal tissue concentrations of calcium and P were higher in group-B than in group-A dogs. Differences were not detected between groups in urinary excretion of protein and in the type or severity of renal lesions.(ABSTRACT TRUNCATED AT 250 WORDS)