Phosphocalcic metabolism and its potential association with biomarkers of kidney disease in dogs with spontaneous hyperadrenocorticism.

The pathogenesis of increased serum phosphate concentration and proteinuria in dogs with spontaneous hyperadrenocorticism (HAC) is unclear. A potential link between proteinuria and calcium/phosphate metabolism has never been studied in dogs with HAC. The aims of the study were: (1) To evaluate calcium/phosphate metabolism in dogs with spontaneous HAC and compare to healthy dogs as well as to dogs with non-HAC illness; (2) to look for associations between markers of calcium/phosphate metabolism and biomarkers of kidney disease in dogs with HAC. Fifty-four dogs were included in the study, classified as HAC (n=27), non-HAC disease (n=17), and healthy (n=10). Serum calcium, phosphate, 25(OH)Vitamin D, 1,25(OH)2Vitamin D, plasma intact parathyroid hormone concentration (iPTH), FGF23, and urinary fractional excretion of calcium and phosphate were evaluated in all dogs at diagnosis and compared between each group. The correlation between these variables and urine protein-to-creatinine ratio (UPC) and urinary N-acetylglucosaminidase-to-creatinine ratio (uNAG/C) was evaluated in the HAC group. Medians [range] of serum phosphate concentration, urinary fractional excretion of calcium (FE(Ca)), and iPTH were significantly higher in dogs with HAC than in dogs with non-HAC illness (P<0.01) and healthy dogs (P<0.01). Increased 1,25(OH)2Vitamin D/25(OH)Vitamin D was also observed (P<0.001). In HAC group, UPC was significantly negatively correlated with 25(OH)Vitamin D (r(s): -0.54; P<0.01). Urinary NAG/C was significantly positively correlated with serum phosphate (r(s): 0.46; P=0.019). Increased serum phosphate, urinary excretion of calcium, and hyperparathyroidism were observed in dogs with HAC. Vitamin D metabolism may be shifted towards increased 1-alpha hydroxylation.

Description of serum symmetric dimethylarginine concentration and of urinary SDS-AGE pattern in dogs with ACTH dependent hyperadrenocorticism.

Serum symmetric dimethylarginine (SDMA) and patterns of urinary protein separated by sodium dodecyl sulfate agarose gel electrophoresis (SDS-AGE) have not been investigated as biomarkers in dogs with ACTH-dependent hyperadrenocorticism (ADHAC). This exploratory prospective study aimed to evaluate SDMA, serum creatinine (sCR), and SDS-AGE in dogs with ADHAC with and without proteinuria (ADHAC-P and ADHAC-nP, respectively). Thirty-five pet dogs classified as ADHAC-P (n=16), ADHAC-nP (n=6) and healthy (n=13) were included. Renal biomarkers were evaluated in all dogs at diagnosis. Baseline concentration of SDMA was not significantly different between the three groups (P = 0.15) whereas sCr was significantly lower in dogs in ADHAC dogs compared to healthy dogs (88.0 µmol/L [70.4-132.6; 79.2-114.4]) whether they had proteinuria or not (P = 0.014 and 0.002, respectively). However, baseline concentrations of sCr and SDMA were not significantly different between dogs with ADHAC-P dogs (SDMA, 8 µg/dL [5-12; 7-9]; sCr, 57.2 µmol/L [35.2-212.2; 52.8-92.4]) and ADHAC-nP dogs (SDMA, 8.5 µg/dL [7-13; 8-10]; sCr, 70.4 µmol/L [61.6-79.2; 61.6-70.4]) (P = 0.35 and P = 0.41, respectively). Proteinuria in dogs with ADHAC-P was mainly of glomerular origin (SDS-AGE pattern: glomerular in 10/16 dogs; mixed glomerular/tubular in four dogs). In our study, SDMA was neither significantly different in dogs with ADHAC whether they were proteinuric or not, nor between ADHAC and healthy dogs. Urinary electrophoresis provides additional information to the UPC and further investigations are needed to determine whether it may help identify dogs with ADHAC-P requiring specific antiproteinuric treatment.

Primary hypoaldosteronism in a dog with pituitary and adrenal T-cell lymphoma.

A 7-year-old mixed breed dog was presented with a 2-week history of vomiting, diarrhoea, weakness and loss of appetite. Initial laboratory tests revealed hyponatraemia and hyperkalaemia consistent with hypoadrenocorticism. Basal plasma cortisol and adrenocorticotropic hormone concentrations were not suggestive of primary hypoadrenocorticism but the aldosterone concentration was undetectable. Abdominal ultrasound scan showed a mass within the left kidney and a nodular enlargement of the left adrenal gland. Cytological analysis revealed a large granular lymphoma. The dog died 17 days later. Post mortem histological and immunohistochemical examinations revealed a diffuse large granular T-cell lymphoma involving the mediastinal lymph node, kidneys, pancreas, adrenal and pituitary glands.

Clinical description of Corynebacterium urealyticum urinary tract infections in 11 dogs and 10 cats.

OBJECTIVES: To examine the clinical signs, laboratory findings, possible predisposing factors, antimicrobial sensitivity and outcomes of dogs and cats diagnosed with urinary tract infection caused by Corynebacterium urealyticum. MATERIALS AND METHODS: Retrospective, observational study of dogs and cats that were diagnosed with urinary tract infection caused by C. urealyticum. RESULTS: This report concerns a total of 11 dogs and 10 cats. All cats had a history of urethral catheterisation, and six cats had undergone urological surgery before diagnosis of this specific infection. Dogs had history of neurogenic urinary incontinence (n=5), urethral catheterisation (n=4), urological surgery (n=4), lower urinary tract disease (n=2) or prostatic disease (n=1). In seven animals, previous urine culture was negative. Median urine pH was 8.6 (interquartile range 7.5 to 9), and 19 animals had pyuria and struvite crystalluria. Encrusted cystitis was diagnosed by abdominal ultrasound in nine animals. All but one isolate were resistant to at least three antimicrobial classes. Thirteen isolates were sensitive to tetracyclines. There was bacterial and clinical resolution of the infection in nine dogs and seven cats. Two animals died from sepsis and two cats were euthanased because of clinical deterioration. CLINICAL SIGNIFICANCE: Urinary tract infection caused by C. urealyticum should be suspected in animals with pre-existing urinary disorders, especially if urine is alkaline and there is struvite crystalluria, negative routine urine culture and encrusted cystitis.

Proopiomelanocortin processing and prohormone convertase 1 level in dogs with pituitary corticotroph tumors.

Preliminary data suggest that prohormone convertase 1 (PC1/3) protein expression and proopiomelanocortin (POMC) processing are altered in large corticotroph tumors. The aim of this study was to characterize the levels of ACTH precursors and PC1/3 protein in small and large corticotroph tumors of dogs with Cushing's disease. Pituitary tumors of dogs with Cushing's disease were collected postmortem 30 min to 12h after natural death or euthanasia, and classified as small or large. POMC, pro-ACTH, and PC1/3 were detected by Western blotting. Five small and 6 large corticotroph tumors were collected. POMC and pro-ACTH signals were visualized in 5/6 large tumors and in 4/5 small tumors. The strongest signal intensity was observed in 2 large tumors. The PC1/3 signal was weak to undetectable in 6/6 large tumors but strong in 5/5 small tumors. These results suggest differences in PC1/3 protein levels and patterns of POMC processing between large and small corticotroph tumors. If confirmed in larger groups of tumors, further studies will be required to characterize the mechanism involved in these differences.

Diagnostic contribution of cytological specimens obtained from biopsies during gastrointestinal endoscopy in dogs and cats.

OBJECTIVES: The aims of this study were to compare cytological samples obtained from endoscopic biopsies using "imprint" and "squash" techniques, and to evaluate the potential value of cytology compared to histology in reaching the diagnosis. MATERIALS AND METHODS: Eighteen dogs and five cats undergoing endoscopy for chronic gastrointestinal signs were prospectively included. Imprint and squash samples were obtained from one biopsy and then analysed. Comparison between cytology and histology was performed using Cohen's j coefficient. RESULTS: Appropriate samples for cytological evaluation were more often obtained with the squash technique (96% of the cases versus 68% with the imprint technique). The diagnoses obtained with cytological samples and by histology, considered as the gold standard, were compared. The same diagnosis was obtained with the squash technique in 65% of the cases. Furthermore, cytology was considered complementary to histology for gastric spiral organisms and mast cells identification. CLINICAL SIGNIFICANCE: These results suggest that squash cytology obtained from endoscopic biopsies of the gastrointestinal tract can provide relevant and additional information to histology in dogs and cats.

Initial evaluation of canine urinary cystatin C as a marker of renal tubular function.

OBJECTIVES: To evaluate the performance of a particle-enhanced turbidimetric assay for measuring canine urinary cystatin C and to investigate if the urinary cystatin C to creatinine ratio is higher in dogs with renal disease than in non-renal disease dogs. METHODS: Urinary cystatin C was measured by particle-enhanced turbidimetric assay using an avian antihuman cystatin C antibody and the performance of this assay was evaluated. Clinical relevance was tested in 46 dogs that were divided into three groups: healthy dogs (n=14), non-renal disease dogs (n=17) and dogs with renal disease (n=15). RESULTS: The assay was linear (R(2)=0·99) and precise (mean intra- and inter-assay coefficients of variation were 2·3 and 2·9%, respectively). The recovery was 111·5% and the limit of blank was 0·02 mg/L. Urinary cystatin C and urinary cystatin C to creatinine ratio differed significantly (P<0·001) between the three cohorts of dogs. CLINICAL SIGNIFICANCE: Measurement of cystatin C by particle-enhanced turbidimetric assay performed with high precision and linearity. This assay can be processed on automated clinical chemistry analysers making it widely available to commercial laboratories. Urinary cystatin C to creatinine ratio can differentiate dogs with renal disease from dogs without renal disease. These preliminary results suggest that urinary cystatin C to creatinine ratio is a promising marker for evaluating renal tubular function.

Plasma ACTH precursors in cats with pituitary-dependent hyperadrenocorticism.

BACKGROUND: Diagnosis of pituitary-dependent hyperadrenocorticism (PDH) in cats is challenging because there is no specific diagnostic test. HYPOTHESIS/OBJECTIVE: The determination of plasma ACTH precursor (POMC and pro-ACTH) concentration might facilitate the diagnosis of PDH in cats. The aim of the study was to evaluate prospectively the plasma concentrations of ACTH precursors in a small cohort of cats with PDH and to estimate the value of this approach for diagnosis. ANIMALS: Four groups of cats were included: group 1 (cats with PDH), group 2 (cats with diabetes mellitus but not hyperadrenocorticism (HAC)), group 3 (cats with diabetes mellitus and confirmed acromegaly but not HAC), and group 4 (healthy cats). METHODS: PDH diagnosis was based on clinical data, low-dose dexamethasone suppression test (LDDST), and adrenal and pituitary gland computed tomography (CT) scan. For groups 2, 3, and 4, hyperadrenocorticism was excluded by LDDST or urine cortisol:creatinine ratio (UCCR). An immunoluminometric assay was used to determine plasma concentrations of ACTH precursors in the 4 groups of cats. RESULTS: Group 1 contained 9 cats (enlarged pituitary gland in 7/9). Plasma ACTH precursor concentrations ranged from <53 to >1010 pmol/L with 8/9 concentrations ≥ 229 pmol/L. Groups 2, 3, and 4 included 13, 7, and 13 cats, respectively. Plasma ACTH precursor concentrations ranged from <53 to 96 pmol/L in group 2, <53 to 72 pmol/L in group 3, and <53 to 99 pmol/L in group 4. CONCLUSION AND CLINICAL IMPORTANCE: High plasma concentration of ACTH precursors in cats (>100 pmol/L) is highly suggestive of PDH.

Use of computed tomography adrenal gland measurement for differentiating ACTH dependence from ACTH independence in 64 dogs with hyperadenocorticism.

BACKGROUND: The measurement of adrenal gland size on computed tomography (CT) scan has been proposed for the etiological diagnosis of hyperadrenocorticism (HAC) in dogs. Symmetric adrenal glands are considered to provide evidence for ACTH-dependent hyperadrenocorticism (ADHAC), whereas asymmetry suggests ACTH-independent hyperadrenocorticism (AIHAC). However, there are currently no validated criteria for such differentiation. OBJECTIVE: The aim of this retrospective study was to compare various adrenal CT scan measurements and the derived ratios in ADHAC and AIHAC cases, and to validate criteria for distinguishing between these conditions in a large cohort of dogs. ANIMALS: Sixty-four dogs with HAC (46 ADHAC, 18 AIHAC). METHODS: Dogs with confirmed HAC and unequivocal characterization of its origin were included. Linear measurements of adrenal glands were made on both cross-sectional and reformatted images. RESULTS: An overlap was systematically observed between the AIHAC and ADHAC groups for all measurements tested. Overlaps also were observed for ratios tested. For the maximum adrenal diameter ratio derived from reformatted images (rADR), only 1/18 AIHAC dogs had a rADR within the range for ADHAC. For a threshold of 2.08, the 95% confidence intervals for estimated sensitivity and specificity extended from 0.815 to 1.000 and from 0.885 to 0.999, respectively, for AIHAC diagnosis. CONCLUSION AND CLINICAL IMPORTANCE: Measurements from cross-sectional or reformatted CT scans are of little use for determining the origin of HAC. However, rADR appears to distinguish accurately between ADHAC and AIHAC, with a rADR > 2.08 highly suggestive of AIHAC.

Ultrasonography criteria for differentiating ACTH dependency from ACTH independency in 47 dogs with hyperadrenocorticism and equivocal adrenal asymmetry.

BACKGROUND: Adrenal ultrasonography (US) in dogs with hyperadrenocorticism (HAC) is commonly used to distinguish adrenocorticotropic hormone (ACTH)-independent (AIHAC) and ACTH-dependent hyperadrenocorticism (ADHAC). To date, no cut-off values for defining adrenal atrophy in cases of adrenal asymmetry have been determined. Given that asymmetrical hyperplasia is sometimes observed in ADHAC, adrenal asymmetry without ultrasonographic proof of adrenocortical tumor such as vascular invasion or metastasis can be equivocal. OBJECTIVE: The purpose of this study was to compare adrenal US findings between cases of ADHAC and AIHAC in dogs with equivocal adrenal asymmetry (EAA), and to identify useful criteria for their distinction. ANIMALS: Forty dogs with EAA were included. METHODS: Ultrasound reports of HAC dogs with adrenal asymmetry without obvious vascular invasion or metastases were reviewed. Dogs were classified as cases of ADHAC (n = 28) or AIHAC (n = 19), determined by plasma ACTH concentration. The thickness, shape, and echogenicity of both adrenal glands and presence of adjacent vascular compression were compared between AIHAC and ADHAC groups. RESULTS: The maximal dorsoventral thickness of the smaller gland (SDV) ranged from 2.0 to 5.0 mm in AIHAC and from 5.0 to 15.0 mm in ADHAC. The 95% confidence intervals for estimated sensitivity and specificity of a SDV cut-off set at 5.0 mm in the diagnosis of AIHAC were 82-100 and 82-99%, respectively. Other tested US criteria were found to overlap extensively between the 2 groups, precluding their usefulness for distinction. CONCLUSION AND CLINICAL IMPORTANCE: In EAA cases, an SDV ≤5.0 mm is an appropriate cut-off for AIHAC ultrasonographic diagnosis.

Effect of thyroxine supplementation on glomerular filtration rate in hypothyroid dogs.

BACKGROUND: Glomerular filtration rate (GFR) is decreased in humans with hypothyroidism, but information about kidney function in dogs with hypothyroidism is lacking. HYPOTHESIS: Hypothyroidism influences GFR in dogs. The objective of this study was to assess GFR in hypothyroid dogs before implementation of thyroxine supplementation and after re-establishing euthyroidism. ANIMALS: Fourteen hypothyroid dogs without abnormalities on renal ultrasound examination or urinalysis. METHODS: Blood pressure and GFR (measured by exogenous creatinine clearance) were measured before treatment (T0, n=14) and at 1 month (T1, n=14) and at 6 months (T6, n=11) after beginning levothyroxine supplementation therapy (20 microg/kg/d, PO). The response to therapy was monitored at T1 by measuring serum total thyroxine and thyroid stimulating hormone concentrations. If needed, levothyroxine dosage was adjusted and reassessed after 1 month. Statistical analysis was performed using a general linear model. Results are expressed as mean+/-standard deviation. RESULTS: At T0, the average age of dogs in the study group was 6.3+/-1.4 years. Their average body weight decreased from 35+/-18 kg at T0 to 27+/-14 kg at T6 (P<.05). All dogs remained normotensive throughout the study. GFR increased significantly with levothyroxine supplementation; the corresponding results were 1.6+/-0.4 mL/min/kg at T0, 2.1+/-0.4 at T1, and 2.0+/-0.4 at T6 (P<.01). CONCLUSION: GFR was <2 mL/min/kg in untreated hypothyroid dogs. Re-establishment of a euthyroid state increased GFR significantly.

Accuracy of an adrenocorticotropic hormone (ACTH) immunoluminometric assay for differentiating ACTH-dependent from ACTH-independent hyperadrenocorticism in dogs.

BACKGROUND: Adrenocorticotropic hormone (ACTH) determination has been used for 30 years to distinguish ACTH-dependent hyperadrenocorticism (ADHAC) from ACTH-independent hyperadrenocorticism (AIHAC) in dogs. However, the few studies that have evaluated its diagnostic accuracy, based in the majority of cases on older assays, have been associated with systematic, but highly variable proportions of misclassified or unclassified cases. OBJECTIVE: The purpose of the present study is to evaluate the accuracy of a validated ACTH immunoluminometric assay (ILMA) for differentiating between ADHAC and AIHAC. ANIMALS: One hundred and nine dogs with hyperadrenocorticism were included: 91 with ADHAC and 18 with AIHAC. METHODS: Retrospective study. Dogs displaying feedback inhibition after the dexamethasone suppression test, adrenal symmetry, or both were considered to have ADHAC. AIHAC was demonstrated by adrenal tumor histology. For each group, ACTH determination by ILMA was reviewed. RESULTS: In the ADHAC group, plasma ACTH measurements ranged between 6 and 1250 pg/mL (median, 30 pg/mL). In the AIHAC group, all ACTH concentrations were below the lower quantification limit of the assay (<5 pg/mL). The 95% confidence interval was 85-100% for sensitivity and 97-100% for specificity in AIHAC diagnosis. CONCLUSION AND CLINICAL IMPORTANCE: No overlap in ACTH concentrations was observed between dogs with ADHAC and dogs with AIHAC. The use of a new technique with high analytical sensitivity made it possible to use a low threshold (5 pg/mL), avoiding the misclassification of some ADHAC cases with low, but quantifiable concentrations of ACTH. The assessment of ACTH concentrations by ILMA is an accurate tool for differentiating between ADHAC and AIHAC.