Lien et al. Acta Veterinaria Scandinavica 2010, 52:61 http://www.actavetscand.com/content/52/1/61 (Coat-A-Count Cortisol, Diagnostic Products Corp., U.S.A.) [27]. In this study, 70 dogs were affected with HAC, with the mean age of 11.7 ± 2.7 years and ranged from 4.5 to 18 years. Dogs had a mean body weight of 9.0 ± 7.6 kg ranging from 2.1 to 43.4 kg. Twenty-seven were spayed females, 21 were intact males, and 11 each of castrated males and intact females. Among these 70 cases, 40 dogs affected with PDH and 30 dogs affected with ADH (functional adrenal tumor). The mean age of the 40 dogs affected with PDH was 10.7 ± 2.7 years, ranging from 4.5 to 16 years with a mean body weight of 9.9 ± 8.6 kg, ranging from 2.1 to 43.4 kg. Sixteen were spayed females, 6 were intact females, 12 were intact males and 6 were castrated males. The mean age of the dogs affected with ADH was 12.9 ± 2.2 years, ranging from 8 to 18 years. This group had a mean body weight of 7.9 ± 5.6 kg, ranging from 3.0 to 24.5 kg. Eleven were spayed females, 5 were intact females, 9 were intact males, and 5 were castrated males. Breeds of 70 dogs affected with HAC are shown in Table 2. Systemic blood pressure measurement Systemic blood pressure values of 100 clinically healthy dogs and 70 dogs affected with (pre-treated) HAC were evaluated by the same protocol–Doppler sphygmomano- metry. A Doppler Ultrasonic Flow Detector (Model 811-B, Parks Medical Electronics Inc., U.S.A.) with an inflatable cuff width of 1.9, 2.5, 3, 4, or 5 cm (depending on the circumference of the antebrachium) was used. The cuff was wrapped around the middle part of the antebrachium, and a Doppler probe coated with ultraso- nic transmission gel was positioned over the palmar area to detect blood flow from the arteria digitalis pal- maris communis. The cuff was then inflated and deflated to obtain a systemic blood pressure reading via an aneroid pressure gauge. During systemic blood pres- sure measurement, the antebrachium was maintained at the level of the heart. A series of 5 readings (with 10 to Page 3 of 6 20 seconds between consecutive measurements) was obtained for each dog, and all measurements were com- pleted within 6 minutes. To minimize procedural stress, all dogs were allowed to assume a comfortable position with only gentle restraint by their owners. The dogs remained in the same position throughout systemic blood pressure measurement. The final systemic blood pressure value was calculated as the mean of 5 readings [28]. The heart rate was manually recorded by pulse Doppler ultrasound detection over a period of 20 sec- onds after the systemic blood pressure value was mea- sured. For this study, hypotension, normotension and hypertension were defined as <100 mmHg, 100-160 mmHg, and >160 mmHg, respectively. Microalbuminuria assay and urinary albumin:creatinine ratio Freshly voided urine samples from 100 clinically healthy dogs and 70 dogs affected with (pre-treated) HAC were collected. Urine albumin analysis was carried out within four hours after the freshly voided urine samples were collected. Urine samples with hematuria (≥10 RBC/high power of field), pyuria (≥5 WBC/hpf), or bacteriuria was excluded after a light-microscopic examination of urine sediment. The urine albumin and creatinine concentra- tions were semi-quantitated (Clinitek Microalbumin Reagent Strips and Clinitek Status Analyser, Bayer Diag- nostic Mfg. Ltd., UK) and urinary albumin:creatinine ratio (UACR) was calculated accordingly. In this study, no microalbuminuria, microalbuminuria and albuminuria were defined as UACR < 30 mg/g (0.03), 30-300 mg/g (0.03-0.3), and >300 mg/g (0.3), respectively. Statistical analysis Comparison of systemic blood pressure between clini- cally healthy dogs and dogs affected with either PDH or ADH were evaluated by means of ANOVA. Relationship between post-ACTH cortisol concentrations and sys- temic blood pressure was analyzed by use of linear Table 2 Systemic blood pressure (SBP) and status of urine albumin excretion in clinically healthy dogs and dogs affected with pituitary-dependent hyperadrenocorticism (PDH) or adrenal-dependent hyperadrenocorticism (ADH) Mean SBP (mmHg) Hypotension % (n) Normotension % (n) Hypertension % (n) Status of urine albumin No microalbuminuria % (n) Microalbuminuria % (n) Albuminuria % (n) Clinically healthy dogs (n = 100) 152.8 ± 27.3 1% (1/100) 87% (87/100) 12% (12/100) 53% (53/100) 45% (45/100) 2% (2/100) PDH (n = 40) 142.2 ± 24.9 0 80% (32/40) 20% (8/40) 15% (6/40) 52.5% (21/40) * 32.5% (13/40) * ADH (n = 30) 164.1 ± 36.7† 0 53.3% (16/30) 46.7% (14/30)*† 10% (3/30) 20% (6/30) *† 70% (21/30) *† * p < 0.05 relative to clinically healthy dogs. † p < 0.05 relative to PDH.