Isoamylases in clinically normal dogs

Isoenzymes of canine serum amylase were evaluated by electrophoresis on cellulose acetate membranes in a discontinuous buffer system. Two isoamylase groups were identified in the serum of clinically normal dogs. Most of the serum amylase activity was the more cathodal isoamylase. The anodal isoamylase was rarely observed in serum from normal dogs and, when present, accounted for little of the enzymatic activity. Amylase activities of various tissues were determined and isoenzymes were identified. The anodal isoenzyme was found in the pancreas and uterus. The cathodal isoamylase had its origin mainly from the intestinal tract. Activities of other tissues were not greater than was serum amylase activity. Effects of feeding upon total serum amylase activity and isoenzyme composition also were examined over a period of 5 minutes to 7 hours. Feeding did not markedly alter the serum amylase activity.     

Diseased dogs isoamylases in clinically normal and diseased dogs

Isoamylases in normal canine sera were separated on cellulose acetate membranes using a discontinuous buffer system without EDTA. Four peaks of amylase activity were present in 17 of 24 sera. Normal values were established. The majority of activity was present in Peak 4 (cathodal isoamylase). Tissue extracts of pancreas, duodenum, kidney, lung, testis, spleen and uterus-ovaries contained Peak 4 isoamylase. Liver and salivary gland lacked all isoamylase activity. Pancreas contained Peak 3 in addition to Peak 4 isoamylase. A tissue origin for Peaks 1 and 2 was not identified. An overall lack of resolution resulted from the inclusion of EDTA in the electrophoresis buffer system. This may account for previous findings suggesting that pancreatic amylase is not present in normal canine serum. An increase in the Peak 3 isoamylase was present in dogs with pancreatitis while dogs with pancreatic atrophy had a decrease in all isoamylases. Total amylase activity was significantly (p < 0.05) decreased in dogs with pancreatic atrophy.     

Impedance audiometric measurements in clinically normal dogs

OBJECTIVE: To measure impedance audiometric values in clinically normal dogs that were sedated or anesthetized, evaluate effects of ear flushing on tympanometric measurements, and determine effects of performing acoustic reflex testing in a sound-attenuated room. ANIMALS: 35 mixed-breed and purebred client-owned dogs and 21 laboratory-bred Beagles. PROCEDURES: Tympanometry and acoustic reflex testing were performed on 27 mixed-breed and purebred dogs under isoflurane anesthesia in a non-sound-attenuated room and 21 Beagles under sedation in a sound-attenuated room. Tympanometry was performed on 8 mixed-breed dogs under halothane anesthesia before and after ear canal flushing. RESULTS: Among impedance audiometric values, ear canal volume and compliance peak were smaller in Beagles than in mixed-breed dogs; differences among other values were not detected. Ear canal volume was dependent on body weight. Differences were not found for tympanometric values measured before and after ear canal flushing. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study established reference range values for impedance audiometric measurements in clinically normal dogs under isoflurane anesthesia or sedation. Acoustic reflex testing does not need to be performed in a sound-attenuated room. The ear canals of clinically normal dogs can be flushed prior to performing tympanometry without altering the results. Impedance audiometry may be a useful noninvasive procedure for the diagnosis of otitis media in dogs.     

Telomerase activity in clinically normal dogs and dogs with malignant lymphoma

OBJECTIVES: To determine whether telomerase activity was present in lymph nodes, buffy coat, and serum samples from dogs with malignant lymphoma (ML) and in liver, lymph node, buffy coat, and serum samples from clinically normal dogs SAMPLE POPULATION: Tissue specimens and blood samples were obtained from 11 clinically normal adult dogs (age range, 1 to 4 years) and 14 client-owned dogs with ML. PROCEDURE: The telomere repeat amplification protocol assay was used to quantify telomerase activity in the tissues from clinically normal dogs and dogs with ML. RESULTS: Of 11 clinically normal dogs, 8 had lymph node samples, 5 had liver samples, and 1 had buffy coat samples with detectable telomerase activity. None of the serum samples from the clinically normal dogs had detectable telomerase activity. Of 14 dogs with ML, 9 had lymph node samples, 3 had buffy coat samples, and 1 had serum samples with measurable telomerase activity. CONCLUSIONS AND CLINICAL RELEVANCE: Telomerase activity was not specific to tumor cells and overlapped with that found in cells from clinically normal dogs. Telomerase activity in neoplastic lymph nodes was not substantially different from that found in lymph nodes from clinically normal dogs. The determination of telomerase activity cannot be used as a sole diagnostic test for cancer. Therapeutic modalities directed toward the telomerase enzyme may not be feasible in dogs, because somatic tissues from clinically normal dogs possess variable amounts of telomerase activity.     

Ureterocolonic anastomosis in a dog with transitional cell carcinoma of the urinary bladder

Ureterocolonic anastomosis and cystectomy were performed in a dog for treatment of transitional cell carcinoma of the urinary bladder. The dog remained an acceptable house pet for 10 months after surgery. However, the tumor recurred 10 months after surgery, and the dog was euthanatized. Our results indicated that the combination of ureterocolonic anastomosis and cystectomy can be an acceptable form of palliative treatment for transitional cell carcinoma of the urinary bladder in the dog.     

Osmotic fragility of erythrocytes in clinically normal dogs and dogs infected with parasites

The osmotic fragility of the erythrocytes was measured in blood samples collected from randomly selected healthy and infected dogs at a dogs' rescue shelter. The dogs were classified into six groups on the basis of the final diagnoses from clinical, post mortem and laboratory findings. The minimum (less than 5 per cent) and maximum (more than 90 per cent) haemolysis of the erythrocytes of the clinically normal dogs (group 1), occurred in 0·60 per cent and 0·30 per cent solutions of sodium chloride (NaCI). For the non-anaemic hookworm-infected dogs (group 2a) the respective values were 0·8 per cent and 0·4 per cent NaCl, and for the anaemic hookworm-infected dogs (group 2b) they were 0·85 per cent and 0·5 per cent NaCl, respectively. The erythrocytes from dogs with Babesia canis (group 3), concurrent hookworm and B canis (group 4) and Ehrlichia canis infections (group 5) had minimum haemolysis in 0·75 per cent NaCl and maximum haemolysis at between 0·20 per cent and 0·35 per cent NaCI solutions. The derivative fragiligrams for groups 2a, 2b, 3 and 4 were shifted to the left, whereas the fragiligram for group 5 was similar to that for the clinically normal dogs (group 1). The left shift for the hookworm-infected dogs was due to the increased osmotic fragility of a minor sub-population of the erythrocytes, but for the dogs, infected with B canis major sub-populations of the erythrocytes had an increased osmotic fragility.     

Nuclear scintigraphic assessment of liver size in clinically normal dogs

Measurement of liver size was made on nuclear scintigraphic images obtained from 16 clinically normal, anesthetized dogs in ventral, dorsal, right and left lateral, and left dorsal oblique positions after administration of technetium 99m-sulfur colloid. Linear measurements of liver length and width were made from each scintiscan, and liver surface area was determined by setting a region of interest manually and by use of a computer count of pixels above a minimal intensity (threshold method). All linear measurements had a statistically significant (P less than 0.05) correlation with liver and body weight, with the exception of the measurements of liver width made on dogs in dorsal and left lateral dorsal oblique positions. The highest correlation (r = 0.89) between the scintigraphic measurements and liver weight was the multiplication of measurements of liver width and length from the right lateral view. Although all area determinations were significantly (P less than 0.05) correlated with liver and body weight, for most views, the manual method of determining the region of interest had a slightly better correlation with the liver weight than did the threshold method.     

Nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions

OBJECTIVE: To determine nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions. ANIMALS: 8 clinically normal female Beagles. PROCEDURE: Dogs were randomly assigned to receive 4 treatments in random order. Treatment A consisted of IV administration of nonlactated Ringer's solution. Treatments B, C, and D consisted of IV administration of isocaloric parenteral solutions containing 0, 1.36, and 2.04 g of amino acids/kg of body weight/d, respectively, for 7 consecutive days. Urine and feces were collected on days 5, 6, and 7 of each treatment period, and Kjeldahl analysis was used to determine nitrogen balance. RESULTS: Mean nitrogen balance was negative with treatments A and B but was not significantly different from 0 with treatments C and D. Dogs had the lowest nitrogen balance values and lost the most weight while receiving treatment A. Dogs were able to conserve protein and had higher nitrogen balance values when receiving treatment B, compared with treatment A. Dogs lost the least amount of weight while receiving treatment D. Regression analysis indicated that an IV amino acid intake of 2.32 g/kg/d (95% confidence interval, 2.00 to 2.81 g/kg/d), as supplied by the commercial product used in this study, would result in zero nitrogen balance in clinically normal dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IV amino acid requirement of clinically normal dogs is approximately 2.3 g/kg/d.     

Effect of mitotane on pituitary corticotrophs in clinically normal dogs

OBJECTIVE: To evaluate the effects of mitotane administration on the function and morphology of pituitary corticotrophs in clinically normal dogs. ANIMALS: 12 clinically normal adult Beagles. PROCEDURES: Dogs were randomly assigned to the control group or the mitotane treatment group. In mitotane treatment group dogs, mitotane was administered for 1 month. In both groups, ACTH stimulation testing and corticotrophin-releasing hormone (CRH) stimulation testing were performed. Magnetic resonance imaging (MRI) of the pituitary gland and brain was performed in mitotane treatment group dogs before and after administration of mitotane. After CRH stimulation testing and MRI, dogs were euthanatized and the pituitary gland and adrenal glands were excised for gross and histologic examination. RESULTS: ACTH concentrations in mitotane treatment group dogs were significantly higher than in the control group dogs following CRH stimulation. Magnetic resonance imaging revealed that pituitary glands were significantly larger in treatment group dogs after administration of mitotane, compared with before administration. On gross and histologic examinations, the adrenal cortex was markedly atrophied. Immunohistochemistry revealed hypertrophy of corticotrophs in pituitary glands of mitotane treatment group dogs. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicate that inhibition of the adrenal cortex by continuous administration of mitotane leads to functional amplification and morphologic enhancement of corticotrophs in clinically normal dogs. In instances of corticotroph adenoma, hypertrophy of individual corticotrophs induced by mitotane may greatly facilitate enlargement of the pituitary gland and increases in ACTH secretion.     

Quantitative noninvasive assessment of liver size in clinically normal dogs

Measurements of liver size were made from radiographs of 16 clinically normal anesthetized dogs. Two measurements were made from each of 3 views: right and left lateral and ventrodorsal. Each measurement was correlated with liver weight and volume. Liver weight and volume were also correlated with both measurements from the same radiograph multiplied together. All measurements, with the exception of ventrodorsal depth, correlated significantly (P less than 0.05) with liver weight and volume. The multiplication of the 2 measurements made from the right lateral radiograph correlated most highly with liver weight. These data were then compared with those from similar studies involving liver measurements obtained from nuclear scintigraphic and ultrasonographic scans on the same dogs. The radiographic measurements made from the right lateral view had the highest correlation with liver weight, followed by the nuclear scintigraphic measurements made from the same position.     

High-frequency ultrasonography of the skin of clinically normal dogs

OBJECTIVE: To assess the applicability of high-frequency diagnostic ultrasonography for evaluation and accurate measurement of the skin thickness of clinically normal dogs. ANIMALS: 26 healthy dogs (12 sexually intact males, 13 sexually intact females, and 1 spayed female) of various breeds and ages. PROCEDURE: Ultrasonographic examination of the skin and histomorphometric analysis of skin biopsy specimens obtained from the same site were performed. A 13-MHz linear-array transducer was used to obtain a series of ultrasonographic images of the skin in the flank region; images were analyzed and measured by use of imaging software. Cutaneous biopsy specimens were placed in fixative and then stained with H&E and Masson trichrome stains. Histomorphometric analysis was performed by use of an image analyzer. Thickness of the epidermis and dermis of each specimen was evaluated by use of a semiautomatic procedure of quantification. Data obtained from ultrasonographic and histologic measurements were compared by use of the Pearson correlation test. RESULTS: The ultrasonographic pattern of canine skin was consistently characterized by 3 distinct, defined echogenic layers corresponding to the epidermal entry echo, epidermis and dermis, and subcutaneous tissues. A positive correlation was found between ultrasonographic and histologic measurements of skin thickness. CONCLUSIONS AND CLINICAL RELEVANCE: Comparison between ultrasonographic and histologic appearance of the skin revealed that layering of canine skin (ie, epidermis and dermis) and the subcutaneous tissues may be recognized and measured by use of high-frequency ultrasonography. Thus, diagnostic ultrasonography may be a useful tool for the noninvasive evaluation of cutaneous disorders in dogs.     

Electromyographic and esophagomanometric findings in clinically normal dogs and in dogs with idiopathic megaesophagus.

Electromyography of 12 clinically normal dogs and 7 dogs with idiopathic megaesophagus revealed trains of positive sharp waves in the muscles of facial expression and in the lingual muscles of both groups. Positive waves are usually indicative of motor-unit disease; however, they are clinically insignificant in these muscles. Positive sharp waves were detected in the esophageal muscle of one dog with congenital megaesophagus. Esophageal electromyograms obtained in a dog with congenital megaesophagus and in 2 clinically normal dogs were normal. Resting caudal esophageal sphincter pressure was similar in both clinically normal dogs (mean, 22.3 mm of Hg; range, 15--37 mm of Hg) and in dogs with congenital or acquired idiopathic megaesophagus (mean, 29.6 mm of Hg; range, 20--50 mm of Hg).     

Fluorophotometric determination of aqueous humor flow rate in clinically normal dogs

OBJECTIVE: To determine aqueous humor flow rate in clinically normal dogs, using fluorophotometry. ANIMALS: 20 clinically normal Beagles. PROCEDURE: A study was performed on 5 dogs to establish an optimal protocol for fluorophotometric determination of aqueous humor flow rate. This protocol then was used to measure aqueous humor flow rate in 15 dogs. Corneas were loaded with fluorescein by topical application, and corneal and aqueous humor fluorescein concentrations were measured 5, 6.5, and 8 hours after application. Concentration-versus-time plots were generated, and slopes and ratios of the fluorescein concentration in the cornea and aqueous humor from these graphs were used to calculate flow rates. Calculations were performed by use of automated software provided with the fluorophotometer and by manual computation, and the 2 calculation methods were compared. RESULTS: The protocol established for the 5 dogs resulted in semilogarithmic and parallel decay of corneal and aqueous humor concentrations. Manually calculated mean +/- SD aqueous humor flow rates for left, right, and both eyes were 5.58 +/- 2.42, 4.86 +/- 2.49, and 5.22 +/- 1.87 microl/min, respectively, whereas corresponding flow rates calculated by use of the automated software were 4.54 +/- 3.08, 4.54 +/- 3.10, and 4.54 +/- 2.57 microl/min, respectively. Values for the left eye were significantly different between the 2 computation methods. CONCLUSIONS AND CLINICAL RELEVANCE: Aqueous humor flow rates can be determined in dogs, using fluorophotometry. This technique can be used to assess pathologic states and medical and surgical treatments that alter aqueous humor dynamics.     

Ultrasonographic and laboratory screening in clinically normal mature golden retriever dogs

Wellness and pre-anesthetic screening of blood and urine of geriatric companion animals are routinely recommended. In addition, there are occasional references to the use of imaging in clinically normal geriatric patients. However, the utility of wellness testing is not known, and there is limited information regarding the value of pre-anesthetic testing. Wellness testing, including complete blood cell count, biochemical profile, urinalysis, and abdominal ultrasound, was performed on 53 clinically normal, mature golden retriever dogs. Laboratory analysis revealed abnormalities in 54.7% (29/53) of the dogs. Abdominal ultrasound screening demonstrated abnormalities in 64.2% (34/53) of the dogs. As only a small number of dogs had follow-up diagnostic testing available, the significance of these abnormalities is unknown. Further study involving a larger cohort of animals and analysis of follow-up data is necessary to determine the utility of laboratory and imaging studies in clinically normal geriatric patients.     

Computed tomographic osteoabsorptiometry of the elbow joint in clinically normal dogs

OBJECTIVE: To evaluate subchondral bone density patterns in elbow joints of clinically normal dogs by use of computed tomographic (CT) osteoabsorptiometry. SAMPLE POPULATION: 20 cadaver forelimbs from 10 clinically normal dogs. PROCEDURE: Each elbow joint was imaged in parasagittal and transverse planes of 1.5-mm thickness. Slice data were converted to dipotassium phosphate equivalent density (PPED) values. Sagittal, parasagittal, and transverse medial coronoid process topographic maps were constructed. Defined zones were created for each of the 3 CT planes, and confluence and peak PPED values were determined. RESULTS: The lowest PPED value was 340 mg/ml (articular and subchondral confluence), and the highest was 1780 mg/ml (peak subchondral density). Detectable effects of joint laterality were not found in the confluence or peak PPED measurements or in the peak-to-confluence PPED ratio for all 3 CT planes. Significant differences were found among zones in all 3 planes for confluence and peak PPED measurements and between sagittal and transverse planes for peak-to-confluence PPED ratios. Subjectively, the pattern of density distribution among dogs was fairly consistent for the sagittal and parasagittal slices. Three specific patterns of density distribution were apparent on the transverse topographic maps of the medial coronoid process that corresponded to conformational differences. CONCLUSIONS AND CLINICAL RELEVANCE: The use of CT osteoabsorptiometry provides a repeatable technique that can be used to noninvasively examine bone density and the effects of stress acting on joints in vivo. Variability in density values for any of the CT planes was not identified among clinically normal dogs.     

Study of long-term survival after transsphenoidal hypophysectomy in clinically normal dogs

Experimental hypophysectomies were performed in 7 clinically normal dogs, using a new modification of the transsphenoidal approach. This approach facilitated centering of the sphenoid bone trephination and allowed safe exposure of the hypophysis regardless of the size or shape of a dog's skull. Complications did not occur during surgery and all dogs recovered well from surgery. Growth hormone secretory capacity was measured over a 3-month period to assess completeness of hypophysectomies. One dog was euthanatized 2 months after surgery, 4 dogs were euthanatized at 3 months after surgery, and 2 dogs were allowed to survive and their progress was followed for 2.5 years. Soft palate dehiscence and keratoconjunctivitis sicca developed in 2 of the dogs. The technical deficiencies responsible for these complications were corrected shortly after the beginning of the study. In 4 of the 5 necropsied dogs, minute remnants of adenohypophyseal tissue were found in the sellae turcica. Measurement of in vivo growth hormone secretory capacity revealed that these remnants had an altered stage of functional activity. Although complete hypophysectomy was not achieved consistently, the main technical obstacle of hypophysectomy, the reliable identification and the avoidance of the vascular structures surrounding the hypophysis, has seemingly been overcome. The surgical technique proved to enhance the safety of hypophysectomy, and the procedure can be recommended to treat clinical cases of canine pituitary-dependent hyperadrenocorticism. The clinical significance of potential subtotal hypophysectomy remains yet to be evaluated.