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- Author or Editor: Craig G. Ruaux x
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Abstract
Objective—To develop and validate a gas chromatography–mass spectrometry (GC-MS) method for determination of Nτ-methylhistamine (NMH) concentration in canine urine and fecal extracts and to assess urinary NMH concentrations in dogs with mast cell neoplasia and fecal NMH concentrations in dogs with protein-losing enteropathy.
Sample Population—Urine specimens were collected from 6 healthy dogs and 7 dogs with mast cell neoplasia. Fecal extracts were obtained from fecal specimens of 28 dogs with various severities of protein-losing enteropathy, as indicated by fecal concentration of α1-proteinase inhibitor.
Procedures—NMH was extracted directly from urine, and fecal specimens were first extracted into 5 volumes of PBSS containing 1% newborn calf serum. Nτ-methylhistamine in specimens was quantified via stable isotope dilution GC-MS. The assay was validated via determination of percentage recovery of known amounts of NMH and interassay coefficients of variation. Urinary excretion of NMH was evaluated by means of NMH-to-creatinine concentration ratios.
Results—Recovery of NMH in urine and fecal extracts averaged 104.6% and 104.5%, respectively. Interassay coefficients of variation ranged from 5.4% to 11.7% in urine and 12.6% to 18.1% in fecal extracts. Urinary NMH excretion was significantly increased in dogs with mast cell neoplasia, compared with that in healthy dogs. No correlation was detected between severity of protein-losing enteropathy and fecal NMH concentration.
Conclusions and Clinical Relevance—This method provided a sensitive, reproducible means of measuring NMH in canine urine and fecal extracts. High urinary NMH-to-creatinine concentration ratios in dogs with mast cell neoplasia are consistent with increased histamine release in this disease.
Abstract
Objective—To investigate postprandial changes in serum concentrations of unconjugated bile acids in healthy Beagles.
Animals—7 healthy Beagles.
Procedure—Blood samples were obtained from dogs at regular intervals up to 8 hours after consumption of a meal. Serum concentrations of 5 unconjugated bile acids were determined at each time point, using gas chromatography-mass spectrometry with selected ion monitoring.
Results—Total serum unconjugated bile acid concentration was significantly increased, relative to baseline values, at 360, 420, and 480 minutes after feeding. Unconjugated cholic acid was significantly increased at 360, 420, and 480 minutes. The proportion of total unconjugated bile acids represented by cholic acid was significantly increased at 240 to 480 minutes. Deoxycholic acid was significantly increased at 360 and 420 minutes. Chenodeoxycholic acid was significantly increased at 360 to 480 minutes. Lithocholic acid was significantly increased at 180 minutes, whereas no significant changes in ursodeoxycholic acid were detected at any time point.
Conclusion and Clinical Relevance—Healthy Beagles had significant increases in serum concentrations and changes in the profile of unconjugated bile acids after a meal. These increases persisted > 8 hours, indicating that prolonged withholding of food is necessary when to avoid the risk of a false-positive diagnosis when assessing serum unconjugated bile acid concentrations in dogs. (Am J Vet Res 2002;63:789–793
Abstract
Objective—To validate an automated chemiluminescent immunoassay for measuring serum cobalamin concentration in cats, to establish and validate gas chromatography-mass spectrometry techniques for use in quantification of methylmalonic acid, homocysteine, cysteine, cystathionine, and methionine in sera from cats, and to investigate serum concentrations of methylmalonic acid, methionine, homocysteine, cystathionine, and cysteine as indicators of biochemical abnormalities accompanying severe cobalamin (vitamin B12) deficiency in cats.
Sample Population—Serum samples of 40 cats with severe cobalamin deficiency (serum cobalamin concentration < 100 ng/L) and 24 control cats with serum cobalamin concentration within the reference range.
Procedure—Serum concentrations of cobalamin were measured, using a commercial automated chemiluminescent immunoassay. Serum concentrations of methylmalonic acid, methionine, homocysteine, cystathionine, and cysteine were measured, using gas chromatography-mass spectrometry, selected ion monitoring, stable-isotope dilution assays.
Results—Cats with cobalamin deficiency had significant increases in mean serum concentrations of methylmalonic acid (9,607 nmol/L), compared with healthy cats (448 nmol/L). Affected cats also had substantial disturbances in amino acid metabolism, compared with healthy cats, with significantly increased serum concentrations of methionine (133.8 vs 101.1 µmol/L) and significantly decreased serum concentrations of cystathionine (449.6 vs 573.2 nmol/L) and cysteine (142.3 vs 163.9 µmol/L). There was not a significant difference in serum concentrations of homocysteine between the 2 groups.
Conclusions and Clinical Relevance—Cats with gastrointestinal tract disease may have abnormalities in amino acid metabolism consistent with cobalamin deficiency. Parenteral administration of cobalamin may be necessary to correct these biochemical abnormalities. (Am J Vet Res 2001;62:1852–1858)
Abstract
Objective—To characterize historical, clinicopathologic, ultrasonographic, microbiological, surgical, and histopathologic features of bacterial cholecystitis and bactibilia in dogs and evaluate response to treatment and outcomes in these patients.
Design—Retrospective case-control study.
Animals—40 client-owned dogs (10 with bacterial cholecystitis on histologic analysis or bactibilia on cytologic examination [case dogs] and 30 without bactibilia [controls]) evaluated at a veterinary teaching hospital between 2010 and 2014.
Procedures—Signalment, history, clinicopathologic findings, ultrasonographic features, microbiological results, surgical findings, histopathologic changes, treatments, and outcomes of case dogs were derived from medical records and summarized. Demographic and clinicopathologic data and ultrasonographic findings were compared between case and control dogs. Relationships among prior antimicrobial treatment, sediment formation in the gallbladder, presence of immobile biliary sludge, and presence of bactibilia or bacterial cholecystitis were assessed.
Results—No finding was pathognomonic for bactibilia or bacterial cholecystitis in dogs. Case dogs were significantly more likely to have immobile biliary sludge and had a greater degree of biliary sediment formation than did control dogs. All case dogs for which gallbladders were examined histologically (6/6) had bacterial cholecystitis. Five of 10 case dogs were Dachshunds. Medical or surgical treatment resulted in good outcomes.
Conclusions and Clinical Relevance—Bactibilia and bacterial cholecystitis were important differential diagnoses in dogs with signs referable to biliary tract disease. Dachshunds were overrepresented, which may suggest a breed predisposition. Cytologic evaluation of bile should be considered in the routine assessment of dogs with hepatobiliary disease if immobile biliary sludge is present. (J Am Vet Med Assoc 2015;246:982–989)
Abstract
OBJECTIVE To assess clinical effects of an omega-3 fatty acid and protein-enriched diet, physical rehabilitation, or both in dogs following tibial plateau leveling osteotomy (TPLO) and arthroscopic surgery for cranial cruciate ligament (CCL) disease.
DESIGN Randomized, prospective clinical trial.
ANIMALS 48 dogs with unilateral CCL disease.
PROCEDURES Dogs were randomly assigned to receive a dry omega-3 fatty acid and protein-enriched dog food formulated to support joint health (test food [TF]), a dry food formulated for maintenance of adult dogs (control food [CF]), TF plus rehabilitation (TF-R), or CF plus rehabilitation (CF-R). Data collected over 6 months included body weight, body condition score, ground reaction force data, tibial plateau angle, limb circumference measurements, subjective pain and lameness scores assigned by surgeons and dog owners, and daily activity measured by accelerometry.
RESULTS Peak vertical force and vertical impulse were greater after surgery for dogs in the TF groups than in the CF groups; peak vertical force was greater after surgery in dogs that underwent rehabilitation than in those that did not. Owner scores indicated lower frequencies of lameness and signs of pain during some activities for the TF group, compared with other groups, and for the TF-R and CF-R groups, compared with the CF group. Sedentary time decreased and time spent in light-to-moderate or vigorous activity increased in all groups over time. Rehabilitation was significantly associated with greater time spent in light-to-moderate activity, regardless of diet.
CONCLUSIONS AND CLINICAL RELEVANCE Feeding the TF and providing physical rehabilitation during the first 6 months after TPLO were associated with improvements in some indices of clinical outcome and function in dogs. Significant interactions between time and some outcome variables were observed, indicating further research is warranted.
Abstract
Objective—To evaluate stability of canine pancreatic lipase immunoreactivity (cPLI) in serum samples and to determine the effect of long-term administration of prednisone on serum cPLI concentrations.
Sample Population—8 canine serum samples for the stability evaluation and serum samples obtained from 6 healthy young adult heterozygous (carrier) dogs with X-linked hereditary nephritis for determining the effect of prednisone administration.
Procedures—To evaluate stability of serum cPLI concentration, an aliquot of each serum sample was stored at each of 4 temperatures between −80° and 24°C; samples were analyzed on days 0, 3, 7, 14, and 21. To determine the effect of long-term prednisone administration, pretreatment serum samples were obtained (days 0 and 14) and prednisone was administered (2.2 mg/kg, q 24 h, PO) on days 15 through 42, with serum samples obtained on days 28 and 42. Additional serum samples were obtained on days 56 and 70.
Results—Mean serum cPLI concentrations did not change significantly from day 0 to day 21 regardless of storage temperature. Serum cPLI concentrations in dogs after prednisone administration were within the reference range for all dogs at all time points, and results of repeated-measures ANOVA revealed that serum cPLI concentrations did not change significantly over time.
Conclusions and Clinical Relevance—Serum cPLI concentrations measured in canine serum samples stored at room temperature, in a refrigerator, or in a freezer at −20° or −80°C were stable for at least 21 days. Also, long-term prednisone administration to dogs did not significantly affect serum cPLI concentrations.
Abstract
Objective—To evaluate the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique.
Animals—14 dogs (similarly housed and fed identical diets).
Procedure—Samples of intestinal contents were collected from the duodenum, jejunum, ileum, colon, and rectum of each dog. Bacterial DNA was extracted from the samples, and the variable V6 to V8 region of 16S ribosomal DNA (gene coding for 16S ribosomal RNA) was amplified by use of universal bacterial primers; polymerase chain reaction amplicons were separated via denaturing gradient gel electrophoresis (DGGE). Similarity indices of DGGE banding patterns were used to assess variation in the bacterial microflora among different compartments of the intestine within and among dogs. Bacterial diversity was assessed by calculating the Simpson diversity index, the Shannon-Weaver diversity index, and evenness.
Results—DGGE profiles indicated marked differences in bacterial composition of intestinal compartments among dogs (range of similarity, 25.6% to 36.6%) and considerable variation among compartments within individual dogs (range of similarity, 36.7% to 57.9%). Similarities between neighboring intestinal compartments were significantly greater than those between non-neighboring compartments. Diversity indices for the colon and rectum were significantly higher than those of the duodenum, jejunum, and ileum.
Conclusions and Clinical Relevance—Results indicated that the different intestinal compartments of individual dogs appear to host different bacterial populations, and these compartmental populations vary among dogs. In dogs, fecal sample analysis may not yield accurate information regarding the composition of bacterial populations in compartments of the gastrointestinal tract. (Am J Vet Res 2005;66:1556–1562)
Abstract
Objective—To measure 11-dehydro-thromboxane B2 (11-dTXB2) in urine of healthy control dogs, dogs undergoing ovariohysterectomy, and dogs with gastric dilatation-volvulus (GDV) and assess the relationship between urinary 11-dTXB2 concentrations in dogs with GDV and postoperative outcomes.
Sample Population—Urine samples from 15 nonsurgical control dogs, 12 surgical control dogs, and 32 dogs with GVD.
Procedure—Urine samples were obtained from healthy pet dogs (ie, nonsurgical control dogs), dogs undergoing ovariohysterectomy at anesthetic induction and 1 hour following surgery (ie, surgical control dogs), and dogs with GDV at hospital admission and 1 hour following surgical derotation of the stomach (ie, GDV dogs). Urinary 11-dTXB2 concentrations were determined with an ELISA and normalized to urinary creatinine (Cr) concentrations by calculation of the 11-dTXB2 -to-Cr ratio. Differences in median 11-dTXB2 -to-Cr ratios among dogs and before and after surgery were analyzed.
Results—Urinary 11-dTXB2-to-Cr ratios did not differ between nonsurgical control dogs and surgical control dogs before or after surgery. Urinary 11-dTXB2-to-Cr ratios were significantly higher in GDV dogs at the time of hospital admission and 1 hour after surgery, compared with those of nonsurgical control dogs. Postoperative urine samples from GDV dogs had significantly higher 11-dTXB2-to-Cr ratios than postoperative urine samples from surgical control dogs. Median urinary 11-dTXB2-to-Cr ratios increased significantly in GDV dogs that developed postoperative complications.
Conclusions and Clinical Relevance—Urinary 11-dTXB2 concentration is increased in GDV dogs at the time of hospital admission and after surgical derotation of the stomach, compared with that of healthy dogs. An increased urinary 11-dTXB2-to-Cr ratio following surgery is associated with an increased incidence of postoperative complications in dogs with GDV.
Abstract
Objective—To detect matrix metalloproteinase (MMP)-9 in serum and CSF and determine relationships between MMP activity and severity of disease, duration of clinical signs, and duration of hospitalization in dogs with acute intervertebral disk disease (IVDD).
Animals—35 dogs with acute IVDD and 8 clinically normal control dogs.
Procedure—CSF and serum were collected from affected and control dogs. Zymography was used to detect MMP-9.
Results—Activity of MMP-9 in CSF was detected in 6 of 35 dogs with IVDD; activity was significantly more common in dogs with duration of signs < 24 hours. Paraplegic dogs were more likely to have MMP-9 activity in the CSF than non-paraplegic dogs. No significant difference in hospitalization time was detected in dogs with IVDD between those with and without activity of MMP-9 in the CSF. Serum MMP-9 was detected more frequently in dogs with IVDD than in control dogs.
Conclusions and Clinical Relevance—Data were consistent with results of experimental rodent spinal cord injury studies that indicate that MMP-9 is expressed early during secondary injury.