Effect of tissue processing on assessment of endoscopic intestinal biopsies in dogs and cats.

BACKGROUND: Prior studies failed to detect significant association between hypoalbuminemia and small intestinal lesions. HYPOTHESIS: Use of pictorial templates will enhance consistency of interpathologist interpretation and identification of intestinal lesions associated with hypoalbuminemia. ANIMALS: Tissues from 62 dogs and 25 cats examined as clinical cases at 7 referral veterinary practices in 4 countries. METHODS: Retrospective, observational study. Histopathology slides from sequential cases undergoing endoscopic biopsy were examined by 4 pathologists by pictorial templates. Changes for 9 microscopic features were recorded as normal, mild, moderate or severe, and 2- and 4-point scales were tested for consistency of interpretation. Logistic regression models determined odds ratios (OR) of histologic lesions being associated with hypoalbuminemia while kappa statistics determined agreement between pathologists on histologic lesions. RESULTS: There was poor agreement (kappa = -0.013 to 0.3) between pathologists, and institution of origin of slides had effect (kappa = 1.0 for 3 of 4 lesions on slides from Institution 5) on agreement between pathologists on selected histologic features. Using 2 point as opposed to 4-point grading scale increased agreement between pathologists (maximum kappa = 0.69 using 4-point scale versus maximum kappa = 1.0 using 2-point scale). Significant association (P = .019- .04; 95% OR = 3.14-10.84) between lacteal dilation and hypoalbuminemia was found by 3 pathologists. CONCLUSIONS AND CLINICAL IMPORTANCE: Substantial inconsistency between pathologists remains despite use of pictorial template because of differences in slide processing. Distinguishing between mild and moderate lesions might be important source of the disagreement among pathologists.

Effect of sample quality on the sensitivity of endoscopic biopsy for detecting gastric and duodenal lesions in dogs and cats.

BACKGROUND: The quality of histopathology slides of endoscopic biopsies from different laboratories varies, but the effect of biopsy quality on outcome is unknown. HYPOTHESIS: The ability to demonstrate a histologic lesion in the stomach or duodenum of a dog or cat is affected by the quality of endoscopic biopsy samples submitted. More endoscopic samples are needed to find a lesion in poor-quality tissue specimens. ANIMALS: Tissues from 99 dogs and 51 cats were examined as clinical cases at 8 veterinary institutions or practices in 5 countries. METHODS: Histopathology slides from sequential cases that underwent endoscopic biopsy were submitted by participating institutions. Quality of the histologic section of tissue (inadequate, marginal, adequate), type of lesion (lymphangiectasia, crypt lesion, villus blunting, cellular infiltrate), and severity of lesion (normal, mild, moderate, severe) were determined. Sensitivity of different quality tissue samples for finding different lesions was determined. RESULTS: Fewer samples were required from dogs for diagnosis as the quality of the sample improved from inadequate to marginal to adequate. Duodenal lesions in cats displayed the same trend except for moderate duodenal infiltrates for which quality of tissue sample made no difference. Gastric lesions in dogs and mild gastric lesions in cats had the same trend, whereas the number of tissue samples needed to diagnose moderately severe gastric lesions in cats was not affected by the quality of tissue sample. CONCLUSIONS AND CLINICAL IMPORTANCE: The quality of endoscopically obtained tissue samples has a profound effect on their sensitivity for identifying certain lesions, and there are differences between biopsies of canine and feline tissues.

Gene expression analysis by transcript profiling coupled to a gene database query.

We describe an mRNA profiling technique for determining differential gene expression that utilizes, but does not require, prior knowledge of gene sequences. This method permits high-throughput reproducible detection of most expressed sequences with a sensitivity of greater than 1 part in 100,000. Gene identification by database query of a restriction endonuclease fingerprint, confirmed by competitive PCR using gene-specific oligonucleotides, facilitates gene discovery by minimizing isolation procedures. This process, called GeneCalling, was validated by analysis of the gene expression profiles of normal and hypertrophic rat hearts following in vivo pressure overload.

First report of canine African trypanosomosis in the UK.

A six-year-old neutered male Jack Russell terrier was presented two years after importation into the UK from southern Africa with severe anaemia and abdominal distension. Abdominal ultrasonography revealed the presence of hepato-splenomegaly and ascites. A diagnosis of trypanosomosis was made by blood smear examination. Shortly after admission the dog collapsed and died. PCR analysis revealed a single infection with Trypanosoma congolense savannah type. This is the first reported case of canine African trypanosomosis in Europe and suggests that chronic trypanosomosis may allow importation of the disease in apparently asymptomatic animals, even with extended quarantine periods.

Evaluation of disease activity markers in dogs with idiopathic inflammatory bowel disease.

OBJECTIVES: To evaluate the clinical utility of serum tumour necrosis factor-alpha, C-reactive protein and microalbuminuria as disease activity markers in canine idiopathic inflammatory bowel disease. METHODS: Dogs with chronic gastrointestinal disease for which no underlying cause could be identified were considered to have idiopathic inflammatory bowel disease and were included in the study. Serum tumour necrosis factor-alpha was assessed using a canine-specific ELISA, C-reactive protein by immunoturbidometric assay and quantitative microalbuminuria was analysed using a monoclonal antibody directed against canine albumin. The canine inflammatory bowel disease activity index and histopathologic grade were used to assess disease severity; biologic markers were then compared with the canine inflammatory bowel disease activity index and histopathologic grade. RESULTS: Sixteen dogs were included in the study. C-reactive protein level was mildly elevated in 15 dogs. Microalbuminuria was elevated in two of 15 dogs, and tumour necrosis factor-alpha was not detected in any dog tested. No correlation was found between the canine inflammatory bowel disease activity index and C-reactive protein or microalbuminuria or between histopathologic grade and C-reactive protein or microalbuminuria. There was no correlation between histopathologic grade and the canine inflammatory bowel disease activity index. CLINICAL SIGNIFICANCE: Although only a small number of dogs were evaluated, this study does not support the use of serum tumour necrosis factor-alpha measured by canine-specific ELISA or microalbuminuria in the evaluation of disease activity in dogs with idiopathic inflammatory bowel disease. Although mildly elevated in most dogs, C-reactive protein did not reflect disease severity as assessed by the canine inflammatory bowel disease activity index or histopathologic grade.

Synthesis of a collagenase inhibitor by gingival fibroblasts in culture.

Human collagenase was inhibited by test solutions of human gingival fibroblast culture media. The fibroblast-derived collagenase inhibitor was only slightly affected by 10 micrograms trypsin but was inactivated with 100 micrograms trypsin. The chaotropic agent KSCN (3 M) completely inactivated the inhibitor, whereas the thiol-blocking reagent, p-aminophenylmercuric acetate, partially inactivated the inhibitor. Inhibitory activity was retained at 60 degrees C but was abolished at 100 degrees C. Following ammonium sulfate fractionation, the fibroblast inhibitor was recovered in the supernatant at concentrations of at least 70% saturation. It is suggested that collagenase latency in soft connective tissues may derive from a collagenase-inhibitor complex formed by interaction of collagenase and a fibroblast-derived inhibitor.

Responsibility of the obstetrician to the fetus. II. Influence of prepregnancy weight and pregnancy weight gain on birthweight.

The diet of pregnant women has been restricted in various ways since earliest history. Dietary restriction may have severely deleterious effects on the fetus. Our study revealed a positive correlation of prepregnancy weight and pregnancy weight gain to term infant birthweight. Their influences are independent and additive. It is of the utmost importance to the fetus and the future of the human race that the diet of pregnant women contain the caloric value and essential nutrients recommended by the National Research Council.

Left atrial aneurysm.

Aneurysms of the left atrium are rare abnormalities. They can be congenital or acquired. Whereas a true congenital aneurysm presents as isolated pathology, inflammatory or degenerative processes involving the endocardium are associated with the acquired type. The clinical records of 2 patients with the diagnosis of left atrial aneurysm were reviewed, along with the surgical strategies, current literature, and patient outcomes. Because of the risk of life-threatening complications, surgery is recommended even in asymptomatic cases. Resection and mitral valvuloplasty should be the treatment of choice.

Ectopia cordis and other midline defects.

BACKGROUND: Thoracic ectopia cordis and other midline defects are rare congenital anomalies that often occur with other intracardiac defects. Despite significant improvements in neonatal and infant cardiac surgeries, operations for thoracoabdominal ectopia cordis carry an extremely high mortality with only a few reported survivors of thoracic ectopia cordis. METHODS: The clinical charts of 4 patients with ectopia cordis over a 6-year period were reviewed. Three of the patients showed varying degrees of Cantrell's Pentalogy; thoracic ectopia cordis was found in 1. We have reviewed our surgical strategies and reported the patients' clinical outcomes. RESULTS: All 4 patients are alive at follow-up. Two infants with double-outlet right ventricle have been fully corrected, and extracorporeal membrane oxygenation was necessary in 1 infant for cardiac failure following the cardiac repair. A newborn with thoracoabdominal ectopia cordis underwent primary repair of his diaphragmatic defect, and a silo was used to progressively reduce the omphalocele. He is currently awaiting elective repair of tetralogy of Fallot. Lastly, the patient with thoracic ectopia cordis underwent successful soft tissue coverage, and she is being followed in the clinic with restrictive muscular ventricular septal defects and a left ventricular diverticulum. CONCLUSIONS: Our experience along with other reports in the literature demonstrates that patients with thoracic and thoracoabdominal ectopia cordis can undergo and survive full cardiac, neurologic, and abdominal repair during infancy. Furthermore we advocate different approaches determined by the severity of the presentation and the presence of other complicating factors.

Identification of collagenase in cultured blood mononuclear cells.

Mononuclear cells isolated from peripheral whole blood were fractionated by erythrocyte-rosetting into T cells and non-T cell subpopulations which were tested for collagenase-producing capacity. Mononuclear cells and purified T lymphocytes, as well as non-T (B) lymphocytes, produced collagenase during serum-free culture. Removal of the macrophages from the non-T cells did not affect collagenase production by lymphocytes.

Quantitative assessment of fat absorption and its diagnostic value in exocrine pancreatic insufficiency.

A new method of quantifying fat absorption based on the estimation of serum triglyceride levels is described. When long chain triglyceride (LCT) was fed to normal dogs a significant elevation of serum triglyceride concentration was recorded which was not observed in dogs with exocrine pancreatic insufficiency. When the test was repeated on the dogs with pancreatic insufficiency using the LCT together with lipase the serum triglyceride concentration increased, suggesting the failure in the initial test was caused by a deficiency in pancreatic lipase. Feeding medium chain triglyceride (MCT) did not raise the serum triglyceride levels in normal dogs.

Assessment of fat absorption in normal dogs and in dogs with hyperadrenocorticalism.

Fat absorption was determined quantitatively in clinically normal dogs, dogs with confirmed hyperadrenocorticalism before and after treatment with mitotane (op' DDD) and in pruritic dogs before and after administration of prednisolone. Fat absorption was significantly higher (P less than 0.001) in dogs with untreated hyperadrenocorticalism and pruritic dogs on prednisolone therapy than in normal dogs. It was normal in pruritic dogs before prednisolone therapy and approached normality in dogs with treated hyperadrenocorticalism. It is concluded that high circulating cortisol or prednisolone levels result in increased intestinal absorption of dietary fat possibly mediated by increased glucocorticoid induced activity of enterocytes or reduced hepatic clearance of triglycerides.

Fat absorption in dogs with demodicosis or zinc-responsive dermatosis.

Quantitative absorption of long-chain triglycerides was studied in normal dogs, dogs with demodicosis and dogs which had been successfully treated for zinc-responsive dermatosis. The mean serum triglyceride concentration of dogs treated for zinc-responsive dermatosis was significantly lower than that of normal dogs before (P less than 0.01) and at one hour (P less than 0.01) and two, three and four hours (P less than 0.001) after feeding vegetable oil. No significant difference was detected in the mean serum triglyceride concentration of dogs with demodicosis and normal dogs at any of the sampling times.

Fat absorption in dogs with diabetes mellitus or hypothyroidism.

Quantitative fat absorption was studied in normal dogs and in dogs with hypothyroidism and diabetes mellitus. The serum triglyceride concentrations of diabetic and hypothyroid dogs were significantly higher at each sampling time than those of normal dogs. The fat absorption curve of hypothyroid dogs peaked at 180 minutes and though significantly raised was parallel to that of normal dogs whereas the fat absorption curve of diabetic dogs continued to rise up to 240 minutes. These results provide evidence for impaired plasma clearance of triglyceride in canine diabetes mellitus and hypothyroidism.

Evaluation of B and T lymphocytes and plasma cells in colonic mucosa from healthy dogs and from dogs with inflammatory bowel disease.

The aim of this study was to investigate the subpopulations of lymphocytes in the colonic mucosa of healthy dogs and dogs with inflammatory bowel disease (IBD). Fourteen normal dogs and 13 dogs with IBD were examined. Endoscopic biopsy specimens of colonic mucosa from each dog were stained specifically for pan T lymphocytes (CD3) and pan B lymphocytes (CD79a), and for plasma cells with methyl green pyronin (MGP) stain. Cells were counted by means of a grid and statistical analysis was performed on the data collected. B and T lymphocytes were also counted in the glandular epithelium of normal dogs and dogs with IBD and the normal and abnormal groups compared statistically. Healthy dogs had significantly lower numbers of T cells in the lamina propria and glandular epithelium and significantly lower numbers of B cells in the lamina propria. Significant group differences for plasma cells were not evident. Our results indicate that in IBD a chronic cellular immune reaction is present in the diseased gut involving increased numbers of B and T lymphocytes.

Effect of pancreatectomy on plasma activities of amylase, isoamylase, lipase and trypsin-like immunoreactivity in dogs.

The contribution of the pancreas to the plasma activities of amylase, isoamylase, lipase and the concentration of trypsin-like immunoreactivity (TLI) in the dog was examined by measuring the activities of these enzymes before and after total pancreatectomy. Pancreatectomy was followed by a decrease in the concentration of TLI (from 6.2 +/- 0.3 micrograms litre-1 to 1.2 +/- 0.3 micrograms litre-1; P less than 0.001) and activity of isoamylase peak 4 (from 1257 +/- 105 iu litre-1 to 894 +/- 171 iu litre-1; P less than 0.05). Though significantly reduced, the activities of peak 4 isoamylase were still within the normal range for control dogs. Pancreatectomy did not significantly alter the activities of amylase, lipase or isoamylase peaks 1, 2 and 3. These findings provide strong evidence that the pancreas is not the sole source of circulating amylase, isoamylase and lipase activities. In contrast the marked reductions in TLI to values close to the limits of assay sensitivity suggest that TLI is derived from the pancreas alone. The results indicate that assay of circulating TLI provides a more sensitive and specific indicator of pancreatic exocrine mass than plasma amylase, lipase or isoamylase activities.

Endocardial fibroelastosis occurring in a mother and daughter.

Endocardial fibroelastosis (EFE) in a mother and daughter are reported. This entity has a primary and secondary form. In this case, the 23-year-old mother died 11 months after delivery with findings of EFE. At age 3(1/2) years, the daughter was admitted in congestive heart failure and did not respond to medical management. The autopsy findings revealed EFE. Endocardial fibroelastosis in both mother and daughter is a very rare finding.

Bacterial overgrowth causing intestinal malabsorption in a dog.

The case history of a six-year-old female chow chow with intractable diarrhoea is described. The clinical history and symptoms, together with the methods of diagnosis, indicated that the cause was bacterial overgrowth of the intestine leading to a malabsorption syndrome. The method of treatment and the response is also recorded.

Diagnostic value of exfoliative cytology of body fluids in dogs and cats.

A method for collecting body fluids for cytological examination and their subsequent preparation and staining is described. The results of the cytological examination of 80 dogs and 26 cats over a period of three years are reported, indicating the number and types of condition which can be diagnosed, together with the clinicopathological correlations.

Diagnostic value of tissue biopsy in gastrointestinal and liver disease.

Methods are described for the endoscopic examination and biopsy of the gastrointestinal tract and liver of the dog, cat, ox and horse. The results of the examination of 41 biopsy samples are shown, followed by a discussion of the value of biopsy in the diagnosis of diseases of the digestive tract.