Serum lysozyme (muramidase) activity in dogs with neoplastic disease

The lysozyme (muramidase) activity was measured in the sera of 84 dogs with neoplastic disease. Neoplasms included 32 lymphomas, 13 primary bone neoplasms, 5 melanomas, 5 thyroid neoplasms, 9 soft tissue sarcomas, 5 mast cell sarcomas, and 15 carcinomas. The sera from 21 healthy dogs served as control. Dogs with neoplastic disease had significantly (P less than 0.005) higher serum lysozyme activity than did the healthy controls. For lymphosarcoma, dogs with clinical signs of systemic disease had significantly higher serum lysozyme activity than did dogs without clinical signs. For bone neoplasms, dogs with metastatic disease had higher serum lysozyme activity than did dogs without metastasis. Increased lysozyme activity may be a useful marker of macrophage-mediated host responses to neoplasms in dogs.     

Serum concentrations of immunoglobulins G, A, and M in dogs with neoplastic disease

The concentrations of the three major classes of immunoglobulins (Ig) were determined in canine serum by single radial immunodiffusion against calibrated standards. Serum samples were collected from 121 dogs categorized into 3 groups: normal dogs (n = 34), those with lymphosarcoma (n =41), and those with malignant, solid neoplasms other than lymphosarcoma (n = 46). The mean value for serum IgM concentration in the group of dogs with lymphosarcoma was significantly (P less than or equal to 0.02) higher than was the mean IgM concentration of the normal dogs. Dogs with neoplasms other than lymphosarcoma had significantly increased serum concentrations of IgG and IgM antibodies. There was no significant difference in serum IgA concentration among the three groups. A wide range of Ig concentrations was in the serum of clinically normal dogs, as well as in dogs with neoplastic diseases. Although individual dogs with neoplasms had low serum Ig concentrations, the 81 dogs with neoplastic disease were generally able to synthesize Ig.     

Usefulness of measuring serum lysozyme activity in dogs with neoplastic disease

Serum lysozyme activity (SLA) was measured in a turbidimetric assay with a microcentrifugal analyzer.In a control group of 53 healthy dogs of both sexes and ranging in age from 4 to 10 years, SLA had a mean value of 1.2 mg/l with a range (±2 SD) of 0.6–1.8 mg/l.In 80 dogs with a variety of neoplastic diseases the histopathological diagnosis was compared with the SLA value. SLA value was increased in 83% of the cases with malignant tumors and in 29% of the cases with benign tumors. Proper clinical examination is essential in differentiating between neoplastic disease and some interfering diseases, e.g. chronic dermatitis, pyometra and chronic nephritis.Measuring of SLA in dogs may be helpful in screening those animals with suspected malignancies.     

Usefulness of measuring serum lysozyme activity in dogs with neoplastic disease

Serum lysozyme activity (SLA) was measured in a turbidimetric assay with a microcentrifugal analyzer. In a control group of 53 healthy dogs of both sexes and ranging in age from 4 to 10 years, SLA had a mean value of 1.2 mg/l with a range (+/- 2 SD) of 0.6 - 1.8 mg/l. In 80 dogs with a variety of neoplastic diseases the histopathological diagnosis was compared with the SLA value. SLA value was increased in 83% of the cases with malignant tumors and in 29% of the cases with benign tumors. Proper clinical examination is essential in differentiating between neoplastic disease and some interfering diseases, e.g. chronic dermatitis, pyometra and chronic nephritis. Measuring of SLA in dogs may be helpful in screening those animals with suspected malignancies.     

Serum concentrations of the third component of complement in healthy dogs and dogs with protein-losing nephropathy

OBJECTIVE: To develop a method for determining the concentration of the third component of complement (C3) in canine serum, to establish a reference range for C3 in healthy dogs, and to evaluate dogs with protein-losing nephropathy (PLN) to determine whether PLN is associated with decreased serum C3 concentrations. ANIMALS: 30 healthy dogs and 49 dogs with PLN. PROCEDURES: Serum samples were obtained from healthy dogs at the time of examination, whereas serum samples were obtained from dogs with PLN at the time of diagnosis. All samples were frozen at -70 degrees C until analyzed. Serum C3 concentrations were determined by use of a sandwich ELISA. Concentrations were expressed as the number of dilutions in which C3 could be detected. RESULTS: C3 was detectable in healthy control dogs (range, 1,920,000 to 15,400,000 dilutions; median, 9,600,000 dilutions). This represented a range of four 2-fold serum dilutions. In addition, C3 was detectable in dogs with PLN (range, 1,460,000 to 30,070,000 dilutions; median, 7,680,000 dilutions), which represented a range of six 2-fold serum dilutions. There was no significant difference in C3 concentrations between the 2 groups. CONCLUSIONS AND CLINICAL RELEVANCE: C3 is a critical part of the immune defense system that has not been extensively examined in veterinary medicine. An ELISA was developed for measuring C3 concentrations, and a reference range for healthy dogs was established. Significant decreases in C3 concentrations were not detected in any dog with PLN. Additional studies will be required to definitively determine the importance of serum C3 concentrations in PLN.     

Serum level of apoptosis inhibitor of macrophage in dogs with histiocytic sarcoma and its association with the disease

Histiocytic sarcoma (HS) is a rare neoplasm of macrophages or dendritic cells with a poor prognosis in dogs. As the apoptosis inhibitor of macrophage (AIM) is characteristically expressed in canine macrophages, we hypothesised that AIM is involved in the development or progression of HS in dogs. In this study, AIM expression in the tumour region and serum AIM levels in dogs with HS was assessed. Additionally, the effects of AIM overexpression on HS cell viability were investigated using a HS cell line that was selected from five validated HS cell lines. Immunohistochemistry showed that AIM expression was observed in the cytoplasm of the HS cells. CD36, a candidate AIM receptor, was also observed on the cell membrane of HS cells. When the serum AIM level was detected in 36 dogs with HS and 10 healthy dogs via western blot analysis, the AIM levels in the HS dogs were significantly higher than those in the controls. AIM mRNA expression in the 5 HS cell lines varied but was higher than that in the other tumour-derived lines. Among the five HS cell lines, DH82 originally had lower AIM and the highest CD36 expression. When AIM was overexpressed in DH82, therein cell growth speed and invasion, apoptosis inhibition and phagocytic activity were strongly upregulated. These data suggest that elevated intra-tumour expression of AIM could induce the progression of HS cells in dogs. Moreover, elevated serum AIM levels in dogs with HS could serve as a biomarker of HS.     

Serum nitrate and nitrite in dogs with spontaneous cardiac disease

The purpose of this study was to determine whether nitric oxide (NO) concentrations are high in dogs with chronic valvular disease (CVD) and dilated cardiomyopathy (DCM) compared to healthy controls and to determine whether NO concentrations are correlated with type of cardiac disease, disease severity, medical therapy, or serum tumor necrosis factor (TNF) and interleukin-1 (IL-1). Blood was collected from 32 dogs with DCM, from 10 dogs with CVD, and from 10 healthy controls. Indirect determination of NO concentrations was performed by a commercial photoabsorbance assay that uses a Greiss reagent to measure the concentration of nitrite and nitrate (NN), end products of NO metabolism. TNF and IL-1 activities were measured by bioassay. Mean NN concentrations were significantly higher in dogs with heart disease (median, 4.57 microM; range, 0.00-31.05 microM) than in controls (median, 0.00 microM; range, 0.00-6.16 microM; P = .04). NN concentrations in dogs with cardiac disease were not correlated with type or severity of cardiac disease, medication type, or TNF and IL-1 concentrations. NN concentrations were inversely correlated with fractional shortening. The results of this study suggest that metabolites of NO are increased in some dogs with cardiac disease, but these increases appear to be independent of disease severity, TNF and IL-1 concentrations, and type of pharmacologic intervention.     

Serum amylase and isoamylase values in dogs with pancreatic disease

Serum amylase and isoamylase values were determined in three groups of dogs. The first group contained control dogs while the other groups contained dogs with confirmed exocrine pancreatic insufficiency and diabetes mellitus respectively. The trypsin-like immunoreactivity test was also carried out on sera from dogs with exocrine pancreatic disease (EPI). A significant difference was detected in the serum amylase values between the three groups which may be of limited diagnostic value. Dogs with EPI had values lower than normal while those with diabetes mellitus had values higher than control dogs. No evidence of exocrine pancreatic insufficiency was found in dogs with diabetes mellitus.     

Serum concentrations of leptin and adiponectin in dogs with chronic kidney disease

BackgroundAn imbalance in adipokines is associated with the progression of chronic kidney disease (CKD) in humans. However, alterations in adipokines in dogs with CKD remain unclear.ObjectivesTo examine whether adipokine concentrations in serum differ between healthy dogs and dogs with CKD and to determine the correlation between serum adipokine concentrations and CKD severity in dogs.AnimalsTwenty dogs with CKD and 10 healthy dogs.MethodsIn this cross‐sectional study, serum concentrations of leptin, adiponectin, interleukin (IL)‐6, IL‐10, IL‐18, and tumor necrosis factor (TNF)‐α were measured in healthy dogs and dogs with CKD, which were classified according to the International Renal Interest Society guidelines.ResultsSerum leptin concentrations were positively correlated with systolic arterial blood pressure (r = .41), creatinine concentrations (r = .39), and symmetric dimethylarginine concentrations (r = .73). Serum adiponectin concentrations (median [range]) in CKD dogs with borderline or non‐proteinuric (20.25 [14.9‐45.8] ng/mL) were significantly higher than those in proteinuric CKD dogs (13.95 [6.4‐22.1] ng/mL; P = .01). Serum IL‐6 (median [range]; 43.27 [24.30‐537.30] vs 25.63 [6.83‐61.03] pg/mL; P = .02), IL‐18 (median [range]; 25.98 [11.52‐280.55] vs 10.77 [3.53‐38.45] pg/mL; P = .01), and TNF‐α (median [range]) concentrations (11.44 [8.54‐38.45] vs 6.105 [3.97‐30.68] pg/mL; P = .02) were significantly different between proteinuric and borderline or non‐proteinuric CKD dogs.Conclusions and Clinical Importanceleptin and adiponectin concentrations in serum might be associated with severity of CKD and proteinuria in dogs with CKD, respectively.     

Serum thyroid hormone concentrations and thyrotropin responsiveness in dogs with generalized dermatologic disease.

Fifty-eight dogs with generalized dermatologic disease that had not been given glucocorticoids systemically or topically within 6 weeks of entering the study were evaluated for thyroid function by use of the thyrotropin-response test. Dogs were classified as euthyroid or hypothyroid on the basis of test results and response to thyroid hormone replacement therapy. Baseline serum thyroxine (T4), free T4 (fT4), and triiodothyronine (T3) concentrations were evaluated in the 58 dogs. Serum T4, fT4, and T3 concentrations were evaluated in 200 healthy dogs to establish normal values. Hormone concentrations were considered low if they were less than the mean -2 SD of the values for control dogs. Specificity of T4 and fT4 concentrations was 100% in predicting hypothyroidism; none of the euthyroid dogs with generalized skin disease had baseline serum T4 or fT4 concentration in the low range. Sensitivity was better for fT4 (89%) than for T4 (44%) concentration. Significant difference was not observed in serum T4 and fT4 concentrations between euthyroid dogs with generalized skin disease and healthy control dogs without skin disease. Serum T3 concentration was not accurate in predicting thyroid function; most of the euthyroid and hypothyroid dogs with skin disease had serum T3 concentration within the normal range.     

Serum enzyme determination in the study of liver disease in dogs

Glutamate dehydrogenase (GLDH), sorbitol dehydrogenase (SDH), 5''-nucIeotidase (5''-ND) and cholin esterase (CHE) were determined in the sera of 37 dogs with various liver diseases. The values for these parameters were compared with the values for aspartate and alanin aminotransferase (ASAT, ALAT), alkaline phosphatase (ALP), γ-glutamyl transferase (γ-GT) and albumin (ALB). GLDH was found to be the most sensitive enzyme parameter (sensitivity 92 %), its values also reflecting the degree of liver cell necrosis well. Next in sensitivity followed ASAT and ALP. SDH values were correlated to ALAT values but had a low sensitivity (43 %). 5''-ND discriminated markedly better between biliary tract disturbance and other lesions than ALP and somewhat better than γ-GT. CHE had a wide reference range, low sensitivity, and was not correlated to ALB. Single determinations of CHE may thus not be of diagnostic value in dogs.     

Genomic hypomethylation in neoplastic cells from dogs with malignant lymphoproliferative disorders

DNA methylation is an epigenetic modification in which a methyl group is added usually to the fifth carbon position of a cytosine residue. Dysregulation of this process is an important molecular event which has been shown to be associated with neoplastic transformation and tumour progression in humans and mice. Features of methylation dysregulation in many different types of neoplasms include general genomic hypomethylation, focal hypermethylation, and altered expression of genes which encode a series of DNA (cytosine-5) methyltransferases. Interestingly, many types of neoplasia that are recognised in humans also develop spontaneously in the dog. By comparing the restriction patterns of MspI and HpaII, this study demonstrates that as in human, genomic hypomethylation is a feature of neoplastic cells in the majority of canine lymphoma cases and approximately one-third of canine leukemia cases confirming that dysregulation of the DNA methylating machinery is implicated in malignant transformation of lymphoid cells in some dogs.     

Serum IgE and IgG responses to food antigens in normal and atopic dogs,and dogs with gastrointestinal disease

In human food allergy, with or without concurrent atopy, there may be significant increases in serum allergen-specific IgE. Serological methods have been tried but are not currently recommended for diagnosis of suspected food allergy in dogs. The aim of this study was to investigate humoral immune responses to food antigens in dogs. Serum IgG and IgE antibodies specific for food antigens were measured by enzyme linked immunosorbent assay (ELISA) using polyclonal anti-dog IgG and IgE reagents. Antigens tested were beef, chicken, pork, lamb, chicken, turkey, white fish, whole egg, wheat, soybean, barley, rice, maize corn, potato, yeast and cow's milk. Three groups were examined: normal dogs, dogs with atopic dermatitis (AD); and dogs with one of four types of gastrointestinal (GI) disease: small intestinal bacterial overgrowth (SIBO), inflammatory bowel disease (IBD), food-responsive disease, and infectious diarrhoea. Statistically significant differences in food-specific antibodies were not detected between the GI subgroups. There were statistically significant differences in the IgE concentration between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested. There were statistically significant differences in the average IgG concentrations between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested, except egg and yeast. The relationship of antigen responses for pooled data was analysed using principle component analysis and cluster plots. Some clustering of variables was apparent for both IgE and IgG. For example, all dogs (normal and diseased) made a similar IgG antibody response to chicken and turkey. Compared with other groups, atopic dogs had more food allergen-specific IgE and this would be consistent with a Th(2) humoral response to food antigens. Dogs with GI disease had more food allergen-specific IgG compared with the other groups. This may reflect increased antigen exposure due to increased mucosal permeability which is a recognised feature of canine intestinal disease.     

Serum bile acids as an indicator of liver disease in dogs

Total serum bile acids were determined in 62 dogs with different primary or secondary liver diseases, using 3α-hydroxysteroid dehydrogenase coupled to nitrobluetetrazolium in a centrifugal analyzer. A reaction time of 4 min was sufficient, yielding a within run coefficient of variation of 7% at 6 µmol/1 and 3% at 27 µmol/1. A reference range of 0–4.4 µmol/1 2 h post prandially was observed. The sensitivity of bile acids as a liver function test was superior to that of alanine and aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase and combinations of two of these. The bile acids test detected 36 of 39 patients with a morphological or clinical liver diagnosis. For dogs with heart failure the bile acids test was a markedly more sensitive indicator of secondary liver involvement than alanine aminotransferase or alkaline phosphatase. For secondary liver affections associated with pyometra or epilepsy medication the opposite was the case. Bile acid values in the pooled patient material was not correlated to any of the 4 enzymes measured. For cirrhosis there was positive correlation, however, with the amino transferase values.     

Serum butyrylcholinesterase activity in dogs with diabetes mellitus

Increased serum butyrylcholinesterase (BChE) activity is a feature of diabetes mellitus (DM) in humans and rats. The objective of this study was to evaluate serum BChE activity in diabetic dogs. The activity of the enzyme was assessed in three cohorts of animals: (1) dogs with naturally occurring DM (n=74); (2) clinically normal dogs (n=74); and (3) dogs with various other diseases (n=74). A statistically significant increase in BChE activity was found in the diabetic dogs (7.59 ± 2.9 kUI/L) compared with the clinically normal animals (6.12 ± 1.94 kUI/L; P<0.05), and with the dogs with other diseases (5.55 ± 2.06 kUI/L; P<0.01). Such increased activity could be the result of the altered glucose and lipid metabolism that occurs in DM.     

Serum isoamylase values in normal dogs and dogs with exocrine pancreatic insufficiency

Estimations were made of the serum isoamylase values of normal dogs and of dogs with confirmed exocrine pancreatic insufficiency. A statistically significant difference was demonstrated between the two groups in respect of the values of one of the isoamylase fractions measured. Further study has confirmed that canine salivary tissue lacks amylase activity and that the source of the isoamylase fractions was the pancreas.This knowledge has potential value in the diagnosis of canine exocrine pancreatic insufficiency.