Dysregulation of gastrointestinal RAGE (receptor for advanced glycation end products) expression in dogs with chronic inflammatory enteropathy.

The pathogenesis of chronic inflammatory enteropathies (CIE) in dogs involves dysregulated innate immune responses. The receptor for advanced glycation end products (RAGE), a pattern recognition receptor, plays a role in chronic inflammation. Abrogation of proinflammatory RAGE signaling by ligand binding (e.g., S100/calgranulins) to soluble RAGE (sRAGE) might also be a novel therapeutic avenue. Serum sRAGE levels are decreased in canine CIE, but gastrointestinal tissue RAGE expression has not been investigated in dogs. Thus, the study aimed to evaluate the gastrointestinal mucosal RAGE expression in dogs with CIE. Further, the potential binding of RAGE to canine S100/calgranulin ligands was investigated. Epithelial RAGE expression was quantified in gastrointestinal (gastric, duodenal, ileal, and colonic) biopsies from 12 dogs with CIE and 9 healthy control dogs using confocal laser scanning microscopy. RAGE expression was compared between both groups of dogs and was tested for an association with patient characteristics, clinical variables, histologic lesion severity, and biomarkers of extra-gastrointestinal disease, systemic or gastrointestinal inflammation, function, or protein loss. Statistical significance was set at p < 0.05. RAGE:S100/calgranulin binding was assessed by immunoassay and electrophoretic techniques. RAGE expression was detected in all 59 biopsies from diseased and healthy control dogs evaluated. Epithelial RAGE expression in the duodenum and colon was significantly higher in dogs with CIE than in healthy controls (p < 0.04). Compared to healthy controls, RAGE expression in dogs with CIE also tended to be higher in the ileum but lower in the stomach. A slight (statistically not significant) shift towards more basal intestinal epithelial RAGE expression was detected in CIE dogs. Serum sRAGE was proportional to epithelial RAGE expression in the duodenum (p < 0.04), and RAGE expression in the colon inversely correlated with biomarkers of protein loss in serum (both p < 0.04). Several histologic morphologic and inflammatory lesion criteria and markers of inflammation (serum C-reactive protein and fecal calprotectin concentration) were related to epithelial RAGE expression in the duodenum, ileum, and/or colon. in vitro canine RAGE:S100A12 binding appeared more pronounced than RAGE:S100A8/A9 binding. This study showed a dysregulation of epithelial RAGE expression along the gastrointestinal tract in dogs with CIE. Compensatory regulations in the sRAGE/RAGE axis are an alternative explanation for these findings. The results suggest that RAGE signaling plays a role in dogs with CIE, but higher anti-inflammatory decoy receptor sRAGE levels paralleled RAGE overexpression. Canine S100/calgranulins were demonstrated to be ligands for RAGE.

Diagnostic contribution of individual components of adrenal function tests to diagnose canine hyperadrenocorticism.

There is limited information regarding the value of constitutive components of the ACTH stimulation test (ACTHST) and low-dose dexamethasone suppression test (LDDST) including serum baseline cortisol (BC), difference between post-ACTH stimulation cortisol (PC) and BC (ΔACTHC), cortisol concentration 4h after dexamethasone administration (4HC), difference between 4HC and BC (Δ4C), and the difference between cortisol concentration 8h after dexamethasone administration and 4HC (Δ8C). Therefore, the objective of this study was to determine if these components can predict hyperadrenocorticism, pituitary-dependent hyperadrenocorticism (PDH), or functional adrenocortical tumor (FAT) in dogs. Cortisol concentrations were normalized, as fold change (FC), to the PC reference interval upper limit. A total of 1267 dogs were included, with hyperadrenocorticism diagnosed in 537 (PDH, n=356; FAT, n=28; undetermined, n=153) and excluded in 730. The area under the receiver operating curves for BC, ΔACTHC, 4HC, Δ4C, and Δ8C to predict hyperadrenocorticism were 0.76 (95% confidence interval (CI), 0.73-0.79), 0.91 (95% CI, 0.89-0.93), 0.83 (95% CI, 0.80-0.87), 0.55 (95% CI, 0.50-0.60), and 0.67 (95% CI, 0.62-0.72), respectively. A diagnostic limit of ≥0.78 FC for ΔACTHC had excellent sensitivity (1.00; 95% CI, 0.74-1.00), but poor specificity (0.67; 95% CI, 0.64-0.71), to predict FAT in dogs with a positive ACTHST. A diagnostic limit of ≥-0.26 FC for Δ4C had excellent sensitivity (1.00; 95% CI, 0.79-1.00), but poor specificity (0.21; 95% CI, 0.18-0.26), to predict FAT in dogs with a positive LDDST. In hyperadrenocorticoid dogs that have positive ACTHST or LDDST results, ΔACTHC or Δ4C, respectively, could be used to exclude FAT.

Biological variation of serum canine calprotectin concentrations as measured by ELISA in healthy dogs.

Calprotectin is a useful biomarker of inflammation in dogs. However, the biological variation of serum canine calprotectin is unknown. Indices of biological variation were determined in serial serum samples (n=147) from 11 healthy dogs (males/females: 4/7, median age: 5 years): analytical (3.0%), intra-individual (29.9%), and inter-individual variation (33.2%), reciprocal index of individuality (1.1), and index of heterogeneity (4.9). Serum calprotectin concentrations measured by ELISA and by the previous radioimmunoassay were highly correlated, but a constant and proportional bias exists between both assays. A de novo ELISA-reference interval (RI) for serum calprotectin concentration was established (0.6-11.8mg/L). Moderate changes in serum calprotectin (minimum critical difference: 6.4mg/L) between sequential measurements are needed to be considered relevant, and a population-based RI may or may not be appropriate for serum calprotectin.

Serum α1-proteinase inhibitor concentrations in dogs with exocrine pancreatic disease, chronic hepatitis or proteinuric chronic kidney disease.

Serum canine α1-proteinase inhibitor (cα1-PI) concentrations were evaluated in dogs with pancreatitis (n=24), exocrine pancreatic insufficiency (EPI; n=29), chronic hepatitis (CH; n=11) or proteinuric chronic kidney disease (CKD-P; n=61) to determine whether systemic proteinase/proteinase-inhibitor balance is altered in these conditions. Dogs with CKD-P had significantly lower cα1-PI concentrations than dogs with pancreatitis, EPI or CH; 16% of dogs with CKD-P had serum cα1-PI concentrations below the reference interval. Serum and urine cα1-PI concentrations were inversely correlated in dogs with CKD-P, but not in dogs with CH. This suggests that renal loss of cα1-PI contributes to decreased serum concentrations in dogs with CKD-P, while hepatic cα1-PI synthesis with CH either is not compromised or is counterbalanced by extrahepatic production.

Hyperhomocysteinemia in Greyhounds and its Association with Hypofolatemia and Other Clinicopathologic Variables.

BACKGROUND: Folate and cobalamin are essential cofactors for homocysteine (HCY) metabolism. Hyperhomocysteinemia, a multifactorial condition, may reflect B vitamin deficiency and is associated with increased risk of cardiovascular disease, thrombosis, and neurodegenerative and chronic gastrointestinal diseases in humans. Hyperhomocysteinemia has been reported in Greyhounds with suspected chronic enteropathy. OBJECTIVES: To evaluate the frequencies of and the association between hypofolatemia and hyperhomocysteinemia in Greyhounds. ANIMALS: Data and serum samples from 559 Greyhounds. METHODS: Nested case-control study. The frequency of hypofolatemia in Greyhounds was determined by a laboratory database search. The relationship between hyperhomocysteinemia (measured by gas chromatography-mass spectrometry) and hypocobalaminemia and hypofolatemia was evaluated, and its frequency compared between healthy Greyhounds and Greyhounds with thrombosis or chronic diarrhea. RESULTS: Hypofolatemia was identified in 172 of 423 (41%) Greyhounds and was more common in hypo- than in normocobalaminemic dogs (49% vs. 35%; P = .0064). Hyperhomocysteinemia was detected in 53 of 78 (68%) of Greyhounds, being more common in hypo- than in normofolatemic dogs (88% vs. 59%; P = .0175). All healthy Greyhounds, 21 of 30 (70%) of dogs with chronic diarrhea and 6 of 8 (75%) of those with thrombosis, were hyperhomocysteinemic. Serum HCY concentrations were inversely correlated with serum folate concentration (ρ = -0.28; P = .0386) and were positively associated with serum albumin concentration (ρ = 0.66; P = .0022). CONCLUSIONS AND CLINICAL RELEVANCE: Hyperhomocysteinemia occurs frequently in the Greyhound population. Its association with hypofolatemia suggests decreased intracellular availability of B vitamins, but the functional implications warrant further investigation. Hyperhomocysteinemia in Greyhounds potentially may serve as a spontaneous canine model to further investigate hyperhomocysteinemia in humans.

Influence of Breed Size, Age, Fecal Quality, and Enteropathogen Shedding on Fecal Calprotectin and Immunoglobulin A Concentrations in Puppies During the Weaning Period.

BACKGROUND: Fecal calprotectin and immunoglobulin A (IgA) are markers of intestinal inflammation and immunity in adult dogs. HYPOTHESIS: Fecal calprotectin and IgA concentrations in puppies are not influenced by fecal moisture in puppies but by enteropathogen shedding. ANIMALS: Three hundred and twenty-four puppies. METHODS: Fecal consistency was assessed by gross examination. Fecal moisture was evaluated before and after lyophilization. Canine parvovirus and coronavirus were detected in feces by qPCR and qRT-PCR respectively. Giardia intestinalis antigen was quantified by ELISA. The standard McMaster flotation technique was used to detect eggs and oocysts in feces. Fecal calprotectin and IgA concentrations were quantified by in-house radioimmunoassays. RESULTS: For each marker (IgA and calprotectin), a strong positive correlation was observed between concentration in fresh feces and concentration in fecal dry matter. 75.6% of the puppies were found to be infected by at ≥1 of the enteropathogens evaluated. Fecal calprotectin concentration was significantly influenced by age (P = .001), with higher concentrations in younger puppies, but not by viral (P = .863) or parasitic infection (P = .791). Fecal IgA concentration was significantly influenced by enteropathogen shedding (P = .01), with a lower fecal IgA concentration in puppies shedding at ≥1 enteropathogen compared to puppies without any enteropathogen shedding, but not by age. CONCLUSIONS: Fecal calprotectin and IgA are of no diagnostic value to detect presence of enteropathogens in clinically healthy puppies or puppies with abnormal feces, but could help to better understand the maturation of digestive tract.

Serum C-reactive protein and S100A12 concentrations in dogs with hepatic disease.

OBJECTIVES: To describe serum C-reactive protein and S100A12 concentrations in dogs with hepatic disease and to determine whether there is a relationship between the concentration of either and the severity of hepatic necroinflammation. METHODS: Serum C-reactive protein and S100A12 concentrations were measured in 46 dogs undergoing hepatic biopsy. Dogs were divided into three groups: congenital portosystemic shunts, chronic hepatitis and hepatic neoplasia. The histological severity of hepatic necroinflammation was scored. RESULTS: C-reactive protein and S100A12 concentrations were greater than the upper limit of the reference intervals in 39 and 26% of dogs, respectively. There was no association of disease group with C-reactive protein (P=0·1733) or S100A12 (P=0·1513) concentrations. There was a positive correlation between serum C-reactive protein concentration and hepatic necroinflammatory activity (rs =0·428, P=0·006). CLINICAL SIGNIFICANCE: Increased serum C-reactive protein and S100A12 concentrations were observed in a subpopulation of dogs with various types of hepatic diseases, suggesting acute-phase inflammation and activation of phagocytic cells, respectively. Dogs with higher hepatic necroinflammatory activity scores tended to have higher serum C-reactive protein concentrations. Further studies are needed to confirm this finding in a larger group of dogs.

[Minimally invasive interventional techniques involving the urogenital tract in dogs and cats]. / Minimal-invasive interventionelle Verfahren am Urogenitaltrakt von Hund und Katze.

Minimally invasive interventional techniques are advancing fast in small animal medicine. These techniques utilize state-of-the-art diagnostic methods, including fluoroscopy, ultrasonography, endoscopy, and laparoscopy. Minimally invasive procedures are particularly attractive in the field of small animal urology because, in the past, treatment options for diseases of the urogenital tract were rather limited or associated with a high rate of complications. Most endourological interventions have a steep learning curve. With the appropriate equipment and practical training some of these procedures can be performed in most veterinary practices. However, most interventions require referral to a specialty clinic. This article summarizes the standard endourological equipment and materials as well as the different endourological interventions performed in dogs and cats with diseases of the kidneys/renal pelves, ureters, or lower urinary tract (urinary bladder and urethra).

[Bilateral renal sclerotherapy as a treatment option in a mixed breed male dog with idiopathic renal haematuria]. / Bilaterale Sklerotherapie zur Behandlung der idiopathischen renalen Hämaturie bei einem Mischlingsrüden.

A 2-year-old male castrated German Shepherd dog mix was presented with chronic macroscopic haematuria. Further diagnostics included abdominal ultrasound and urethrocystoscopy and led to a diagnosis of severe bilateral idiopathic renal haematuria (IRH). Medical treatment with Yunnan Baiyao was unsuccessful. Bilateral renal-sparing sclerotherapy was performed and, despite distal migration of both ureteral stents within 12 days, permanently resolved the macroscopic haematuria.

Impact of interactive voice response technology on primary adherence to bisphosphonate therapy: a randomized controlled trial.

UNLABELLED: Osteoporosis (weak bones) is a disorder that has high morbidity, mortality, and healthcare utilization. Effective treatment is available for this disorder, but many patients choose not to start therapy. This is the first study showing an intervention that increases the initiation rates to medications for osteoporosis. INTRODUCTION: One out of six patients prescribed an oral bisphosphonate does not initiate therapy, a phenomenon known as primary non-adherence. Reasons for bisphosphonate primary non-adherence have been identified, but not interventions that positively impact primary adherence rates. The purpose of this study is to determine the effectiveness of interactive voice response technology to improve oral bisphosphonate primary adherence. METHODS: This was a prospective, randomized controlled trial conducted in January-December 2014 at Kaiser Permanente Colorado, an integrated healthcare system. Adults with a new oral bisphosphonate prescription for osteoporosis or osteopenia which was not purchased within 14-20 days of being ordered were included. There were 127 and 118 patients in the intervention group and control groups, respectively. The intervention group received an interactive voice response phone call followed by a letter 1 week later if primary non-adherence continued, whereas the control group did not receive any outreach. The primary outcome was the proportion of patients who purchased their oral bisphosphonate within 25 days of randomization. RESULTS: There were 62/127 (48.8%) intervention patients and 36/118 (30.5%) control patients who purchased their bisphosphonate prescription within 25 days of randomization (OR = 2.17, 95% CI 1.29-3.67). When adjusted for age, sex, history of bone mineral density scan and fracture, the odds ratio for intervention versus control group was 2.3 (95% CI 1.34-3.94). CONCLUSION: An interactive voice response phone call and follow-up letter significantly improved primary adherence to oral bisphosphonate therapy. Such an intervention could be considered for improving primary adherence rates to other medication classes.

Purification and partial characterization of canine calprotectin.

Calprotectin (CP) is an abundant protein in human neutrophilic granulocytes and macrophages. In humans, serum, urine, and fecal concentrations of neutrophil-derived proteins, such as CP are used as markers of disease activity for conditions associated with increased neutrophil activity, such as inflammatory bowel disease. The aims of the present study were to purify and partially characterize CP in the dog (Canis familiaris) as a prelude to the development of an immunoassay for the quantification of canine serum, urine, and fecal CP in dogs with inflammatory conditions. Leukocytes were isolated from whole blood by dextran sedimentation, and canine CP (cCP) was extracted from the cytosol fraction by repeated freezing--thawing--sonication, followed by further purification using anion- and cation-exchange column chromatography. The overall yield of the purification protocol was 3.7mg cCP per 600ml whole blood. The relative molecular masses of the two proteins representing cCP (cS100A8 and cS100A9) were estimated at 10,340 and 14,628, respectively. Isoelectric focusing revealed two bands with isoelectric points of 6.4 and 6.2 for the heterodimeric protein. The approximate specific absorbance of cCP at 280nm was 0.872 for a 1mg/ml solution. The amino acid sequence of the first 13 N-terminal residues of cS100A8 was Met-Leu-Thr-Glu-Leu-Glu-Ser-Ala-Ile-Asn-Ser-Leu-Ile, whereas the N-terminus of cS100A9 was blocked. Identity of both cS100A8 and cS100A9 was confirmed by tryptic peptide mass fingerprinting followed by peptide sequencing. Antibacterial activity of cCP against Escherichia coli was shown to be concentration-dependent and was reversible upon addition of micromolar amounts of zinc. We conclude that cCP can be successfully purified from canine whole blood using this reproducible, rapid and efficient method.