Comparative pathophysiology and management of protein-losing enteropathy.

Protein-losing enteropathy, or PLE, is not a disease but a syndrome that develops in numerous disease states of differing etiologies and often involving the lymphatic system, such as lymphangiectasia and lymphangitis in dogs. The pathophysiology of lymphatic disease is incompletely understood, and the disease is challenging to manage. Understanding of PLE mechanisms requires knowledge of lymphatic system structure and function, which are reviewed here. The mechanisms of enteric protein loss in PLE are identical in dogs and people, irrespective of the underlying cause. In people, PLE is usually associated with primary intestinal lymphangiectasia, suspected to arise from genetic susceptibility, or "idiopathic" lymphatic vascular obstruction. In dogs, PLE is most often a feature of inflammatory bowel disease (IBD), and less frequently intestinal lymphangiectasia, although it is not proven which process is the true driving defect. In cats, PLE is relatively rare. Review of the veterinary literature (1977-2018) reveals that PLE was life-ending in 54.2% of dogs compared to published disease-associated deaths in IBD of <20%, implying that PLE is not merely a continuum of IBD spectrum pathophysiology. In people, diet is the cornerstone of management, whereas dogs are often treated with immunosuppression for causes of PLE including lymphangiectasia, lymphangitis, and crypt disease. Currently, however, there is no scientific, extrapolated, or evidence-based support for an autoimmune or immune-mediated mechanism. Moreover, people with PLE have disease-associated loss of immune function, including lymphopenia, severe CD4+ T-cell depletion, and negative vaccinal titers. Comparison of PLE in people and dogs is undertaken here, and theories in treatment of PLE are presented.

Retrospective characterization of canine coagulopathies using the turbidimetric ACL-TOP 300 analyzer (2014-2015): Forty-seven dogs.

OBJECTIVE: To characterize disseminated intravascular coagulation (DIC), liver failure (LF), post-hepatic cholestasis (PHC), and anticoagulant rodenticide intoxication (ROD) in dogs using an immunoturbidimetric coagulation analyzer and to characterize the relationship between clinical bleeding and bleeding parameters. DESIGN: Retrospective study (August 2014-July 2015). SETTING: University teaching hospital. ANIMALS: Forty-seven client-owned dogs diagnosed with DIC (n = 24), LF (n = 9), PHC (n = 5), or ROD (n = 9) based on history, clinical pathology, cytology, histopathology. or exploratory surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Median prothrombin time (PT), activated partial thromboplastin time (aPTT), and quantitative fribrinogen assay (QFA) were above the reference interval for DIC, LF, PHC and ROD with the exception of a normal QFA for LF. Clot curve analysis for DIC was characterized by elevated PT Delta, PT first derivative, and aPTT Delta, and normal for aPTT second derivative; all LF parameters were within the RI; all PHC parameters were above the RI; and ROD had elevated aPTT delta, but low aPTT second derivative. Coagulopathic bleeding recognized within the DIC group was characterized by median PT delta in mABS (milliabsorbance), first derivative and aPTT delta values in mABS within the RI at 35.0, 55.5 and 38, respectively. The nonbleeding DIC group median values of these same parameters were 189.5, 586.5 and 288, respectively. CONCLUSIONS: The classically utilized indicators of secondary hemostasis, PT and aPTT, were prolonged within all 4 groups; DIC, LF, PHC and ROD as expected. Fibrinogen concentration was increased in both PHC and ROD, decreased in LF and increased but with a bimodal distribution in DIC that correlated with clinical bleeding. The degree of PT and aPTT prolongation did not correlate with clinical bleeding in the DIC group, however clot curve analysis, did reveal an association.

Assessment of eosinophils in gastrointestinal inflammatory disease of dogs.

BACKGROUND: Accurate identification of eosinophils in the gastrointestinal (GI) tract of dogs with eosinophilic GI disease (EGID) by histological evaluation is challenging. The currently used hematoxylin and eosin (H&E) staining method detects intact eosinophils but does not detect degranulated eosinophils, thus potentially underrepresenting the number of infiltrating eosinophils. OBJECTIVE: To develop a more sensitive method for identifying and quantifying both intact and degranulated eosinophils to diagnose EGID more accurately. METHODS: Endoscopically obtained paraffin-embedded intestinal biopsy specimens from dogs with GI signs were examined. The study groups were dogs with eosinophilic enteritis (EE), lymphoplasmacytic and mixed enteritis, and control dogs with GI signs but no histologic changes on tissue sections. Consecutive sections were immunolabeled with monoclonal antibodies (mAbs) against the eosinophil granule protein eosinophil peroxidase (Epx) and stained by H&E, respectively. The number of eosinophils was manually quantified and classified as intact or degranulated. RESULTS: The number of intact eosinophils detected in Epx mAb-labeled duodenal sections was significantly higher compared with that in H&E-stained sections, with a similar relationship noted in the colon and stomach. The Epx mAb allowed the unique assessment of eosinophil degranulation. The number of intact and degranulated eosinophils was significantly higher in duodenal lamina propria of the EE and mixed group compared to the control group. CONCLUSION: Immunohistochemical detection of Epx provides a more precise method to detect GI tract eosinophils compared to H&E staining and could be used as an alternative and reliable diagnostic tool for assessment of biopsy tissues from dogs with EGID.

Inhibition of soluble epoxide hydrolase attenuates eosinophil recruitment and food allergen-induced gastrointestinal inflammation.

Prevalence of food allergies in the United States is on the rise. Eosinophils are recruited to the intestinal mucosa in substantial numbers in food allergen-driven gastrointestinal (GI) inflammation. Soluble epoxide hydrolase (sEH) is known to play a pro-inflammatory role during inflammation by metabolizing anti-inflammatory epoxyeicosatrienoic acids (EETs) to pro-inflammatory diols. We investigated the role of sEH in a murine model of food allergy and evaluated the potential therapeutic effect of a highly selective sEH inhibitor (trans-4-{4-[3-(4-trifluoromethoxyphenyl)-ureido]-cyclohexyloxy}-benzoic acid [t-TUCB]). Oral exposure of mice on a soy-free diet to soy protein isolate (SPI) induced expression of intestinal sEH, increased circulating total and antigen-specific IgE levels, and caused significant weight loss. Administration of t-TUCB to SPI-challenged mice inhibited IgE levels and prevented SPI-induced weight loss. Additionally, SPI-induced GI inflammation characterized by increased recruitment of eosinophils and mast cells, elevated eotaxin 1 levels, mucus hypersecretion, and decreased epithelial junction protein expression. In t-TUCB-treated mice, eosinophilia, mast cell recruitment, and mucus secretion were significantly lower than in untreated mice and SPI-induced loss of junction protein expression was prevented to variable levels. sEH expression in eosinophils was induced by inflammatory mediators TNF-α and eotaxin-1. Treatment of eosinophils with t-TUCB significantly inhibited eosinophil migration, an effect that was mirrored by treatment with 11,12-EET, by inhibiting intracellular signaling events such as ERK (1/2) activation and eotaxin-1-induced calcium flux. These studies suggest that sEH induced by soy proteins promotes allergic responses and GI inflammation including eosinophilia and that inhibition of sEH can attenuate these responses.

Assessment of eosinophil peroxidase as a potential diagnostic and prognostic marker in dogs with inflammatory bowel disease.

OBJECTIVE To evaluate a method for identifying intact and degranulated eosinophils in the small intestine of dogs with inflammatory bowel disease (IBD) by use of a monoclonal antibody (mAb) against eosinophil peroxidase (EPX). ANIMALS 11 untreated dogs with IBD, 5 dogs with IBD treated with prednisolone, and 8 control dogs with no clinical evidence of gastrointestinal tract disease and no immunosuppressive treatment. PROCEDURES 4-µm-thick sections of paraffin-embedded tissues from necropsy specimens were immunostained with EPX mAb. Stained intact and degranulated eosinophils in consecutive microscopic fields (400X magnification) of the upper (villus tips) and lower (between the muscularis mucosae and crypts) regions of the lamina propria of the jejunum were manually counted. RESULTS Compared with control and treated IBD dogs, untreated IBD dogs had a significantly higher number of degranulated eosinophils in the lower region of the lamina propria. However, no significant differences were detected in the number of intact eosinophils in this region among groups. In the upper region of the lamina propria, untreated IBD dogs had a significantly higher number of degranulated and intact eosinophils, compared with control and treated IBD dogs. Number of degranulated and intact eosinophils did not differ significantly between control and treated IBD dogs. CONCLUSIONS AND CLINICAL RELEVANCE Immunohistologic analysis with EPX mAb yielded prominent granule staining that allowed reliable morphological identification of degranulated and intact eosinophils, which may provide a strategy for quantitative and selective evaluation of eosinophils in gastrointestinal biopsy specimens and a potential method to diagnose IBD and evaluate treatment outcome.

Signalment, history, and outcome of cats with gastrointestinal tract intussusception: 20 cases (1986-2000).

OBJECTIVE: To determine signalment, history, and outcome of cats with gastrointestinal tract intussusception and to identify physical examination, diagnostic imaging, surgical, histologic, and necropsy findings in affected cats. DESIGN: Retrospective case series. ANIMALS: 20 cats with intussusception. PROCEDURES: Medical records were evaluated for information on signalment; history; physical examination, diagnostic imaging, surgical, histologic, and necropsy findings; and outcome. RESULTS: Ten cats were < 1 year old, and 9 were >or= 6 years old. Anorexia (14/17), lethargy (12/17), and vomiting (12/17) were the most common reasons for examination. Dehydration (13/18), poor body condition (12/18), signs of abdominal pain (8/18), and an abdominal mass (8/18) were the most common physical examination findings. Abdominal radiography revealed intestinal obstruction in all 10 cats in which it was performed; abdominal ultrasonography revealed intussusception in all 7 cats in which it was performed. The most common intussusception was jejuno-jejunal (8/20), and no intussusceptions were found proximal to the duodenum. Eleven of 13 cats that underwent laparotomy required intestinal resection and anastomosis. Histologic examination revealed intestinal lymphoma or inflammatory bowel disease in 7 of 8 cats >/= 6 years old and idiopathic intussusception in 7 of 8 cats < 1 year old. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that in cats, intussusception has a bimodal age distribution, is most commonly jejuno-jejunal, often requires surgical resection and anastomosis, is often associated with alimentary lymphoma or inflammatory bowel disease in older cats, and is readily diagnosed by means of ultrasonography.

Presumed primary immune-mediated thrombocytopenia in four cats.

Feline primary immune-mediated thrombocytopenia (pIMT) is a rare hematological disorder. Platelet-bound antibody assays for cats have variable specificity and sensitivity and are not widely available. Diagnosis of pIMT is made on the basis of exclusion of other identifiable causes of thrombocytopenia and the response to immunosuppressive therapy. This report describes four cats with severe thrombocytopenia and no detectable underlying disease. One cat was euthanased because of pulmonary hemorrhage, while the other cats had frequent relapses, two of these cats developed diabetes mellitus due to long-term corticosteroid therapy. In these cats IMT had a chronic course and responded poorly to therapy with prednisolone. Alternative immunomodulatory drugs may be considered in the treatment of feline IMT.

Treatment of severe immune-mediated thrombocytopenia with human IV immunoglobulin in 5 dogs.

BACKGROUND: Glucocorticoids with or without other immunotherapy are the initial treatment of choice for dogs with severe immune-mediated thrombocytopenia (IMT). The majority of treated dogs will have improvements in platelet counts within 5 to 7 days of starting therapy, but complications from hemorrhage often occur before a response is seen. Human IV immunoglobulin (hIVIG) blocks Fc receptors on mononuclear phagocytic cells in dogs; it is used in people with idiopathic thrombocytopenic purpura. HYPOTHESIS: The purpose of this study was to describe adverse effects and benefit of hIVIG in addition to conventional immunosuppressive therapy in dogs with severe IMT. ANIMALS: Five client-owned dogs with severe primary IMT. METHODS: Case series. The hospital database was searched for dogs with primary IMT treated with hIVIG. RESULTS: No adverse effects were noted during or after hIVIG infusion in any treated dog. Over a 6-month follow-up, all dogs were clinically normal when using conventional immunosuppressive therapy. Human IVIG was administered 3 days after initiation of immunosuppressive therapy in 4 dogs, and, after 2 days, in 1 dog. In all dogs, the mean platelet counts pre- and 24 hours post-hIVIG infusion (0.28-0.76 g/kg) were 2,500/pL and 50,600/microL (62,750/microL for the 4 responders), respectively. One dog failed to respond as promptly to hIVIG (0.34 g/kg), and the platelet count increased to 66,000/microL after 9 days of immunosuppressive therapy. The mean duration of hospitalization post-hIVIG in all 5 dogs was 1.8 days (12 hours for responders), and the mean total length of hospitalization was 4.6 days (3.5 days for responders). Active hemorrhage resolved and no packed red blood cell transfusions were required after hIVIG infusion for responders. CONCLUSIONS AND CLINICAL IMPORTANCE: Human IVIG was well tolerated and appeared to be associated with rapid platelet count recovery and amelioration of clinical signs in most dogs with IMT.

Detection of Bartonella henselae DNA in two dogs with pyogranulomatous lymphadenitis.

CASE DESCRIPTION: 1 dog evaluated because of inappetence and lameness of the left hind limb of 1 day's duration and 1 dog evaluated because of inappetence, fever, and lymphadenopathy of 2 weeks' duration. CLINICAL FINDINGS: Histologic examination of excisional biopsy specimens from lymph nodes revealed pyogranulomatous lymphadenitis in both dogs. Quantitative real-time PCR assays detected Bartonella henselae DNA in blood samples and affected lymph node specimens from both dogs. Antibodies against B. henselae were not detected via immunofluorescent antibody testing during active disease in either dog. TREATMENT AND OUTCOME: 1 dog recovered after 6 weeks of treatment with doxycycline (5 mg/kg [2.3 mg/lb], p.o., q 12 h), whereas the other dog recovered after receiving a combination of azithromycin (14.5 mg/kg [6.6 mg/lb], p.o., q 24 h for 21 days), doxycycline (17.3 mg/kg [7.9 mg/lb], p.o., q 24 h for 4 weeks), and immunosuppressive corticosteroid (prednisone [3 mg/kg {1.4 mg/lb}, p.o., q 24 h], tapered by decreasing the daily dose by 25% every 2 weeks) treatment. CLINICAL RELEVANCE: B. henselae is implicated as a possible cause or a cofactor in the development of pyogranulomatous lymphadenitis in dogs. In dogs with pyogranulomatous lymphadenitis, immunofluorescent assays may not detect antibodies against B. henselae. Molecular testing, including PCR assay of affected tissues, may provide an alternative diagnostic method for detection of B. henselae DNA in pyogranulomatous lymph nodes.

Expression and function of TLR2, TLR4, and Nod2 in primary canine colonic epithelial cells.

The gut maintains a delicate balance between the downregulation of inflammatory reactions to commensal bacteria and the capacity to respond to pathogens with vigorous cellular and humoral immune responses. Intestinal epithelial cells, including colonic epithelial cells (CECs) possess many properties of cells of the innate immune system, in particular the ability to recognize and respond to microbial antigens. Recognition of microorganisms by CECs is based upon their recognition of signature molecules, called microbe-associated molecular patterns (MAMP), by pattern recognition receptors (PRR) including membrane toll-like receptors (TLR) and cytosolic Nod2, an intracellular counterpart of TLRs. The purpose of this study was to determine whether primary CECs from normal dogs express a functional TLR2, TLR4, and Nod2 and whether they are regulated by inflammatory mediators. We show that canine primary CECs express TLR2, TLR4, and Nod2 that can be modulated in response to their respective MAMPs, lipopolysaccharides (LPS) or peptidoglycans (PGN). Furthermore, we demonstrate that these receptors are functional as evidenced by the induction of cytokine gene expression in response to LPS or PGN.

Suppurative, nonseptic polyarthropathy in dogs.

The goals of this study were to determine the historical, physical examination, and clinicopathologic findings in dogs with suppurative, nonseptic polyarthropathy and to identify concurrent disorders associated with this syndrome. Medical records of 52 dogs with cytologic evidence of suppurative inflammation in two or more joints were examined retrospectively. Age of dogs was 4.8 years (median, range: 0.5-12 years). There was no clear breed or sex predilection, but most were large-breed dogs (body weight > or = 20 kg [44.4 lbs] in 40/52). Body temperature was 103.0 degrees F (39.4 degrees C) (median, range: 100.0-105.9 degrees F), with 29 of 52 dogs having a body temperature > or = 103 degrees F (39.4 degrees C). Lameness was identified in 42 of 52 dogs. Erosive changes were found in only 1 of 37 dogs that had radiography performed. A clear underlying disease process was not identified in 34 of 52 dogs. Seven dogs had evidence of infectious or inflammatory processes at extra-articular sites; 4 dogs were diagnosed with systemic lupus erythematosus (SLE); 2 dogs had gastrointestinal disease; 2 dogs had been vaccinated within 1 month before onset of polyarthritis; 1 dog had cancer; 1 dog had polyarthritis and meningitis; and 1 dog had erosive polyarthritis. Of the 44 dogs tested, 25 had antibodies to Borrelia burgdorferi, detected by an ELISA assay, which was significantly greater than the general hospital population (P = .007). Antibodies against Rickettsia rickettsiae and Ehrlichia canis were not definitively identified in the sera of any dog tested in this study (45 and 44 dogs, respectively). We conclude that an underlying disease process is not identified in most cases of suppurative polyarthropathy in dogs and that intestinal disease, neoplasia, and SLE are uncommon causes of polyarthritis. While seropositivity against the causative agent of Lyme disease was common and possibly a cause of polyarthritis in some dogs of our study, evidence of other vector-borne infection was not identified.

Six-month prophylactic efficacy of moxidectin sustained release (SR) injectable for dogs against experimental heartworm infection in growing puppies.

The purpose of this study was to assess the efficacy of moxidectin sustained release injectable for dogs (moxidectin SR, Fort Dodge Animal Health) in protecting growing puppies from experimental infection with the heartworm, Dirofilaria immitis, six months after treatment. The study involved 27 puppies, approximately 12 weeks of age at the beginning of the study, with nine puppies in each of three size classes. The small breed class included eight Pekingese and one purpose-bred small breed mongrel; the medium breed class included nine purpose-bred mongrels, and the large breed class included nine puppies with an anticipated adult weight >or=30-35 kg. Both genders were included with no attempt made to have equal numbers of male and female puppies. Puppies were blocked by weight within each size class and randomly assigned to three treatment groups of nine dogs. On Day 0, pups in two groups were injected subcutaneously with moxidectin SR, dosed to deliver 0.17 mg moxidectin/kg b.w. The third group was injected with sterile saline. Personnel making observations were blinded to the treatment status of the animals. Following treatment, puppies were observed for signs of adverse local and systemic reactions. Puppy weights and serum moxidectin levels were also monitored. On Day 180, puppies in all treatment groups were inoculated subcutaneously with 50 third-stage larvae of D. immitis. On Days 348 and 349, puppies were euthanatized and necropsied. Hearts and lungs were examined for adult heartworms. All animals in the saline control group were infected with an arithmetic mean of 39.22 adult heartworms each. Seventeen of 18 dogs in the moxidectin SR-treated groups were uninfected. One treated puppy was infected with a single adult heartworm. This infected individual was from the large breed size class and had the second highest percent increase in body weight. Based on arithmetic means, the heartworm recovery from all treated puppies represents a 99.86% reduction relative to the saline control. There were no adverse local or systemic reactions to treatment in any animal.

Gastrointestinal motility disorders and gastrointestinal prokinetic therapy.

Gastrointestinal motility disorders represent a diagnostic and therapeutic challenge. Disorders of gastrointestinal motility may result in accelerated transit, delayed transit, impaired relaxation, or inappropriate relaxation. The delayed transit disorders are the most important motility disorders of companion animals and may involve the esophagus (hypomotility and megaesophagus), stomach (delayed gastric emptying), small intestine (postoperative ileus and intestinal pseudo-obstruction), or colon (constipation and megacolon).

Clinical differentiation of acute necrotizing from chronic nonsuppurative pancreatitis in cats: 63 cases (1996-2001).

OBJECTIVE: To characterize clinical, clinicopathologic, radiographic, and ultrasonographic findings in cats with histologically confirmed acute necrotizing pancreatitis (ANP) or chronic nonsuppurative pancreatitis (CP) and identify features that may be useful in the antemortem differentiation of these disorders. DESIGN: Retrospective study. ANIMALS: 63 cats with histologically confirmed ANP (n = 30) or CP (33). PROCEDURE: Medical records were reviewed for signalment, clinical signs, concurrent diseases, clinicopathologic findings, and results of radiography and ultrasonography. RESULTS: Cats in both groups had similar nonspecific clinical signs, physical examination findings, and radiographic and ultrasonographic abnormalities. Abdominal ultrasonographic abnormalities, including hypoechoic pancreas, hyperechoic mesentery, and abdominal effusion, were found in cats in both groups and, therefore, were not specific for ANP. Cats with CP were significantly more likely to have concurrent diseases than were cats with ANP (100 and 83%, respectively). Clinicopathologic abnormalities were similar between groups; however, serum alanine aminotransferase and alkaline phosphatase activities were significantly higher in cats with CP. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that ANP and CP in cats cannot be distinguished from each other solely on the basis of history, physical examination findings, results of clinicopathologic testing, radiographic abnormalities, or ultrasonographic abnormalities.

Short chain fatty acids stimulate feline colonic smooth muscle contraction.

The effect of short chain fatty acids (SCFA) on feline colonic smooth muscle contraction was evaluated in vitro. Colonic tissue was obtained from seven healthy male and female adult cats and seven healthy male and female kittens. Longitudinal and circular colonic smooth muscle strips from proximal and distal colon were incubated with SCFA (acetate, butyrate and propionate; 1-100mM). SCFA-induced contractions were compared to responses obtained using maximal concentrations (10(-4)M) of acetylcholine (ACh). The calcium dependence of the SCFA response was investigated by incubating with nifedipine (1 microM) or verapamil (1 microM). Acetate, butyrate and propionate elicited isometric stress responses (0.25-1.98 x 10(4)N/m(2)) in longitudinal, but not circular, smooth muscle from both the proximal and distal colon of adult cats. Maximal responses were attained at 50 and 100mM SCFA. Maximal butyrate and propionate responses were 29 and 19% of the maximal ACh response (10(-4)M), respectively. Acetate was least effective in stimulating contractile responses. Nifedipine and verapamil abolished all responses. Contractile responses in kittens were similar to those observed in adult cats, but were smaller in amplitude. Results of these studies have shown that SCFA stimulate longitudinal colonic smooth muscle contractions in kittens and adult cats in vitro. These SCFA-induced contractions involve activation of calcium influx. These in vitro findings may account for some of the effects of dietary fiber on feline colonic motility in vivo.

Detection of Bartonella henselae and Bartonella clarridgeiae DNA in hepatic specimens from two dogs with hepatic disease.

A 4-year-old Basset Hound and a 6-year-old Doberman Pinscher were referred for diagnostic evaluation following documentation of persistently increased hepatic enzyme activities and hepatic dysfunction. Histologic evaluation of hepatic biopsy specimens from the 2 dogs revealed granulomatous hepatitis in the Basset Hound and lymphocytic hepatitis with fibrosis and copper accumulation in the Doberman Pinscher. No etiologic agents were identified histologically. Bartonella henselae DNA was subsequently amplified from hepatic tissue from the Basset Hound and Bartonella clarridgeiae was amplified from hepatic tissue from the Doberman Pinscher. Amplification was performed with a polymerase chain reaction assay incorporating primers that target a portion of the 16S-23S rRNA intergenic spacer region. Both dogs were treated with azithromycin, in combination with a variety of other medications and herbal treatments, and improved clinically. Identification of Bartonella DNA in these dogs indicates the need for future prospective studies to determine the clinical relevance of Bartonella spp infection in dogs with hepatic disease.

Ultrasonographic findings in cats with clinical, gross pathologic, and histologic evidence of acute pancreatic necrosis: 20 cases (1994-2001).

OBJECTIVE: To determine ultrasonographic findings in cats with clinical, gross pathologic, and histologic evidence of acute pancreatic necrosis. DESIGN: Retrospective study. ANIMALS: 20 cats. PROCEDURE: Ultrasound reports and permanent ultrasonographic images were reviewed, and ultrasonographic findings were recorded. Thoracic and abdominal radiographs were also reviewed, when available. Anatomic localization of pancreatic necrosis was determined from the gross pathology report; duration and severity of pancreatic necrosis were determined by reviewing histologic specimens. The presence of concurrent disease was recorded from the final pathology report. RESULTS: The pancreas was considered ultrasonographically normal in 10 cats and was not observed in 3. Ultrasonographic findings were considered compatible with pancreatitis in the remaining 7 cats. Gross pathologic findings indicated that pancreatitis was multifocal in all 7 of these cats; histologically, pancreatitis was acute or subacute in 5 and associated with severe or moderate necrosis in 6. In the remaining 13 cats, gross pathologic findings indicated that pancreatitis was multifocal (n = 8) or focal (2), or gross pathologic findings were normal (3). Histologically, pancreatitis was peracute or acute in 11 of these 13 cats and associated with severe or moderate necrosis in 8. Thoracic and abdominal radiographic findings were nonspecific. CONCLUSIONS AND CLINICAL RELEVANCE: Results of ultrasonography were consistent with a diagnosis of pancreatitis in only 7 of 20 cats with acute pancreatic necrosis in the present study. This suggests that new diagnostic criteria must be established if abdominal ultrasonography is to be an effective tool in the diagnosis of pancreatitis in cats.

Cerebrospinal fluid glutamine, tryptophan, and tryptophan metabolite concentrations in dogs with portosystemic shunts.

OBJECTIVE: To determine whether glutamine (GLN), tryptophan (TRP), and tryptophan metabolite concentrations are higher in cerebralspinal fluid (CSF) dogs with naturally occurring portosystemic shunts (PSS), compared with control dogs. ANIMALS: 11 dogs with confirmed PSS and 12 control dogs fed low- and high-protein diets. PROCEDURE: Cerebrospinal fluid and blood samples were collected from all dogs. Serum and CSF concentrations of GLN, alanine, serine, TRP, 5-hydroxyindoleacetic acid (5-HIAA), and quinolinic acid (QUIN) were measured. RESULTS: Cerebrospinal fluid concentrations of GLN, TRP, and 5-HIAA were significantly higher in PSS dogs, compared with control dogs fed high- or low-protein diets. Cerebrospinal fluid QUIN concentration was significantly higher in PSS dogs, compared with control dogs fed the low-protein diet. Serum QUIN concentration was significantly lower in PSS dogs, compared with control dogs fed either high- or low-protein diets. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in CNS GLN concentration is associated with high CSF concentrations of TRP and TRP metabolites in dogs with PSS. High CSF 5-HIAA concentrations indicate an increased flux of TRP through the CNS serotonin metabolic pathway, whereas high CSF QUIN concentrations indicate an increased metabolism of TRP through the indolamine-2,3-dioxygenase pathway. The high CSF QUIN concentrations in the face of low serum QUIN concentrations in dogs with PSS indicates that QUIN production from TRP is occurring in the CNS. High concentrations of QUIN and other TRP metabolites in the CNS may contribute to neurologic abnormalities found in dogs with PSS and hepatic encephalopathy.

Mediation of acetylcholine and substance P induced contractions by myosin light chain phosphorylation in feline colonic smooth muscle.

OBJECTIVES: To determine the role of myosin light chain phosphorylation in feline colonic smooth muscle contraction. SAMPLE POPULATION: Colonic tissue was obtained from eight 12- to 24-month-old cats. PROCEDURE: Colonic longitudinal smooth muscle strips were attached to isometric force transducers for measurements of isometric stress. Myosin light chain phosphorylation was determined by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Stress and phosphorylation were determined following stimulation with ACh or SP, in the absence or presence of a calmodulin antagonist (W-7; 0.1 to 1.0 mM), myosin light chain kinase inhibitor (ML-9; 1 to 10 microM), or extracellular calcium free solutions. RESULTS: Unstimulated longitudinal colonic smooth muscle contained low amounts (6.9+/-3.2%) of phosphorylated myosin light chain. Phosphorylation of the myosin light chains was dose and time dependent with maximal values of 58.5% at 30 seconds of stimulation with 100 microM Ach and 60.2% at 45 seconds of stimulation with 100 nM SP Active isometric stress development closely paralleled phosphorylation of the myosin light chains in ACh- or SP-stimulated muscle. W-7 and ML-9 dose dependently inhibited myosin light chain phosphorylation and isometric stress development associated with ACh or SP stimulation. Removal of extracellular calcium inhibited myosin light chain phosphorylation and isometric stress development in ACh-stimulated smooth muscle. CONCLUSIONS AND CLINICAL RELEVANCE: Feline longitudinal colonic smooth muscle contraction is calcium-, calmodulin-, and myosin light chain kinase-dependent. Myosin light chain phosphorylation is necessary for the initiation of contraction in feline longitudinal colonic smooth muscle. These findings may prove useful in determining the biochemical and molecular defects that accompany feline colonic motility disorders.

Effect of short-chain fatty acids on contraction of smooth muscle in the canine colon.

OBJECTIVE: To determine effects of short-chain fatty acids (SCFA) on canine colonic smooth muscle. SAMPLE POPULATION: Colonic tissue obtained from 14 healthy dogs. PROCEDURE: Short-chain fatty acid (SCFA; acetate, propionate, and butyrate; 1 to 100 mmol/L)-induced contractions were compared with responses obtained with acetylmethylcholine (AMCh; 10(-4) mol/L). Roles of enteric neurons, cholinergic receptors, calcium stores in the sarcoplasmic reticulum, and extracellular calcium in the SCFA-induced responses were investigated by incubating muscle strips with tetrodotoxin (1 micromol/L), atropine (1 micromol/L), ryanodine (10 micromol/L), nifedipine (1 micromol/L), ethylene glycol-bis (beta-aminoethylether)-N,N,N',N'-tetra-acetate (EGTA; 0.1 mmol/L), or an extracellular calcium-depleted (zero extracellular calcium) solution prior to the addition of propionate or butyrate. RESULTS: Incubation with SCFA elicited isometric stress responses (0.25 to 2.15 x 10(4) N/m2) in colonic longitudinal smooth muscle. Maximal responses to butyrate and propionate (50 mmol/L) were 37 and 23%, respectively, of the maximal AMCh response. Acetate was least effective in stimulating contractile responses. Tetrodotoxin and atropine did not affect SCFA-induced contractions. Nifedipine and zero extracellular calcium solution abolished responses to butyrate and propionate, whereas EGTA attenuated (> 60%) but did not abolish those responses. Ryanodine did not affect SCFA-induced contractile responses. The SCFA did not affect colonic circular smooth muscle. CONCLUSIONS AND CLINICAL RESPONSE: The SCFA stimulate longitudinal but not circular colonic smooth muscle contractions via a direct effect on smooth muscle. The mechanism of the SCFA effect appears to involve the influx of extracellular calcium. These findings may account for some of the effects of fiber on canine colonic motility [corrected].