Adenosine A2A receptor agonists inhibit lipopolysaccharide-induced production of tumor necrosis factor-alpha by equine monocytes

Adenosine is an endogenous nucleoside that regulates many physiological processes by activating one or more adenosine receptor subtypes, namely A1, A2A, A2B and A3. The results of previous studies indicate that adenosine analogues inhibit lipopolysaccharide (LPS)-induced production of reactive oxygen species (ROS) by equine neutrophils primarily through activation of A2A receptors. Because peripheral blood monocytes produce cytokines that are responsible for many of the deleterious effects of LPS, the current study was performed to evaluate the effects of an array of novel adenosine receptor agonists on LPS-induced production of tumor necrosis factor-alpha (TNF-alpha), and to assess the selectively of these agonists for equine adenosine A2A over the A1 receptor. Radioligand binding studies performed with equine tissues expressing adenosine A1 and A2A receptor subtypes yielded a rank order of affinity for the equine A2A receptor of ATL307>ATL309 approximately ATL310 approximately ATL313>ATL202 approximately ATL361 approximately ATL376>ATL372>CGS21680>NECA. Co-incubation of equine peripheral blood monocytes with LPS and these agonists resulted in inhibition of TNF-alpha production with a rank order of potency that strongly correlated with their binding affinities for equine adenosine A2A receptors. Results of experiments performed with one of the adenosine receptor agonists (ATL313) and selective adenosine receptor antagonists confirmed that inhibition of LPS-induced production of TNF-alpha occurred via stimulation of A2A receptors. Although incubation of monocytes with IB-MECA, a compound purported to act as an adenosine A3 receptor agonist, reduced LPS-induced TNF-alpha production, this effect of IB-MECA was inhibited by the A2A selective antagonist ZM241385 but not by the A3 receptor antagonist MRS1220. These results indicate that the adenosine receptor subtype responsible for regulation of LPS-induced cytokine production by equine monocytes is the A2A receptor. To address the signal transduction mechanism responsible for the anti-inflammatory effects of ATL313 in equine monocytes, production of cAMP was compared in the presence and absence of either the adenosine A2A receptor antagonist ZM241385 or the adenosine A2B receptor antagonist MRS1706. In the absence of the antagonists, ATL313 increased production of cAMP; ZM241385 inhibited this effect of ATL313, whereas MRS1706 did not. Furthermore, incubation of monocytes with either the stable analogue of cAMP, dibutyryl cAMP, or forskolin, an activator of adenylyl cyclase, also inhibited LPS-induced production of TNF-alpha production by equine monocytes. Collectively, the results of the current study indicate that adenosine analogues inhibit LPS-induced production of TNF-alpha by equine monocytes primarily via activation of adenosine A2A receptors and do so in a cAMP-dependent manner. The results of this study indicate that stable adenosine analogues that are selective for adenosine A2A receptors may be suitable for development as anti-inflammatory drugs in horses.     

Intestinal myxosarcoma in a thoroughbred mare.

A large fibrotic mass originating from the cecal base was discovered upon surgical exploration of the abdomen in a Thoroughbred mare with a history of chronic colic and weight loss. The mass protruded intraluminally resulting in partial obstruction. Surgical excision was not feasible due to the location of the mass and the inability to exteriorize it adequately from the abdominal cavity. The mass was fibrous with a shiny, gelatinous material present throughout the neoplasm. Histologically, large confluent spaces filled with mucopolysaccharides were identified by staining with Alcian blue. The diagnosis of myxosarcoma was based upon finding of atypical fibroblastic cells, mucinous stroma, local invasiveness, and metastasis to the regional lymph nodes. Myxomatous tumors have not previously been documented to occur in the equine intestinal tract.     

Evaluation of leukapheresis and thrombocytapheresis in the horse

Continuous-flow centrifugation leukapheresis techniques were used to collect 300-ml volumes of leukocyte-rich plasma from 5 nonmedicated horses and from 5 corticosteroid-stimulated horses. White blood cell counts and differential counts were performed on the horses before (base line) and up to 48 hours after leukapheresis. Systemic administration of hydrocortisone increased numbers of total WBC and neutrophils and improved harvest of these cells. Nonmedicated horses had a mean yield of 3.38 X 10(10) leukocytes in the 300-ml volume. Stimulated horses yielded a mean of 6.88 X 10(10) leukocytes. After leukapheresis, WBC counts decreased a mean of 38% in nonstimulated horses and decreased a mean of 30% in stimulated horses. By 6 hours after leukapheresis, circulating WBC counts of horses in both groups had returned to preleukapheresis values. The relationship between neutrophil yield and the 4 variables (preleukapheresis WBC count, preleukapheresis neutrophil count, preleukapheresis lymphocyte count, and the PCV of the leukocyte-rich plasma) were examined, using simple (pair-wise) correlation and multiple linear regression. A significant positive correlation was found between neutrophil yield and preleukapheresis WBC and neutrophil counts. Because sodium citrate was used in the collection system to prevent extracorporeal blood coagulation, ionized and total serum calcium concentrations were monitored before and after leukapheresis. Although total serum calcium concentrations remained unchanged, ionized calcium concentrations decreased approximately 33% from base-line values during the 2-hour leukapheresis procedures. Occasional mild muscle fasciculations were the only adverse clinical signs of citrate toxicity exhibited by the horses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Current concepts of navicular disease

Navicular disease is unlikely to be a single disease. The close anatomical and functional relationships between the navicular bone, collateral sesamoidean ligament, distal sesamoidean impar ligament, deep digital flexor tendon, navicular bursa and distal interphalangeal joint result in the frequent occurrence of combinations of injuries. There are a number of different pathological processes that affect the navicular bone, which probably have different aetiopathogeneses. While there is strong evidence that biomechanical forces may result in failure of functional adaptation and excessive modelling, it is likely that each disease or injury type has a multifactorial cause.     

Radiographic and scintigraphic evaluation of spondylosis in the equine thoracolumbar spine: A retrospective study

Reasons for performing study: Clinical, radiographic and scintigraphic signs associated with spondylosis of the equine thoracolumbar spine have been poorly documented. Objectives: To establish an objective radiographic and scintigraphic grading system for spondylosis lesions; to estimate the prevalence of spondylosis in a population of horses with back pain; and to compare the results of radiography and scintigraphy Methods: Radiographic images of the thoracolumbar spine from 670 horses with clinical signs of back pain were graded. Scintigraphic images from horses with spondylosis lesions underwent subjective and objective analysis. Sensitivity and specificity of scintigraphy for detection of spondylosis relative to radiography for identification of spondylosis were calculated, and Chi‐squared analysis was performed to test for an association between location and severity of lesions. Results: Twenty‐three of 670 horses (3.4%) with back pain had radiographic evidence of spondylosis. Of these horses, 14 (61%) had more than one lesion and 44% (n = 22) of lesions occurred between T11‐T13 vertebral bodies. Only 33% (n = 28) of locations with radiographic changes had increased radiopharmaceutical uptake. Conclusion: Spondylosis occurs at a low prevalence in horses with back pain. It may be present alone or in association with other osseous abnormalities. The clinical significance of spondylosis needs further investigation. Potential relevance: Spondylosis is uncommon but may be a contributor to back pain in the horse.     

Bovine endometritis: comparative efficacy of alfaprostol and intrauterine therapies, and other factors influencing clinical success

A comparative study of the treatment of 306 severe, moderate, or mild cases of bovine endometritis was carried out over two calving seasons. The cases were treated with alfaprostol, or an intrauterine antibacterial preparation, or with a combination of both therapies. There was no significant difference between the efficacies of these treatments, and a single injection of alfaprostol was effective in 74 per cent of the cases treated. The effectiveness of the treatments was related to the degree of self-cure of the endometritis after parturition, the luteal activity at the time of treatment, and farm management factors affecting the health and condition of the calving cows.