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.     

Diagnostic exercise: papulovesicular dermatitis in rhesus macaques (Macaca mulatta)

Eleven rhesus monkeys developed multifocal erythematous and a vesicular rash. Most recovered spontaneously, but a 21-year-old female became moribund and was euthanized. Findings were of vesicular dermatitis and widespread multifocal hemorrhagic necrosis of the lungs and other viscera, with intralesional intranuclear inclusions. Simian varicella virus was identified as the cause by polymerase chain reaction analysis and serologic testing.     

Sheep performance under intensive continuous grazing of native grasslands of <Emphasis Type="Italic">Paspalum notatum</Emphasis> and <Emphasis Type="Italic">Axonopus compressus</Emphasis> in the subtropical region of the Highlands of Central Mexico

Liveweight gain was evaluated in tropical Dorper X Pelibuey lambs under intensive continuous grazing of native grasslands dominated by Paspalum notatum (PN) or Axonopus compressus (AC) in the subtropics of Central Mexico. Two trials were undertaken. Trial 1 lasted 12 weeks with 10 lambs (initial weight 18 +/- 2.57 kg, 3 months old) per treatment in 2002, and Trial 2 for 13 weeks with 8 lambs (initial weight 24.0 +/- 2.0 kg, 4 months old) per treatment. Lambs were weighed once per week, and liveweight change was estimated by linear regression over day of the experiment, using individual regression coefficients as unbiased estimates of daily liveweight change; analysed in a random block design. Lambs on Trial 1 gained 0.061 kg/lamb/day on PN and 0.047 kg/lamb/day on AC (P > 0.05) at an overall mean stocking rate of 25 lambs/ha. In Trial 2, liveweight gain was significantly larger in PN (0.060 kg/lamb/day) than on AC (0.043 kg/lamb/day) (P < 0.05), at a mean stocking rate of 21.5 lambs/ha. It is concluded that intensive continuous grazing of native grasslands in the subtropics of the highlands of Central Mexico enables moderate liveweight gains for weaned lambs during the rainy season; with better results in grasslands dominated by Paspalum notatum.     

Treatment with rhBMP-2 of extreme radial bone atrophy secondary to fracture management in an Italian Greyhound

rhBMP-2 solution on a collagen sponge was placed along the diaphysis of an atrophicradius, which had a history of recurring fractures. Two months after rhBMP-2 treatment, new mineralized bone was present, which significantly increased the diameter of the radius and allowed the removal of the external skeletal fixator (ESF). Due to carpo-metacarpal joint compromise, a pancarpal arthrodesis was performed seven months later. At follow-up evaluation two years later the dog was only very mildly lame.     

Injection of the insertion of the deep digital flexor tendon in horses using radiographic guidance

Insertional tendinopathies of the DDFT have been reported both as the sole lesion and as part of a multifocal lesion (Dyson et al. 2003). Computed tomography (CT) and magnetic resonance imaging allow specific diagnosis of deep digital flexor tendon lesions within the hoof capsule; however, direct intralesional treatment of such lesions is difficult because of the hoof's rigid structure. A technique designed to mimic intralesional injection of insertional tendinopathies of the DDFT in the standing horse using radiographic guidance was assessed. Radiographic and contrast CT imaging and sectioning of the limbs confirmed accurate injection in all cases although inadvertant administration of injectate into adjacent structures was also evident.     

Retrospective evaluation of sildenafil citrate as a therapy for pulmonary hypertension in dogs

Pulmonary arterial hypertension (PH) is a pathologic condition in dogs characterized by abnormally high pressures in the pulmonary circulation and has been associated with a poor outcome. Sildenafil is a type V phosphodiesterase inhibitor that produces nitric oxide mediated vasodilatation. Sildenafil treatment decreases pulmonary arterial pressure and pulmonary vascular resistance in people with PH. The purpose of this study was to describe the clinical characteristics and outcome of dogs with PH treated with sildenafil. The cardiology database was searched for dogs with PH treated with sildenafil. PH was defined as systolic pulmonary arterial pressure (PAPs) > or = 25 mmHg at rest. Medical records were reviewed for the following information: signalment, duration and type of clinical signs before treatment, underlying disease, estimated or measured PAPs, dosage and dosing interval of sildenafil, and the effect of treatment on clinical signs and pulmonary arterial pressure and survival time. Thirteen affected dogs were identified. Clinical signs included collapse, syncope, respiratory distress, and cough. Duration of clinical signs before presentation ranged from 3 days to 5 months. An underlying cause was identified in 8 dogs. The median sildenafil dosage was 1.9 mg/kg. Ten dogs received concurrent medications. Median PAPs was 90 mmHg; 8 dogs were reevaluated after therapy, and the median decrease in PAPs was 16.5 mmHg. The median survival time of all dogs was 91 days. Sildenafil appeared to be well tolerated in dogs with PH and was associated with decreased PAPs and amelioration of clinical signs in most. Sildenafil represents a reasonable treatment option for dogs with pulmonary hypertension.     

Evaluation of systemic administration of gentamicin for treatment of coliform mastitis in cows

Recovery of cows (n = 61) with mastitis caused by gram-negative bacteria and treated systemically with an antibiotic (gentamicin) to which the bacteria were susceptible in vitro, was compared with recovery of cows (n = 13) with similar infections treated with a systemically administered antibiotic (erythromycin) to which the bacteria were resistant in vitro or with recovery of cows (n = 12) not given an antibiotic systemically. In the first part of the study, cows were selected for treatment groups by use of a diagnostic scheme designed to predict whether the mastitis was caused by gram-negative or gram-positive bacteria. In the second part of the study, all cows were treated without systemic administration of an antibiotic. Significant difference was not observed in the outcome of the disease between cows given gentamicin and cows of the other 2 treatment groups at 24 hours or at 4 weeks after treatment. At 24 hours after initial treatment, 71.9% of cows treated with gentamicin, 92.3% of those treated with erythromycin, and 45.5% not treated systemically had improved appetite. At 4 weeks after initial treatment, of the cows treated with gentamicin, 11.5% died; in 32.8%, lactation ceased in the affected mammary gland; in 21.3%, lactation was decreased in the affected gland; and 34.4% returned to normal lactation and health. Of cows treated with erythromycin, none died; in 23%, lactation ceased in the affected mammary gland; in 23%, lactation decreased in the affected gland; and 54% returned to normal lactation and health.(ABSTRACT TRUNCATED AT 250 WORDS)     

The use of biotin-labeled cDNA probes for the detection of infectious bursal disease viruses

A cDNA library was prepared from the double-stranded RNA genome of the infectious bursal disease virus (IBDV) strain ST-C. The cDNA molecules were annealed into the plasmid pUC9 and used to transform Escherichia coli strain JM107. A cDNA clone that contained IBDV-specific nucleotide sequences was selected and designated STC-1. Radiolabeled probes were prepared from STC-1 and hybridized to genome segment A of ST-C in a northern blot hybridization assay. The STC-1 cDNA was 448 base pairs in length, and its nucleotide sequence indicated that it is located near the VP-2/VP-4 junction in IBDV genome segment A. Biotin-labeled probes were prepared from STC-1 and used in a dot-blot hybridization assay to detect IBDV. Under relatively low stringency conditions of hybridization, the biotinylated probes detected four subtypes of IBDV serotype 1 and a serotype 2 IBDV isolate.