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.     

Selected Aspects of Advanced Porcine Reproductive Technology

In vitro fertilization (IVF) of in vitro matured (IVM) oocytes in pigs has become the most popular method of studying gametogenesis and embryogenesis in this species. Furthermore, because of recent advances in in vitro culture (IVC) of IVM–IVF embryos, in vitro production (IVP) of embryos now enables us to generate viable embryos as successfully as for in vivo -derived embryos and with less cost and in less time. These technologies contribute not only to developments in reproductive physiology and agriculture but also to the conservation of porcine genetic resources and the production of cloned or genetically modified pigs. However, in IVP, there still remains the problem of abnormal ploidy, which is caused by performing procedures under non-physiological conditions. In recent years, unique technologies such as intracytoplasmic sperm injection (ICSI) or xenografting of gonadal tissue into immunodeficient experimental animals have been developed to help conserve gamete resources. These technologies combined with IVP are expected to be useful for the conservation of gametes from important genetic resources. Here, we discuss the developmental ability and normality of porcine IVP embryos and also the utilization of ICSI and xenografting in advancing biotechnology in pigs.     

Ocular distribution and toxicity of intravitreal injection of triamcinolone acetonide in normal equine eyes

Objective  To determine ocular distribution and toxicity of a single injection of intravitreal triamcinolone acetonide (TA) in normal horses.
Animals studied  Six adult horses, donated to North Carolina State University.
Procedures  Six horses were injected intravitreally with either 10, 20, or 40 mg ( n  = 2 each) of TA. The opposite eye of each horse was injected with balanced salt solution (BSS). Ocular toxicity was assessed by biomicroscopy, tonometry, indirect ophthalmoscopy, and electroretinogram. Aqueous humor (AH), vitreous humor (VH), and plasma samples were collected. Horses were euthanized 7 or 21 days after injection and eyes enucleated for histopathology. TA concentrations in AH, VH, and plasma were measured by HPLC.
Results  Three control eyes and one TA eye developed inflammation after injection or collection of AH. Positive bacterial cultures ( Corynebacterium spp., Staphylococcus spp., and Streptococcus spp.) were obtained from three of these eyes. Other than transient corneal edema in TA injected eyes, which resolved by 7 days after injection, no other changes were observed. TA crystals were visible within the vitreous body. No evidence of TA toxic effect was noted on histopathology. TA was detected in all AH and VH samples from treated eyes following injection. Drug was not detected in the plasma.
Conclusions  There was no evidence of overt toxicity from intravitreal TA in normal horses and a single intravitreal injection resulted in TA ocular levels for 21 days. However, the risk for bacterial infections with intravitreal injection or anterior chamber aspirations in horses is high. Use of topical and systemic antibiotics after injection is recommended.     

Time budgets and activity patterns of sub-Antarctic fur seals at Gough Island

The diurnal activity patterns of sub-Antarctic fur seals, Arctocephalus tropicalis, were observed at a non- breeding colony site at Gough Island (40°20′S, 9°54′W) during summer. Time budgets of adult males were also studied at idle and breeding colony sites, Levels of activity were highest during the early morning and late afternoon. High ambient temperatures depressed the interaction rate on the dry hauling ground, but activity increased as a result of the movement of heat-stressed seals to and from the sea where favourable conditions for heat loss exist. The fur seals were largely inactive, in particular adult males which spent 93,2% of the time inactive in breeding colonies and 97,9% of Ihe time inactive at idle colony beaches. The daily change in numbers ashore, the relative contribution of the different age and sex classes, their location and distribution on site, and the prevailing weather conditions influence the pattern of interaction and allocation of time to the various activities in A. tropicalis. The predominance of inactivity is considered to be a behavioural thermoregulatory response to limit endogenous heat production as is energy conservation. Both views translate into improved tenure for territorial males, in particular those without access to water for cooling during the breeding season.     

Hydrothermal Assessment of Temporal Variability in Seedbed Microclimate

The microclimatic requirements for successful seedling establishment are much more restrictive than those required for adult plant survival. The purpose of the current study was to use hydrothermal germination models and a soil energy and water flux model to evaluate intra- and interannual variability in seedbed microclimate relative to potential germination response of six perennial grasses and cheatgrass. We used a 44-yr weather record to parameterize a seedbed microclimate model for estimation of hourly temperature and moisture at seeding depth for a sandy loam soil type at the Orchard Field Test Site in southwestern Ada County, Idaho. Hydrothermal germination response was measured in the laboratory for two seed lots of cheatgrass (Bromus tectorum L.), four seed lots of bluebunch wheatgrass (Pseudoroegneria spicata [Pursh] Löve), three seed lots of bottlebrush squirreltail (Elymus elymoides [Raf] Swezey), and one seed lot each of Sandberg bluegrass (Poa secunda J. Presl.), big squirreltail (Elymus multisetus [J.G. Smith] M.E. Jones), thickspike wheatgrass (Elymus lanceolatus [Scribn. And J.G. Smith] Gould) and Idaho fescue (Festuca idahoensis Elmer). Germination response models were developed to estimate potential germination rate for 13 subpopulations of each seed lot for every hour of the 44-yr simulation. Seedbed microclimate was assessed seasonally and for each day, month, and year, and germination rate-sum estimates integrated for a numerical index of relative site favorability for germination for each time period. The rate-sum favorability index showed a consistent pattern among seed lots for different years, and provides a relatively sensitive indicator of annual and seasonal variability in seedbed microclimate. This index could be used with field data to define minimum weather thresholds for successful establishment of alternative plant materials, in conjunction with weather forecast models for making restoration and fire-rehabilitation management decisions in the fall season, for evaluation of potential climate-change impacts on plant community trajectories, and in optimization schemes for selecting among alternative restoration/rehabilitation management scenarios.     

Endometrial polyp in an African wild dog (Lycaon pictus)

An 8-year-old female African wild dog (Lycaon pictus) from a zoo in Gyeonggi province, Republic of Korea presented with a 3.0 x 2.0 x 2.5 cm in size, smooth-surfaced, solitary pedunculated mass protruding into the uterine lumen. Microscopically, the mass was covered with epithelium, contained endometrial gland tissue, and was dilated in the vascularised stroma. Within the mass, there was extensive diffuse haemorrhage with several blood vessels apparently plugged with fibrin. At the base of the mass, the spaces lined with epithelium near the attachment of the stalk were interpreted to be glandular structures. There were segments of cuboidal epithelium found on the surface of the mass, which was similar to the lining the uterus. A diagnosis of an endometrial polyp was made based on the gross and histology findings. This is the first case report of a spontaneous endometrial polyp in an African wild dog.