Ethyl pyruvate decreases proinflammatory gene expression in lipopolysaccharide-stimulated equine monocytes

Monocytes are among the initial cells that interact with circulating LPS. Binding of LPS to monocyte surface receptors triggers an intracellular signaling cascade and results in the production of proinflammatory cytokines. Ethyl pyruvate, a stable derivative of pyruvate, has been effective in mitigating LPS induced alterations in isolated human monocytes. We hypothesized that ethyl pyruvate would suppress proinflammatory gene expression in LPS-stimulated equine monocytes without affecting cell viability. Equine monocytes were isolated from whole blood using a sediment-gradient centrifugation protocol and enriched to 76% purity by adhesion to tissue culture dishes. Isolated monocytes were incubated with 0, 1, 5, 10 and 50 mM ethyl pyruvate. Cell viability, production of caspase 3/7, and caspase-3 gene expression were determined. In a separate experiment, monocytes were stimulated with LPS (0.1 ng/ml for 1h) followed by incubation with 0, 1, 5, or 10 mM ethyl pyruvate for 1 h. Proinflammatory gene expression was determined by real-time PCR. Ethyl pyruvate at 50 mM adversely affected monocyte viability. Ethyl pyruvate at 10mM or less had no significant effect on monocyte viability, and did not increase activity of caspase 3/7 nor caspase-3 gene expression. Incubation with LPS alone induced a significant upregulation in proinflammatory gene expression. Subsequent treatment of monocytes with ethyl pyruvate significantly reduced IL-8 expression in LPS stimulated monocytes at 5 mM, and IL-8, TNF-α and COX-2 at 10 mM. No beneficial effect on expression of IL-1β or IL-6 was detected. Overall, 10 mM ethyl pyruvate did not adversely affect monocyte viability and suppressed LPS-induced proinflammatory gene expression. Ethyl pyruvate may be a beneficial anti-inflammatory therapy in equine endotoxemia.     

Further insights into the characterization of equine adipose tissue-derived mesenchymal stem cells

Adipose tissue-derived stem cells (ADSCs) represent a promising subpopulation of adult stem cells for tissue engineering applications in veterinary medicine. In this study we focused on the morphological and molecular biological properties of the ADSCs. The expression of stem cell markers Oct4, Nanog and the surface markers CD90 and CD105 were detected using RT-PCR. ADSCs showed a proliferative potential and were capable of adipogenic and osteogenic differentiation. Expression of Alkaline phosphatase (AP), phosphoprotein (SPP1), Runx2 and osteocalcin (OC) mRNA were positive in osteogenic lineages and peroxisome proliferator activated receptor (Pparγ2) mRNA was positive in adipogenic lineages. ADSCs show stem cell and surface marker profiles and differentiation characteristics that are similar to but distinct from other adult stem cells, such as bone marrow-derived mesenchymal stem cells (BM-MSCs). The availability of an easily accessible and reproducible cell source may greatly facilitate the development of stem cell based tissue engineering and therapies for regenerative equine medicine.     

Active surveillance of bat rabies in France: a 5-year study (2004-2009)

Active surveillance of bats in France started in 2004 with an analysis of 18 of the 45 bat species reported in Europe. Rabies antibodies were detected in six indigenous species, mainly in Eptesicus serotinus and Myotis myotis, suggesting previous contact with the EBLV-1 rabies virus. Nineteen of the 177 tested bats were shown serologically positive in seven sites, particularly in central and south-western France. Neither infectious viral particles nor viral genomes were detected in 173 and 308 tested oral swabs, respectively. The presence of neutralising antibodies in female bats (18.6%) was significantly higher than in males (5.6%).     

Serum antibody responses in horses and mice following immunization with Actinobacillus equuli outer membrane proteins and recombinant Aqx toxin

The immune responsiveness of mice (without prior natural exposure) and mares (with naturally acquired antibodies) was determined following vaccination with Actinobacillus equuli outer membrane proteins (OMPs) and/or recombinant A. equuli toxin (rAqx). Mice were vaccinated subcutaneously on days 0 and 21 with one of three doses (5, 25 or 50μg) of A. equuli OMPs, rAqx or both, together with Freund's incomplete adjuvant (FIA). Antibodies against formalin-killed whole bacterial cells (WBCs), OMPs and Aqx were determined on days 0, 21 and 42. Mares were vaccinated subcutaneously on days 0 and 21 with 100μg OMPs, 100μg rAqx or a combination of 50μg of each antigen, together with FIA. Antibodies against WBCs, OMPs and Aqx were determined at 7day intervals for the first 42days, as well as on days 56, 70, 154 and 238. Vaccination of mice stimulated an apparent dose response to OMPs and Aqx. Antibodies against OMPs and Aqx were enhanced following vaccination of mares that had naturally acquired pre-existing antibodies. There was no evidence of interference with antibody responses to the individual antigens when OMPs and rAqx were combined prior to vaccination.     

Choroidal melanocytoma in a cat

An 11-year-old male, neutered European cat was presented for anisocoria due to pupillary dilation in the right eye. Ophthalmic findings were restricted to this eye and consisted of a raised, darkly pigmented, retrolental mass associated with retinal detachment. Ultrasonography identified the mass lesion protruding into the vitreous cavity from the posterior pole of the eyeball and confirmed the detachment of the retina. A tentative diagnosis of an intraocular tumor was made. Radiographic evaluation and retromandibular lymph node cytology did not reveal evidence of distant metastasis. Orbital exenteration of the affected eye was performed and the tumor was diagnosed as a choroidal melanocytic tumor with no criteria of malignancy (melanocytoma). The cat died 5 months later from renal lymphoma, and necropsy did not detect metastasis of the melanocytic tumor. To our knowledge this is the first reported case of feline choroidal melanocytoma.     

Spiroplasma found in the eyes of scrapie affected sheep

Objective Scrapie, a transmissible spongiform encephalopathy (TSE) occurring naturally in sheep, characteristically shows a severe retinopathy that is well developed in the terminal phases of the disease. In this study, we set out to demonstrate similar retinal changes in our ruminant spiroplasmosis TSE model. Procedure The eyes from deer, sheep, and goats that were inoculated intracranially with the laboratory strain of spiroplasma (suckling mouse cataract [SMCA] strain of Spiroplasma mirum) or with Spiroplasma sp. isolated from the brains affected with scrapie or with chronic wasting disease were examined by light microscopy for pathologic changes and by immunocytochemistry for distribution of spiroplasma antigen. The eyes were also obtained from a research flock of sheep with terminal scrapie, from which the intraocular tissues were submitted aseptically for culture assay in M1D broth or as explants on bovine corneal endothelia (BCE). Results The eyes from the spiroplasmosis ruminant models showed retinopathy remarkably similar to eye lesions seen in sheep with scrapie. The spiroplasma antigen accrued in the ruminant model eye tissues, particularly in the retina, the vitreous humor, and the corneal endothelia. A Spiroplasma sp. grew out of the scrapie‐affected eyes both in the M1D broth and in the BCE cultures but did not expand. These new spiroplasma isolates differed immunologically from SMCA. Conclusion These data showed a clear association of spiroplasma with scrapie suggesting that these bacteria have a role in the pathogenesis of TSE and that the eye should be a research focus for future studies of TSE.     

Application of thermal sums concept to estimate the time to harvest new banana hybrids for export

Application of the thermal sum concept was developed to determine the optimal harvesting stage of new banana hybrids to be grown for export. It was tested on two triploid hybrid bananas, FlhorBan 916 (F916) and FlhorBan 918 (F918), created by CIRAD's banana breeding programme, using two different approaches. The first approach was used with F916 and involved calculating the base temperature of bunches sampled at two sites at the ripening stage, and then determining the thermal sum at which the stage of maturity would be identical to that of the control Cavendish export banana. The second approach was used to assess the harvest stage of F918 and involved calculating the two thermal parameters directly, but using more plants and a longer period. Using the linear regression model, the estimated thermal parameters were a thermal sum of 680 degree-days (dd) at a base temperature of 17.0 °C for cv. F916, and 970 dd at 13.9 °C for cv. F918. This easy-to-use method provides quick and reliable calculations of the two thermal parameters required at a specific harvesting stage for a given banana variety in tropical climate conditions. Determining these two values is an essential step for gaining insight into the agronomic features of a new variety and its potential for export.     

Late season harvest and storage of Rubus berries—Major antioxidant and sugar levels

Late season production of raspberries and blackberries has become more common due to increased consumer demand and new production techniques, but information is scarce regarding the effect on nutritional quality, especially during storage. Antioxidant levels in berries can be influenced by pre-harvest climate conditions (light intensity, day length and temperature) and post-harvest storage time. This study analysed the effect of these factors on the content of different antioxidants (vitamin C, ellagic acid, anthocyanins and total phenolics) in raspberries (Rubus idaeus L. cv. Polka) and blackberries (Rubus fructicosus L. cv. Loch Ness) during storage. The analyses were performed using high performance liquid chromatography (HPLC) with photodiode array detection. Ellagic acid content during storage ranged between 104 and 114 mg/100 g fresh weight in raspberries and 172 and 182 mg/100 g fresh weight in blackberries. Anthocyanin content increased during storage from 64 to 77 mg/100 g fresh weight in raspberries, but decreased from 199 to 162 mg/100 g fresh weight in blackberries. Vitamin C content in raspberries ranged during storage between 22 and 28 mg/100 g fresh weight, but in blackberries values were lower (10–12 mg/100 g fresh weight). Significant changes in total phenolics and sugars occurred during different storage periods (0–9 or 10 days). The results suggest that late season raspberries and blackberries maintain high quality in terms of antioxidant content.