Quantitation of adenine nucleotides in equine colonic mucosal tissue using high performance liquid chromatography.

The objectives were to use high performance liquid chromatography (HPLC) to validate an established method for adenine nucleotide separation in equine colonic mucosal tissue, to determine the inherent variability in the tissue and extraction method, and to determine the stability of ATP, ADP, and AMP in the tissue with time. Equine colonic mucosal tissue obtained from a single horse was immediately submersed in liquid nitrogen, and stored at -70 degrees C. Samples were lyophilized, extracted, and separated by HPLC. The limit of quantitation was 0.05 microg/mL. The coefficient of variation for the instrument was less than 10% for all nucleotides measured. When the tissue was not homogenized prior to sampling, there were significant differences in adenine nucleotide content between samples. However, when the tissue was homogenized prior to analysis, these differences were no longer significant. There was no significant decrease in ATP, ADP, or AMP content over a 54-day analysis period.     

Research note: Postnatal development of electrolyte transport in calf rumen as affected by weaning time.

In a previous study we found a positive correlation between early weaning in calves and morphological parameters which were indicative of ruminal development, i.e. the length and width of the papillae. The objective of the present study was to determine to what extent this observation could be reflected by modulations of absorptive and secretory functions of the rumen mucosa. For this purpose the short-circuit currents (Isc) as a measure of electrogenic net ion fluxes and the transepithelial conductances (G(T)) as a measure of the overall tissue permeability were measured in vitro applying the Ussing-chamber technique. Simultaneously, the unidirectional flux rates of sodium and chloride across rumen wall epithelia were determined in the absence of electrochemical gradients. Under these conditions, significant positive net flux rates (Jnet) clearly indicate active mechanisms for electrolyte absorption. For the experiments 12 male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I, slaughtered after 6 weeks of age), late weaning group (II, slaughtered after 9 weeks of age) and early weaning group (II, weaned after 6 weeks of age and slaughtered after 9 weeks of age). Whereas G(T) values remained unaffected by different age and feeding, Isc values were significantly affected by early weaning but were not influenced by age. Irrespective of weaning time active absorption of Na+ tended to be higher by about 60% in 9 weeks old animals. Active absorption of chloride was significantly increased in milk fed 9 weeke old calves and this effect was further stimulated by early weaning. In conclusion, the data show an increasing active Na+ absorption with age in calf rumen that could not be influenced by early weaning. Similarly, active Cl- absorption was initially increased during postnatal development and this effect could be stimulated further by early weaning.     

The Effect of Diet on the Biochemical Composition of Juvenile Artemia: Potential Formulations for Rock Lobster Aquaculture

The lipid class and fatty acid (FA) composition of juvenile Artemia fed continuously on four diets—the microalga Tetraselmis suecica , a mix of oat bran-wheat germ-lecithin (OWL), OwL-eicosapentaenoic acid (EPA), and OWL-EPA-arachidonic acid (AA)—were examined over a 9-d experiment in an attempt to approximate the FA profile of phyllosoma larvae of wild southern rock lobster Jasus edwardrii . The main difference in lipid class composition of Artemia fed the four diets was the relative level of polar lipid (PL) and triacylglycerol (TAG). By day 9, the algal-fed Artemia were highest in PL (95% of total lipid) and lowest in TAG (2%), whereas the remaining diets resulted in Artemia with 16–30% PL and 41–82% TAG. After 2 d, the relative FA composition of all Artemia treatments closely reflected those of the diets, with no marked change after further feeding (to day 9). In terms of the content of essential polyunsaturated fatty acids (PUFA), by day 5 Artemia fed: 1) with the algal diet contained 7 mg/g FA dry mass (0.3% DHA, 6.3% EPA, 3.4% AA of total FA); 2) with the OWL diet contained 3 mg/g (0.3% DHA, 0.9% EPA, 0.7% AA); 3) with the OWL-EPA diet contained 55 mg/g (6.2% DHA, 11.6% EPA, 1.1% AA); and 4) with the OWL-EPA-AA contained 83 mg/g (3.8% DHA, 7.5% EPA, 17.4% AA). The PUFA profiles of Artemia using the OWL-oil diets were similar to wild rock lobster phyllmmata, although levels of doco-sahexaenoic acid (DHA) were lower (10% DHA) than in J. edwardsii larvae. On the basis of PUFA composition data alone, the results suggest the suitability of the OWL-oil mixed diets for consideration for feeding to Artemia used in the culture of southern rock lobster larvae, particularly if the level of DHA can be further enhanced.     

Radiographic and ultrasonographic features of uroliths and other urinary tract filling defects

Radiopaque uroliths and nonradiopaque (water density) uroliths are filling defects encountered in the urinary tracts of dogs and cats. Other free luminal and attached soft tissue density filling defects encountered during uroradiographic special procedures include blood clots, air bubbles, hematomas, granulomas, abscesses, inflammatory and neoplastic polyps. Nonradiopaque uroliths cannot be identified on survey radiographs from other soft tissue dense structures. Gray scale ultrasonography can be used to differentiate nonradiopaque (water dense) uroliths from other soft tissue attached or free luminal filling defects of the excretory pathway. The differential radiographic features of filling defects encountered during cystography and urethrography are described and illustrated.     

Immunocompetence of sheep experimentally infected with bovine leukemia virus

The humoral and cellular immunological reactivity of sheep were studied throughout the first 32 weeks following experimental infection with bovine leukemia virus (BLV). Seroconversion of BLV-inoculated sheep occurred within 4 weeks, but infection was not transmitted to contact control sheep. Despite the persistence of the viral infection, no differences were demonstrated in leukograms, serum IgG concentrations, humoral response to immunization with an irrelevant antigen (rabbit red blood cells), phytomitogen (Concanavalin A and Pokeweek mitogen)-induced lymphocyte blastogenesis, or chemical (1-chloro, 2-4 dinitrobenzene) skin contact hypersensitivity, between BLV-infected and uninfected contact control sheep. These results demonstrate the absence of a nonspecific immunosuppressive effect of BLV and further negate the influence of a generalized immunological deficit on the development of clinical disease in BLV-infected animals.     

Molecular structure of troponin C from chicken skeletal muscle at 3-angstrom resolution

The x-ray structure of chicken skeletal muscle troponin C (TnC), the Ca2+-binding subunit of the troponin complex, shows that the protein is about 70 angstroms long with an unusual dumbbell shape. The carboxyl and amino domains are separated by a single long alpha helix of about nine turns. Only the two high-affinity Ca2+-Mg2+ sites of the COOH-domain are occupied by metal ions resulting in conformational differences between the COOH- and NH2-domains. These differences are probably important in the triggering of muscle contraction by TnC. Also the structure of TnC is relevant in understanding the function of other calcium-regulated proteins, in particular that of calmodulin because of its strong similarity in amino acid sequence.     

MAGNETIC RESONANCE IMAGING FEATURES OF EQUINE NIGROPALLIDAL ENCEPHALOMALACIA

Magnetic resonance imaging (MR) was used to make a diagnosis of equine nigropallidal encephalomalacia in a horse. Equine nigropallidal encephalomalacia is a neurodegenerative disease that has many characteristics with Parkinson-like diseases in humans. Historically, horses were euthanized based on clinical signs and exposure to the toxic weed, yellow star thistle (Centaurea solstitialis). Previously, the disease has only been confirmed on necropsy. MR imaging can provide accurate and sensitive visualization of typical lesions seen in the brain of horses affected with equine nigropallidal encephalomalacia. Lesions were seen on T1-weighted, T2-weighted and proton density images. There was no contrast enhancement following Gd-DTPA administration. Lesions seen on MR were confirmed at necropsy. Using MR to confirm a diagnosis of equine nigropallidal encephalomalacia will prevent unnecessary suffering of horses and expense to owners that would otherwise incur, while further diagnostics are performed.