Regulation of bovine pyruvate carboxylase mRNA and promoter expression by thermal stress

Pyruvate carboxylase (PC) catalyzes the rate-limiting step in gluconeogenesis from lactate and is a determinant of tricarboxylic acid cycle carbon flux. Bovine PC 5' untranslated region (UTR) mRNA variants are the products of a single PC gene containing 3 promoter regions (P3, P2, and P1, 5′ to 3′) that are responsive to physiological and nutritional stressors. The objective of this study was to determine the direct effects of thermal stress on PC mRNA and gene expression in bovine hepatocyte monolayer cultures, rat hepatoma (H4IIE) cells, and Madin-Darby bovine kidney epithelial (MDBK) cells. Hepatocytes were isolated from 3 Holstein bull calves and used to prepare monolayer cultures. Rat hepatoma cells and MDBK cells were obtained from American Type Culture Collection, Manassas, VA. Beginning 24 h after initial seeding, cells were subjected to either 37°C (control) or 42°C (thermal stress) for 24 h. Treatments were applied in triplicate in a minimum of 3 independent cell preparations. For bovine primary hepatocytes, endogenous expression of bovine PC mRNA increased (P < 0.1) with 24 h of thermal stress (1.31 vs. 2.79 ± 0.49, arbitrary units, control vs. thermal stress, respectively), but there was no change (P ≥ 0.1) in cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) mRNA expression. Similarly, exposure of MDBK cells to thermal stress increased (P < 0.1) expression of bovine PC mRNA without altering (P ≥ 0.1) PEPCK-C mRNA expression. Conversely, there was no effect (P ≥ 0.1) of thermal stress on endogenous rat PC (0.47 vs. 0.30 ± 0.08, control vs. thermal stress) or PEPCK-C (1.61 vs. 1.20 ± 0.48, arbitrary units, control vs. thermal stress, respectively) mRNA expressions in H4IIE cells. To further investigate the regulation of PC, H4IIE cells were transiently transfected with bovine promoter-luciferase constructs containing either P1, P2, or P3, and exposed to thermal stress for 23 h. Activity of P1 was suppressed (P < 0.1) 5-fold, activity of P2 was unchanged (P ≥ 0.1), and activity of P3 was increased (P < 0.1) by 5.4-fold. These data indicate that response of bovine PC gene to thermal stress is through promoter regulation and suggest that there are unique characteristics of bovine PC promoters that may contribute to the physiological response to thermal stress.     

European consensus document on mast cell tumours in dogs and cats

In preparing this document the Authors aimed to pool current information on canine and feline mast cell disease. The information was gathered from international studies and a emphasis was placed on material and opinion with a strong evidence base. We intend it to form the basis of our understanding in this disease at the current time and we anticipate that it will be particularly useful for the general practitioner. It should be emphasized that the authors are presenting this work from a European perspective.     

Effects of forage level in feedlot finishing diets on carcass characteristics and palatability of Jersey beef

Jersey cattle are known for producing carcasses with a greater amount of marbling, but they require more days on feed to achieve acceptable market weights compared with other breeds. The objective of this study was to evaluate the effect of dietary forage (12 vs. 24% sudangrass:alfalfa hay, DM basis) in steam-flaked, corn-based finishing diets on carcass characteristics, beef palatability, and retail color stability of steaks from Jersey beef compared with conventionally fed commodity beef strip loins (COM) of identified quality (Choice(-) and Select(+)). Jersey steers (n = 77) were blocked by BW and randomly assigned to 1 of the following treatments for a 383-d trial period: Jersey low 12% (JL; n = 38) or Jersey high 24% (JH; n = 39) forage (DM basis). A comparison group was selected from conventionally fed cattle on the same day of slaughter as the Jersey treatments, and strip loins from USDA Select(+) (COM; n = 20) and Choice(-) (COM; n = 20) were removed for data analysis. Seventy-two hours postmortem, strip loins were removed, vacuum-packaged, and aged at 3°C for 18 d postmortem. After the aging period, steaks from the LM were sliced, vacuum-packaged, and frozen (-20°C) until analyzed. Jersey steaks had reduced (P < 0.05) Warner-Bratzler shear force values compared with COM steaks. Trained sensory panelists rated JL greater (P < 0.05) for initial and sustained tenderness and initial juiciness than COM, whereas JH was intermediate. As expected, marbling was greater (P < 0.05) for both JL and JH compared with COM, and trained sensory panel sustained juiciness, beef flavor intensity, and overall acceptability scores were greater (P < 0.05) for both JL and JH compared with COM; however, no differences (P = 0.14) were reported for consumer tenderness and flavor. Objective color (L*, a*, b*) measurements decreased (P < 0.05) over time across treatments. There were no differences among treatments for lightness (L*); however, overall during retail display JL were less (P < 0.05) red (a*) and yellow (b*) than JH and COM. Subjective color scores indicated both JL and JH were less red (P < 0.05) than COM. Steaks from Jersey were equal to and on some measurements more desirable than steaks from COM carcasses for both color stability and palatability. These results suggest that dietary forage level had minimal effects on carcass characteristics and beef palatability. However, feeding a low-forage diet decreases input cost and potentially results in a greater valued carcass. Finishing long-fed (383 d) Jersey steers can meet beef industry expectations with respect to quality grade.     

Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

Prednisolone is associated with an increase in serum insulin but not serum fructosamine concentrations in dogs with atopic dermatitis

The aim of this study was to assess the effects of a standard therapeutic protocol of prednisolone (Pred) on glucose homeostasis in atopic dogs and compare it with previously published data for ciclosporin A (CsA). The central aim of the study was to assess and compare the effects of standard therapeutic protocols of prednisolone (Pred) and ciclosporin A (CsA) on glucose homeostasis in dogs with atopic dermatitis (CAD). Both treatments significantly reduced the physical signs of CAD, as determined by the canine atopic dermatitis extent and severity index version 3 (CADESI-03) and the Edinburgh Pruritus Scale (EPS). Post-treatment plasma glucose concentrations were not significantly different in the two groups, but serum insulin concentrations were significantly higher following Pred therapy (P<0.05). Serum fructosamine concentrations were not significantly different pre- and post-treatment with Pred, although previous studies had shown that CsA treatment increased fructosamine concentrations (P<0.005). The two treatment groups were recruited in a similar timeframe, were numerically matched and there were no differences in CADESI-03 and EPS scores between the CsA and Pred groups either before or after treatment. Thus, both CsA and Pred treatment were associated with mild disturbances in glucose metabolism, but only CsA therapy resulted in a significant increase in fructosamine concentrations. This information may be relevant to clinicians when considering therapeutic options for dogs with atopic dermatitis which already have impaired glucose homeostasis.