Direct isolation of satellite cells for skeletal muscle regeneration

Muscle satellite cells contribute to muscle regeneration. We have used a Pax3(GFP/+) mouse line to directly isolate (Pax3)(green fluorescent protein)-expressing muscle satellite cells, by flow cytometry from adult skeletal muscles, as a homogeneous population of small, nongranular, Pax7+, CD34+, CD45-, Sca1- cells. The flow cytometry parameters thus established enabled us to isolate satellite cells from wild-type muscles. Such cells, grafted into muscles of mdx nu/nu mice, contributed both to fiber repair and to the muscle satellite cell compartment. Expansion of these cells in culture before engraftment reduced their regenerative capacity.     

Chemical composition of Acid fog

Fog water collected at three sites in Los Angeles and Bakersfield, California, was found to have higher acidity and higher concentrations of sulfate, nitrate, and ammonium than previously observed in atmospheric water droplets. The pH of the fog water was in the range of 2.2 to 4.0. The dominant processes controlling the fog water chemistry appear to be the condensation and evaporation of water vapor on preexisting aerosol and the scavenging of gas-phase nitric acid.     

Extensification of grassland use in the Welsh uplands: sheep performance in years 1–6

An experiment was established in 1991 on a 25-year-old perennial ryegrass/bent ( Lolium perenne L. /Agrostis capillaris L.)-dominated pasture in Wales to study the effects of reducing nutrient inputs to previously fertilized upland pasture. The effects of the removal of applications of (1) N (denoted by CaPK) (2) N, P and K (Ca) and (3) N, P, K, and Ca (Nil) were compared with a treatment which received applications of all four nutrients (CaPKN) over a 6-year period (1991–96) in a randomized block design replicated three times. The experiment was managed under a continuous variable stocking regime (ewes and lambs until weaning and ewes thereafter) maintaining a sward surface height of 4·0 cm throughout the grazing season. Although individual liveweight gain of the lambs was unaffected by the treatments, there was a significant reduction ( P  < 0·05) in total lamb liveweight gain, ewe stocking rate and length of grazing season as a result of the withdrawal of nutrients. Over the 6 years total lamb liveweight gain was reduced by 17%, 32% and 45% and ewe stocking rate by 21%, 36% and 49% on treatments CaPK, Ca and Nil, respectively, compared with treatment CaPKN. The effect of withdrawing nutrient inputs on ewe stocking rate was progressive and by 1996 the Nil input treatment displayed a 63% reduction compared with the CaPKN treatment and this was also coupled with a 21-day reduction in length of the grazing season. During the post-weaning period, ewes from the Nil input treatment recorded a liveweight loss in 1995 and only a modest liveweight gain in 1996. This coupled with significantly lower body condition scores ( P  < 0·01) of these ewes in the autumn indicated that the Nil input treatment could lead to reductions in reproductive performance.     

Caspase activation in equine influenza virus induced apoptotic cell death.

Equine influenza virus (EIV) is the leading cause of acute respiratory infection in horses worldwide. In recent years, the precise mechanism by which influenza infection kills host cells is being re-evaluated. In this report, we examined whether caspases, a group of intracellular proteases, are activated following EIV infection and contribute to EIV-mediated cell death. Western blotting analysis indicated that a nuclear target of caspase-3, poly(ADP-ribose) polymerase (PARP) was proteolytically cleaved in EIV-infected MDCK cells, but not in mock-infected cells. In comparison with caspase-3 specific inhibitor Ac-DEVD-CHO, a general caspase inhibitor Boc-D-FMK provided much stronger inhibition of EIV-induced cytopathic effect and apoptosis. Our results suggest that EIV may activate more than one caspase. Caspase activation and cleavage of its cellular targets may play a critical role in EIV-mediated cytotoxicity.     

Proteolysis of insulin-like growth factor binding proteins during preovulatory follicular development in cattle

The present study was conducted to evaluate changes in follicular fluid (FF) insulin-like growth factor binding protein (IGFBP) proteolytic activity and levels of steroids and IGFBP during follicular development in cattle. Estrous cycles of cows were synchronized with two injections of prostaglandin F2alpha (PGF) 11 d apart and follicular growth monitored via daily rectal ultrasonography in order to identify the dominant follicle. All cows were ovariectomized 48 hr after the second injection of PGF. Follicular fluid was collected individually for all follicles > 5 mm and pooled for small (1 to 5 mm) follicles. Follicular fluid estradiol and androstenedione levels were greater (P < 0.05) and progesterone and IGFBP-3 levels not different (P > 0.10) in large dominant than in small (1 to 5 mm) or large (>5 mm) subordinate follicles, whereas IGFBP-2, -4 and -5 levels were less (P < 0.05) in large dominant than in small or large subordinate follicles. To evaluate proteolysis of IGFBPs, FF was incubated with recombinant human (125) I-labeled IGFBP-2, -3, -4, and -5 and proteins separated by 12% SDS-PAGE. Follicular fluid caused little or no proteolysis of (125)I-lableled IGFBP-2 or -3. However, cleavage of (125)I-labeled IGFBP-4 and -5 by FF from large dominant follicles was greater (P < 0.05) than by FF from small or large subordinate follicles indicating that a protease to IGFBP-4 and -5 exists in estrogen dominant follicles. We conclude that lower levels of IGFBP-2 in estrogen dominant follicles of cattle are not due to increased proteolysis, whereas decreases in IGFBP-4 and -5 levels are likely due, in part, to increased protease activity. Changes in IGFBP may alter levels of bioavailable IGFs that stimulate steroidogenesis and mitogenesis in developing bovine follicles.     

Stocktake study of current fertilisation recommendations across Europe and discussion towards a more harmonised approach

The European Commission has set targets for a reduction in nutrient losses by at least 50% and a reduction in fertiliser use by at least 20% by 2030 while ensuring no deterioration in soil fertility. Within the mandate of the European Joint Programme EJP Soil ‘Towards climate-smart sustainable management of agricultural soils’, the objective of this study was to assess current fertilisation practices across Europe and discuss the potential for harmonisation of fertilisation methodologies as a strategy to reduce nutrient loss and overall fertiliser use. A stocktake study of current methods of delivering fertilisation advice took place across 23 European countries. The stocktake was in the form of a questionnaire, comprising 46 questions. Information was gathered on a large range of factors, including soil analysis methods, along with soil, crop and climatic factors taken into consideration within fertilisation calculations. The questionnaire was completed by experts, who are involved in compiling fertilisation recommendations within their country. Substantial differences exist in the content, format and delivery of fertilisation guidelines across Europe. The barriers, constraints and potential benefits of a harmonised approach to fertilisation across Europe are discussed. The general consensus from all participating countries was that harmonisation of fertilisation guidelines should be increased, but it was unclear in what format this could be achieved. Shared learning in the delivery and format of fertilisation guidelines and mechanisms to adhere to environmental legislation were viewed as being beneficial. However, it would be very difficult, if not impossible, to harmonise all soil test data and fertilisation methodologies at EU level due to diverse soil types and agro-ecosystem influences. Nevertheless, increased future collaboration, especially between neighbouring countries within the same environmental zone, was seen as potentially very beneficial. This study is unique in providing current detail on fertilisation practices across European countries in a side-by-side comparison. The gathered data can provide a baseline for the development of scientifically based EU policy targets for nutrient loss and soil fertility evaluation.     

Comparison of the Biobullet versus traditional pharmaceutical injection techniques on injection-site tissue damage and tenderness in beef subprimals

The incidence and severity of injection-site lesions has decreased since the development of the Beef Quality Assurance program. The objective was to evaluate the route of administration and the pharmaceutical product on the impact on tenderness, collagen concentration, and lesion occurrence in muscles of chucks and rounds. A total of 144 yearling steers (initial BW = 383 +/- 29.4 kg) were selected and transported to Oklahoma State University. Steers were blocked into 2 groups of 72 based on initial BW and were randomly allocated, within block, into pens of 6 head per pen (12 pens per block). Each pen was randomly assigned an injection protocol. On May 19, 2006 (d 0), steers were administered one of the following treatment injections: a standard Biobullet containing 100 mg of ceftiofur sodium (Naxel, Pfizer Inc., New York, NY); a traditional needle and syringe dose of ceftiofur sodium; a standard Biobullet containing BallistiVac infectious bovine rhinotracheitis (IBR; Titanium 5, SolidTech Animal Health, Newcastle, OK); a traditional needle and syringe dose of IBR; a traditional needle and syringe dose of Vira Shield 5 (Grand Laboratories Inc., Freeman, SD); a standard placebo Biobullet; or a traditional needle and syringe dose of sterile water. Percentage of samples with an identifiable lesion did not differ by drug administered or injection method. Warner-Bratzler shear force values of lesion center cores in chucks tended to be different (P = 0.07) from cores from the control steaks and at 2.54 and 5.08 cm away from the lesion center. Lesion centers from the Biobullet-BallistiVac IBR had a Warner-Bratzler shear force value of 7.01 kg, which was greater (P < 0.05) than values for lesion centers from chucks injected with a Biobullet-placebo (6.27 kg) or needle-ceftiofur sodium (5.08 kg). No significant differences (P > 0.10) were observed in the total collagenous connective tissue in samples extracted from the chuck or round. The comparison of lesion site and control (nonlesion site) samples for lipid concentration showed no significant difference (P > 0.10) among treatments in the round. It was concluded the Biobullet did not create a greater occurrence of lesions in the muscles of the chuck and round. The Biobullet is not an appropriate injection method for the round of beef cattle because it caused tissue damage similar to a needle injection. However, the Biobullet can be used effectively in neck (chuck) applications without additional negative effects on tenderness.     

Rapid Evaporative Ionization Mass Spectrometry (REIMS): a Potential and Rapid Tool for the Identification of Insecticide Resistance in Mosquito Larvae

Insecticide resistance is a significant challenge facing the successful control of mosquito vectors globally. Bioassays are currently the only method for phenotyping resistance. They require large numbers of mosquitoes for testing, the availability of a susceptible comparator strain, and often insectary facilities. This study aimed to trial the novel use of rapid evaporative ionization mass spectrometry (REIMS) for the identification of insecticide resistance in mosquitoes. No sample preparation is required for REIMS and analysis can be rapidly conducted within hours. Temephos resistant Aedes aegypti (Linnaeus) larvae from Cúcuta, Colombia and temephos susceptible larvae from two origins (Bello, Colombia, and the lab reference strain New Orleans) were analyzed using REIMS. We tested the ability of REIMS to differentiate three relevant variants: population source, lab versus field origin, and response to insecticide. The classification of these data was undertaken using linear discriminant analysis (LDA) and random forest. Classification models built using REIMS data were able to differentiate between Ae. aegypti larvae from different populations with 82% (±0.01) accuracy, between mosquitoes of field and lab origin with 89% (±0.01) accuracy and between susceptible and resistant larvae with 85% (±0.01) accuracy. LDA classifiers had higher efficiency than random forest with this data set. The high accuracy observed here identifies REIMS as a potential new tool for rapid identification of resistance in mosquitoes. We argue that REIMS and similar modern phenotyping alternatives should complement existing insecticide resistance management tools.     

Dietary soluble non-starch polysaccharide level and xylanase supplementation influence performance,egg quality and nutrient utilization in laying hens fed wheat-based diets

The aim of this study was to evaluate the effects of dietary soluble non-starch polysaccharide (sNSP) content and xylanase supplementation on production performance, egg quality parameters, and nutrient digestibility in Hy-line Brown layers from 25 to 32 wk of age. A total of 144 Hy-line Brown laying hens (25 wk of age) were randomly allocated to 1 of 4 wheat-based dietary treatments in a 2 × 2 factorial experimental design, with 36 replicates of individual hens per treatment. The diets were formulated to contain either a high or low sNSP level (at 13.3 or 10.8 g/kg) and were supplemented with either 0 or 12,000 BXU/kg exogenous xylanase. Birds were fed these treatment diets for an 8-wk period, and hen production performance, including daily egg production, average egg weight, daily egg mass, feed conversion ratio and proportion of dirty and abnormal eggs were measured at bird age 25 to 28 wk and 29 to 32 wk. An interaction between sNSP content of the diet and xylanase supplementation was observed on daily egg production from 25 to 28 wk of age (P = 0.018); birds fed the high sNSP diet without xylanase had lower egg production than those fed any other treatment. An interaction between the 2 dietary factors was also observed on hen weight gain at 29 to 32 wk of age (P = 0.014), with birds fed the low sNSP diet with 12,000 BXU/kg xylanase presenting greater weight gain compared to those fed the high sNSP diet with 12,000 BXU/kg xylanase. Feed intake at 29 to 32 wk of age was reduced by xylanase supplementation (P = 0.047). Xylanase supplementation also increased yolk colour score at both 28 and 32 wk of age, and decreased yolk weight at 32 wk of age (P = 0.014, 0.037 and 0.013, respectively). Birds fed the low sNSP diet presented lower protein digestibility (P = 0.024) than those fed the high sNSP diet. Additionally, birds fed high sNSP presented higher shell reflectivity at both 28 and 32 wk of age (P = 0.05 and 0.036, respectively). The influence of duration of feeding the treatment diets on egg quality was also determined. It was observed that egg weight, yolk weight and yolk colour score consistently increased over time, regardless of experimental treatment effects. In contrast, Haugh Unit and albumen height significantly decreased throughout the study period in all treatments, although this was less pronounced in hens fed the treatment with high sNSP and no supplemental xylanase. A reduction in shell breaking strength over time was observed only in hens fed the treatments without xylanase addition, and shell thickness was improved over time only in birds fed the low sNSP diet with xylanase. The impacts of the dietary treatments were largely inconsistent in this study, so a solid conclusion cannot be drawn. However, these findings do indicate that dietary NSP level influences layer production performance, and thus should be considered when formulating laying hen diets. It also proved that further research is warranted into how to optimize the benefits of xylanase application in laying hens.