Cloning and functional characterization of recombinant equine P-selectin

The recent molecular characterization and sequencing of equine P-selectin (ePsel), and its glycoprotein ligand, P-selectin glycoprotein ligand-1 (PSGL-1), have provided the tools for further investigation into their role in leukocyte trafficking. Here, we report the generation of a genetically engineered chimeric protein (ePsel-IgG) in which the equine P-selectin lectin and epithelial growth factor (EGF) domains were covalently linked to the equine IgG1 heavy chain constant region. The soluble ePsel-IgG was observed to bind to equine monocytes by confocal microscopy and flow cytometry. Furthermore, equine monocytes bound to immobilized ePsel-IgG in a time course and dose dependent manner. Not only did ePsel-IgG act as an adhesion molecule, it was also found to activate ERK1/2 kinase and induce IL-8 mRNA expression in equine monocytes. That all of the aforementioned ePsel-IgG-induced cell binding and cell signaling were abolished by the addition of EDTA, suggested that ePsel-IgG chimera mediated events occurred via the P-selectin ligand, PSGL-1. We were able to demonstrate that 78% of equine monocytes cross-reacted with anti-human HECA-452 antibody, which recognizes the sialy-Lewis X (sLex) epitope, a well-known carbohydrate binding site on human PSGL-1. Pre-incubation of equine PBMC with neuraminidase or O-sialoglycoprotein endopeptidase (OSGP) reduced ePsel-IgG monocyte binding to 36% or 60%, respectively. Taken together, these data suggest that there might be two ligand recognition sites on P-selectin, one of which recognizes sLex and another which recognizes P-selectin ligand core protein. The ePsel-IgG chimera can be a useful as a reagent for further studies on the role of equine P-selectin and signal transduction in inflammatory events in horse.     

Silage yield and quality traits in elite maize hybrids and their relationship to elemental concentrations in juvenile plants

European silage maize is cultivated for animal feed and biogas production. We evaluated 210 factorial crosses of elite dent and flint lines in multilocation trials for agronomic and quality traits together with biomass and shoot concentrations of 10 elements in juvenile plants. Significant genotypic variances, mainly due to general combining ability variance of the dent lines, and high heritabilities were observed for dry matter yield (DMY) and quality traits. DMY was not correlated with quality traits, but methane fermentation yield (MFY) and metabolizable energy content (MEC) showed significant correlations with starch and fibre content. Concentrations of elements N,P,S,K were positively correlated with each other but only in few cases correlated with DMY and quality traits. Parent lines with contrasting P concentrations differed in root morphology traits. Results support DMY as primary trait for selection of silage maize hybrids, but MFY is of negligible importance in breeding for biogas contrary to MEC for animal feed. Neither biomass nor elemental composition of juvenile plants were of predictive value for final DMY or quality traits.     

Early timing and new combinations to increase the efficacy of neonicotinoid-entomopathogenic nematode (Rhabditida: Heterorhabditidae) combinations against white grubs (Coleoptera: Scarabaeidae)

BACKGROUND: An investigation was carried out to determine whether new neonicotinoid-nematode combinations and earlier applications against younger larval stages could increase the efficacy of synergistic neonicotinoid-entomopathogenic nematode combinations against white grubs. RESULTS: In the laboratory, combinations of the neonicotinoids imidacloprid and clothianidin and the nematodes Heterorhabditis bacteriophora Poinar and H. zealandica Poinar against third instars were compared. In Anomala orientalis (Waterhouse) and Popillia japonica Newman, H. bacteriophora-imidacloprid combinations showed the most consistent synergism but did not cause significantly higher mortality than H. zealandica-imidacloprid combinations. In Cyclocephala borealis Arrow, there was no clear trend as to which combinations caused the most consistent synergism, but H. zealandica-imidacloprid combinations tended to cause the highest mortalities. In the laboratory, imidacloprid-H. bacteriophora combinations provided more consistent synergism against third-instar than against second-instar A. orientalis, but mortality was higher in second instars. In field experiments, imidacloprid-H. bacteriophora combinations against A. orientalis and P. japonica provided more consistent synergism when applied in mid-September but more consistent control when applied in late August. CONCLUSIONS: Imidacloprid is a better synergist for entomopathogenic nematodes than clothianidin. Imidacloprid-nematode combinations are more effective against second instars than against third instars, allowing rate reductions of both agents to make this approach more competitive with full-rate applications of neonicotinoid alone.     

Alterations in the jejunum of young goats caused by feeding soy protein‐based diets

This study was designed to investigate whether soy protein or soy protein supplemented with indispensable amino acids (AA) change the protein expression pattern and utilization of pre‐cursors for RNA biosynthesis in jejunal mucosa in relation to casein and whether these changes affect mucosal cell growth. Kids were fed comparable diets based on cow`s milk, of which 50% of crude protein were replaced by either casein (CAS), soy protein (SP) or soy protein supplemented with indispensible AA (SPA) for 34 days (n = 4/group). Jejunal tissue was collected 5 h after adding a single dose of ¹⁵N‐RNA to the diet, in order to determine morphology, protein repertoire by two‐dimensional gel electrophoresis and matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry, and RNA biosynthesis by isotope ratio‐mass spectrometry. In mid‐jejunum, morphological alterations induced by partial replacement of casein with soy protein were accompanied by changes in mucosal proteins related to generation of the cytoskeleton and in pathways for mucosal RNA biosynthesis, resulting in a smaller re‐utilization of dietary RNA pre‐cursors and in an increased activity of enzymes involved in nucleic acid breakdown. Soy protein supplemented with indispensible aminoacids tended to revise mucosal growth retardation with no impact on salvage of dietary RNA pre‐cursors for mucosal RNA biosynthesis, but changes in cytoskeleton generation. Feeding soy protein with supplementation of indispensible AA does not ameliorate soy protein effects on mucosal morphology and RNA metabolism in the jejunum in a significant manner.     

Aggregate associated-C and physical protection in a tropical clayey soil under Malagasy conventional and no-tillage systems

Share-ploughed tillage with residue removed (CT-R) is the traditional tillage practice in the Highlands of Madagascar. No-tillage with residue mulching (NT+R) is nowadays often used as an alternative cultivation practice. Soils (0–5 cm layer) were sampled in Spring 2003 from both management systems after 11 years of soybean–maize annual rotation on a clayey Ferralsol. Soil aggregate stability can influence soil organic carbon (SOC) storage by its protection from microbial decomposition. The soil organic carbon (SOC) content was significantly impacted by systems and crop residues derived-carbon represented 64% of the annual benefit in SOC of NT+R system. The carbon associated with soil water stable macro- (200–2000 μm), meso- (20–200 μm) and microaggregates (<20 μm) from both systems, and their physical protection was studied by an incubation experiment of intact vs. crushed aggregates. Results showed macroaggregate content was significantly higher in NT+R than in CT-R system and mesoaggregate content was significantly higher in CT-R than in NT+R. Macroaggregates associated-C were 1.8 time higher in NT+R than in CT-R (31.9 and 17.9 g C g⁻¹ soil, respectively) and made up the largest percentage (>80%) of the difference of SOC content between NT+R and CT-R systems. The amount of mineralized C over 28 days was higher in NT+R than in CT-R, and higher in meso- than in macroaggregates. However, crushing aggregates did not significantly affect the amount of mineralized C in macro- and mesoaggregates for both management systems. The macro- and mesoaggregates protected-C was lower than 54 μg g⁻¹ soil for both NT+R and CT-R systems. Hence, the physical protection of C in aggregate larger than 50 μm was not the main process of C protection in the studied systems. Thus, C protection might occur in aggregates larger or smaller than 50 μm via physico-chemical protection mechanisms by association of organic matter to clay and silt fractions, or by protection due to chemical composition.     

Population genetic simulation and data analysis with Plabsoft

Computer simulations are a useful tool to solve problems in population genetics for which no analytical solutions are available. We developed Plabsoft, a powerful and flexible software for population genetic simulation and data analysis. Various mating systems can be simulated, comprising planned crosses, random mating, partial selfing, selfing, single-seed descent, double haploids, topcrosses, and factorials. Selection can be simulated according to selection indices based on phenotypic values and/or molecular marker scores. Data analysis routines are provided to analyze simulated and experimental datasets for allele and genotype frequencies, genotypic and phenotypic values and variances, molecular genetic diversity, linkage disequilibrium, and parameters to optimize marker-assisted backcrossing programs. Plabsoft has already been employed in numerous studies, we chose some of them to illustrate the functionality of the software.     

HPLC Determination of Stability and Distribution of Added Folic Acid and Some Endogenous Folates During Breadmaking

Bread flour was spiked with folic acid (1.40 mg/lb or 3.08 μg/g of flour) and processed into bread by the sponge and dough method. Changes that occurred to added folic acid and endogenous folate contents through different processing stages, including sponge formation, proofing, and baking, were assessed by reversed‐phase ion‐pair HPLC combined with UV and fluorometric detection. Sample extraction required α‐amylase and rat plasma deconjugase digestion, and sample preparation required purification by solid‐phase extraction. Added folic acid was measured by monitoring UV absorption at 280 nm. Four selected forms of endogenous folates including tetrahydrofolate (THF), 5‐formyl‐THF, 10‐formylfolate, and 5‐methyl‐THF were identified and quantified throughout the bread processing using a fluorescence excitation wavelength of 290 nm and emission wavelength of 350 or 450 nm. Data indicate a relatively good stability of added folic acid and native folates to the baking process, and increased endogenous folate contents in dough and bread as compared with the flour from which they were made.     

Post-emergence application of herbicides and growth regulators on soybean growth and agronomic performance

Journal of Crop Science and Biotechnology - Plant lodging is among the factors that limit the productive potential of the soybean crop. Growth regulators or even herbicides may be used in pre- and...     

Growth- and breed-related changes of muscle bundle structure in cattle

The objective of this study was to investigate the changes in muscle fiber bundles of cattle of different breeds during growth. Different numbers of muscle fibers are surrounded by connective tissue to form bundles macroscopically visible as meat fibers or meat grain, a common meat quality trait. To determine the influence of breed and age on morphological characteristics of muscle fiber bundles, 4 cattle breeds with different growth impetus and muscularity were reared and slaughtered under experimental conditions. German Angus, a typical beef cattle; Galloway, a smaller beef type; Holstein Friesian, a dairy type; and double-muscled Belgian Blue, an extreme type for muscle growth, were used. Between 5 and 15 bulls of each breed were slaughtered at 2, 4, 6, 12, or 24 mo of age, and slices of semitendinosus muscle were removed. Muscle structure characteristics were determined by computerized image analysis. During growth, the muscle cross-sectional area enlarged (P < 0.001) about 5-fold in double-muscled Belgian Blue bulls and about 4-fold in the other breeds. This was a result of the enlargement (P < 0.001) of primary bundles and muscle fibers. The bundle size was similar (P > or = 0.15) in bulls of German Angus and Galloway in all age groups and was doubled (P < 0.001) in double-muscled Belgian Blue animals from 4 mo of age on. The Holstein Friesian bulls had the smallest (P < 0.001) muscle fiber bundles at 24 mo of age. The number of muscle fibers per bundle and the number of bundles per muscle remained nearly constant (P > 0.05) during growth. This supports the existing view that the structure of the muscle is already fixed in prenatal life. The double-muscled Belgian Blue bulls showed a more than 2.5-fold greater (P < 0.001) number of muscle fibers per primary bundle compared with the other breeds investigated. The larger muscle fiber bundles led to a smaller amount of connective tissue per muscle area in double-muscled cattle. The coarser grain of meat in double-muscled Belgian Blue bulls and in older animals was not related to greater shear force values.