INCORPORATION OF SUMMER ANNUAL MIXTURES INTO GRAZING SYSTEMS IN KENTUCKY

Utilizing summer annual grass-legume forage mixtures has the potential to improve forage yield and nutritive characteristics, and/or animal performance during times when cool-season pasture growth is limited by high temperatures. Legumes can utilize atmospheric nitrogen, which can increase crude protein and forage digestibility in mixtures. As nitrogen application generally improves both the yield and nutritive characteristics of summer annual forages, but can have a negative effect on legume competitiveness, nitrogen fertilizer recommendations for legume-containing summer annual mixtures are not well established. Two experiments were conducted to determine the feasibility of utilizing summer annual mixtures in Kentucky, USA. The first experiment was a small plot study. The objective was to evaluate the effects of increasing botanical diversity and N application rates on the yield, botanical composition, and nutritive characteristics of summer annual forage mixtures. The second experiment was a grazing study that evaluated the effects of increasing summer annual species diversity on forage yield and nutritive value, and animal performance. In the first experiment, N rates of 0, 56, 112, 168, and 224 kg N ha-1 were applied to a sudangrass monoculture, a three-species mixture, and an 11-species mixture. Sward biomass in three out of four environments increased as N application increased (average of 14 kg DM ha-1 per kg N ha-1; p \u3c 0.05). As all treatments were dominated by grass species, mixture complexity had no effect on forage DM accumulation for three out of four environments (4000, 5830, and 7280 kg DM ha-1 averaged over N rates for three environments; p \u3e 0.05). Swards were dominated by sudangrass and pearl millet (73 and 24% in simple mixtures, and 62 and 22% in complex mixtures, respectively), resulting in low functional diversity, likely due to high grass seeding rates. Mixture complexity also did not affect most nutritive characteristics (p \u3e 0.05). Although N application up to 224 kg N ha-1 often had a positive impact on forage quality parameters, forages in three out of four environments would not support the nutritional demands of growing or lactating cattle when averaged across harvests. A sensitivity analyses showed that applying N resulted in positive net returns only when hay prices were very high and N prices were low. When pasture utilization rates and hay feeding/storage losses are accounted for, enterprise budgets determined grazing to have 10% greater expenses than haying. In the second experiment, yearling angus-cross beef calves were assigned to graze one of three summer annual forage treatments, a sorghum-sudangrass monoculture, a simple three-species mixture, or a complex 12-species mixture. Animals grazed for an average of 40 days per year without supplementation. Forage yield was not different between treatments (P \u3e 0.85). Although several forage quality parameters were affected by mixture, none provided useful insight into differences observed in average daily gain (ADG). In 2017 and 2019, calves grazing the monoculture and simple mixture had higher ADG than calves grazing the complex mixture (2017: 0.79 vs. 0.66 kg/day, P \u3c 0.03; 2019: 0.59 vs. 0.43 kg/day, P \u3c 0.03). In 2018, there were no differences in ADG (P \u3e 0.3); however, calves only gained 0.01 kg/day, possibly due to lower nutritive value of more mature forages. Forages in 2018 were abnormally tall and calves were observed to be flightier and more agitated. The added stress of a low-visibility environment may have contributed to poor gains. Taller forages may also have limited dry matter intake and/or sward utilization since calves could not reach the top of the plants. In these studies, increasing species diversity did not improve forage yield, nutritive characteristics, or animal performance. This was likely due to heavy grass competition and poor legume establishment. If sward diversity is of interest, care must be taken to select compatible species, utilize appropriate seeding rates, and implement management that will promote less well-adapted species. Under the constraints of these experiments, utilizing summer annuals in forage systems in Kentucky would only be economical when hay costs were high, when production costs were low, and when animal performance was enhanced

The Application of FAST-NMR for the Identification of Novel Drug Discovery Targets

The continued success of genome sequencing projects has resulted in a wealth of information, but 40-50% of identified genes correspond to hypothetical proteins or proteins of unknown function. The Functional Annotation Screening Technology by NMR (FAST-NMR) screen was developed to assign a biological function for these unannotated proteins with a structure solved by the Protein Structure Initiative. FAST-NMR is based on the premise that a biological function can be described by a similarity in binding sites and ligand interactions with proteins of known function. The resulting co-structure and functional assignment may provide a starting point for a drug discovery effort

Using Databases and Computational Techniques to Infer the Function of Novel Proteins

The Human Genome Project and similar efforts have resulted in the identification of an abundance of novel proteins. There is a need to expedite the process of assigning function to novel proteins. Nuclear magnetic resonance (NMR) spectroscopy can be used to infer a general biological function for a protein of unknown function by identifying compounds that preferentially bind the protein and comparing these results against proteins with defined structure and function. The Functional NMR screen generates hundreds of data sets and a manual analysis of these data sets is laborious and time- consuming. It is hypothesized that several sub-tasks of the Functional NMR can be automated successfully using an integrated database and data analysis system. Our database system integrates NMR data collection, processing, analysis, and data archiving into a unified user interface. An NMR spectra comparison algorithm is designed and implemented to compare NMR data in the presence and absence of a protein to ascertain if any compound-protein binding occurred

Is There an Economic Advantage to Planting Diverse Summer Annual Forage Mixtures?

This study examined economic implications of planting summer annual mixtures of grasses, legumes, and forbs at varying nitrogen rates. No differences in yield occurred between the three mixtures, indicating that mixtures with lowest seed cost will be most economical. Applying N resulted in yield increases of 12.26 lb DM per lb N applied. Although yield responses to N were positive, sensitivity analyses showed that applying N resulted in positive net returns only when hay prices were high and N prices were low. When utilization rates are accounted for, enterprise budgets determined grazing to be 18% cheaper to implement than haying

Metabolomics Reveals New Mechanisms for Pathogenesis in Barth Syndrome and Introduces Novel Roles for Cardiolipin in Cellular Function

Barth Syndrome is the only known Mendelian disorder of cardiolipin remodeling, with characteristic clinical features of cardiomyopathy, skeletal myopathy, and neutropenia. While the primary biochemical defects of reduced mature cardiolipin and increased monolysocardiolipin are well-described, much of the downstream biochemical dysregulation has not been uncovered, and biomarkers are limited. In order to further expand upon the knowledge of the biochemical abnormalities in Barth Syndrome, we analyzed metabolite profiles in plasma from a cohort of individuals with Barth Syndrome compared to age-matched controls via 1H nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry. A clear distinction between metabolite profiles of individuals with Barth Syndrome and controls was observed, and was defined by an array of metabolite classes including amino acids and lipids. Pathway analysis of these discriminating metabolites revealed involvement of mitochondrial and extra-mitochondrial biochemical pathways including: insulin regulation of fatty acid metabolism, lipid metabolism, biogenic amine metabolism, amino acid metabolism, endothelial nitric oxide synthase signaling, and tRNA biosynthesis. Taken together, this data indicates broad metabolic dysregulation in Barth Syndrome with wide cellular effects

\u3csup\u3e1\u3c/sup\u3eH, \u3csup\u3e13\u3c/sup\u3eC, and \u3csup\u3e15\u3c/sup\u3eN NMR assignments for the \u3ci\u3eBacillus subtilis\u3c/i\u3e yndB START domain

The steroidogenic acute regulatory-related lipid transfer (START) domain is found in both eukaryotes and prokaryotes, with putative functions including signal transduction, transcriptional regulation, GTPase activation and thioester hydrolysis. Here we report the near complete 1H, 15N and 13C backbone and side chain NMR resonance assignments for the Bacillus subtilis START domain protein yndB

FAST-NMR - Functional Annotation Screening Technology Using NMR Spectroscopy

An abundance of protein structures emerging from structural genomics and the Protein Structure Initiative (PSI) are not amenable to ready functional assignment because of a lack of sequence and structural homology to proteins of known function. We describe a high-throughput NMR methodology (FAST-NMR) to annotate the biological function of novel proteins through the structural and sequence analysis of protein-ligand interactions. This is based on basic tenets of biochemistry where proteins with similar functions will have similar active sites and exhibit similar ligand binding interactions, despite global differences in sequence and structure. Protein-ligand interactions are determined through a tiered NMR screen using a library composed of compounds with known biological activity. A rapid co-structure is determined by combining the experimental identification of the ligand-binding site from NMR chemical shift perturbations with the proteinligand docking program AutoDock. Our CPASS (Comparison of Protein Active Site Structures) software and database is then used to compare this active site with proteins of known function. The methodology is demonstrated using unannotated protein SAV1430 from Staphylococcus aureus

Managing Interspecies Competition to Improve Spring Pasture

Orchardgrass (Dactylis glomerata L.) is one of the earliest maturing pasture grasses utilized in the northeastern United States. However, wet springs can delay forage harvesting resulting in advanced forage maturity and reduction in nutritive value. Chicory (Cichorium intybus L.) is a tall, upright-growing forb that shows promise as a high-energy companion crop to orchardgrass and may delay orchardgrass maturity through shading effects on plant morphology. The objective of this study was to evaluate monocultures and mixtures of orchardgrass, chicory, and white clover (Trifolium repens L.) over two consecutive springs to determine the effects of species diversity on plant maturity, nutritive characteristics, and botanical composition of forage mass. Forage monocultures and mixtures were planted in central Pennsylvania in August 2018 and were observed for two years with three harvests occurring each year (one each in spring, summer, and fall). In the first spring, orchardgrass demonstrated nine days delay in maturity when grown with chicory as compared to when grown in monocultures or in orchardgrass-white clover mixtures. Although orchardgrass was at an earlier developmental stage, fiber concentrations were similar when grown with or without chicory. Additionally, in the first spring, orchardgrass mixtures containing chicory had 1.5x greater forage mass than orchardgrass monocultures and orchardgrass-white clover mixtures. Chicory biomass was low in the second spring, likely due to winterkill following a late fall harvest the previous year, resulting in a negligible effect on orchardgrass. However, orchardgrass-chicory-white clover mixtures (even with low amounts of chicory in the second year) had the greatest forage mass and nutritive value yield over both years, indicating that these mixtures can provide greater agronomic benefits than orchardgrass monocultures

FAST-NMR - Functional Annotation Screening Technology Using NMR Spectroscopy

An abundance of protein structures emerging from structural genomics and the Protein Structure Initiative (PSI) are not amenable to ready functional assignment because of a lack of sequence and structural homology to proteins of known function. We describe a high-throughput NMR methodology (FAST-NMR) to annotate the biological function of novel proteins through the structural and sequence analysis of protein-ligand interactions. This is based on basic tenets of biochemistry where proteins with similar functions will have similar active sites and exhibit similar ligand binding interactions, despite global differences in sequence and structure. Protein-ligand interactions are determined through a tiered NMR screen using a library composed of compounds with known biological activity. A rapid co-structure is determined by combining the experimental identification of the ligand-binding site from NMR chemical shift perturbations with the proteinligand docking program AutoDock. Our CPASS (Comparison of Protein Active Site Structures) software and database is then used to compare this active site with proteins of known function. The methodology is demonstrated using unannotated protein SAV1430 from Staphylococcus aureus

Structure and function of \u3ci\u3ePseudomonas aeruginosa\u3ci\u3e protein PA1324 (21–170)

Pseudomonas aeruginosa is the prototypical biofilm-forming gram-negative opportunistic human pathogen. P. aeruginosa is causatively associated with nosocomial infections and with cystic fibrosis. Antibiotic resistance in some strains adds to the inherent difficulties that result from biofilm formation when treating P. aeruginosa infections. Transcriptional profiling studies suggest widespread changes in the proteome during quorum sensing and biofilm development. Many of the proteins found to be upregulated during these processes are poorly characterized from a functional standpoint. Here, we report the solution NMR structure of PA1324, a protein of unknown function identified in these studies, and provide a putative biological functional assignment based on the observed prealbumin-like fold and FAST-NMR ligand screening studies. PA1324 is postulated to be involved in the binding and transport of sugars or polysaccharides associated with the peptidoglycan matrix during biofilm formation