Structural modeling and functional analysis of the essential ribosomal processing protease Prp from Staphylococcus aureus

In Firmicutes and related bacteria, ribosomal large subunit protein L27 is encoded with a conserved N-terminal extension that is removed to expose residues critical for ribosome function. Bacteria encoding L27 with this N-terminal extension also encode a sequence-specific cysteine protease, Prp, which carries out this cleavage. In this work, we demonstrate that L27 variants with an un-cleavable N-terminal extension, or lacking the extension (pre-cleaved), are unable to complement an L27 deletion in Staphylococcus aureus. This indicates that N-terminal processing of L27 is not only essential but possibly has a regulatory role. Prp represents a new clade of previously uncharacterized cysteine proteases, and the dependence of S. aureus on L27 cleavage by Prp validates the enzyme as a target for potential antibiotic development. To better understand the mechanism of Prp activity, we analyzed Prp enzyme kinetics and substrate preference using a fluorogenic peptide cleavage assay. Molecular modeling and site-directed mutagenesis implicate several residues around the active site in catalysis and substrate binding, and support a structural model in which rearrangement of a flexible loop upon binding of the correct peptide substrate is required for the active site to assume the proper conformation. These findings lay the foundation for the development of antimicrobials that target this novel, essential pathway

Effects of Degree and Configuration of Hearing Loss on the Contribution of High- and Low-Frequency Speech Information to Bilateral Speech Understanding

Objectives: The purpose of this study was to examine the effects of degree and configuration of hearing loss on the use of, and benefit from, information in amplified high- and low-frequency speech presented in background noise. Design: Sixty-two adults with a wide range of high- and low-frequency sensorineural hearing loss (5 to 115+ dB HL) participated in the study. To examine the contribution of speech information in different frequency regions, speech understanding in noise was assessed in multiple low- and high-pass filter conditions, as well as a band-pass (713 to 3534 Hz) and wideband (143 to 8976 Hz) condition. To increase audibility over a wide frequency range, speech and noise were amplified based on each individual\u27s hearing loss. A stepwise multiple linear regression approach was used to examine the contribution of several factors to (1) absolute performance in each filter condition and (2) the change in performance with the addition of amplified high- and low-frequency speech components. Results: Results from the regression analysis showed that degree of hearing loss was the strongest predictor of absolute performance for low- and high-pass filtered speech materials. In addition, configuration of hearing loss affected both absolute performance for severely low-pass filtered speech and benefit from extending high-frequency (3534 to 8976 Hz) bandwidth. Specifically, individuals with steeply sloping high-frequency losses made better use of low-pass filtered speech information than individuals with similar low-frequency thresholds but less high-frequency loss. In contrast, given similar high-frequency thresholds, individuals with flat hearing losses received more benefit from extending high-frequency bandwidth than individuals with more sloping losses. Conclusions: Consistent with previous work, benefit from speech information in a given frequency region generally decreases as degree of hearing loss in that frequency region increases. However, given a similar degree of loss, the configuration of hearing loss also affects the ability to use speech information in different frequency regions. Except for individuals with steeply sloping high-frequency losses, providing high-frequency amplification (3534 to 8976 Hz) had either a beneficial effect on, or did not significantly degrade, speech understanding. These findings highlight the importance of extended high-frequency amplification for listeners with a wide range of high-frequency hearing losses, when seeking to maximize intelligibility

Sequestered Alkaloid Defenses in the Dendrobatid Poison Frog Oophaga pumilio Provide Variable Protection from Microbial Pathogens

Most amphibians produce their own defensive chemicals; however, poison frogs sequester their alkaloid-based defenses from dietary arthropods. Alkaloids function as a defense against predators, and certain types appear to inhibit microbial growth. Alkaloid defenses vary considerably among populations of poison frogs, reflecting geographic differences in availability of dietary arthropods. Consequently, environmentally driven differences in frog defenses may have significant implications regarding their protection against pathogens. While natural alkaloid mixtures in dendrobatid poison frogs have recently been shown to inhibit growth of non-pathogenic microbes, no studies have examined the effectiveness of alkaloids against microbes that infect these frogs. Herein, we examined how alkaloid defenses in the dendrobatid poison frog, Oophaga pumilio, affect growth of the known anuran pathogens Aeromonas hydrophila and Klebsiella pneumoniae. Frogs were collected from five locations throughout Costa Rica that are known to vary in their alkaloid profiles. Alkaloids were isolated from individual skins, and extracts were assayed against both pathogens. Microbe subcultures were inoculated with extracted alkaloids to create dose-response curves. Subsequent spectrophotometry and cell counting assays were used to assess growth inhibition. GC-MS was used to characterize and quantify alkaloids in frog extracts, and our results suggest that variation in alkaloid defenses lead to differences in inhibition of these pathogens. The present study provides the first evidence that alkaloid variation in a dendrobatid poison frog is associated with differences in inhibition of anuran pathogens, and offers further support that alkaloid defenses in poison frogs confer protection against both pathogens and predators

A new chemistry for managing soybean aphid

Since the arrival of soybean aphid, Aphis glycines Matsumura, to North America, entomologists have generated considerable research to effectively protect yield in soybean, Glycine max (L.). Population dynamics of this relatively new pest have been erratic between fields and among years, and timely insecticide applications must be used to prevent severe economic loss

Seed Source May Determine Field-Specific Germination and Emergence: The Source by Planting Environment Interaction

Farm environmental conditions and management practices can result in within-cultivar differences in seed quality and lead to transgenerational plasticity (farm-specific effects on offspring, or TGP) that affect germination and emergence in transplant fields. We used three perennial bunchgrasses, [green needlegrass (Nassella viridula) ‘Lodorm’, slender wheatgrass (Elymus trachycaulus) ‘Pryor’, and bluebunch wheatgrass (Pseudoregneria spicata) prevariety registered germplasm P-7] to determine if seeds exhibited TGP. We also determined if TGP was affected by the interaction between production farms and planting environments (farm × environment interaction, or context-dependent TGP), using four laboratory temperature regimes to test germination response and four field environments to test emergence response in 2013. We stored seeds in four different environments for 10 mo before repeating the experiment to test if recent seed storage conditions mitigated TGP. Contextdependent TGP affected emergence for Pryor and Lodorm both years, however, only Pryor exhibited context-dependent TGP for germination in 2013. Sources with low germination and emergence in the field were less likely to exhibit context-dependent TGP. Some transplant fields did not show differences among sources, but in other transplant fields, emergence increased as much as 24% depending on farm source. The effect of recent seed storage conditions, significant only for Pryor, was opposite for germination and emergence, with room-temperature stored seeds exhibiting the highest emergence and lowest germination. Context-dependent TGP in emergence could not be predicted by our coarse information regarding seed production environments and storage conditions or by germination in the lab. Nonetheless, context-dependent TGP significantly determined emergence in two of the three study species. Mechanisms underlying this phenomenon need further study to understand potential benefits and pitfalls for producers and seed buyers

Avian Foraging Patterns in Crop Field Edges Adjacent to Woody Habitat

As natural predators of pest insects, woodland birds provide biological pest suppression in crop fields adjacent to woody edges. Although many birds using these habitats forage widely, earlier studies have found that most foraging activity occurs within 50 m of the woody edge. The goals of this study were to determine the primary area of use, or functional edge, for birds foraging in crop fields adjacent to woody edges, and to evaluate their foraging distance patterns. During the summers of 2005 and 2006, avian foraging behavior was observed at 12 research sites in east central Nebraska that contained either a shelterbelt or woody riparian edge. At each site, perches were provided at 10 m intervals out from the edge and insect larvae were placed in feeders at random locations to simulate a pest insect food resource. Birds were recorded foraging in five distance categories out from the edge (0–10, 10–20, 20–30, 30–40, and 40–50 m). Seven species foraged primarily within 20 m of the edge (72% all observations; 79% without perch or feeder observations). Ten species foraged throughout the plots but six of these generally foraged more often (45% and 49%) and four less often (30% and 30%) within 20 m of the edge. The 13 species that tended to forage more often within 20 m of the edge, with 56% of their foraging overall in this area, also tended to forage farther when perch and feeder observations were included, indicating willingness to forage farther when food resources were available. Based on a repeated measures analysis of variance, foraging distances appeared to be greater at sites with soybean as the planted crop, although this apparent trend was significant for only some species. There was no clear difference in foraging distances outward from shelterbelt versus riparian sites. These results indicate that conservation efforts within the 20 m functional edge offer potential to enhance the sustainability of both birds and crops in agricultural

Avian Foraging Patterns in Crop Field Edges Adjacent to Woody Habitat

As natural predators of pest insects, woodland birds provide biological pest suppression in crop fields adjacent to woody edges. Although many birds using these habitats forage widely, earlier studies have found that most foraging activity occurs within 50 m of the woody edge. The goals of this study were to determine the primary area of use, or functional edge, for birds foraging in crop fields adjacent to woody edges, and to evaluate their foraging distance patterns. During the summers of 2005 and 2006, avian foraging behavior was observed at 12 research sites in east central Nebraska that contained either a shelterbelt or woody riparian edge. At each site, perches were provided at 10 m intervals out from the edge and insect larvae were placed in feeders at random locations to simulate a pest insect food resource. Birds were recorded foraging in five distance categories out from the edge (0–10, 10–20, 20–30, 30–40, and 40–50 m). Seven species foraged primarily within 20 m of the edge (72% all observations; 79% without perch or feeder observations). Ten species foraged throughout the plots but six of these generally foraged more often (45% and 49%) and four less often (30% and 30%) within 20 m of the edge. The 13 species that tended to forage more often within 20 m of the edge, with 56% of their foraging overall in this area, also tended to forage farther when perch and feeder observations were included, indicating willingness to forage farther when food resources were available. Based on a repeated measures analysis of variance, foraging distances appeared to be greater at sites with soybean as the planted crop, although this apparent trend was significant for only some species. There was no clear difference in foraging distances outward from shelterbelt versus riparian sites. These results indicate that conservation efforts within the 20 m functional edge offer potential to enhance the sustainability of both birds and crops in agricultural

Effects of Experimentally Elevated Traffic Noise on Nestling White-Crowned Sparrow Stress Physiology, Immune Function and Life History

Roads have been associated with behavioral and physiological changes in wildlife. In birds, roads decrease reproductive success and biodiversity and increase physiological stress. Although the consequences of roads on individuals and communities have been well described, the mechanisms through which roads affect birds remain largely unexplored. Here, we examine one mechanism through which roads could affect birds: traffic noise. We exposed nestling mountain white-crowned sparrows (Zonotrichia leucophrys oriantha) to experimentally elevated traffic noise for 5 days during the nestling period. Following exposure to traffic noise we measured nestling stress physiology, immune function, body size, condition and survival. Based on prior studies, we expected the traffic noise treatment to result in elevated stress hormones (glucocorticoids), and declines in immune function, body size, condition and survival. Surprisingly, nestlings exposed to traffic noise had lower glucocorticoid levels and improved condition relative to control nests. These results indicate that traffic noise does affect physiology and development in white-crowned sparrows, but not at all as predicted. Therefore, when evaluating the mechanisms through which roads affect avian populations, other factors (e.g. edge effects, pollution and mechanical vibration) may be more important than traffic noise in explaining elevated nestling stress responses in this species

Healthy Lifestyles for People with Disabilities

People with disabilities are more susceptible to compromised health status and preventable secondary conditions. A Healthy Lifestyles curriculum was developed as a health promotion program for people with disabilities. Using the curriculum, ten free 2½-day workshops were provided for people with various disabilities in Oregon and Southwest Washington. Workshops were conducted in collaboration with local entities such as Centers for Independent Living. The workshops took an integrated approach to health, addressing connections among physical, social, emotional, and spiritual health, and health through meaningful activities. During workshops, the participants obtained health information and experienced healthy activities such as yoga and non-impact aerobics, both tailored for people with disabilities. At the end of the workshop, each participant identified two healthy lifestyle goals to work toward. Progress and/or barriers in accomplishing those goals were shared in support groups for 6-9 months. Preliminary results indicate early and sustained improvements in health behaviors and health-related attitudes. The Healthy Lifestyles program offers a promising approach to promoting health among people with disabilities

C.H.A.M.P. Families : Description and Theoretical Foundations of a Paediatric Overweight and Obesity Intervention Targeting Parents-A Single-Centre Non-Randomised Feasibility Study.

Childhood obesity represents a significant global health challenge, and treatment interventions are needed. The purpose of this paper is to describe the components and theoretical model that was used in the development and implementation of a unique parent-focussed paediatric overweight/obesity intervention. C.H.A.M.P. Families was a single-centre, prospective intervention offered to parents of children aged between 6⁻14 years with a body mass index (BMI) ≥85th percentile for age and sex. The intervention included: (1) eight group-based (parent-only) education sessions over 13-weeks; (2) eight home-based activities; and (3) two group-based (family) follow-up support sessions. The first section of the manuscript contains a detailed description of each intervention component, as well as an overview of ongoing feasibility analyses. The theoretical portion details the use of evidence-based group dynamics principles and motivational interviewing techniques within the context of a broader social cognitive theory foundation. This paper provides researchers with practical examples of how theoretical constructs and evidence-based strategies can be applied in the development and implementation of parent-focussed paediatric obesity interventions. Given the need for transparent reporting of intervention designs and theoretical foundations, this paper also adds to the areas of implementation science and knowledge translation research