Effects of Selective Substitution of Cysteine Residues on the Conformational Properties of Chlorotoxin Explored by Molecular Dynamics Simulations.

Chlorotoxin (CTX) is a 36⁻amino acid peptide with eight Cys residues that forms four disulfide bonds. It has high affinity for the glioma-specific chloride channel and matrix metalloprotease-2. Structural and binding properties of CTX analogs with various Cys residue substitutions with l-α-aminobutyric acid (Abu) have been previously reported. Using 4.2 µs molecular dynamics, we compared the conformational and essential space sampling of CTX and analogs with selective substitution of the Cys residues and associated disulfide bonds with either Abu or Ser. The native and substituted peptides maintained a high degree of α-helix propensity from residues 8 through 21, with the exception of substitution of the Cys5⁻Cys28 residues with Ser and the Cys16⁻Cys33 residues with Abu. In agreement with previous circular dichroism spectropolarimetry results, the C-terminal ß-sheet content varied less from residues 25 through 29 and 32 through 36 and was well conserved in most analogs. The Cys16⁻Cys33 and Cys20⁻Cys35 disulfide-bonded residues appear to be required to maintain the αß motif of CTX. Selective substitution with the hydrophilic Ser, may mitigate the destabilizing effect of Cys16⁻Cys33 substitution through the formation of an inter residue H-bond from Ser16:OγH to Ser33:OγH bridged by a water molecule. All peptides shared considerable sampled conformational space, which explains the retained receptor binding of the non-native analogs.

Unique methionine-aromatic interactions govern the calmodulin redox sensor.

Calmodulin contains multiple redox sensitive methionines whose oxidation alters the regulation of numerous targets. Molecular dynamics simulations were used to define the molecular principles that govern how calmodulin is structurally poised to detect and respond to methionine oxidation. We found that calmodulin's open and closed states were preferentially stabilized by unique, redox sensitive, methionine-aromatic interactions. Key methionine-aromatic interactions were coupled to reorientation of EF hand helices. Methionine to glutamine substitutions designed to mimic methionine oxidation strongly altered conformational transitions by modulating the strength of methionine-aromatic interactions. Together, these results suggest a broadly applicable redox sensing mechanism though which methionine oxidation by cellular oxidants alters the strength of methionine-aromatic interactions critical for functional protein dynamics.

Structural properties of amyloid ß(1-40) dimer explored by replica exchange molecular dynamics simulations.

Replica exchange molecular dynamics simulations (300 ns) were used to study the dimerization of amyloid ß(1-40) (Aß(1-40)) polypeptide. Configurational entropy calculations revealed that at physiological temperature (310 K, 37°C) dynamic dimers are formed by randomly docked monomers. Free energy of binding of the two chains to each other was -93.56 ± 6.341 kJ mol-1 . Prevalence of random coil conformations was found for both chains with the exceptions of increased ß-sheet content from residues 16-21 and 29-32 of chain A and residues 15-21 and 30-33 of chain B with ß-turn/ß-bend conformations in both chains from residues 1-16, 21-29 of chain A, 1-16, and 21-29 of chain B. There is a mixed ß-turn/ß-sheet region from residues 33-38 of both chains. Analysis of intra- and interchain residue distances shows that, although the individual chains are highly flexible, the dimer system stays in a loosely packed antiparallel ß-sheet configuration with contacts between residues 17-21 of chain A with residues 17-21 and 31-36 of chain B as well as residues 31-36 of chain A with residues 17-21 and 31-36 of chain B. Based on dihedral principal component analysis, the antiparallel ß-sheet-loop-ß-sheet conformational motif is favored for many low energy sampled conformations. Our results show that Aß(1-40) can form dynamic dimers in aqueous solution that have significant conformational flexibility and are stabilized by collapse of the central and C-terminal hydrophobic cores with the expected ß-sheet-loop-ß-sheet conformational motif. Proteins 2017; 85:1024-1045. © 2017 Wiley Periodicals, Inc.

Response variables for evaluation of the effectiveness of conservation corridors.

Many studies have evaluated effectiveness of corridors by measuring species presence in and movement through small structural corridors. However, few studies have assessed whether these response variables are adequate for assessing whether the conservation goals of the corridors have been achieved or considered the costs or lag times involved in measuring the response variables. We examined 4 response variables-presence of the focal species in the corridor, interpatch movement via the corridor, gene flow, and patch occupancy--with respect to 3 criteria--relevance to conservation goals, lag time (fewest generations at which a positive response to the corridor might be evident with a particular variable), and the cost of a study when applying a particular variable. The presence variable had the least relevance to conservation goals, no lag time advantage compared with interpatch movement, and only a moderate cost advantage over interpatch movement or gene flow. Movement of individual animals between patches was the most appropriate response variable for a corridor intended to provide seasonal migration, but it was not an appropriate response variable for corridor dwellers, and for passage species it was only moderately relevant to the goals of gene flow, demographic rescue, and recolonization. Response variables related to gene flow provided a good trade-off among cost, relevance to conservation goals, and lag time. Nonetheless, the lag time of 10-20 generations means that evaluation of conservation corridors cannot occur until a few decades after a corridor has been established. Response variables related to occupancy were most relevant to conservation goals, but the lag time and costs to detect corridor effects on occupancy were much greater than the lag time and costs to detect corridor effects on gene flow.

Effects of wind energy development on nesting ecology of greater prairie-chickens in fragmented grasslands.

Wind energy is targeted to meet 20% of U.S. energy needs by 2030, but new sites for development of renewable energy may overlap with important habitats of declining populations of grassland birds. Greater Prairie-Chickens (Tympanuchus cupido) are an obligate grassland bird species predicted to respond negatively to energy development. We used a modified before-after control-impact design to test for impacts of a wind energy development on the reproductive ecology of prairie-chickens in a 5-year study. We located 59 and 185 nests before and after development, respectively, of a 201 MW wind energy facility in Greater Prairie-Chicken nesting habitat and assessed nest site selection and nest survival relative to proximity to wind energy infrastructure and habitat conditions. Proximity to turbines did not negatively affect nest site selection (ß = 0.03, 95% CI = -1.2-1.3) or nest survival (ß = -0.3, 95% CI = -0.6-0.1). Instead, nest site selection and survival were strongly related to vegetative cover and other local conditions determined by management for cattle production. Integration of our project results with previous reports of behavioral avoidance of oil and gas facilities by other species of prairie grouse suggests new avenues for research to mitigate impacts of energy development.

An Introduction to the Native and Non-Native Plant-Insect Interactions and Potential Pollinators of Puerto Williams and Yendegaia, Cabo de Hornos, Chile.

The Magellanic sub-Antarctic ecoregion of southern Chile represents one of the last remaining pristine areas on Earth, but there are knowledge gaps concerning the biodiversity and interactions of the regions' flora and fauna. Non-native insect species like Bombus terrestris and Vespula vulgaris are known to have detrimental influence on native populations through competition for resources/nesting habitat, larvae predation, and foreign pathogen introduction. However, their interactions with the native and non-native plants in the region and between introduced species are unknown. This study highlights the importance of further investigations documenting the region's biodiversity, native and non-native species interactions, and local pollinators.

Mechanical Dissociation of Tissues for Single Cell Analysis Using a Motorized Device.

Being able to isolate and prepare single cells for the analysis of tissue samples has rapidly become crucial for new biomedical discoveries and research. Manual protocols for single-cell isolations are highly time-consuming and prone to user variability. Automated mechanical protocols are able to reduce processing time and sample variability but aren't easily accessible or cost-effective in lower-resourced research settings. The device described here was designed for semi-automated tissue dissociation using commercially available materials as a low-cost alternative for academic laboratories. Instructions to fabricate, assemble, and operate the device design have been provided. The dissociation protocol reliably produces single-cell suspensions with comparable cell yields and sample viability to manual preparations across multiple mouse tissues. The protocol provides the ability to process up to 12 tissue samples simultaneously per device, making studies requiring large sample sizes more manageable. The accompanying software also allows for customization of the device protocol to accommodate varying tissues and experimental constraints.

Outcomes of Activity-Related Lower Extremity Muscle Tears After Application of the British Athletics Muscle Injury Classification: A Systematic Review.

CONTEXT: Muscle injury classification and grading systems have been reported for >100 years; yet it offer limited evidence relating the clinical or radiological qualities of a muscle injury to the pathology or clinical outcome. The British Athletics Muscle Injury Classification (BAMIC) incorporates recent predictive features of muscle injuries and provides a precise radiographic framework for clinical prediction and management. OBJECTIVE: To investigate clinical outcomes, particularly time to return to play (RTP), reinjury rate (RIR), and prognostic value of specific magnetic resonance imaging (MRI) findings, of activity-related muscle injuries (tears) in athletes after application of the BAMIC. DATA SOURCES: A search of PubMed (NLM), EMBASE (Ovid), Web of Science (Clarivate), Cochrane Library (Wiley), and ClinicalTrials.gov from the inception date of each database through August 31, 2022, was conducted. Keywords included the BAMIC. STUDY SELECTION: All English language studies evaluating clinical outcomes of RTP and RIR after activity-related muscle injuries and where BAMIC was applied were included. A total of 136 articles were identified, and 11 studies met inclusion criteria. STUDY DESIGN: Systematic review (PROSPERO: CRD42022353801). LEVEL OF EVIDENCE: Level 2. DATA EXTRACTION: Two reviewers independently screened studies for eligibility and extracted data. Methodological quality of included study was assessed independently by 2 reviewers with the Newcastle-Ottawa Quality Scale (NOS); 11 good quality studies (4 prospective cohort studies, 7 retrospective cohort studies) with 468 athletes (57 female) and 574 muscle injuries were included. RESULTS: All studies reported a statistically significant relationship between BAMIC grade, BAMIC injury site, and/or combined BAMIC grade and injury site with RTP. A statistically significant increased RIR was reported by BAMIC grade and BAMIC injury site in 2 of 4 and 3 of 4 studies, respectively. The prognostic value of individual MRI criteria was limited. CONCLUSION: Consistent evidence suggests that BAMIC offers prognostic and therapeutic guidance for clinical outcomes, particularly RTP and RIR, after activity-related muscle injuries in athletes that may be superior to previous muscle injury classification and grading systems.

Habitat amount or landscape configuration: Emerging HotSpot analysis reveals the importance of habitat amount for a grassland bird in South Dakota.

Habitat loss and fragmentation are two important drivers of biodiversity decline. Understanding how species respond to landscape composition and configuration in dynamic landscapes is of great importance for informing the conservation and management of grassland species. With limited conservation resources, prescribed management targeted at the appropriate landscape process is necessary for the effective management of species. We used pheasants (Phasianus colchicus) across South Dakota, USA as a model species to identify environmental factors driving spatiotemporal variation in population productivity. Using an emerging Hotspot analysis, we analyzed annual count data from 105 fixed pheasant brood routes over a 24-year period to identify high (HotSpot) and low (ColdSpot) pheasant population productivity areas. We then applied classification and regression tree modeling to evaluate landscape attributes associated with pheasant productivity among spatial scales (500 m and 1000 m). We found that the amount of grassland at a local spatial scale was the primary factor influencing an area being a HotSpot. Our results also demonstrated non-significant or weak effects of fragmentation per se on pheasant populations. These findings are in accordance with the habitat amount hypothesis highlighting the importance of habitat amount in the landscape for maintaining and increasing the pheasant population. We, therefore, recommend that managers should focus on increasing the total habitat area in the landscape and restoring degraded habitats. Our method of identifying areas of high productivity across the landscape can be applied to other species with count data.

Sideline emergencies: an evidence-based approach.

As participation in athletics continues to increase, so too will the occurrence of on-field injuries and medical emergencies. The field of sports medicine continues to advance and many events will have on-site medical staff present. This article reviews the most catastrophic injuries and medical emergencies that are encountered in sports and presents the highest level evidence in regards to on-field approach and management of the athlete.

Contrasting landscape epidemiology of two sympatric rabies virus strains.

Viral strain evolution and disease emergence are influenced by anthropogenic change to the environment. We investigated viral characteristics, host ecology, and landscape features in the rabies-striped skunk disease system of the central Great Plains to determine how these factors interact to influence disease emergence. We amplified portions of the N and G genes of rabies viral RNA from 269 samples extracted from striped skunk brains throughout the distribution of two different rabies strains for which striped skunks were the reservoir. Because the distribution of these two strains overlapped on the landscape and were present in the same host population, we could evaluate how viral properties influenced epidemiological patterns in the area of sympatry. We found that South Central Skunk rabies (SCSK) exhibited intense purifying selection and high infectivity, which are both characteristics of an epizootic virus. Conversely, North Central Skunk rabies (NCSK) exhibited relaxed purifying selection and comparatively lower infectivity, suggesting the presence of an enzootic virus. The host population in the area of sympatry was highly admixed, and skunks among allopatric and sympatric areas had similar effective population sizes. Spatial analysis indicated that landscape features had minimal influence on NCSK movement across the landscape, but those same features were partial barriers to the spread of SCSK. We conclude that NCSK and SCSK have different epidemiological properties that interact differently with both host and landscape features to influence rabies spread in the central Great Plains. We suggest a holistic approach for future studies of emerging infectious diseases that includes studies of viral properties, host characteristics, and spatial features.

The efficacy and duration of intra-articular corticosteroid injection for knee osteoarthritis: a systematic review of level I studies.

We performed a systematic review of the current literature to determine the efficacy and duration of intra-articular corticosteroid injection in reducing pain caused by knee osteoarthritis and to determine whether the type of corticosteroid used affected these results. Following an electronic search of multiple databases and a review of reference lists from various articles, we found six trials in five papers that compared corticosteroid versus placebo and four papers that compared different corticosteroids. Results of corticosteroid compared with placebo showed both a statistically and clinically significant reduction in pain at 1 week, with an average difference between groups of 22%. Two of four trials showed triamcinolone to be more effective in pain reduction than other corticosteroids. We concluded that intra-articular corticosteroids reduce knee pain for at least 1 week and that intra-articular corticosteroid injection is a short-term treatment of a chronic problem.

What Pediatricians Need to Know About the CDC Guideline on the Diagnosis and Management of mTBI.

Pediatric traumatic brain injury (TBI) is a growing health concern, with over half a million TBI-related emergency department (ED) visits annually. However, this is likely an underestimate of the true incidence, with many children presenting to their pediatrician. The Centers for Disease Control and Prevention (CDC) published a guideline on the diagnosis and management of pediatric mild traumatic brain injury (mTBI). We outline key points and a decision checklist for pediatricians based on this evidence-based guideline.

Descriptive Epidemiology of Injuries Sustained in National Collegiate Athletic Association Men's and Women's Volleyball, 2013-2014 to 2014-2015.

BACKGROUND: There were 18,844 volleyball players in the National Collegiate Athletic Association (NCAA) in the 2014-2015 academic year. Little research has examined sex-based differences among these athletes. PURPOSE: To examine injury epidemiology in NCAA men's and women's volleyball athletes. STUDY DESIGN: Descriptive epidemiology study. LEVEL OF EVIDENCE: Level 3. METHODS: Injury surveillance data from the 2013-2014 through 2014-2015 academic years were obtained from the NCAA Injury Surveillance Program for 6 men's and 33 women's collegiate volleyball teams. Injury rates per 1000 athlete-exposures (AEs) and injury rate ratios (IRRs) with 95% CIs were calculated. Time-loss (TL) injuries resulted in participation restriction for at least 24 hours, and non-time-loss (NTL) injuries resulted in participation restriction of less than 24 hours. RESULTS: Overall, 83 and 510 injuries were reported in men and women, respectively, leading to injury rates of 4.69 and 7.07 per 1000 AEs. The injury rate was greater in women than men (IRR, 1.51; 95% CI, 1.19-1.90). TL injury rates were 1.75 and 2.62 per 1000 AEs for men and women, respectively. The ankle was the most commonly injured body part among TL injuries (men, 25.8%; women, 24.3%); the knee was the most commonly injured body part among NTL injuries (men, 25.5%; women, 16.3%). Among TL injuries, common diagnoses included sprains (men, 25.8%; women, 31.2%) and concussions (men, 19.4%; women, 14.8%). Most TL concussions were due to ball contact (men, 83.3%; women, 53.6%). Compared with men, women had a greater NTL overuse injury rate (IRR, 3.47; 95% CI, 1.61-7.46). Compared with women, men had a greater TL injury rate associated with ball contact (IRR, 2.24; 95% CI, 1.07-4.68). CONCLUSION: There are differences in injury patterns and rates between male and female intercollegiate volleyball players. Although a limited-contact sport, a notable number of concussions were sustained, mostly from ball contact. CLINICAL RELEVANCE: Understanding injury patterns may aid clinicians in injury diagnosis, management, and prevention.

The First Decade of Web-Based Sports Injury Surveillance: Descriptive Epidemiology of Injuries in US High School Girls' Volleyball (2005-2006 Through 2013-2014) and National Collegiate Athletic Association Women's Volleyball (2004-2005 Through 2013-2014).

CONTEXT:: The advent of Web-based sports injury surveillance via programs such as the High School Reporting Information Online system and the National Collegiate Athletic Association Injury Surveillance Program has aided in the acquisition of girls' and women's volleyball injury data. OBJECTIVE:: To describe the epidemiology of injuries sustained in high school girls' volleyball in the 2005-2006 through 2013-2014 academic years and collegiate women's volleyball in the 2004-2005 through 2013-2014 academic years using Web-based sports injury surveillance. DESIGN:: Descriptive epidemiology study. SETTING:: Online injury surveillance from high school girls' (annual average = 100) and collegiate women's (annual average = 50) volleyball teams. PATIENTS OR OTHER PARTICIPANTS:: Girls' and women's volleyball players who participated in practices and competitions during the 2005-2006 through 2013-2014 academic years in high school and the 2004-2005 through 2013-2014 academic years in college. MAIN OUTCOME MEASURE(S):: Athletic trainers collected time-loss (≥24 hours) injury and exposure data. Injury rates per 1000 athlete-exposures (AEs), injury rate ratios (IRRs) with 95% confidence intervals (CIs), and injury proportions by body site and diagnosis were calculated. RESULTS:: The High School Reporting Information Online system documented 1634 time-loss injuries during 1 471 872 AEs; the National Collegiate Athletic Association Injury Surveillance Program documented 2149 time-loss injuries during 563 845 AEs. The injury rate was higher in college than in high school (3.81/1000 versus 1.11/1000 AEs; IRR = 3.43; 95% CI = 3.22, 3.66), and higher in high schools with ≤1000 students than in those with >1000 students (IRR = 1.35; 95% CI = 1.23, 1.49). Injury rates did not vary by collegiate division. The injury rate was higher during competitions than practices for high school (IRR = 1.23; 95% CI = 1.12, 1.36) but not for college (IRR = 1.01; 95% CI = 0.92, 1.10). Ankle sprains were common in both the high school and collegiate setting. However, liberos had a high incidence of concussion. CONCLUSIONS:: Injury rates were higher among collegiate than high school players. However, injury rates differed by event type in high school, unlike college. Concussion injury patterns among liberos varied from those for other positions. These findings highlight the need for injury-prevention interventions specific to setting and position.

Sports medicine: performance-enhancing drugs.

Performance-enhancing drugs, ergogenic aids, or sports supplements have been a part of sports since sporting competition began and likely always will be. Considered cheating by purists and necessary by some athletes, we must accept the fact that they are used, understand why they are used, and study how to prevent their use to institute change. This article summarizes current information regarding the use of performance-enhancing drugs in young athletes and provides proven prevention strategies for instituting a program in your local schools.

Probability of lek collapse is lower inside sage-grouse Core Areas: Effectiveness of conservation policy for a landscape species.

Greater sage-grouse (Centrocercus urophasianus) occupy sagebrush (Artemisia spp.) habitats in 11 western states and 2 Canadian provinces. In September 2015, the U.S. Fish and Wildlife Service announced the listing status for sage-grouse had changed from warranted but precluded to not warranted. The primary reason cited for this change of status was that the enactment of new regulatory mechanisms was sufficient to protect sage-grouse populations. One such plan is the 2008, Wyoming Sage Grouse Executive Order (SGEO), enacted by Governor Freudenthal. The SGEO identifies "Core Areas" that are to be protected by keeping them relatively free from further energy development and limiting other forms of anthropogenic disturbances near active sage-grouse leks. Using the Wyoming Game and Fish Department's sage-grouse lek count database and the Wyoming Oil and Gas Conservation Commission database of oil and gas well locations, we investigated the effectiveness of Wyoming's Core Areas, specifically: 1) how well Core Areas encompass the distribution of sage-grouse in Wyoming, 2) whether Core Area leks have a reduced probability of lek collapse, and 3) what, if any, edge effects intensification of oil and gas development adjacent to Core Areas may be having on Core Area populations. Core Areas contained 77% of male sage-grouse attending leks and 64% of active leks. Using Bayesian binomial probability analysis, we found an average 10.9% probability of lek collapse in Core Areas and an average 20.4% probability of lek collapse outside Core Areas. Using linear regression, we found development density outside Core Areas was related to the probability of lek collapse inside Core Areas. Specifically, probability of collapse among leks >4.83 km from inside Core Area boundaries was significantly related to well density within 1.61 km (1-mi) and 4.83 km (3-mi) outside of Core Area boundaries. Collectively, these data suggest that the Wyoming Core Area Strategy has benefited sage-grouse and sage-grouse habitat conservation; however, additional guidelines limiting development densities adjacent to Core Areas may be necessary to effectively protect Core Area populations.

Fine-scale distribution modeling of avian malaria vectors in north-central Kansas.

Infectious diseases increasingly play a role in the decline of wildlife populations. Vector-borne diseases, in particular, have been implicated in mass mortality events and localized population declines are threatening some species with extinction. Transmission patterns for vector-borne diseases are influenced by the spatial distribution of vectors and are therefore not uniform across the landscape. Avian malaria is a globally distributed vector-borne disease that has been shown to affect endemic bird populations of North America. We evaluated shared habitat use between avian malaria vectors, mosquitoes in the genus Culex and a native grassland bird, the Greater Prairie-Chicken (Tympanuchus cupido), by (1) modeling the distribution of Culex spp. occurrence across the Smoky Hills of north-central Kansas using detection data and habitat variables, (2) assessing the occurrence of these vectors at nests of female Greater Prairie-Chickens, and (3) evaluating if shared habitat use between vectors and hosts is correlated with malarial infection status of the Greater Prairie-Chicken. Our results indicate that Culex occurrence increased at nest locations compared to other available but unoccupied grassland habitats; however the shared habitat use between vectors and hosts did not result in an increased prevalence of malarial parasites in Greater Prairie-Chickens that occupied habitats with high vector occurrence. We developed a predictive map to illustrate the associations between Culex occurrence and infection status with malarial parasites in an obligate grassland bird that may be used to guide management decisions to limit the spread of vector-borne diseases.

Alterations in default-mode network connectivity may be influenced by cerebrovascular changes within 1 week of sports related concussion in college varsity athletes: a pilot study.

The goal of this pilot study is to use complementary MRI strategies to quantify and relate cerebrovascular reactivity, resting cerebral blood flow and functional connectivity alterations in the first week following sports concussion in college varsity athletes. Seven college athletes (3F/4M, age = 19.7 ± 1.2 years) were imaged 3-6 days following a diagnosed sports related concussion and compared to eleven healthy controls with no history of concussion (5M/6F, 18-23 years, 7 athletes). Cerebrovascular reactivity and functional connectivity were measured using functional MRI during a hypercapnia challenge and via resting-state regional partial correlations, respectively. Resting cerebral blood flow was quantified using arterial spin labeling MRI methods. Group comparisons were made within and between 18 regions of interest. Cerebrovascular reactivity was increased after concussion when averaged across all regions of interest (p = 0.04), and within some default-mode network regions, the anterior cingulate and the right thalamus (p < 0.05) independently. The FC was increased in the concussed athletes within the default-mode network including the left and right hippocampus, precuneus and ventromedial prefrontal cortex (p < 0.01), with measures being linearly related to cerebrovascular reactivity in the hippocampus in the concussed athletes. Significant resting cerebral blood flow changes were not detected between the two groups. This study provides evidence for increased cerebrovascular reactivity and functional connectivity in the medial regions of the default-mode network within days of a single sports related concussion in college athletes. Our findings emphasize the utility of complementary cerebrovascular measures in the interpretation of alterations in functional connectivity following concussion.