Distribution of LeConte\u27s Free-tailed Bat (Tadarida brasiliensis cynocephala) in Arkansas, with Notes on Reproduction and Natural History

During the past 20 years (1982-2001) we have studied the biology, occurrence and distribution of LeConte\u27s (Brazilian) free-tailed bat, Tadarida brasiliensis cynocephala, in Arkansas. Colonies and individuals were reported from man made structures only. Four new county records have been documented since 1988, extending the range from the central part of the state to Arkansas\u27s northern-most tier of counties. Numerous nuisance maternity colonies were investigated during exclusion activities and one, year-round colony provided the majority of reproductive data. A total of 152 free-tailed bats was submitted to the Arkansas Department of Health Rabies Laboratory (1982-2001); most during February through April, a period that corresponded to annual mating activity. Pregnant bats had single embryos only in the right uterine horn and parturition occurred in mid-June. Seven specimens tested positive for rabies

Epitope Targeting of Tertiary Protein Structure Enables Target-Guided Synthesis of a Potent In-Cell Inhibitor of Botulinum Neurotoxin

Botulinum neurotoxin (BoNT) serotype A is the most lethal known toxin and has an occluded structure, which prevents direct inhibition of its active site before it enters the cytosol. Target-guided synthesis by in situ click chemistry is combined with synthetic epitope targeting to exploit the tertiary structure of the BoNT protein as a landscape for assembling a competitive inhibitor. A substrate-mimicking peptide macrocycle is used as a direct inhibitor of BoNT. An epitope-targeting in situ click screen is utilized to identify a second peptide macrocycle ligand that binds to an epitope that, in the folded BoNT structure, is active-site-adjacent. A second in situ click screen identifies a molecular bridge between the two macrocycles. The resulting divalent inhibitor exhibits an in vitro inhibition constant of 165 pM against the BoNT/A catalytic chain. The inhibitor is carried into cells by the intact holotoxin, and demonstrates protection and rescue of BoNT intoxication in a human neuron model

A protein-targeting strategy used to develop a selective inhibitor of the E17K point mutation in the PH domain of Akt1

Ligands that can bind selectively to proteins with single amino-acid point mutations offer the potential to detect or treat an abnormal protein in the presence of the wild type (WT). However, it is difficult to develop a selective ligand if the point mutation is not associated with an addressable location, such as a binding pocket. Here we report an all-chemical synthetic epitope-targeting strategy that we used to discover a 5-mer peptide with selectivity for the E17K-transforming point mutation in the pleckstrin homology domain of the Akt1 oncoprotein. A fragment of Akt1 that contained the E17K mutation and an I19[propargylglycine] substitution was synthesized to form an addressable synthetic epitope. Azide-presenting peptides that clicked covalently onto this alkyne-presenting epitope were selected from a library using in situ screening. One peptide exhibits a 10:1 in vitro selectivity for the oncoprotein relative to the WT, with a similar selectivity in cells. This 5-mer peptide was expanded into a larger ligand that selectively blocks the E17K Akt1 interaction with its PIP3 (phosphatidylinositol (3,4,5)-trisphosphate) substrate

Toward a conceptual framework of emotional relationship marketing: an examination of two UK political parties

The purpose of this paper is to review the notion of branding and evaluate its applicability to political parties. As ideological politics is in decline, branding may provide a consistent narrative where voters feel a sense of warmth and belonging. The paper aims to build an understanding of the complexity of building a political brand where a combination of image, logo, leadership, and values can all contribute to a compelling brand narrative. It investigates how competing positive and negative messages attempt to build and distort the brand identity. A critical review of bran ding, relationship marketing, and political science literature articulates the conceptual development of branding and its applicability to political parties. The success or failure of negative campaigning is due to the authenticity of a political party’s brand values — creating a coherent brand story — if there is no distance between the brand values articulated by the political party and the values their community perceives then this creates an "authentic" brand. However, if there is a gap this paper illustrates how negative campaigning can be used to build a "doppelganger brand," which undermines the credibility of the authentic political brand. The paper argues that political parties need to understand how brand stories are developed but also how they can be used to protect against negative advertising. This has implications for political marketing strategists and political parties. This paper draws together branding theory and relationship marketing and incorporates them into a framework that makes a contribution to the political marketing literature

A rapid in vivo screen for pancreatic ductal adenocarcinoma therapeutics

Pancreatic ductal adenocarcinoma (PDA) is the fourth leading cause of cancer-related deaths in the United States, and is projected to be second by 2025. It has the worst survival rate among all major cancers. Two pressing needs for extending life expectancy of affected individuals are the development of new approaches to identify improved therapeutics, addressed herein, and the identification of early markers. PDA advances through a complex series of intercellular and physiological interactions that drive cancer progression in response to organ stress, organ failure, malnutrition, and infiltrating immune and stromal cells. Candidate drugs identified in organ culture or cell-based screens must be validated in preclinical models such as KIC (p48Cre;LSL-KrasG12D;Cdkn2af/f) mice, a genetically engineered model of PDA in which large aggressive tumors develop by 4 weeks of age. We report a rapid, systematic and robust in vivo screen for effective drug combinations to treat Kras-dependent PDA. Kras mutations occur early in tumor progression in over 90% of human PDA cases. Protein kinase and G-protein coupled receptor (GPCR) signaling activates Kras. Regulators of G-protein signaling (RGS) proteins are coincidence detectors that can be induced by multiple inputs to feedback-regulate GPCR signaling. We crossed Rgs16::GFP bacterial artificial chromosome (BAC) transgenic mice withKIC mice and show that the Rgs16::GFP transgene is a KrasG12D-dependent marker of all stages of PDA, and increases proportionally to tumor burden in KIC mice. RNA sequencing (RNA-Seq) analysis of cultured primary PDA cells reveals characteristics of embryonic progenitors of pancreatic ducts and endocrine cells, and extraordinarily high expression of the receptor tyrosine kinase Axl, an emerging cancer drug target. In proof-of-principle drug screens, we find that weanling KIC mice with PDA treated for 2 weeks with gemcitabine (with or without Abraxane) plus inhibitors of Axl signaling (warfarin and BGB324) have fewer tumor initiation sites and reduced tumor size compared with the standard-of-care treatment. Rgs16::GFP is therefore an in vivo reporter of PDA progression and sensitivity to new chemotherapeutic drug regimens such as Axl-targeted agents. This screening strategy can potentially be applied to identify improved therapeutics for other cancers

A Cocktail of Thermally Stable, Chemically Synthesized Capture Agents for the Efficient Detection of Anti-Gp41 Antibodies from Human Sera

We report on a method to improve in vitro diagnostic assays that detect immune response, with specific application to HIV-1. The inherent polyclonal diversity of the humoral immune response was addressed by using sequential in situ click chemistry to develop a cocktail of peptide-based capture agents, the components of which were raised against different, representative anti-HIV antibodies that bind to a conserved epitope of the HIV-1 envelope protein gp41. The cocktail was used to detect anti-HIV-1 antibodies from a panel of sera collected from HIV-positive patients, with improved signal-to-noise ratio relative to the gold standard commercial recombinant protein antigen. The capture agents were stable when stored as a powder for two months at temperatures close to 60°C

Lamellipodia are crucial for haptotactic sensing and response

Haptotaxis is the process by which cells respond to gradients of substrate-bound cues, such as extracellular matrix proteins (ECM); however, the cellular mechanism of this response remains poorly understood and has mainly been studied by comparing cell behavior on uniform ECMs with different concentrations of components. To study haptotaxis in response to gradients, we utilized microfluidic chambers to generate gradients of the ECM protein fibronectin, and imaged the cell migration response. Lamellipodia are fan-shaped protrusions that are common in migrating cells. Here, we define a new function for lamellipodia and the cellular mechanism required for haptotaxis – differential actin and lamellipodial protrusion dynamics lead to biased cell migration. Modest differences in lamellipodial dynamics occurring over time periods of seconds to minutes are summed over hours to produce differential whole cell movement towards higher concentrations of fibronectin. We identify a specific subset of lamellipodia regulators as being crucial for haptotaxis. Numerous studies have linked components of this pathway to cancer metastasis and, consistent with this, we find that expression of the oncogenic Rac1 P29S mutation abrogates haptotaxis. Finally, we show that haptotaxis also operates through this pathway in 3D environments

Wetland Management Strategies that Maximize Marsh Bird Use in the Midwest: Final Performance Report: F14AP00485

We sought to investigate marsh bird occupancy and abundance across a wide range of representative wetlands types, hydrologic regimes, management practices, and former disturbance regimes in Illinois. We hypothesized that characteristics of wetlands that were actively and passively managed for waterfowl would be positively correlated with marsh bird occupancy and abundance in Illinois during the migration and breeding seasons. Our specific objectives were to: 1) compare marsh bird use of wetland impoundments managed for waterfowl across a continuum of management intensities and strategies to predict how impoundment management actions can increase use by both groups; 2) compare marsh bird use of restored and natural wetlands; and 3) determine characteristics of wetlands and the surrounding landscape that influence marsh bird use of restored wetlands. Our results are important to understanding spatiotemporal, hydrological, and vegetative conditions suitable for multi-species management of wetlands. Moreover, our research provided information regarding the effectiveness of conservation actions, particularly wetland restoration in meeting conservation priorities for migrating birds.U.S. Fish & Wildlife Service Wildlife and Sport Fish Restoration Programsunpublishednot peer reviewedOpe

Protein Catalyzed Capture Agents with Tailored Performance for In Vitro and In Vivo Applications

We report on peptide-based ligands matured through the protein catalyzed capture (PCC) agent method to tailor molecular binders for in vitro sensing/diagnostics and in vivo pharmacokinetics parameters. A vascular endothelial growth factor (VEGF) binding peptide and a peptide against the protective antigen (PA) protein of Bacillus anthracis discovered through phage and bacterial display panning technologies, respectively, were modified with click handles and subjected to iterative in situ click chemistry screens using synthetic peptide libraries. Each azide-alkyne cycloaddition iteration, promoted by the respective target proteins, yielded improvements in metrics for the application of interest. The anti-VEGF PCC was explored as a stable in vivo imaging probe. It exhibited excellent stability against proteases and a mean elimination in vivo half-life (T_(1/2)) of 36 min. Intraperitoneal injection of the reagent results in slow clearance from the peritoneal cavity and kidney retention at extended times, while intravenous injection translates to rapid renal clearance. The ligand competed with the commercial antibody for binding to VEGF in vivo. The anti-PA ligand was developed for detection assays that perform in demanding physical environments. The matured anti-PA PCC exhibited no solution aggregation, no fragmentation when heated to 100°C, and  > 81% binding activity for PA after heating at 90°C for 1 h. We discuss the potential of the PCC agent screening process for the discovery and enrichment of next generation antibody alternatives

The 6dF Galaxy Survey: bulk flows on 50-70 h

We measure the bulk flow of the local Universe using the 6dF Galaxy Survey peculiar velocity sample (6dFGSv), the largest and most homogeneous peculiar velocity sample to date. 6dFGSv is a Fundamental Plane sample of ∼104 peculiar velocities covering the whole Southern hemisphere for galactic latitude |b| > 10°, out to redshift z = 0.0537. We apply the ‘minimum variance’ bulk flow weighting method, which allows us to make a robust measurement of the bulk flow on scales of 50 and 70 h−1 Mpc. We investigate and correct for potential bias due to the lognormal velocity uncertainties, and verify our method by constructing Λ cold dark matter (ΛCDM) 6dFGSv mock catalogues incorporating the survey selection function. For a hemisphere of radius 50 h−1 Mpc we find a bulk flow amplitude of U = 248 ± 58 km s−1 in the direction (l, b) = (318° ± 20°, 40° ± 13°), and for 70 h−1 Mpc we find U = 243 ± 58 km s−1, in the same direction. Our measurement gives us a constraint on σ8 of 1.01+1.07−0.58. Our results are in agreement with other recent measurements of the direction of the bulk flow, and our measured amplitude is consistent with a ΛCDM prediction