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A strategy for the evaluation of different HAT (hemagglutinin variants) ratios on the FluMos-v1 mosaic nanoparticle influenza vaccine by LC-MS/MS-based PRM.

Many animals use sound as a medium for detecting or locating potential prey items or predation threats. Northern saw-whet owls (Aegolius acadicus) are particularly interesting in this regard, as they primarily rely on sound for hunting in darkness, but are also subject to predation pressure from larger raptors. We hypothesized that these opposing tasks should favor sensitivity to low-frequency sounds arriving from many locations (potential predators) and high-frequency sounds below the animal (ground-dwelling prey items). Furthermore, based on the morphology of the saw-whet owl skull and the head-related transfer functions of related species, we expected that the magnitude of changes in sensitivity across spatial locations would be greater for higher frequencies than low frequencies (i.e., more “directional” at high frequencies). We used auditory-evoked potentials to investigate the frequency-specific directional sensitivity of Northern saw-whet owls to acoustic signals. We found some support for our hypothesis, with smaller-magnitude changes in sensitivity across spatial locations at lower frequencies and larger-magnitude changes at higher frequencies. In general, owls were most sensitive to sounds originating in front of and above their heads, but at 8 kHz there was also an area of high sensitivity below the animals. Our results suggest that the directional hearing of saw-whet owls should allow for both predator and prey detection.

Multiple active vaccination approaches have proven ineffective in reducing the substantial morbidity and mortality caused by respiratory syncytial virus (RSV) in infants and older adults (aged ≥65 years). A vaccine conferring a substantial and sustainable boost in neutralising activity is required to protect against severe RSV disease. To that end, we evaluated the safety and immunogenicity of DS-Cav1, a prefusion F subunit vaccine.In this randomised, open-label, phase 1 clinical trial, the stabilised prefusion F vaccine DS-Cav1 was evaluated for dose, safety, tolerability, and immunogenicity in healthy adults aged 18-50 years at a single US site. Participants were assigned to receive escalating doses of either 50 μg, 150 μg, or 500 μg DS-Cav1 at weeks 0 and 12, and were randomly allocated in a 1:1 ratio within each dose group to receive the vaccine with or without aluminium hydroxide (AlOH) adjuvant. After 71 participants had been randomised, the protocol was amended to allow some participants to receive a single vaccination at week 0. The primary objectives evaluated the safety and tolerability at every dose within 28 days following each injection. Neutralising activity and RSV F-binding antibodies were evaluated from week 0 to week 44 as secondary and exploratory objectives. Safety was assessed in all participants who received at least one vaccine dose; secondary and exploratory immunogenicity analysis included all participants with available data at a given visit. The trial is registered with ClinicalTrials.gov, NCT03049488, and is complete and no longer recruiting.Between Feb 21, 2017, and Nov 29, 2018, 244 participants were screened for eligibility and 95 were enrolled to receive DS-Cav1 at the 50 μg (n=30, of which n=15 with AlOH), 150 μg (n=35, of which n=15 with AlOH), or 500 μg (n=30, of which n=15 with AlOH) doses. DS-Cav1 was safe and well tolerated and no serious vaccine-associated adverse events deemed related to the vaccine were identified. DS-Cav1 vaccination elicited robust neutralising activity and binding antibodies by 4 weeks after a single vaccination (p0·0001 for F-binding and neutralising antibodies). In analyses of exploratory endpoints at week 44, pre-F-binding IgG and neutralising activity were significantly increased compared with baseline in all groups. At week 44, RSV A neutralising activity was 3·1 fold above baseline in the 50 μg group, 3·8 fold in the 150 μg group, and 4·5 fold in the 500 μg group (p0·0001). RSV B neutralising activity was 2·8 fold above baseline in the 50 μg group, 3·4 fold in the 150 μg group, and 3·7 fold in the 500 μg group (p0·0001). Pre-F-binding IgG remained significantly 3·2 fold above baseline in the 50 μg group, 3·4 fold in the 150 μg group, and 4·0 fold in the 500 μg group (p0·0001). Pre-F-binding serum IgA remained 4·1 fold above baseline in the 50 μg group, 4·3 fold in the 150 μg group, and 4·8 fold in the 500 μg group (p0·0001). Although a higher vaccine dose or second immunisation elicited a transient advantage compared with lower doses or a single immunisation, neither significantly impacted long-term neutralisation. There was no long-term effect of dose, number of vaccinations, or adjuvant on neutralising activity.In this phase 1 study, DS-Cav1 vaccination was safe and well tolerated. DS-Cav1 vaccination elicited a robust boost in RSV F-specific antibodies and neutralising activity that was sustained above baseline for at least 44 weeks. A single low-dose of pre-F immunisation of antigen-experienced individuals might confer protection that extends throughout an entire RSV season.The National Institutes of Allergy and Infectious Diseases.

Broadly neutralizing antibody (bNAb) CAP256-VRC26.25 (abbreviated CAP256LS), a human IgGI monoclonal antibody targeting the V1V2 site of the HIV-1 envelope, has demonstrated high therapeutic potential as a broadly neutralizing monoclonal antibody against HIV-1. During the process development, a heavy chain fragmentation (clipping) was observed, that led to a relative potency reduction. In this report, we highlighted a series of process and product mitigation strategies deployed to advance this product. We have detailed how analytical characterization tools, especially the microchip reduced capillary gel electrophoresis (CGE-SDS), played a pivotal role in identifying the development issues and in providing measurements to guide implementation of mitigation strategies.

Both behavioral receptivity and neural sensitivity to acoustic mate attraction signals vary across the reproductive cycle, particularly in seasonally breeding animals. Across a variety of taxa receptivity to signals increases, as does peripheral auditory sensitivity, as females transition from a non-breeding to breeding condition. We recently documented decreases in receptivity to acoustic mate attraction signals and circulating hormone levels, but an increase in peripheral auditory sensitivity to call-like stimuli following oviposition in Cope's gray treefrogs (Hyla chrysoscelis). However, it is not known if changes in auditory sensitivity are confined to the frequency range of calls, or if they result from more generalized changes in the auditory periphery. Here, we used auditory brainstem responses (ABRs) to evaluate peripheral frequency sensitivity in female Cope's gray treefrogs before and after oviposition. We found lower ABR thresholds, greater ABR amplitudes, and shorter ABR latencies following oviposition. Changes were most pronounced and consistent at lower frequencies associated with the amphibian papilla, but were also detectable at higher frequencies corresponding to the tuning of the basilar papilla. Furthermore, only ABR latencies were correlated with circulating steroid hormones (testosterone). Changes in peripheral processing may result from changes in metabolic function or sensorineural adaptation to chorus noise.

In light of a modern understanding of early Eocene greenhouse climate fluctuations and new highly seasonal fluvial system faces models, the role of climate in the evolution of one classically-cited continental, terminal lake system is re-examined. Detailed stratigraphic description and elemental abundance data from fifteen cores and seven outcrop regions of the Green River Formation were used to construct a ∼150 km cross section across the Uinta Basin, Utah, USA. Lake Uinta in the Uinta Basin is divided into five lake phases: (1) post-Paleocene Eocene Thermal Maximum, (2) peak Eocene hyperthermal, (3) waning hyperthermal, Early Eocene Climatic Optimum (EECO), (4) post-hyperthermal, and (5) post-EECO regimes, based primarily on climatically driven changes in fluvial style in combination with sedimentary indicators of lacustrine carbonate deposition, organic matter preservation, salinity, and lake depth. Basinwide siliciclastic dominated intervals were deposited by highly seasonal fluvial systems and record negative organic carbon isotope excursions associated with early Eocene abrupt, transient global warming (hyperthermal) events. Carbonate dominated or organic rich intervals record stable, less seasonal climate periods between hyperthermals, with lower siliciclastic sediment supply allowing the development of carbonate and organic matter preservation. The stratigraphic progression from alternating organic rich and lean zones to the overlying organic rich Mahogany and R8 zones represents the global transition out of the pulsed early Eocene hyperthermal climate regime to a time of sediment starvation and lake stratification, sequestering sedimentary organic carbon. This study provides a novel approach to terrestrial paleoclimate reconstruction that relies largely on unique sedimentary indicators and secondarily on isotopic proxy records within the context of a large basin-wide sedimentologic and stratigraphic data set, thus setting the stage for future detailed geochemical terrestrial paleoclimate proxy development.

Study question Modeling the feedback loop controlling sperm capacitation to study the reversibility/irreversibility of the process Summary answer We demonstrate the existence of a feedback loop between pH increase and hyperpolarization, inducing bistability and possibly the reversibility of the capacitation process. What is known already The process of mammalian capacitation does need an influx of calcium ions through the transmembrane Catsper Channel Complex. This plasmic membrane channel, which is activated by an increase in either the intracellular pH or the membrane potential, is a sperm-specific protein that is localized in the sperm flagella. Two other sperm specific transporters, sNHE and SLO3, found in the flagella, have been shown to control protons influx and transmembrane voltage respectively, and could work together in a feedback loop keeping the sperm in a high pH state with hyperpolarized membrane. Study design, size, duration Mathematical model Participants/materials, setting, methods Not applicable Main results and the role of chance The results given by this minimal theoretical model are robust as the predicted qualitative behavior can accommodate to a wide span of physiological parameters variations. Limitations, reasons for caution The model and its parameters are mainly based on in vitro results of the literature, so that the in vivo implications should be inferred carefully. Wider implications of the findings: The possible reversibility of the capacitation process could have major clinical implications, relevant for the optimization of sperm preparation in assisted reproductive techniques and cryopreservation procedures. Trial registration number Not applicable

Synopsis Most animals experience reproductive transitions in their lives; for example, reaching reproductive maturity or cycling in and out of breeding condition. Some reproductive transitions are abrupt, while others are more gradual. In most cases, changes in communication between the sexes follow the time course of these reproductive transitions and are typically thought to be coordinated by steroid hormones. We know a great deal about hormonal control of communication behaviors in birds and frogs, as well as the central neural control of these behaviors. There has also been significant interest in the effects of steroid hormones on central nervous system structures that control both the production and reception of communication signals associated with reproductive behaviors. However, peripheral sensory structures have typically received less attention, although there has been growing interest in recent years. It is becoming clear that peripheral sensory systems play an important role in reproductive communication, are plastic across reproductive conditions, and, in some cases, this plasticity may be mediated by steroid hormones. In this article, we discuss recent evidence for the role of peripheral auditory structures in reproductive communication in birds and frogs, the plasticity of the peripheral auditory system, and the role of steroid hormones in mediating the effects of the peripheral auditory system on reproductive communication. We focus on both seasonal and acute reproductive transitions, introduce new data on the role of hormones in modulating seasonal patterns, and make recommendations for future work.

Glucocorticoids (GCs) are rarely studied in the context of female mate choice, despite the expression of receptors for these products in sexual, sensory and decision-making brain areas. Here we investigated the effects of GC concentrations on three aspects of female sexual behavior in breeding Cope's gray treefrogs (Hyla chrysoscelis): proceptivity—a measure of sexual motivation, intraspecific mate preferences, and mate choosiness. To our knowledge this is the first experimental study on the endocrine basis of mate choosiness. We predicted that mate choosiness—forfeiting an initial mate preference to pursue a suddenly more attractive mate—would be particularly impacted by elevated GCs with moderate GC levels associated with greater choosiness. We found support for this predicted inverted-U relationship. Females in the control group (no injection) showed no change in choosiness across timepoints. In contrast, females in the vehicle, Low (20 ng g−1) and High (180 ng g−1) corticosterone groups exhibited a nominal decline in choosiness after injection, suggesting that the experience of injection has little or perhaps slightly suppressive effects on female choosiness. Females in the moderate dose group (60 ng g−1), however, exhibited a significant increase (>100%) in choosiness. Further, we found no effect of elevated GCs on sexual proceptivity or the species-typical preference for longer calls. These findings may reflect a buffering of primary sensory areas in the brain against elevated GCs. The recruitment of other cognitive processes during active decision-making, however, may facilitate GC modulation of mate choosiness, thereby promoting tactical plasticity at this critical life history juncture.

Northern saw-whet owls (Aegolius acadicus) are nocturnal predators that are able to acoustically localize prey with great accuracy; an ability that is attributed to their unique asymmetrical ear structure. While a great deal of research has focused on open loop sound localization prior to flight in owls (primarily barn owls), directional sensitivity of the ears may also be important in locating moving prey on the wing. Furthermore, directionally sensitive ears may also reduce the effects of masking noise, either from the owls’ wings during flight or environmental noise (e.g., wind and leaf rustling), by enhancing spatial segregation of target sounds and noise sources. Here, we investigated auditory processing of Northern saw-whet owls in three-dimensional space using auditory evoked potentials (AEPs). We simultaneously evoked auditory responses in two channels (right and left ear) with broadband clicks from a sound source that could be manipulated in space. Responses were evoked from 66 spatial locations, separated by 30° increments in both azimuth and elevation. We found that Northern saw-whet owls had increased sensitivity to sound sources directly in front of and above their beaks and decreased sensitivity to sound sources below and behind their heads. The spatial region of highest sensitivity extends from the lower beak to the crown of the head and 30° left or right of the median plane, dropping off beyond those margins. Directional sensitivity is undoubtedly useful during foraging and predator evasion, and may also reduce the effect of masking noise from the wings during flight due to the spatial segregation of the noise and targets of interest.

The early to middle Eocene Green River Formation consists of continental strata deposited in Laramide ponded basins in Utah, Colorado, and Wyoming. This study (1) documents fluvial and lacustrine strata from the Douglas Creek and Parachute Creek members of the middle Green River Formation, southeastern Uinta Basin, Utah, and (2) uses new interpretations of the link between climate and fluvial sedimentary expression to interpret the terrestrial evolution of early Eocene climate. The stratigraphy was analyzed via outcrops along a 10 km transect in Main Canyon on the Tavaputs Plateau, and is divided into three distinct, stratigraphically separated depositional settings: (1) the lowermost Interval 1 is dominated by amalgamated sandstone channels that contain 70–100% upper-flow-regime sedimentary structures. The channels are interpreted to represent fluvial deposits controlled by a highly seasonal climate, where most deposition was limited to seasonal flooding events. (2) Interval 2 is dominated by alternating siliciclastic and carbonate lacustrine deposits, interpreted as local pulsed fluvial siliciclastic input into shallow Lake Uinta, and periods of fluvial quiescence represented by littoral carbonate deposition. (3) The uppermost Interval 3 is dominated by erosionally-based, trough cross bedded sandstone channels interbedded with littoral lacustrine and deltaic deposits. The Interval 3 sandstone channels are interpreted as perennial fluvial deposits with relatively little variation in annual discharge, akin to modern humid-temperate fluvial systems. The stratigraphic transition from seasonally controlled (Interval 1) to perennial (Interval 3) fluvial deposits is interpreted to represent a fundamental shift in Eocene climate, from the peak hyperthermal regime of the Early Eocene Climatic Optimum (EECO) to a more stable post-EECO climate.

Monitoring auditory brainstem responses (ABRs) is a common method of assessing auditory processing in non-model species. Although ABRs are widely used to compare auditory abilities across taxa, the extent to which different features of acoustic stimuli affect the ABR is largely unknown in most non-mammalian species. The authors investigated the effects of varying presentation rate and onset time to determine how different features of acoustic stimuli influence the ABR in Northern saw-whet owls (Aegolius acadicus), a species known for their unique auditory adaptations and hunting abilities. At presentation rates ranging from 21.1 to 51.1 s−1, there were no differences in the size or synchrony of ABRs, suggesting that stimuli can be presented at a relatively rapid rate to maximize the number of observations recorded for analysis. While increasing onset time was associated with a decrement in response size and synchrony, tonebursts with 1 ms onset times produced overgeneralized neural responses as a result of spectral splatter. This suggests that 2 to 3 ms onset times may balance the trade-off between response synchrony and frequency specificity when comparing relative neural recruitment across frequencies. These findings highlight the importance of considering stimulus parameters when interpreting ABR data.Monitoring auditory brainstem responses (ABRs) is a common method of assessing auditory processing in non-model species. Although ABRs are widely used to compare auditory abilities across taxa, the extent to which different features of acoustic stimuli affect the ABR is largely unknown in most non-mammalian species. The authors investigated the effects of varying presentation rate and onset time to determine how different features of acoustic stimuli influence the ABR in Northern saw-whet owls (Aegolius acadicus), a species known for their unique auditory adaptations and hunting abilities. At presentation rates ranging from 21.1 to 51.1 s−1, there were no differences in the size or synchrony of ABRs, suggesting that stimuli can be presented at a relatively rapid rate to maximize the number of observations recorded for analysis. While increasing onset time was associated with a decrement in response size and synchrony, tonebursts with 1 ms onset times produced overgeneralized neural responses as a result o...

Successful communication between a sender and a receiver is critical for coordinating behaviors between organisms. This coordination can be disturbed by anthropogenic noise, which has been shown to alter vocal signal production in many species of birds. In addition to affecting senders, noise may also alter reception and behavioral response. Here we investigated the effects of anthropogenic noise on behavioral response to acoustic signals in mixed-species flocks of songbirds. We used playbacks of Tufted Titmouse (Baeolophus bicolor) chick-a-dee calls and local anthropogenic noise to determine how receivers respond to calls with and without added noise. We found that the addition of noise caused a significant decrease of ∼80% in the number of birds that approached the speaker during a chick-a-dee call playback; however, we saw no effect of noise on feeding behavior. Our data support the hypothesis that anthropogenic noise can alter behavioral responses to chick-a-dee calls. This finding is of part...

Summary Background Human monoclonal antibodies that potently and broadly neutralise HIV-1 are under development to prevent and treat HIV-1 infection. In this phase 1 clinical trial we aimed to determine the safety, tolerability, and pharmacokinetic profile of the broadly neutralising monoclonal antibody VRC07-523LS, an engineered variant of VRC01 that targets the CD4 binding site of the HIV-1 envelope protein. Methods This phase 1, open-label, dose-escalation clinical trial was done at the National Institutes of Health Clinical Center in Bethesda, MD, USA. Individuals were recruited from the greater Washington, DC, area by IRB-approved written and electronic media. We enrolled healthy, HIV-1-negative adults aged 18–50 years. Inclusion criteria were good general health, measured through clinical laboratory tests, medical history, and physical examination. Participants self-selected into one of seven open groups during enrolment without randomisation. Four groups received a single intravenous dose of 1, 5, 20, or 40 mg/kg of VRC07-523LS, and one group received a single 5 mg/kg subcutaneous dose. Two groups received three doses of either 20 mg/kg intravenous VRC07-523LS, or 5 mg/kg subcutaneous VRC07-523LS at 12-week intervals. The primary outcome was the safety and tolerability of VRC07-523LS, assessed by dose, route, and number of administrations. This study is registered with ClinicalTrials.gov, NCT03015181. Findings Between Feb 21, 2017, and September 13, 2017, we enrolled 26 participants, including 11 (42%) men and 15 (58%) women. Two (8%) participants withdrew from the study early: one participant in group 1 enrolled in the study but never received VRC07-523LS, and one participant in group 6 chose to withdraw after a single administration. One (4%) participant in group 7 received only one of the three scheduled administrations. 17 participants received intravenous administrations and 8 participants received subcutaneous administrations. VRC07-523LS was safe and well tolerated, we observed no serious adverse events or dose-limiting toxic effects. All reported local and systemic reactogenicity was mild to moderate in severity. The most commonly reported symptoms following intravenous administration were malaise or myalgia in three (18%) participants and headache or chills in two (12%) participants. The most commonly reported symptoms following subcutaneous administration were pain and tenderness in four participants (50%) and malaise or headache in three (38%) participants. Interpretation Safe and well tolerated, VRC07-523LS is a strong and practical candidate for inclusion in HIV-1 prevention and therapeutic strategies. The results from this trial also indicate that an HIV-1 broadly neutralising monoclonal antibody engineered for improved pharmacokinetic and neutralisation properties can be safe for clinical use. Funding National Institutes of Health.

Importance Chikungunya virus (CHIKV) is a mosquito-borneAlphavirusprevalent worldwide. There are currently no licensed vaccines or therapies. Objective To evaluate the safety and tolerability of an investigational CHIKV virus–like particle (VLP) vaccine in endemic regions. Design, Setting, and Participants This was a randomized, placebo-controlled, double-blind, phase 2 clinical trial to assess the vaccine VRC-CHKVLP059-00-VP (CHIKV VLP). The trial was conducted at 6 outpatient clinical research sites located in Haiti, Dominican Republic, Martinique, Guadeloupe, and Puerto Rico. A total of 400 healthy adults aged 18 through 60 years were enrolled after meeting eligibility criteria. The first study enrollment occurred on November 18, 2015; the final study visit, March 6, 2018. Interventions Participants were randomized 1:1 to receive 2 intramuscular injections 28 days apart (20 µg, n = 201) or placebo (n = 199) and were followed up for 72 weeks. Main Outcomes and Measures The primary outcome was the safety (laboratory parameters, adverse events, and CHIKV infection) and tolerability (local and systemic reactogenicity) of the vaccine, and the secondary outcome was immune response by neutralization assay 4 weeks after second vaccination. Results Of the 400 randomized participants (mean age, 35 years; 199 [50%] women), 393 (98%) completed the primary safety analysis. All injections were well tolerated. Of the 16 serious adverse events unrelated to the study drugs, 4 (25%) occurred among 4 patients in the vaccine group and 12 (75%) occurred among 11 patients in the placebo group. Of the 16 mild to moderate unsolicited adverse events that were potentially related to the drug, 12 (75%) occurred among 8 patients in the vaccine group and 4 (25%) occurred among 3 patients in the placebo group. All potentially related adverse events resolved without clinical sequelae. At baseline, there was no significant difference between the effective concentration (EC50)—which is the dilution of sera that inhibits 50% infection in viral neutralization assay—geometric mean titers (GMTs) of neutralizing antibodies of the vaccine group (46; 95% CI, 34-63) and the placebo group (43; 95% CI, 32-57). Eight weeks following the first administration, the EC50 GMT in the vaccine group was 2005 (95% CI, 1680-2392) vs 43 (95% CI, 32-58;P Conclusions and Relevance Among healthy adults in a chikungunya endemic population, a virus-like particle vaccine compared with placebo demonstrated safety and tolerability. Phase 3 trials are needed to assess clinical efficacy. Trial Registration ClinicalTrials.gov Identifier:NCT02562482

Building a better RSV vaccine Respiratory syncytial virus (RSV) causes severe respiratory disease, especially in infants and the elderly. However, attempts to produce effective human vaccines have largely been unsuccessful. Structure-based design has been used to generate an RSV fusion glycoprotein stabilized in its prefusion conformation (DS-Cav1). This immunogen is highly effective in mice and macaques. Crank et al. now report the results of a phase I vaccine clinical trial using the stabilized prefusion DS-Cav1 molecule. Four weeks after immunization, these vaccines elicited substantially more high-quality antibody titers than those typically generated using earlier RSV immunogens. The findings provide a proof of concept for how structural biology can contribute to precision vaccine design. Science , this issue p. 505

In seasonal breeders, there are behavioral, endocrine, and neural adaptations that promote the sexual receptivity of females and tune their sensory systems to detect and discriminate among advertising males and to successfully copulate. What happens immediately after this key life history event is unclear, but this transitional moment offers a window into the mechanisms that remodel sexual phenotypes. In this study of wild female Cope's gray treefrogs (Hyla chrysoscelis), we tested the hypothesis that oviposition results in a suite of coordinated changes in the sexual phenotype. Specifically, we predicted that sexual receptivity and discrimination behaviors would decline along with circulating concentrations of steroid hormones (corticosterone, estradiol, testosterone) and auditory sensitivity to the acoustic frequencies emphasized in male advertisement calls. We conducted these trait measurements before and after oviposition (ca. 24-h period). There was a 100% decrease in behavioral responsiveness after oviposition, and the concentrations of all three steroids plummeted during this brief window of time, especially testosterone. Moreover, higher concentrations of corticosterone—an important component of the endocrine stress response—were associated with longer response latencies, suggesting that adrenal hormones should be considered in future studies on the hormonal basis of mate choice. Counter to our prediction, auditory sensitivity increased following oviposition, and the amplitude of the auditory brainstem response was influenced by concentrations of estradiol. In pre-oviposition females auditory sensitivity diminished with increasing estradiol concentrations, while sensitivity increased with increasing estradiol concentrations in post-oviposition females, suggesting non-linear estrogenic modulation of peripheral auditory neural recruitment. Overall, our results indicate that there is considerable remodeling of behavioral output following oviposition that co-occurs with changes in both endocrine and sensory physiology.

The black-capped chickadee is a songbird that has been used extensively as a model of animal communication in field and laboratory settings. Although many studies have focused on the complex call and song systems of the black-capped chickadee, relatively fewer studies have focused on chickadee audition. However, we do know from behavioral and molecular work that chickadees (and auditory processing areas in their brains) discriminate between artificially generated tones, between conspecific and heterospecific vocalizations, and among different types of conspecific vocalizations. In this paper we investigate peripheral auditory processing of frequency in the black-capped chickadee and the potential influence of sex on frequency sensitivity using a technique called auditory evoked potentials. We found that male and female black-capped chickadees did not differ in any measure of frequency sensitivity. Both sexes had the greatest sensitivity to frequencies between 2 and 4 kHz. This range of frequencies is well represented in black-capped chickadee song, partially supporting the idea that sender and receiver coevolve. Finally, we suggest that the call and song system of North American parids make them an ideal taxonomic group for comparative work exploring the relationship between call systems and the evolution of auditory processing.

Songs mediate mate attraction and territorial defence in songbirds during the breeding season. Outside of the breeding season, the avian vocal repertoire often includes calls that function in foraging, antipredator and social behaviours. Songs and calls can differ substantially in their spectral and temporal content. Given seasonal variation in the vocal signals, the sender–receiver matching hypothesis predicts seasonal changes in auditory processing that match the physical properties of songs during the breeding season and calls outside of it. We tested this hypothesis in white-breasted nuthatches, Sitta carolinensis, tufted titmice, Baeolophus bicolor, and Carolina chickadees, Poecile carolinensis. We measured the envelope-following response (EFR), which quantifies phase locking to the amplitude envelope, and the frequency-following response (FFR), which quantifies phase locking to the temporal fine structure of sounds. Because songs and calls of nuthatches are amplitude modulated at different rates, we predicted seasonal changes in EFRs that match the rates of amplitude fluctuation in songs and calls. In chickadees and titmice, we predicted stronger FFRs during the spring and stronger EFRs during the winter because songs are tonal and calls include amplitude-modulated elements. In all three species, we found seasonal changes in EFRs and FFRs. EFRs varied across seasons and matched the amplitude modulations of songs and calls in nuthatches. In addition, female chickadees had stronger EFRs in the winter than in the spring. In all three species, FFRs during the spring tended to be stronger in females than in males. We also found species differences in EFRs and FFRs in both seasons; EFRs and FFRs tended to be higher in nuthatches than in chickadees and titmice. We discuss the potential mechanisms underlying seasonality in EFRs and FFRs and the implications of our results for communication during the breeding season and outside of it, when these three species form mixed-species flocks.

Recombinant adenoviral vectors (rAds) are lead vaccine candidates for protection against a variety of pathogens, including Ebola, HIV, tuberculosis, and malaria, due to their ability to potently induce T cell immunity in humans. However, the ability to induce protective cellular immunity varies among rAds. Here, we assessed the mechanisms that control the potency of CD8 T cell responses in murine models following vaccination with human-, chimpanzee-, and simian-derived rAds encoding SIV-Gag antigen (Ag). After rAd vaccination, we quantified Ag expression and performed expression profiling of innate immune response genes in the draining lymph node. Human-derived rAd5 and chimpanzee-derived chAd3 were the most potent rAds and induced high and persistent Ag expression with low innate gene activation, while less potent rAds induced less Ag expression and robustly induced innate immunity genes that were primarily associated with IFN signaling. Abrogation of type I IFN or stimulator of IFN genes (STING) signaling increased Ag expression and accelerated CD8 T cell response kinetics but did not alter memory responses or protection. These findings reveal that the magnitude of rAd-induced memory CD8 T cell immune responses correlates with Ag expression but is independent of IFN and STING and provide criteria for optimizing protective CD8 T cell immunity with rAd vaccines.

Northern saw-whet owls (Aegolius acadicus) are known for their unique asymmetrical ear structure and ability to localize prey acoustically, yet few attempts have been made to explore the auditory capabilities of this species. In this study, we evoked auditory brainstem responses (ABRs) with tonebursts to assess three main hypotheses regarding the evolution of auditory sensitivity: sender-receiver matching, ecological constraints, and phylogenetic/morphological constraints. We found that ABR amplitude increased with increasing stimulus level, which is consistent with results in other avian species. ABR amplitudes, latencies, and thresholds indicate that the hearing range of Northern saw-whet owls extends from 0.7 to 8.6 kHz, with an extended frequency range of best sensitivity between 1.6 and 7.1 kHz. Sensitivity fell off rapidly above and below these frequencies. The average audiogram was structurally similar to those found in other species of owls, suggesting that phylogeny or morphology may be constraining the frequency range of auditory sensitivity. However, ABR thresholds were 10–25 dB lower than those of Eastern screech-owls (Megascops asio), with thresholds below 0 dB SPL in some individuals. The lowest thresholds were at frequencies not found in the vocalizations of Northern saw-whet owls, suggesting ecological constraints rather than conspecific vocalizations are driving absolute sensitivity.

Summary The evolution of vocal signals can be constrained by a host of factors including habitat effects on sound propagation, morphology of sound-producing structures and phylogenetic relationships among species. Here, we asked whether auditory sensitivity over a broad range of frequencies correlates with the spectral content of conspecific vocalizations, or whether it is constrained by the overall structure of vocalizations, habitat effects on sound propagation or relatedness among species. We studied nine New World sparrows (Passeriformes: Emberizidae) including three open-habitat species, three scrub-like habitat species and three forest species. For each habitat, one species had pure-trilled songs, another had tonal songs and another had complex songs with tones, trills and amplitude-modulated buzzes. As predicted by the acoustic adaptation hypothesis, song spectral properties (specifically frequency and entropy) had the highest values in open-habitat species and the lowest values in forest species. Based on our results from song analyses, and the sender–receiver matching hypothesis, we predicted that open-habitat species would be more sensitive to high-frequency sounds compared to forest species. Contrary to this prediction, habitat and high-frequency song content had little effect on audiogram shape. Song type, however, had a strong effect, with species that produce complex songs showing higher sensitivity to high-frequency sounds than all other species. Our results suggest that the use of song frequency by receivers depends on song structure and not necessarily on song spectral content. Therefore, our current understanding of how signal-processing mechanisms should match signal properties appears to be too simple. When thinking about the evolution of signal-processing mechanisms, the multidimensionality of signals, and how the different dimensions can interact, should be considered.

We have generated hexon-modified adenovirus serotype 5 (Ad5) vectors that are not neutralized by Ad5-specific neutralizing antibodies in mice. These vectors are attractive for the advancement of vaccine products because of their potential for inducing robust antigen-specific immune responses in people with prior exposure to Ad5. However, hexon-modified Ad5 vectors displayed an approximate 10-fold growth defect in complementing cells, making potential vaccine costs unacceptably high. Replacing hypervariable regions (HVRs) 1, 2, 4, and 5 with the equivalent HVRs from Ad43 was sufficient to avoid Ad5 preexisting immunity and retain full vaccine potential. However, the resulting vector displayed the same growth defect as the hexon-modified vector carrying all 9 HVRs from Ad43. The growth defect is likely due to a defect in capsid assembly, since DNA replication and late protein accumulation were normal in these vectors. We determined that the hexon-modified vectors have a 32°C cold-sensitive phenotype and selected revertants that restored vector productivity. Genome sequencing identified a single base change resulting in a threonine-to-methionine amino acid substitution at the position equivalent to residue 342 of the wild-type protein. This mutation has a suppressor phenotype (SP), since cloning it into our Ad5 vector containing all nine hypervariable regions from Ad43, Ad5.H(43m-43), increased yields over the version without the SP mutation. This growth improvement was also shown for an Ad5-based hexon-modified vector that carried the hexon hypervariable regions of Ad48, indicating that the SP mutation may have broad applicability for improving the productivity of different hexon-modified vectors.

Inosine triphosphate (ITP) pyrophosphohydrolase, or ITPase, is an intracellular enzyme that is responsible for the hydrolysis of the acidic anhydride bond between the alpha and beta phosphates in ITP, and other noncanonical nucleoside triphosphates, producing the corresponding nucleoside monophosphate and pyrophosphate. This activity protects the cell by preventing noncanonical nucleoside triphosphates from accumulating in (deoxy) nucleoside triphosphate ((d)NTP) pools and/or being integrated into nucleic acids. This enzyme is encoded by the ITPA gene in mammals. It has been reported that Itpa homozygous-null knock-out mice die before weaning and have gross cardiac abnormalities. Additionally, certain variations in the human ITPA gene have been linked to adverse reactions to the immunosuppressive prodrugs azathioprine and 6-mercaptopurine and protection against ribavirin-induced hemolytic anemia. These drugs are bioactivated to form noncanonical nucleoside triphosphates. Human ITPase enzymes engineered to modulate nucleobase specificity may be valuable tools for studying the role of ITPase in heart development and drug metabolism or developing gain-of-function mutants or inhibitory molecules. Based on x-ray crystallography and amino acid sequence data, a panel of putative human ITPase nucleobase specificity mutants has been generated. We targeted eight highly conserved amino acid positions within the ITPase sequence that correspond to amino acids predicted to directly interact with the nucleobase or help organize the nucleobase binding pocket. The ability of the mutants to protect against exogenous and endogenous noncanonical purines was tested with two Escherichia coli complementation assays. Nucleobase specificity of the mutants was investigated with an in vitro biochemical assay using ITP, GTP and ATP as substrates. This methodology allowed us to identify gain-of-function mutants and categorize the eight amino acid positions according to their ability to protect against noncanonical purines as follows: Glu-22, Trp-151 and Arg-178, essential for protection; Phe-149, Asp-152, Lys-172 and Ser-176, intermediate protection; His-177, dispensable for protection against noncanonical purines.

Recombinant adenoviral vectors (rAds) are the most potent recombinant vaccines for eliciting CD8+ T cell–mediated immunity in humans; however, prior exposure from natural adenoviral infection can decrease such responses. In this study we show low seroreactivity in humans against simian- (sAd11, sAd16) or chimpanzee-derived (chAd3, chAd63) compared with human-derived (rAd5, rAd28, rAd35) vectors across multiple geographic regions. We then compared the magnitude, quality, phenotype, and protective capacity of CD8+ T cell responses in mice vaccinated with rAds encoding SIV Gag. Using a dose range (1 × 107–109 particle units), we defined a hierarchy among rAd vectors based on the magnitude and protective capacity of CD8+ T cell responses, from most to least, as: rAd5 and chAd3, rAd28 and sAd11, chAd63, sAd16, and rAd35. Selection of rAd vector or dose could modulate the proportion and/or frequency of IFN-γ+TNF-α+IL-2+ and KLRG1+CD127−CD8+ T cells, but strikingly ∼30–80% of memory CD8+ T cells coexpressed CD127 and KLRG1. To further optimize CD8+ T cell responses, we assessed rAds as part of prime-boost regimens. Mice primed with rAds and boosted with NYVAC generated Gag-specific responses that approached ∼60% of total CD8+ T cells at peak. Alternatively, priming with DNA or rAd28 and boosting with rAd5 or chAd3 induced robust and equivalent CD8+ T cell responses compared with prime or boost alone. Collectively, these data provide the immunologic basis for using specific rAd vectors alone or as part of prime-boost regimens to induce CD8+ T cells for rapid effector function or robust long-term memory, respectively.

We studied how age affects selection of foraging microhabitats, foraging behaviors, and rates of vigilance in the black phoebe (Sayornis nigricans). Juveniles generally perched higher in trees than adults and in areas with higher intensity of light, which may increase the difficulty of visually tracking prey. Juveniles allocated more effort to visual searching prior to a foraging attempt and flew farther than adults to capture a prey item. When at rest on a perch, juveniles monitored the environment more slowly than adults, suggesting lower levels of vigilance. In general, our results suggest that differences between ages in foraging behavior of visually guided predators may be influenced by intensity of ambient light in their foraging territories.

The ability of a receiver to detect a signal is a product of the signal characteristics at the sender, habitat-specific degradation of the signal, and properties of the receiver's sensory system. Active space describes the maximum distance at which a receiver with a given sensory system can detect a signal in a given habitat. Here the effect of habitat structure and urbanization on brown-headed cowbird (Molothrus ater) perched song active space was explored. The active space of the cowbird song was affected by both habitat type and level of urbanization. High frequency (4 to 6 kHz) portions of song resulted in the maximum active space. Surprisingly, the active space was the largest in open urban environments. The hard surfaces found in open urban areas (e.g., sidewalks, buildings) may provide a sound channel that enhances song propagation. When the introductory phrase and final phrase were analyzed separately, the active space of the introductory phrase was found to decrease in open urban environments but the active space of the final phrase increased in open urban environments. This suggests that different portions of the vocalization may be differentially influenced by habitat and level of urbanization.

The RV144 trial demonstrated that an experimental AIDS vaccine can prevent human immunodeficiency virus type 1 (HIV-1) infection in humans. Because of its limited efficacy, further understanding of the mechanisms of preventive AIDS vaccines remains a priority, and nonhuman primate (NHP) models of lentiviral infection provide an opportunity to define immunogens, vectors, and correlates of immunity. In this study, we show that prime-boost vaccination with a mismatched SIV envelope (Env) gene, derived from simian immunodeficiency virus SIVmac239, prevents infection by SIVsmE660 intrarectally. Analysis of different gene-based prime-boost immunization regimens revealed that recombinant adenovirus type 5 (rAd5) prime followed by replication-defective lymphocytic choriomeningitis virus (rLCMV) boost elicited robust CD4 and CD8 T-cell and humoral immune responses. This vaccine protected against infection after repetitive mucosal challenge with efficacies of 82% per exposure and 62% cumulatively. No effect was seen on viremia in infected vaccinated monkeys compared to controls. Protection correlated with the presence of neutralizing antibodies to the challenge viruses tested in peripheral blood mononuclear cells. These data indicate that a vaccine expressing a mismatched Env gene alone can prevent SIV infection in NHPs and identifies an immune correlate that may guide immunogen selection and immune monitoring for clinical efficacy trials.

The aircraft industry, as a whole, has been deeply concerned with improving the aerodynamic efficiency of current and future flight vehicles, particularly in the commercial and military markets. However, of particular interest to the field of aerodynamics is the elusive concept of a workable flow control mechanism. Effective flow control is a concept which if properly applied can increase aerodynamic efficiency. Various concepts and ideas to obtain successful flow control have been studied in an attempt to reap these rewards. Some examples include boundary layer blowing (steady and periodic), suction, and synthetic jets. The overall goal of flow control is to increase performance. The specific objectives of flow control include: 1) delay or eliminate flow separation, 2) delay boundary layer transition or 3) reduce skin friction drag. The purpose of this research is to investigate dynamic surface roughness as a novel method of flow control technology for external boundary layer flows. As opposed to standard surface roughness, dynamic roughness incorporates small time dependent perturbations to the surface of the airfoil. These surface perturbations are actual humps and/or ridges that are on the scale of the laminar boundary layer, and oscillate with an unsteady motion. Research has shown that this can provide a means to modify the instantaneous and mean velocity profile near the wall and favorably control the existing state of the boundary layer. The results of this study have shown that dynamic roughness can be a viable alternative in delaying and/or eliminating the leading edge laminar separation bubble and hence reaping some of the rewards of an effective flow control system, while also maintaining some physical advantages over other techniques.

Brown-headed cowbirds, Molothrus ater , and red-winged blackbirds, Agelaius phoeniceus , are frequently used as model systems for communication and vocal learning; however, relatively little is known about their auditory processing. Although auditory space and acoustic signal space are assumed to coevolve, it is less clear when, or even if, sex differences are expected in auditory processing. Here we explored frequency-specific auditory responses using auditory brainstem responses (ABRs). We generated sex-specific audiograms using two methods: visual detection and cross-correlation. We also measured the amplitude and latency of ABRs to stimuli over a range of frequencies (0.5–6 kHz) and intensities (8–72 dB sound pressure level). Thresholds generated by both methods were good predictors of behavioural thresholds in the best frequency range (2–4 kHz). The visual detection method resulted in a slightly better correspondence with behavioural thresholds than the cross-correlation method above and below best frequencies. We found no significant differences between red-winged blackbird and brown-headed cowbird ABR thresholds; however, overall, females had lower thresholds than males. Sex differences were larger than species differences for ABR amplitude, while latency differences were greater between species than between sexes. Taken together our results suggest that despite high-frequency vocal elements, brown-headed cowbirds do not have enhanced high-frequency auditory sensitivity when measured with auditory evoked potentials. Moreover, we show that females have greater auditory sensitivity than males, which is generally not seen in other passerine species.

Recombinant adenoviruses (rAds) based on types 5 (rAd5) and 35 (rAd35) have emerged as important vaccine delivery vectors in clinical testing for a variety of pathogens. A major difference between these vectors is their binding to cellular receptors used for infection. Whereas rAd5 binds coxsackie-adenovirus receptor (CAR), rAd35 binds the complement regulatory protein CD46. Although rAd35 infected and phenotypically matured human blood dendritic cells (DCs) more efficiently than rAd5, we show here that rAd35 markedly suppressed DC-induced activation of naive CD4 + T cells. rAd35 specifically blocked both DCs and anti-CD3/CD28 mAb-induced naive T-cell proliferation and IL-2 production. This effect was also observed in CD4 + memory T cells but to a lesser extent. The suppression occurred by rAd35 binding to CD46 on T cells and was independent of infection. CD46 engagement with mAb mimicked the effects of rAd35 and also led to deficient NF-κB nuclear translocation. In contrast, rAd5 and rAd35 vectors with ablated CD46 binding did not inhibit T-cell activation. Our findings provide insights into the basic biology of adenoviruses and indicate that CD46 binding may have an impact on the generation of primary CD4 + T-cell responses by Ad35.

Predator–prey interactions are regulated by the ability of individuals to detect, and then approach or avoid, each other. In visually guided organisms, the prevalent view is that predators have large binocular visual fields and high acuity, whereas prey have wide lateral areas and low acuity, which could affect vigilance behaviour. We characterized the configuration of the visual system (visual fields, retinal topography, visual acuity) and vigilance behaviour (head movement rate) of two ground-foraging avian prey (white-crowned sparrow, Zonotrichia leucophrys, California towhee, Pipilo crissalis) with laterally placed eyes. We found that the binocular field of both species (45°) was actually wider than those of some of their avian predators. Both species also had a single retinal specialization (high ganglion cell density area) located in the centro-temporal sector of the retina, which projected into the lateral and frontal part of the head. Wide binocular fields may increase binocular contrast to detect and visually guide the bill towards prey items. Both species had wider lateral visual fields and faster head movement rates than some of their predators, probably to enhance detection and visual tracking of predators. California towhees made faster sideways movements of the head than did white-crowned sparrows, probably to cover visual space more quickly with their retinal specialization because of the comparatively lower spatial resolution of their retinal periphery. Alternatively, California towhees might move their heads more rapidly to monitor for potential risks (e.g. competitors, predators), as they rely mostly on personal information because of their degree of territoriality. Our findings suggest that the visual system and vigilance behaviour of these two avian prey species combine traits to enhance predator detection through large visual coverage and fast head movements, but also to enhance food detection at close range through enhanced binocular vision.