High-Content Imaging for Large-Scale Detection of Low-Affinity Extracellular Protein Interactions.

Extracellular protein interactions coordinate cellular responses with their local environment and have important roles in pathogen invasion and disease. Due to technical challenges associated with studying binding events at the cell surface, the systematic and reliable identification of novel ligand-receptor pairs remains difficult. Here, we describe the development of a cell-based assay using large-scale transient transfections and high-content imaging (HCI) to detect extracellular binding events. We optimized the parameters for efficient transfection of human cells with cDNA plasmids encoding full-length cell surface receptors in 384-well plates. Using a range of well-characterized structurally diverse low-affinity cell surface interactions, we show that transfected cells probed with highly avid ligands can be used to successfully identify ligand-receptor pairs using an HCI platform and automated image analysis software. To establish the high-throughput potential of this approach, we also screened a pool of ligands against a collection of 2455 cell surface expression clones and found that known ligand-receptor interactions could be robustly and consistently detected across the library using this technology

Using Spectral and Cross-Spectral Analysis to Identify Patterns and Synchrony in Couples\u27 Sexual Desire

Sexual desire discrepancy is one of the most frequently reported sexual concerns for individuals and couples and has been shown to be negatively associated with sexual and relationship satisfaction. Sexual desire has increasingly been examined as a state-like construct that ebbs and flows, but little is known about whether there are patterns in the fluctuation of sexual desire. Utilizing spectral and cross-spectral analysis, we transformed 30 days of dyadic daily diary data for perceived levels of sexual desire for a non-clinical sample of 133 couples (266 individuals) into the frequency domain to identify shared periodic state fluctuations in sexual desire. Spectral analysis is a technique commonly used in physics and engineering that allows time series data to be analyzed for the presence of regular cycles of fluctuation. Cross-spectral analysis allows for dyadic data to be analyzed for shared rates of fluctuation between partners as well as the degree of (a)synchrony (or phase shift) between these fluctuations. Men and women were found to exhibit fluctuations in sexual desire at various frequencies including rates of once and twice per month, and to have sexual desire that was unlikely to fluctuate over periods of three days or less and therefore exhibited persistence. Similar patterns of fluctuation were exhibited within couples and these patterns were found to be largely synchronous. While instances of desire discrepancy may arise due to differences in rates of sexual desire fluctuation and random fluctuations, such instances may be normal for romantic relationships. The results have important implications for researchers, clinicians, and educators in that they corroborate the supposition that sexual desire ebbs and flows and suggest that it does so with predictable regularity

Spatial and temporal dynamics and value of nature-based recreation, estimated via social media

Conserved lands provide multiple ecosystem services, including opportunities for naturebased recreation. Managing this service requires understanding the landscape attributes underpinning its provision, and how changes in land management affect its contribution to human wellbeing over time. However, evidence from both spatially explicit and temporally dynamic analyses is scarce, often due to data limitations. In this study, we investigated nature-based recreation within conserved lands in Vermont, USA.We used geotagged photographs uploaded to the photo-sharingwebsite Flickr to quantify visits by in-state and outof- state visitors, and we multiplied visits by mean trip expenditures to show that conserved lands contributed US $1.8 billion (US$0.18\u2720.2 at 95% confidence) to Vermont\u27s tourism industry between 2007 and 2014.We found eight landscape attributes explained the pattern of visits to conserved lands; visits were higher in larger conserved lands, with less forest cover, greater trail density and more opportunities for snow sports. Some of these attributes differed from those found in other locations, but all aligned with our understanding of recreation in Vermont.We also found that using temporally static models to informconservation decisions may have perverse outcomes for nature-based recreation. For example, static models suggest conserved land with less forest cover receive more visits, but temporally dynamic models suggest clearing forests decreases, rather than increases, visits to these sites. Our results illustrate the importance of understanding both the spatial and temporal dynamics of ecosystem services for conservation decision-making

Rebuilding a foundation of trust: A call to action in creating a safe environment for everyone

Well before the COVID-19 pandemic, incivility and physical threats directed toward healthcare employees and often registered nurses was a growing concern by Chief Nurse Executive (CNE) leaders. In 2019, conversations initiated by The Beryl Institute’s Nurse Executive Council (NEC) to consider how best to achieve a much-needed balance between patient/family and staff safety have now become a critical priority to ensure the safety of everyone receiving and providing health care services. The heart of this work was organized around a set of newly developed ethical precepts designed to guide the exploration of key concepts. A call to action grounded in rebuilding a foundation of trust is proposed. In pursuing future steps to deepen this foundation, and to reaffirm the vital role for nurse leaders and of all in healthcare, we must be willing to engage in dialogue, to ask openly, and question respectfully. We believe healthcare systems and nurse leaders both play a vital role in elevating the humanity on which we will find brighter days ahead. Through a sustained commitment to this aim, we seek to strengthen health care delivery environments that shape physically and psychologically safe environments for everyone. Experience Framework This article is associated with the Culture & Leadership lens of The Beryl Institute Experience Framework (https://www.theberylinstitute.org/ExperienceFramework). Access other PXJ articles related to this lens. Access other resources related to this len

Effects of 30-, 60-, and 90-Day Bed Rest on Postural Control in Men and Women

INTRODUCTION Head-down-tilt bed rest (HDT) has been used as a safe gr ound-based analog to mimic and develop countermeasures for the physiological effects of spaceflight, including decrements in postural stability. The purpose of this investigation was to characterize the effects of 30-, 60-, and 90-day bed rest on postural control in men and women. METHODS Twenty-nine subjects (18M,11F) underwent 13 days of ambula tory acclimatization and were placed in 6? HDT for 30 (n=12), 60 (n=8), or 90 (n=9) days, followed by 14 days of ambulatory recovery. Computerized dynamic posturography (CDP) was used to assess changes in sensory and motor components of postural control, and recovery after HDT. Sensory Organization Tests (SOTs) objectively evaluate one?s ability to effectively use or suppress visual, vestibular, and proprioceptive information for postural control. Stability during the SOTs was assessed using peak-to-peak sway and convergence toward stability limits to derive an equilibrium score. Motor Control Tests (MCTs) evaluate one?s ability to recover from unexpected support surface perturbations, with performance determined by center-of-pressure path length. Whole-body kinematic data were collected to determine body-sway strategy used to maintain stability during each condition. Baselines were determined pre-HDT. Recovery was tracked post-HDT on days 0, 1, 2, and 4. RESULTS Immediately after HDT, subjects showed decreased performance on most SOTs, primarily on sway-referenced support conditions, typically returning to baseline levels within 4 days. MCT performance was not significantly affected. There were no significant gender or duration differences in performance. Kinematic data revealed a tendency to use ankle strategy to maintain an upright stance during most SOT conditions. Interestingly, six subjects (2M,4F) experienced orthostatic intolerance and were unable to complete day 0 testing. CONCLUSION HDT mimics some un loading mechanisms of spaceflight and elicits orthostatic issues present post-spaceflight (contributing to instability); however, it does not sufficiently address the vestibular dysfunction which occurs post-spaceflight

Microperimetry hill of vision and volumetric measures of retinal sensitivity

Purpose: Mean retinal sensitivity is the main output measure used in microperimetry. It is, however, of limited use in patients with poor vision because averaging is weighted toward zero in those with significant scotomas creating an artificial floor effect. In contrast, volumetric measures avoid these issues and are displayed graphically as a hill of vision. Methods: An open-source program was created to manipulate raw sensitivity threshold data files obtained from MAIA microperimetry. Thin plate spline interpolated heat maps and three-dimensional hill of vision plots with an associated volume were generated. Retrospective analyses of microperimetry volumes were undertaken in patients with a range of retinal diseases to assess the qualitative benefits of three-dimensional visualization and volumetric measures. Simulated pathology was applied to radial grid patterns to investigate the performance of volumetric sensitivity in nonuniform grids. Results: Volumetric analyses from microperimetry in RPGR-related retinitis pigmentosa, choroideremia, Stargardt disease, and age-related macular degeneration were analyzed. In simulated nonuniform testing grids, volumetric sensitivity was able to detect differences in retinal sensitivity where mean sensitivity could not. Conclusions: Volumetric measures do not suffer from averaging issues and demonstrate superior performance in nonuniform testing grids. Additionally, volume measures enable detection of localized retinal sensitivity changes that might otherwise be undetectable in a mean change. Translational Relevance: As microperimetry has become an outcome measure in several gene-therapy clinical trials, three-dimensional visualization and volumetric sensitivity enables a complementary analysis of baseline disease characteristics and subsequent response to treatment, both as a signal of safety and efficacy

Global change and conservation triage on National Wildlife Refuges

National Wildlife Refuges (NWRs) in the United States play an important role in the adaptation of social-ecological systems to climate change, land-use change, and other global-change processes. Coastal refuges are already experiencing threats from sea-level rise and other change processes that are largely beyond their ability to influence, while at the same time facing tighter budgets and reduced staff. We engaged in workshops with NWR managers along the U.S. Atlantic coast to understand the problems they face from global-change processes and began a multidisciplinary collaboration to use decision science to help address them. We are applying a values-focused approach to base management decisions on the resource objectives of land managers, as well as those of stakeholders who may benefit from the goods and services produced by a refuge. Two insights that emerged from our workshops were a conspicuous mismatch between the scale at which management can influence outcomes and the scale of environmental processes, and the need to consider objectives related to ecosystem goods and services that traditionally have not been explicitly considered by refuges (e.g., protection from storm surge). The broadening of objectives complicates the decision-making process, but also provides opportunities for collaboration with stakeholders who may have agendas different from those of the refuge, as well as an opportunity for addressing problems across scales. From a practical perspective, we recognized the need to (1) efficiently allocate limited staff time and budgets for short-term management of existing programs and resources under the current refuge design and (2) develop long-term priorities for acquiring or protecting new land/habitat to supplement or replace the existing refuge footprint and thus sustain refuge values as the system evolves over time. Structuring the decision-making problem in this manner facilitated a better understanding of the issues of scale and suggested that a long-term solution will require a significant reassessment of objectives to better reflect the comprehensive values of refuges to society. We discuss some future considerations to integrate these two problems into a single framework by developing novel optimization approaches for dynamic problems that account for uncertainty in future conditions

A mixed methods approach to advance the understanding of physical activity behaviour during pregnancy

An active pregnancy encompasses benefits that apply to both mother and baby. However, before investing resources in interventions aimed at changing or supporting expectant mothers’ physical activity behaviours, it is necessary to have a greater fundamental understanding of the modifiable factors influencing women’s decisions during this time. Therefore, the main purpose of this study was to investigate the socio-cognitive determinants of physical activity during pregnancy. A mixed methods research approach using a multiphase design framed by the Theory of Planned Behaviour (TPB; Ajzen, 1991, Organizational Behavior and Human Decision Processes, 50, 179-211) guided the investigation. Following ethical approval and permission to carry out the study, participants were recruited from randomly selected antenatal clinics in the East Kent region of England. Application of the TPB required both qualitative and quantitative cross-sectional data to be collected. Study One (n = 18) involved the elicitation of pregnant women’s beliefs towards being physically active. The modal salient behavioural, normative and control beliefs were used to inform the development of a TPB questionnaire which was implemented during the next phase of the investigation. Study Two (n = 78) examined the predictive utility of the TPB in explaining pregnant women’s physical activity intentions and behaviour and scrutinised the role of past behaviour within this context. Finally, Study Three (n = 10) comprised semi-structured interviews involving community midwives. Meta-inferences maintained the function of improving the validity of the study and advancing the overall aim of the research project. Specifically, triangulation was used to ascertain convergence, corroboration, and correspondence of findings between studies. Whilst results of the individual studies hold merit, meta-inferences point towards the development of three main themes: (1) pregnant women lack access to the necessary information that would allow them to make informed decisions regarding their engagement in physical activities; (2) a co-ordinated effort involving interprofessional collaboration is required to support pregnant women in overcoming barriers associated with regular physical activity participation; and (3) profiling pregnant women according to motivation and behaviour status could serve as a useful and manageable starting point for intervening to produce positive changes in pregnant women’s physical activity behaviour. In normalising physical activity during pregnancy and achieving better health outcomes for mothers and babies, it is necessary to consider the modifiable factors involved in behaviour change, identify opportune moments to intervene, and involve a network of professionals in facilitating and supporting pregnant women’s engagement with physical activities

Multifaceted Role for p53 in Pancreatic Cancer Suppression

The vast majority of human pancreatic ductal adenocarcinomas (PDACs) harbor TP53 mutations, underscoring p53\u27s critical role in PDAC suppression. PDAC can arise when pancreatic acinar cells undergo acinar-to-ductal metaplasia (ADM), giving rise to premalignant pancreatic intraepithelial neoplasias (PanINs), which finally progress to PDAC. The occurrence of TP53 mutations in late-stage PanINs has led to the idea that p53 acts to suppress malignant transformation of PanINs to PDAC. However, the cellular basis for p53 action during PDAC development has not been explored in detail. Here, we leverage a hyperactive p53 variant-p5353,54-which we previously showed is a more robust PDAC suppressor than wild-type p53, to elucidate how p53 acts at the cellular level to dampen PDAC development. Using both inflammation-induced and KRASG12D-driven PDAC models, we find that p5353,54 both limits ADM accumulation and suppresses PanIN cell proliferation and does so more effectively than wild-type p53. Moreover, p5353,54 suppresses KRAS signaling in PanINs and limits effects on the extracellular matrix (ECM) remodeling. While p5353,54 has highlighted these functions, we find that pancreata in wild-type p53 mice similarly show less ADM, as well as reduced PanIN cell proliferation, KRAS signaling, and ECM remodeling relative to Trp53-null mice. We find further that p53 enhances chromatin accessibility at sites controlled by acinar cell identity transcription factors. These findings reveal that p53 acts at multiple stages to suppress PDAC, both by limiting metaplastic transformation of acini and by dampening KRAS signaling in PanINs, thus providing key new understanding of p53 function in PDAC