Three-Dimensional Neurophenotyping of Adult Zebrafish Behavior

The use of adult zebrafish (Danio rerio) in neurobehavioral research is rapidly expanding. The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes. Here, we generated temporal and spatial three-dimensional (3D) reconstructions of zebrafish locomotion, globally assessed behavioral profiles evoked by several anxiogenic and anxiolytic manipulations, mapped individual endpoints to 3D reconstructions, and performed cluster analysis to reconfirm behavioral correlates of high- and low-anxiety states. The application of 3D swim path reconstructions consolidates behavioral data (while increasing data density) and provides a novel way to examine and represent zebrafish behavior. It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature. Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior

Cdk5 Is Required for Memory Function and Hippocampal Plasticity via the cAMP Signaling Pathway

Memory formation is modulated by pre- and post-synaptic signaling events in neurons. The neuronal protein kinase Cyclin-Dependent Kinase 5 (Cdk5) phosphorylates a variety of synaptic substrates and is implicated in memory formation. It has also been shown to play a role in homeostatic regulation of synaptic plasticity in cultured neurons. Surprisingly, we found that Cdk5 loss of function in hippocampal circuits results in severe impairments in memory formation and retrieval. Moreover, Cdk5 loss of function in the hippocampus disrupts cAMP signaling due to an aberrant increase in phosphodiesterase (PDE) proteins. Dysregulation of cAMP is associated with defective CREB phosphorylation and disrupted composition of synaptic proteins in Cdk5-deficient mice. Rolipram, a PDE4 inhibitor that prevents cAMP depletion, restores synaptic plasticity and memory formation in Cdk5-deficient mice. Collectively, our results demonstrate a critical role for Cdk5 in the regulation of cAMP-mediated hippocampal functions essential for synaptic plasticity and memory formation.Norman B. Leventhal FellowshipUnited States. National Institutes of Health (NIH T32 MH074249)United States. National Institutes of Health (NIH RO1 NS051874

Enacting molecular complexity : data and health in the metabonomics laboratory

In this dissertation, I examine how biological data practices enable researchers to interact with and enact biological life in statistical ways, and how this poses challenges to the use and integration of biological knowledge with clinical practices. Instead of considering data as a pre-existing cognitive representation of the world, I combine scholarship on the anthropology of science with scholarship from science and technology studies to consider data as a form of material practice. I consider, in other words, how data is intertwined with technologies, people, and values, such that data is used to make normative and naturalized claims about biology and disease. To explore the generation, interpretation, and use of biological data, I focus on the field of “metabonomics”—the post-genomic study of metabolism—as it is carried out within the Biomolecular Medicine Laboratory (BMM) at Imperial College London. In doing so, I examine how metabonomics researchers use biochemical techniques and multivariate statistics to investigate metabolism and disease. After providing an overview of the literature, central questions, and methodology that frame this dissertation, I examine how multivariate statistical practices are central to the historical identity and epistemic culture of metabonomics research at the BMM. From there, I demonstrate how multivariate statistics require and enable metabonomics to enact metabolism as an inherently complex entity. Consequently, I examine how researchers struggle to assign the categories of “normal” and “abnormal” to dynamic notions of metabolism and health. I then explore how the translation of metabonomics knowledge into clinical practices places value on multivariate forms and large volumes of information, eclipsing the importance of human interpretation and judgment. Finally, I examine how metabonomics research is used to develop personalized medicine, but in ways that make it difficult to address the health of individual patients.</p

Comparison of up-front cash cards and checks as incentives for participation in a clinician survey: a study within a trial.

BackgroundEvidence is needed regarding effective incentive strategies to increase clinician survey response rates. Cash cards are increasingly used as survey incentives; they are appealing because of their convenience and because in some cases their value can be reclaimed by investigators if not used. However, their effectiveness in clinician surveys is not known. In this study within the BRCA Founder OutReach (BFOR) study, a clinical trial of population-based BRCA1/2 mutation screening, we compared the use of upfront cash cards requiring email activation versus checks as clinician survey incentives.MethodsParticipants receiving BRCA1/2 testing in the BFOR study could elect to receive their results from their primary care provider (PCP, named by the patient) or from a geneticist associated with the study. In order to understand PCPs' knowledge, attitudes, experiences and willingness to disclose results we mailed paper surveys to the first 501 primary care providers (PCPs) in New York, Boston, Los Angeles and Philadelphia who were nominated by study participants to disclose their BRCA1/2 mutation results obtained through the study. We used alternating assignment stratified by city to assign the first 303 clinicians to receive a $50 up-front incentive as a cash card (N = 155) or check (N = 148). The cash card required PCPs to send an activation email in order to be used. We compared response rates by incentive type, adjusting for PCP characteristics and study site.ResultsIn unadjusted analyses, PCPs who received checks were more likely to respond to the survey than those who received cash cards (54.1% versus 41.9%, p = 0.046); this remained true when we adjusted for provider characteristics (OR for checks 1.61, 95% CI 1.01, 2.59). No other clinician characteristics had a statistically significant association with response rates in adjusted analyses. When we included an interaction term for incentive type and city, the favorable impact of checks on response rates was evident only in Los Angeles and Philadelphia.ConclusionsAn up-front cash card incentive requiring email activation may be less effective in eliciting clinician responses than up-front checks. However, the benefit of checks for clinician response rates may depend on clinicians' geographic location.Trial registrationClinicalTrials.gov ( NCT03351803 ), November 24, 2017

Comparison of up-front cash cards and checks as incentives for participation in a clinician survey: a study within a trial.

BackgroundEvidence is needed regarding effective incentive strategies to increase clinician survey response rates. Cash cards are increasingly used as survey incentives; they are appealing because of their convenience and because in some cases their value can be reclaimed by investigators if not used. However, their effectiveness in clinician surveys is not known. In this study within the BRCA Founder OutReach (BFOR) study, a clinical trial of population-based BRCA1/2 mutation screening, we compared the use of upfront cash cards requiring email activation versus checks as clinician survey incentives.MethodsParticipants receiving BRCA1/2 testing in the BFOR study could elect to receive their results from their primary care provider (PCP, named by the patient) or from a geneticist associated with the study. In order to understand PCPs' knowledge, attitudes, experiences and willingness to disclose results we mailed paper surveys to the first 501 primary care providers (PCPs) in New York, Boston, Los Angeles and Philadelphia who were nominated by study participants to disclose their BRCA1/2 mutation results obtained through the study. We used alternating assignment stratified by city to assign the first 303 clinicians to receive a $50 up-front incentive as a cash card (N = 155) or check (N = 148). The cash card required PCPs to send an activation email in order to be used. We compared response rates by incentive type, adjusting for PCP characteristics and study site.ResultsIn unadjusted analyses, PCPs who received checks were more likely to respond to the survey than those who received cash cards (54.1% versus 41.9%, p = 0.046); this remained true when we adjusted for provider characteristics (OR for checks 1.61, 95% CI 1.01, 2.59). No other clinician characteristics had a statistically significant association with response rates in adjusted analyses. When we included an interaction term for incentive type and city, the favorable impact of checks on response rates was evident only in Los Angeles and Philadelphia.ConclusionsAn up-front cash card incentive requiring email activation may be less effective in eliciting clinician responses than up-front checks. However, the benefit of checks for clinician response rates may depend on clinicians' geographic location.Trial registrationClinicalTrials.gov ( NCT03351803 ), November 24, 2017

Challenges and Opportunities in Engaging Primary Care Providers in BRCA Testing: Results from the BFOR Study

PurposeEngaging primary care providers (PCPs) in BRCA1/2 testing and results disclosure would increase testing access. The BRCA Founder OutReach (BFOR) study is a prospective study of BRCA1/2 founder mutation screening among individuals of Ashkenazi Jewish descent that sought to involve participants' PCPs in results disclosure. We used quantitative and qualitative methods to evaluate PCPs' perspectives, knowledge, and experience disclosing results in BFOR.MethodsAmong PCPs nominated by BFOR participants to disclose BRCA1/2 results, we assessed the proportion agreeing to disclose. To examine PCP's perspectives, knowledge, and willingness to disclose results, we surveyed 501 nominated PCPs. To examine PCPs' experiences disclosing results in BFOR, we surveyed 101 PCPs and conducted 10 semi-structured interviews.ResultsIn the BFOR study overall, PCPs agreed to disclose their patient's results 40.5% of the time. Two hundred thirty-four PCPs (46.7%) responded to the initial survey. Responding PCPs were more likely to agree to disclose patients' results than non-responders (57.3% vs. 28.6%, p&lt;0.001). Among all respondents, most felt very (19.7%) or somewhat (39.1%) qualified to share results. Among PCPs declining to disclose, insufficient knowledge was the most common reason. In multivariable logistic regression, feeling qualified was the only variable significantly associated with agreeing to disclose results (OR 6.53, 95% CI 3.31, 12.88). In post-disclosure surveys (response rate=55%), PCPs reported largely positive experiences. Interview findings suggested that although PCPs valued the study-provided educational materials, they desired better integration of results and decision support into workflows.ConclusionBarriers exist to incorporating BRCA1/2 testing into primary care. Most PCPs declined to disclose their patients' BFOR results, although survey respondents were motivated and had positive disclosure experiences. PCP training and integrated decision support could be beneficial.Trial registrationClinicalTrials.gov (NCT03351803), November 24, 2017

Integrating Multidisciplinary Observations in Vent Environments (IMOVE): Decadal Progress in Deep-Sea Observatories at Hydrothermal Vents

The unique ecosystems and biodiversity associated with mid-ocean ridge (MOR) hydrothermal vent systems contrast sharply with surrounding deep-sea habitats, however both may be increasingly threatened by anthropogenic activity (e.g., mining activities at massive sulphide deposits). Climate change can alter the deep-sea through increased bottom temperatures, loss of oxygen, and modifications to deep water circulation. Despite the potential of these profound impacts, the mechanisms enabling these systems and their ecosystems to persist, function and respond to oceanic, crustal, and anthropogenic forces remain poorly understood. This is due primarily to technological challenges and difficulties in accessing, observing and monitoring the deep-sea. In this context, the development of deep-sea observatories in the 2000s focused on understanding the coupling between sub-surface flow and oceanic and crustal conditions, and how they influence biological processes. Deep-sea observatories provide long-term, multidisciplinary time-series data comprising repeated observations and sampling at temporal resolutions from seconds to decades, through a combination of cabled, wireless, remotely controlled, and autonomous measurement systems. The three existing vent observatories are located on the Juan de Fuca and Mid-Atlantic Ridges (Ocean Observing Initiative, Ocean Networks Canada and the European Multidisciplinary Seafloor and water column Observatory). These observatories promote stewardship by defining effective environmental monitoring including characterizing biological and environmental baseline states, discriminating changes from natural variations versus those from anthropogenic activities, and assessing degradation, resilience and recovery after disturbance. This highlights the potential of observatories as valuable tools for environmental impact assessment (EIA) in the context of climate change and other anthropogenic activities, primarily ocean mining. This paper provides a synthesis on scientific advancements enabled by the three observatories this last decade, and recommendations to support future studies through international collaboration and coordination. The proposed recommendations include: i) establishing common global scientific questions and identification of Essential Ocean Variables (EOVs) specific to MORs, ii) guidance towards the effective use of observatories to support and inform policies that can impact society, iii) strategies for observatory infrastructure development that will help standardize sensors, data formats and capabilities, and iv) future technology needs and common sampling approaches to answer today’s most urgent and timely questions