A Systematic Review of Community Engagement in the US Environmental Protection Agency's Extramural Research Solicitations: Implications for Research Funders.

OBJECTIVES: We systematically reviewed the Environmental Protection Agency, National Center for Environmental Research's (NCER's) requests for applications (RFAs) and identified strategies that NCER and other funders can take to bolster community engagement. METHODS: We queried NCER's publically available online archive of funding opportunities from fiscal years 1997 to 2013. From an initial list of 211 RFAs that met our inclusion criteria, 33 discussed or incorporated elements of community engagement. We examined these RFAs along 6 dimensions and the degree of alignments between them. RESULTS: We found changes over time in the number of RFAs that included community engagement, variations in how community engagement is defined and expected, inconsistencies between application requirements and peer review criteria, and the inclusion of mechanisms supporting community engagement in research. CONCLUSIONS: The results inform a systematic approach to developing RFAs that support community engagement in research.

Community-Based Review of Research Across Diverse Community Contexts: Key Characteristics, Critical Issues, and Future Directions.

A growing number of community-based organizations and community-academic partnerships are implementing processes to determine whether and how health research is conducted in their communities. These community-based research review processes (CRPs) can provide individual and community-level ethics protections, enhance the cultural relevance of study designs and competence of researchers, build community and academic research capacity, and shape research agendas that benefit diverse communities. To better understand how they are organized and function, representatives of 9 CRPs from across the United States convened in 2012 for a working meeting. In this article, we articulated and analyzed the models presented, offered guidance to communities that seek to establish a CRP, and made recommendations for future research, practice, and policy.

Ovarian imaging by magnetic resonance in adolescent girls with polycystic ovary syndrome and age-matched controls.

PURPOSE: To compare ovarian morphology in adolescent girls with and without polycystic ovary syndrome (PCOS) using magnetic resonance imaging (MRI). MATERIALS AND METHODS: In 21 adolescent girls (age 12-18 years) without and 19 adolescents with PCOS (diagnosis based on excessive hair growth and irregular menstrual cycles) ovarian volume, antral follicle count (AFC) per ovary, and follicle size were evaluated. MRI was performed at 1.5 T or 3 T and axial or angled-axial single-shot echo-train spin echo images of 6 mm slice thickness were acquired. In a subset of subjects, 2-mm images were also obtained. PCOS and non-PCOS groups were compared using mixed affects regression. RESULTS: Mean AFC per ovary and ovarian volume were substantially greater in PCOS subjects compared to non-PCOS subjects. Mean follicle size was similar between groups. Follicles exceeding 10 mm were seen in 2/19 PCOS subjects versus 9/21 non-PCOS subjects. Consistently higher follicle counts were detected in images obtained at 2 mm compared to 6-mm slice thickness. CONCLUSION: In adolescence, MRI of the ovary reveals distinct differences between girls with and without PCOS. MRI may help evaluate young patients in whom transvaginal ultrasound is contraindicated.

In vitromagnetohydrodynamics system for modulating cell migration.

Fluid shear stress (FSS) is an important parameter that regulates various cell functions such as proliferation and migration. While there are a number of techniques to generate FSSin vitro, many of them require physical deformation or movement of solid objects to generate the fluid shear, making it difficult to decouple the effects of FSS and mechanical strains. This work describes the development of a non-mechanical means to generate fluid flow and FSS in a 2Din vitrosetting. This was accomplished with a magnetohydrodynamic (MHD) pump, which creates liquid flow by generating a Lorentz force through the interaction between an electric field and an orthogonal magnetic field. The MHD pump system presented here consisted of trapezoidal prism-shaped magnets, a pair of platinum electrodes, and a modified petri dish. The system was validated and tested on anin vitrowound model, which is based on analyzing the migration of fibroblast cells through an artificially created scratch on a confluent cell culture surface. Experiments were performed to a control group, an electric field only group, and a group that was subject to fluid flow with the application of both electric field and magnetic field. Results show that fibroblast cells that experienced fluid shear have higher wound closure rate compared to the control group and the electric field only group. The data shows that the MHD pump can be a great tool to study FSSin vitro. Furthermore, due to its fluid flow generation without mechanical force, this system can be adapted and implemented to study the role of FSS and electric field on wound healingin vivo.

Bioanalytical Methods for Characterization of CAR-T Cellular Kinetics: Comparison of PCR Assays and Matrices.

Recently, multiple chimeric antigen receptor T-cell (CAR-T)-based therapies have been approved for treating hematological malignancies, targeting CD19 and B-cell maturation antigen. Unlike protein or antibody therapies, CAR-T therapies are "living cell" therapies whose pharmacokinetics are characterized by expansion, distribution, contraction, and persistence. Therefore, this unique modality requires a different approach for quantitation compared with conventional ligand binding assays implemented for most biologics. Cellular (flow cytometry) or molecular assays (polymerase chain reaction (PCR)) can be deployed with each having unique advantages and disadvantages. In this article, we describe the molecular assays utilized: quantitative PCR (qPCR), which was the initial platform used to estimate transgene copy numbers and more recently droplet digital PCR (ddPCR) which quantitates the absolute copy numbers of CAR transgene. The comparability of the two methods in patient samples and of each method across different matrices (isolated CD3+ T-cells or whole blood) was also performed. The results show a good correlation between qPCR and ddPCR for the amplification of same gene in clinical samples from a CAR-T therapy trial. In addition, our studies show that the qPCR-based amplification of transgene levels was well-correlated, independent of DNA sources (either CD3+ T-cells or whole blood). Our results also highlight that ddPCR can be a better platform for monitoring samples at the early phase of CAR-T dosing prior to expansion and during long-term monitoring as they can detect samples with very low copy numbers with high sensitivity, in addition to easier implementation and sample logistics.

Moving From Transactional to Relational: How Funders Can Work in Partnership With Black, Indigenous and People of Color Communities.

BACKGROUND: The Racial Equity Coalition (REC) formed to address persistent educational disparities. The coalition was composed of 14 Black, Indigenous and People of Color (BIPOC) organizations that provide culturally integrative youth services. OBJECTIVES: REC, with support from United Way of King County, engaged in participatory research to identify commonalities and shared struggles to inform collective action. Participatory research aligns with REC's commitment to equitable participatory processes. This article focuses on REC's experiences with funders. The objective was to understand what creates positive and challenging experiences with funders, and to identify recommendations for funders to become more culturally responsive. METHODS: A research committee was formed including representatives of nine REC organizations and United Way of King County staff. The committee conducted interviews with each of the 14 REC organizations and conducted thematic analysis of interview transcripts. Through participatory analysis, the committee drafted narratives that were further refined through a series of research retreats attended by all REC organizations. RESULTS: Recommendations were to incentivize collaboration, listen to communities to create culturally responsive definitions of success and measurement strategies, arrive at mutually agreed upon approaches with organizations, honor the connections BIPOC organizations have with their communities, and provide unrestricted funding to allow BIPOC organizations greater agency. CONCLUSIONS: A major challenge for BIPOC organizations is navigating White dominant culture that too often shows up in funding requirements. Having to fit dominant culture standards stifles BIPOC organizations' abilities to meet community needs and the responsiveness of their approaches. REC identified recommendations for funders to be more culturally responsive and community centered.

Zebrafish Tsc1 reveals functional interactions between the cilium and the TOR pathway.

The cell surface organelle called the cilium is essential for preventing kidney cyst formation and for establishing left-right asymmetry of the vertebrate body plan. Recent advances suggest that the cilium functions as a sensory organelle in vertebrate cells for multiple signaling pathways such as the hedgehog and the Wnt pathways. Prompted by kidney cyst formation in tuberous sclerosis complex (TSC) patients and rodent models, we investigated the role of the cilium in the TSC-target of rapamycin (TOR) pathway using zebrafish. TSC1 and TSC2 genes are causal for TSC, and their protein products form a complex in the TOR pathway that integrates environmental signals to regulate cell growth, proliferation and survival. Two TSC1 homologs were identified in zebrafish, which we refer to as tsc1a and tsc1b. Morpholino knockdown of tsc1a led to a ciliary phenotype including kidney cyst formation and left-right asymmetry defects. Tsc1a was observed to localize to the Golgi, but morpholinos against it, nonetheless, acted synthetically with ciliary genes in producing kidney cysts. Consistent with a role of the cilium in the same pathway as Tsc genes, the TOR pathway is aberrantly activated in ciliary mutants, resembling the effect of tsc1a knockdown. Moreover, kidney cyst formation in ciliary mutants was blocked by the Tor inhibitor, rapamycin. Surprisingly, we observed elongation of cilia in tsc1a knockdown animals. Together, these data suggest a signaling network between the cilium and the TOR pathway in that ciliary signals can feed into the TOR pathway and that Tsc1a regulates the length of the cilium itself.

Intraflagellar transport proteins are essential for cilia formation and for planar cell polarity.

The highly conserved intraflagellar transport (IFT) proteins are essential for cilia formation in multiple organisms, but surprisingly, cilia form in multiple zebrafish ift mutants. Here, we detected maternal deposition of ift gene products in zebrafish and found that ciliary assembly occurs only during early developmental stages, supporting the idea that maternal contribution of ift gene products masks the function of IFT proteins during initial development. In addition, the basal bodies in multiciliated cells of the pronephric duct in ift mutants were disorganized, with a pattern suggestive of defective planar cell polarity (PCP). Depletion of pk1, a core PCP component, similarly led to kidney cyst formation and basal body disorganization. Furthermore, we found that multiple ift genes genetically interact with pk1. Taken together, these data suggest that IFT proteins play a conserved role in cilia formation and planar cell polarity in zebrafish.

Adherence to the 2014 American Academy of Pediatrics palivizumab prophylaxis recommendations.

INTRODUCTION: This study aims to describe adherence rates to the 2014 American Academy of Pediatrics (AAP) Committee on Infectious Disease guidance document recommending which patients should receive palivizumab for prophylaxis against respiratory syncytial virus (RSV). METHODS: A retrospective, single-center analysis of patients who received at least one dose of palivizumab between October 1, 2012, and March 1, 2017 was conducted. Data collected included demographics, medical history, palivizumab administration regimens, and incidence of RSV infection. RESULTS: Data were collected on 457 patients who received palivizumab over five RSV seasons. Approximately half of the patients (45% and 55%, respectively) received palivizumab according to the AAP recommendations in place at the time (2012 or 2014 recommendations, respectively). One percent of patients had a breakthrough RSV infection after receiving at least one dose of palivizumab. There was no significant difference in the number of breakthrough infections before and after the 2014 recommendations were released (3 vs. 2). CONCLUSIONS: Approximately half of the patients received prophylaxis in accordance with the 2014 AAP recommendations and infrequently suffered from a breakthrough RSV infection.

Genetic evidence for a role for protein kinase A in the maintenance of sleep and thalamocortical oscillations.

STUDY OBJECTIVES: Genetic manipulation of cAMP-dependent protein kinase A (PKA) in Drosophila has implicated an important role for PKA in sleeplwake state regulation. Here, we characterize the role of this signaling pathway in the regulation of sleep using electroencephalographic (EEG) and electromyographic (EMG) recordings in R(AB) transgenic mice that express a dominant negative form of the regulatory subunit of PKA in neurons within cortex and hippocampus. Previous studies have revealed that these mutant mice have reduced PKA activity that results in the impairment of hippocampus-dependent long-term memory and long-lasting forms of hippocampal synaptic plasticity. DESIGN: PKA assays, in situ hybridization, immunoblots, and sleep studies were performed in R(AB) transgenic mice and wild-type control mice. MEASUREMENTS AND RESULTS: We have found that R(AB) transgenic mice have reduced PKA activity within cortex and reduced Ser845 phosphorylation of the glutamate receptor subunit GluR1. R(AB) transgenic mice exhibit non-rapid eye movement (NREM) sleep fragmentation and increased amounts of rapid eye movement (REM) sleep relative to wild-type mice. Further, R(AB) transgenic mice have more delta power but less sigma power during NREM sleep relative to wild-type mice. After sleep deprivation, the amounts of NREM and REM sleep were comparable between wild-type and R(AB) transgenic mice. However, the homeostatic rebound of sigma power in R(AB) transgenic mice was reduced. CONCLUSIONS: Alterations in cortical synaptic receptors, impairments in sleep continuity, and alterations in sleep oscillations in R(AB) mice imply that PKA is involved not only in synaptic plasticity and memory storage but also in the regulation of sleep/wake states.