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Chromatin immunoprecipitation sequencing (ChIP-seq) and the Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) have become essential technologies to effectively measure protein-DNA interactions and chromatin accessibility. However, there is a need for a scalable and reproducible pipeline that incorporates proper normalization between samples, correction of copy number variations, and integration of new downstream analysis tools. Here we present Containerized Bioinformatics workflow for Reproducible ChIP/ATAC-seq Analysis (CoBRA), a modularized computational workflow which quantifies ChIP-seq and ATAC-seq peak regions and performs unsupervised and supervised analyses. CoBRA provides a comprehensive state-of-the-art ChIP-seq and ATAC-seq analysis pipeline that can be used by scientists with limited computational experience. This enables researchers to gain rapid insight into protein-DNA interactions and chromatin accessibility through sample clustering, differential peak calling, motif enrichment, comparison of sites to a reference database, and pathway analysis. CoBRA is publicly available online at https://bitbucket.org/cfce/cobra.

Following the discovery of heart regeneration in zebrafish, several more species within the Cyprinidae family have been found to have the same capability, suggesting heart regeneration may be conserved within this family. Although gonad regeneration has been observed in grass carp (Ctenopharyngodon idella), one of the largest cyprinid fish, the species’ response to cardiac injury has not been characterized. Surprisingly, we found cardiomyocytes do not repopulate the injured region following cryoinjury to the ventricle, instead exhibiting unresolved fibrosis and decreased cardiac function that persists for the 8-week duration of this study. Compared to other cyprinid fish studied, infiltration of macrophages is delayed and muted in this model. Additionally, fibroblasts are depleted following injury, a phenomenon not previously described in any cardiac model. This study shows that heart regeneration is not conserved among the Cyprinidae family and suggests the important role of non-fibroblasts in chronic fibrosis. Further study of these phenomenon may reveal the underlying differences between regeneration versus unresolved fibrosis in heart disease.Summary statementGrass carp, a member of the Cyprinidae family that includes regenerative zebrafish, do not regenerate functional cardiac tissue after cryoinjury. Instead, healing progresses through collagen deposition and scar formation.

Mechanistic understanding of the interaction of copper-based nanomaterials with crops is crucial for exploring their application in precision agriculture and their implications on plant health. We investigated the biological response of soybean (Glycine max) plants to the foliar application of copper hydroxide nanowires (CNWs) at realistic exposure concentrations. A commercial copper based-fungicide (Kocide), dissolved copper ions, and untreated controls were used for comparison to identify unique features at physiological, cellular, and molecular levels. After 32 d of exposure to CNW (0.36, 1.8, and 9 mg CNW/plant), the newly developed tissues accumulated significantly high levels of Cu (18-60 μg/g) compared to Kocide (10 μg/g); however, the rate of Cu translocation from the site of CNW treatment to other tissues was slower compared to other Cu treatments. Like Kocide, CNW exposure at medium and high doses altered Co, Mn, Zn, and Fe accumulation in the tissues and enhanced photosynthetic activities. The proteomic and metabolomic analyses of leaves from CNW-treated soybean plants suggest a dose-dependent response, resulting in the activation of major biological processes, including photosynthesis, energy production, fatty acid metabolism, lignin biosynthesis, and carbohydrate metabolism. In contrast to CNW treatments, Kocide exposure resulted in increased oxidative stress response and amino acid metabolism activation.

Nitrogen oxides (NOx = NO + NO2) play a crucial role in the formation of ozone and secondary inorganic and organic aerosols, thus affecting human health, global radiation budget, and climate. The diurnal and spatial variations in NO2 are functions of emissions, advection, deposition, vertical mixing, and chemistry. Their observations, therefore, provide useful constraints in our understanding of these factors. We employ a Regional chEmical and trAnsport model (REAM) to analyze the observed temporal (diurnal cycles) and spatial distributions of NO2 concentrations and tropospheric vertical column densities (TVCDs) using aircraft in situ measurements and surface EPA Air Quality System (AQS) observations as well as the measurements of TVCDs by satellite instruments (OMI: the Ozone Monitoring Instrument; GOME-2A: Global Ozone Monitoring Experiment – 2A), ground-based Pandora, and the Airborne Compact Atmospheric Mapper (ACAM) instrument in July 2011 during the DISCOVER-AQ campaign over the Baltimore–Washington region. The model simulations at 36 and 4 km resolutions are in reasonably good agreement with the regional mean temporospatial NO2 observations in the daytime. However, we find significant overestimations (underestimations) of model-simulated NO2 (O3) surface concentrations during nighttime, which can be mitigated by enhancing nocturnal vertical mixing in the model. Another discrepancy is that Pandora-measured NO2 TVCDs show much less variation in the late afternoon than simulated in the model. The higher-resolution 4 km simulations tend to show larger biases compared to the observations due largely to the larger spatial variations in NOx emissions in the model when the model spatial resolution is increased from 36 to 4 km. OMI, GOME-2A, and the high-resolution aircraft ACAM observations show a more dispersed distribution of NO2 vertical column densities (VCDs) and lower VCDs in urban regions than corresponding 36 and 4 km model simulations, likely reflecting the spatial distribution bias of NOx emissions in the National Emissions Inventory (NEI) 2011.

Nitrogen oxides (NO(x)=NO+NO(2)) play a crucial role in the formation of ozone and secondary inorganic and organic aerosols, thus affecting human health, global radiation budget, and climate. The diurnal and spatial variations in NO(2) are functions of emissions, advection, deposition, vertical mixing, and chemistry. Their observations, therefore, provide useful constraints in our understanding of these factors. We employ a Regional chEmical and trAnsport model (REAM) to analyze the observed temporal (diurnal cycles) and spatial distributions of NO(2) concentrations and tropospheric vertical column densities (TVCDs) using aircraft in situ measurements and surface EPA Air Quality System (AQS) observations as well as the measurements of TVCDs by satellite instruments (OMI: the Ozone Monitoring Instrument; GOME-2A: Global Ozone Monitoring Experiment – 2A), ground-based Pandora, and the Airborne Compact Atmospheric Mapper (ACAM) instrument in July 2011 during the DISCOVER-AQ campaign over the Baltimore–Washington region. The model simulations at 36 and 4 km resolutions are in reasonably good agreement with the regional mean temporospatial NO(2) observations in the daytime. However, we find significant overestimations (underestimations) of model-simulated NO(2) (O(3)) surface concentrations during night-time, which can be mitigated by enhancing nocturnal vertical mixing in the model. Another discrepancy is that Pandora-measured NO(2) TVCDs show much less variation in the late afternoon than simulated in the model. The higher-resolution 4 km simulations tend to show larger biases compared to the observations due largely to the larger spatial variations in NO(x) emissions in the model when the model spatial resolution is increased from 36 to 4 km. OMI, GOME-2A, and the high-resolution aircraft ACAM observations show a more dispersed distribution of NO(2) vertical column densities (VCDs) and lower VCDs in urban regions than corresponding 36 and 4 km model simulations, likely reflecting the spatial distribution bias of NO(x) emissions in the National Emissions Inventory (NEI) 2011.

Abiotic conditions can influence the effect that herbivores have on plant growth. Such biotic and abiotic interactions are of special interest in plant biological control programs because the goal of herbivore suppression of the target weed may not be reached in some abiotic settings. Additionally, target invasive plants typically occur across diverse landscapes raising the possibility that local adaptation to site-specific conditions leads to phenotypic variation that can affect herbivore responses. Here, we used Tamarix, an invasive plant, and its associated biological control agent, Diorhabda carinulata, to investigate how local variation in soil salinity and host plant origin influence interactions between the two taxa. To test if Tamarix was adapted to local conditions, we collected plants from sites with either low or high groundwater salinity, asexually propagated them through multiple generations, and then treated them with their home or reciprocal salinity levels. We found that plants accumulated the most biomass when grown at the salinities of their origin site. The biological control agent, D. carinulata preferred plants grown at source site salinity when given a choice against plants grown in the reciprocal salinity treatment. Although plants compensated for herbivory by regrowing foliage over three defoliation events and maintained similar leaf biomass through regrowth, they ultimately had a reduced basal area and 62% lower root biomass compared to the controls. Thus, herbivory caused a shift in plant allocation of resources from overall growth to compensation, reducing root and stem investment. Overall, D. carinulata caused a significantly greater reduction in total biomass in the high salinity plants than the low salinity ones when grown at their source salinity (averages of 63% and 32% respectively). Thus, the Tamarix biological control program may experience its greatest efficacy in high salinity areas where the impact of the agent is the greatest, likely due to increased water stress and reduced resources to enable regrowth.

CDK4/6 inhibitors are approved to treat breast cancer and are in trials for other malignancies. We examined CDK4/6 inhibition in mouse and human CD8+ T cells during early stages of activation. Mice receiving tumor-specific CD8+ T cells treated with CDK4/6 inhibitors displayed increased T-cell persistence and immunologic memory. CDK4/6 inhibition upregulated MXD4, a negative regulator of MYC, in both mouse and human CD8+ T cells. Silencing of Mxd4 or Myc in mouse CD8+ T cells demonstrated the importance of this axis for memory formation. We used single-cell transcriptional profiling and T-cell receptor clonotype tracking to evaluate recently activated human CD8+ T cells in patients with breast cancer before and during treatment with either palbociclib or abemaciclib. CDK4/6 inhibitor therapy in humans increases the frequency of CD8+ memory precursors and downregulates their expression of MYC target genes, suggesting that CDK4/6 inhibitors in patients with cancer may augment long-term protective immunity. Significance: CDK4/6 inhibition skews newly activated CD8+ T cells toward a memory phenotype in mice and humans with breast cancer. CDK4/6 inhibitors may have broad utility outside breast cancer, particularly in the neoadjuvant setting to augment CD8+ T-cell priming to tumor antigens prior to dosing with checkpoint blockade. This article is highlighted in the In This Issue feature, p. 2355

Small cell lung cancer (SCLC) is a high-grade neuroendocrine cancer that accounts for ~15% of lung cancers. While nearly all SCLCs are genetically driven by near universal loss of function (LOF) mutations in RB1 and TP53; several recent studies show that there are different phenotypic SCLC molecular subtypes characterized by expression of lineage transcription factors. These include the neuroendocrine ASCL1 and NEUROD1 subtypes which together comprise ~70-80% of SCLCs. Initially subtypes were thought to be mutually exclusive, but recent evidence shows intra-tumoral subtype heterogeneity and plasticity between subtypes. A recent study found that 35-40% of human SCLCs express both ASCL1 and NEUROD1, but the mechanisms driving ASCL1 and NEUROD1 intra-tumoral heterogeneity are not well understood. My laboratory previously developed an autochthonous CRISPR-based SCLC genetically-engineered mouse model (GEMM) generated by intratracheally injecting adenoviruses encoding Cre recombinase and sgRNAs targeting Rb1, Trp53, and Rbl2. Cre turns on Cas9 expression and allows for CRISPR/Cas9 editing of Rb1, Trp53, and Rbl2 in somatic cells in the lungs. The unique advantage of this model is that it allows the inclusion of sgRNAs targeting additional genes of interest in the same adenovirus. Using this CRISPR-based autochthonous SCLC GEMM approach, we studied the consequences of inactivating the epigenetic modifier KDM6A during SCLC tumorigenesis. KDM6A functions as an H3K27 histone demethylase and also exists in the COMPASS complex with KMT2C/D to promote H3K4 mono-/di-methylation at enhancers. KDM6A along with its protein binding partner KMT2D are mutated in SCLC and KDM6A has been implicated in controlling differentiation in other lineages. Strikingly, we found that KDM6A inactivation in SCLC GEMMs induced plasticity from ASCL1 to NEUROD1 resulting in SCLC tumors that expressed both ASCL1 and NEUROD1. ATAC-sequencing showed open chromatin at the promoters of NEUROD1 and NEUROD1 target genes in KDM6A inactivated tumors. Interestingly, KDM6A inactivated tumors showed a spectrum of ASCL1 to NEUROD1 heterogeneity where some KDM6A inactivated tumors completely lost ASCL1 and solely expressed NEUROD1, some tumors expressed ASCL1 and NEUROD1 in a mutually exclusive manner, while others primarily expressed ASCL1 with very few NEUROD1 positive cells. Mechanistically, KDM6A binds and maintains ASCL1 target genes in an active chromatin state with its loss increasing H3K27me3 near both promoters and enhancers, and decreasing H3K4me1/2 at enhancers together leading to a cell state primed for ASCL1 to NEUROD1 subtype switching. This work identifies KDM6A as an epigenetic regulator that controls ASCL1 to NEUROD1 subtype plasticity and provides an autochthonous SCLC GEMM to model ASCL1 and NEUROD1 subtype heterogeneity, which is found in 35-40% of human SCLCs. Citation Format: Leslie Duplaquet, Yixiang Li, Matthew A. Booker, Yingtian Xie, Radhika A. Patel, Deli Hong, Thomas Denize, Emily Walton, Yasmin N. Laimon, Roderick Bronson, Jackson Southard, Shuqiang Li, Sabina Signoretti, Michael Y. Tolstorukov, Paloma Cejas, Henry W. Long, Michael C. Haffner, Matthew G. Oser. Small cell lung cancer subtype plasticity is regulated by KDM6A. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5774.

ObjectiveOesophageal squamous cell carcinoma (OSCC), like other squamous carcinomas, harbour highly recurrent cell cycle pathway alterations, especially hyperactivation of the CCND1/CDK4/6 axis, raising the potential for use of existing CDK4/6 inhibitors in these cancers. Although CDK4/6 inhibition has shown striking success when combined with endocrine therapy in oestrogen receptor positive breast cancer, CDK4/6 inhibitor palbociclib monotherapy has not revealed evidence of efficacy to date in OSCC clinical studies. Herein, we sought to elucidate the identification of key dependencies in OSCC as a foundation for the selection of targets whose blockade could be combined with CDK4/6 inhibition.DesignWe combined large-scale genomic dependency and pharmaceutical screening datasets with preclinical cell line models, to identified potential combination therapies in squamous cell cancer.ResultsWe identified sensitivity to inhibitors to the ERBB family of receptor kinases, results clearly extending beyond the previously described minority of tumours with EGFR amplification/dependence, specifically finding a subset of OSCCs with dual dependence on ERBB3 and ERBB2. Subsequently. we demonstrated marked efficacy of combined pan-ERBB and CDK4/6 inhibition in vitro and in vivo. Furthermore, we demonstrated that squamous lineage transcription factor KLF5 facilitated activation of ERBBs in OSCC.ConclusionThese results provide clear rationale for development of combined ERBB and CDK4/6 inhibition in these cancers and raises the potential for KLF5 expression as a candidate biomarker to guide the use of these agents. These data suggested that by combining existing Food and Drug Administration (FDA)-approved agents, we have the capacity to improve therapy for OSCC and other squamous cancer.

Prostate cancers are considered to be immunologically ‘cold’ tumors given the very few patients who respond to checkpoint inhibitor (CPI) therapy. Recently, enrichment of interferon-stimulated genes (ISGs) predicted a favorable response to CPI across various disease sites. The enhancer of zeste homolog-2 (EZH2) is overexpressed in prostate cancer and known to negatively regulate ISGs. In the present study, we demonstrate that EZH2 inhibition in prostate cancer models activates a double-stranded RNA–STING–ISG stress response upregulating genes involved in antigen presentation, Th1 chemokine signaling and interferon response, including programmed cell death protein 1 (PD-L1) that is dependent on STING activation. EZH2 inhibition substantially increased intratumoral trafficking of activated CD8(+) T cells and increased M1 tumor-associated macrophages, overall reversing resistance to PD-1 CPI. Our study identifies EZH2 as a potent inhibitor of antitumor immunity and responsiveness to CPI. These data suggest EZH2 inhibition as a therapeutic direction to enhance prostate cancer response to PD-1 CPI.

Introduction Automated text messaging programs have been studied as a treatment tool, but have not been studied as an outreach tool to increase the reach of smoking cessation treatment. Aims and Methods Two distinct text messaging programs were developed. One was aimed at connecting smokers to quitline phone counseling via text message (Text4Coach [T4C]) and the other was aimed at connecting smokers to a smoking cessation text messaging program (Text&Quit [T&Q]). Adult daily smokers with Medicaid insurance (N = 80) were recruited from the Emergency Department at an urban hospital and randomized to T4C or T&Q. The primary outcome was program reach. Results Outreach text messages were found to have moderately high uptake, with the majority of participants (63.8%) opting into their assigned tobacco treatment program and younger and female participants more likely to opt in (p < .01). Receipt of the treatment portion of the program differed among the programs with 67.5% of T&Q receiving the treatment program and 27.5% of T4C receiving the program (p < .001). Most participants across both groups replied to at least one message (71.3%) and very few unsubscribed from the service over the 3-week trial. The majority of participants reported overall satisfaction with their program (63.8%), found it helpful for quitting smoking (60.0%) and would recommend the program to a friend (62.5%). Overall, 11 (13.8%) participants reported being abstinent from smoking for the past 7 days at follow-up, with no differences between groups. Conclusions Outreach text messages were found to have moderately high reach among Medicaid smokers. Larger trials are needed to evaluate the impact of such programs on helping low-income smokers quit. Implications Automated text messaging programs have been tested as a treatment tool, but have not been tested as an outreach tool to increase the reach of smoking cessation treatment. This study tests a new way of conducting outreach to smokers in a health system through text messages. It tests the effect of outreach on (1) rates of opting in and (2) successful treatment delivery. Results may inform new models of providing outreach for tobacco treatment in health systems.

Immune checkpoint blockade (ICB) therapy revolutionized cancer treatment, but many patients with impaired MHC-I expression remain refractory. Here, we combined FACS-based genome-wide CRISPR screens with a data-mining approach to identify drugs that can upregulate MHC-I without inducing PD-L1. CRISPR screening identified TRAF3, a suppressor of the NFκB pathway, as a negative regulator of MHC-I but not PD-L1. The Traf3-knockout gene expression signature is associated with better survival in ICB-naïve patients with cancer and better ICB response. We then screened for drugs with similar transcriptional effects as this signature and identified Second Mitochondria-derived Activator of Caspase (SMAC) mimetics. We experimentally validated that the SMAC mimetic birinapant upregulates MHC-I, sensitizes cancer cells to T cell–dependent killing, and adds to ICB efficacy. Our findings provide preclinical rationale for treating tumors expressing low MHC-I expression with SMAC mimetics to enhance sensitivity to immunotherapy. The approach used in this study can be generalized to identify other drugs that enhance immunotherapy efficacy. Significance: MHC-I loss or downregulation in cancer cells is a major mechanism of resistance to T cell–based immunotherapies. Our study reveals that birinapant may be used for patients with low baseline MHC-I to enhance ICB response. This represents promising immunotherapy opportunities given the biosafety profile of birinapant from multiple clinical trials. This article is highlighted in the In This Issue feature, p. 1307

We previously described how thousands of the heme protein cytochrome c (cyt.c) self-organize into multilayered, roughly spherical superstructures as initiated by nucleation around one colloidal gold or silver nanoparticle. Within these superstructures, the protein is stabilized to unfolding in buffered media and survives superstructure encapsulation within silica gels and processing to form bioaerogels. We now report that Au∼cyt.c superstructures in buffered media are not simply static groupings of proteins, but that the Au core and protein corona exhibit dynamic electron-transfer reactions within the superstructure as verified by UV-visible and resonance Raman spectroscopy. Within the superstructure, hundreds to thousands of ferricytochrome c (FeIII-cyt.c) are reduced to ferrocytochrome c (FeII-cyt.c) following first-order kinetics with an average apparent forward rate constant of 1.9 ±0.4 × 10-5 s-1. The reducing power in the microheterogeneous medium is derived from two multielectron reductants: tannic acid used to stabilize the commercial gold sol and the Au nanoparticle at the center of the protein superstructure. Fluorescence monitoring of guanidinium chloride-induced unfolding reveals that superstructure-associated cyt.c is stabilized to unfolding before and after chemical reduction of FeIII-cyt.c to form FeII-cyt.c, indicating that the superstructures remain intact during microheterogeneous redox reactions. Smaller nucleating Au nanoparticles or lower ionic strength in the buffered medium yields a greater extent of cyt.c reduction. Partial oxidation of the cyt.c-associated nanoparticulate Au is verified by X-ray photoelectron spectroscopy. The Au nanoparticle at the heart of the superstructure functions as a direct electron donor to the heme with oxidized Au atoms being recycled back to Au(0) as long as residual tannic acid, derived from the Au sol mother liquor, is present in the aqueous microheterogeneous medium.

Batteries that offer high specific energy and energy density coupled with improved safety and lower cost will affect applications ranging from electric vehicles, portable electronic devices, and grid-level energy storage. Alkaline nickel–zinc (Ni–Zn) batteries use nonflammable aqueous electrolyte and nonstrategic, low-cost electrode materials; however with a two-electron anode, a cathode that stores more than one electron per Ni atom would increase energy density. Herein, we report the effect of substituting metal ions (aluminium, cobalt, manganese, or zinc) into α-Ni(OH)2, a phase that can accommodate more than one-electron charge storage, but which typically converts to lower-capacity β-Ni(OH)2 upon cycling in alkaline electrolytes. We adapt a microwave-assisted process that expresses α-Ni(OH)2 as a high surface-area nanosheet morphology and find that we retain this morphology with all metal-ion substituents. The series is characterized using scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and Raman spectroscopy. Metal-ion substitution influences aggregate growth, interlayer distance, and vibrational frequencies. We test powder-composite cathodes prepared using the substituted α-Ni(OH)2 series versus zinc sponge anodes in alkaline electrolyte under device-relevant mass loadings and using an intentionally aggressive charging protocol to determine onset voltage for oxygen evolution. The electrochemical charge-storage behaviour is established using galvanostatic cycling and differential capacity analysis. The substituents significantly influence both Ni-centred redox and oxygen-evolution voltages (vs. Zn/Zn2+). The incorporation of Al3+ within α-Ni(OH)2 nanosheets provides higher capacity and phase stability compared to the divalent substituents and unsubstituted α-Ni(OH)2. The presence of ordered free nitrates in the interlayer of Al3+-substituted α-Ni(OH)2, not seen with Co2+ or Mn2+ substituents, correlates with the improved electrochemical performance.

Rechargeable zinc-ion batteries represent an emerging energy-storage technology that offers the advantages of low cost, use of abundant and nontoxic materials, and competitive energy content in lightly packaged forms. Nanoscale manganese oxides are among the most promising positive-electrode materials for zinc-ion cells, and their performance is further enhanced when these oxides are expressed as conformal deposits on porous carbon architectures, such as carbon nanfoam paper (CNF). We describe an “in-place” conversion of nanometric birnessite Na+-MnOx@CNF to crystalline spinel ZnMn2O4@CNF, a manganese oxide polymorph that nominally contains sites for Zn2+ insertion. The ZnMn2O4@CNF cathodes are electrochemically conditioned in two-terminal cells and ex situ characterized using X-ray diffraction, scanning electron microscopy, energy-dispersive spectroscopy, and X-ray photoelectron spectroscopy. Despite specific Zn2+ insertion sites in ZnMn2O4, we demonstrate that the predominant discharge mechanism involves coupled insertion of protons and precipitation of Zn4(OH)6SO4·xH2O; upon recharge, protons deinsert and Zn4(OH)6SO4 dissolves.

Immune therapies have had limited efficacy in high-grade serous ovarian cancer (HGSC), as the cellular targets and mechanism(s) of action of these agents in HGSC are unknown. Here we performed immune functional and single-cell RNA sequencing transcriptional profiling on novel HGSC organoid/immune cell co-cultures treated with a unique bispecific anti-programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) antibody compared with monospecific anti-PD-1 or anti-PD-L1 controls. Comparing the functions of these agents across all immune cell types in real time identified key immune checkpoint blockade (ICB) targets that have eluded currently available monospecific therapies. The bispecific antibody induced superior cellular state changes in both T and natural killer (NK) cells. It uniquely induced NK cells to transition from inert to more active and cytotoxic phenotypes, implicating NK cells as a key missing component of the current ICB-induced immune response in HGSC. It also induced a subset of CD8 T cells to transition from naïve to more active and cytotoxic progenitor-exhausted phenotypes post-treatment, revealing the small, previously uncharacterized population of CD8 T cells responding to ICB in HGSC. These state changes were driven partially through bispecific antibody-induced downregulation of the bromodomain-containing protein BRD1. Small-molecule inhibition of BRD1 induced similar state changes in vitro and demonstrated efficacy in vivo, validating the co-culture results. Our results demonstrate that state changes in both NK and a subset of T cells may be critical in inducing an effective anti-tumor immune response and suggest that immune therapies able to induce such cellular state changes, such as BRD1 inhibitors, may have increased efficacy in HGSC. Significance: This study indicates that increased efficacy of immune therapies in ovarian cancer is driven by state changes of NK and small subsets of CD8 T cells into active and cytotoxic states.

Cation-defective iron oxides have proven to be effective Li-ion charge-storage hosts in nonaqueous electrolytes, particularly when expressed in disordered, nanoscale forms such as aerogels. Replacing a fraction of Fe sites in ferrites with high-valent cations such as V5+ introduces cation-vacancy defects that increase Li-ion capacity. Herein, we show that compositional substitution with electroinactive Al3+ further increases Li-ion capacity by 30% when incorporated within a disordered VFe2Ox aerogel, as verified by electrochemical tests in a two-terminal Li half-cell. We use electroanalytical techniques to show that both Al-VFe2Ox and VFe2Ox aerogels exhibit many of the hallmarks of pseudocapacitive materials, including fast charge–discharge and surface-controlled charge-storage kinetics. These disordered, substituted ferrites also provide the high specific capacity expected from battery-type electrode materials, up to 130 mA h g−1 for Al-VFe2Ox. Our findings are discussed in the context of related Li-insertion hosts that blur the distinctions between battery-like and capacitor-like behavior.

Patients with heart failure face increased morbidity and mortality when infected with COVID-19. The objective of this study was to evaluate the outcomes of patients with Heart Failure (HF), Left Ventricular Assist Devices (LVADs), or Heart Transplants (HTx) diagnosed with COVID-19 within an advanced HF practice.Out of 2635 patients followed, 96 patients were diagnosed with COVID-19 between March 2020 and January 2021. Median hospital length of stay (LOS), requirement for mechanical ventilation (MV), and mortality rate were evaluated.The distribution of COVID-19 among the 96 patients was: HF = 43 (45 %), LVAD = 16 (17 %) and HTx = 37 (38 %). Among 43 HF patients, 5 (12 %) died, 18 (42 %) required hospitalization with an LOS of 7 days, 5 (12 %) required ICU and 4 (9 %) required MV. Of the 16 LVAD patients, 2 (13 %) died, 8 (50 %) required hospitalization with an LOS of 11 days, 3 (19 %) required ICU and 3 (19 %) required MV. Among 37 HTx patients, 7 (19 %) died, 23 (62 %) required hospitalization with an LOS of 9 days, 6 (16 %) required ICU and 6 (16 %) required MV.This report is among the first to describe the impact of COVID-19 on a diverse advanced HF practice. It highlights the risks associated with COVID-19 faced by the HF, LVAD and HTx patients. A 90-day mortality rate of 19 % with HTx patients acquiring COVID-19 is ominous as is a mortality rate of 12 % each for HF and LVAD patients. This clinical impact should serve as a reminder of unique challenges with these populations.

Cairpol and Aeroqual air quality sensors measuring CO, CO2, NO2, and other species were tested on fresh biomass burning plumes in field and laboratory environments. We evaluated the sensors by comparing 1 min sensor measurements to collocated reference instrument measurements. The sensors were evaluated based on the coefficient of determination (r2) between the sensor and reference measurements, as well as by the accuracy, collocated precision, root mean square error (RMSE), and other metrics. In general, CO and CO2 sensors performed well (in terms of accuracy and r2 values) compared to NO2 sensors. Cairpol CO and NO2 sensors had better sensor-versus-sensor agreement (i.e., collocated precision) than Aeroqual CO and NO2 sensors of the same species. Tests of other sensors (e.g., NH3, H2S, VOC, and NMHC) provided more inconsistent results and need further study. Aeroqual NO2 sensors had an apparent O3 interference that was not observed in the Cairpol NO2 sensors. Although the sensor accuracy lags that of reference-level monitors, with location-specific calibrations they have the potential to provide useful data about community air quality and personal exposure to smoke impacts.

PurposeThe study aims to assess medical engagement levels at two teaching hospitals and a 500 bed private hospital in two states operated by the same health care provider and to describe individual and organisational factors that influence and change medical engagement.Design/methodology/approachA survey was emailed to all junior and senior medical staff, seeking responses to 30 pre-determined items. The survey used a valid and reliable instrument which provided an overall index of medical engagement. Qualitative data were also collected by including an open ended question.FindingsDoctors (n = 810) working at all sites are in the top 20-40 percentile when compared to Australia and the United Kingdom. Two sites in one state were in the highest relative engagement band with the other being in the high relative range when compared to the (UK) and the medium relative band when compared to sites in Australia. Senior doctors working at all three were less engaged on feeling valued and empowered, when compared to having purpose and direction or working in a collaborative culture. This appears to be related to work satisfaction and whether they feel encouraged to develop their skills and progress their careers. Junior doctors at 1 site are much less engaged than colleagues working at another. Since their formal training pathways are identical the informal training experience appears to be an engagement factor.Originality/valueDespite medical engagement being recognised as crucial, little is known about individual and organisational factors that support doctors to be engaged, particularly for juniors and in the private sector.

The androgen receptor (AR) is a master transcription factor that regulates prostate cancer (PC) development and progression. Inhibition of AR signaling by androgen deprivation is the first-line therapy with initial efficacy for advanced and recurrent PC. Paradoxically, supraphysiological levels of testosterone (SPT) also inhibit PC progression. However, as with any therapy, not all patients show a therapeutic benefit, and responses differ widely in magnitude and duration. In this study, we evaluated whether differences in the AR cistrome before treatment can distinguish between SPT-responding (R) and -nonresponding (NR) tumors. We provide the first preclinical evidence to our knowledge that SPT-R tumors exhibit a distinct AR cistrome when compared with SPT-NR tumors, indicating a differential biological role of the AR. We applied an integrated analysis of ChIP-Seq and RNA-Seq to the pretreatment tumors and identified an SPT-R signature that distinguishes R and NR tumors. Because transcriptomes of SPT-treated clinical specimens are not available, we interrogated available castration-resistant PC (CRPC) transcriptomes and showed that the SPT-R signature is associated with improved survival and has the potential to identify patients who would respond to SPT. These findings provide an opportunity to identify the subset of patients with CRPC who would benefit from SPT therapy.

BACKGROUND Frameworks such as the WSCC model provide evidence‐based guidance for addressing school health at the school, district, and regional level. However, frameworks do not implement themselves; they require the mobilization and collaboration of stakeholders within communities and an understanding of the unique resources and barriers within each context. Furthermore, addressing school health presents a complex systems problem. METHODS Community‐based system dynamics (CBSD) is a participatory approach for engaging communities in understanding and changing complex systems. We used a descriptive multiple case study design to evaluate how and why CBSD was used as a tool for stakeholders to engage with the complexity of school health. RESULTS We analyzed 3 cases to understand how these methods were used to enhance collaboration, analysis, and community action at multiple levels, including in 2 school districts, with a city‐wide stakeholder committee, and with a group of high school students. CONCLUSIONS Community‐based system dynamics presents a promising approach for building shared language and ownership among stakeholders, tailoring to local community contexts, and mobilizing stakeholders for action based on new system insights. We close with a discussion of unique opportunities and challenges of expanding the use of CBSD in the field of school health.

The TMPRSS2-ERG fusion is the most common genomic rearrangement in human prostate cancer. However, in established adenocarcinoma, it is unknown how the ERG oncogene promotes a cancerous phenotype and maintains downstream androgen receptor (AR) signaling pathways. In this study, we utilized a murine prostate organoid system to explore the effects of ERG on tumorigenesis and determined the mechanism underlying prostate cancer dependence on ERG. Prostate organoids lacking PTEN and overexpressing ERG (Pten−/− R26-ERG) faithfully recapitulated distinct stages of prostate cancer disease progression. In this model, deletion of ERG significantly dampened AR-dependent gene expression. While ERG was able to reprogram the AR cistrome in the process of prostate carcinogenesis, ERG knockout in established prostate cancer organoids did not drastically alter AR binding, H3K27ac enhancer, or open chromatin profiles at these reprogrammed sites. Proteomic analysis of DNA-bound AR complexes demonstrated that ERG deletion causes a loss of recruitment of critical AR coregulators and basal transcriptional machinery, including NCOA3 and RNA polymerase II, but does not alter AR binding itself. Together, these data reveal a novel mechanism of ERG oncogene addiction in prostate cancer, whereby ERG facilitates AR signaling by maintaining coregulator complexes at AR bound sites across the genome. Significance: These findings exploit murine organoid models to uncover the mechanism of ERG-mediated tumorigenesis and subsequent oncogenic dependencies in prostate cancer.

Download : Download high-res image (193KB) Download : Download full-size image Brandon J. Hopkins is a National Research Council postdoctoral fellow at the U.S. Naval Research Laboratory (NRL) focused on advancing next-generation electrochemical energy systems. Hopkins received his PhD at the Massachusetts Institute of Technology (MIT) where he worked on aqueous metal–air batteries. As a master’s candidate at MIT, he was part of the Joint Center for Energy Storage Research (JCESR) where he created gravity-driven flow batteries using semi-solid suspensions. He received his bachelor’s degree from Harvard University and interned at Akamai Technologies, Inc. and Lawrence Livermore National Laboratory. Download : Download high-res image (181KB) Download : Download full-size image Debra Rolison (left) heads and Jeffrey Long (center) and Joseph Parker (right) are members of the Advanced Electrochemical Materials Section at NRL. They design, synthesize, characterize, and prototype 3D-structured, ultraporous, multifunctional, hold-in-your-hand nanoarchitectures for such rate-critical applications as catalysis, energy storage and conversion, ultrafiltration, and sensors. They recently demonstrated that reformulating zinc into a monolithic sponge form-factor allows Zn-based batteries to be cycled at high rate to high specific energy without forming cell-shorting dendrites. They received their PhDs in Chemistry from the University of North Carolina at Chapel Hill in 1980, 1997, and 2010, respectively.

Commercial primary zinc–air batteries provide 450 Wh kgcell–1 (at the cell level), but the practical specific energy of secondary zinc–air batteries remains unclear. Using a specific-energy model a...

Tuberous sclerosis complex (TSC) is an autosomal dominant tumor suppressor syndrome, characterized by tumor development in multiple organs, including renal angiomyolipoma. Biallelic loss of TSC1 or TSC2 is a known genetic driver of angiomyolipoma development, however, whether an altered transcriptional repertoire contributes to TSC-associated tumorigenesis is unknown. RNA-seq analyses showed that MITF A isoform (MITF-A) was consistently highly expressed in angiomyolipoma, immunohistochemistry showed microphthalmia-associated transcription factor nuclear localization, and Chromatin immuno-Precipitation Sequencing analysis showed that the MITF-A transcriptional start site was highly enriched with H3K27ac marks. Using the angiomyolipoma cell line 621-101, MITF knockout (MITF.KO) and MITF-A overexpressing (MITF.OE) cell lines were generated. MITF.KO cells showed markedly reduced growth and invasion in vitro, and were unable to form xenografted tumors. In contrast, MITF.OE cells grew faster in vitro and as xenografted tumors compared to control cells. RNA-Seq analysis showed that both ID2 and Cysteine-rich angiogenic inducer 61 (CYR61) expression levels were increased in the MITF.OE cells and reduced in the MITF.KO cells, and luciferase assays showed this was due to transcriptional effects. Importantly, CYR61 overexpression rescued MITF.KO cell growth in vitro and tumor growth in vivo. These findings suggest that MITF-A is a transcriptional oncogenic driver of angiomyolipoma tumor development, acting through regulation of CYR61.

OBJECTIVE The most common endoleak after endovascular aneurysm repair is type II. Although type II endoleaks (TIIEL) are generally considered benign, there are reports that they can lead to aortic rupture. In this study, we reviewed the effect of TIIEL on sac size change to determine if sac expansion owing to a TIIEL could result in the development of a type IA endoleak (TIAEL). METHODS After internal review board approval, all aortoiliac endovascular aneurysm repairs performed at a single institution between June 2006 and June 2012 were retrospectively reviewed. Patient demographics, comorbidities, aneurysm diameter, graft type, need for reintervention, and complications were collected. Patients with TIIEL diagnosed on follow-up imaging were categorized as those who underwent intervention for their TIIEL and those who did not. Outcomes were tabulated with attention to sac size change, development of TIAEL, rupture, and survival. RESULTS Six hundred twenty-seven patients underwent aortoiliac stent graft placement at our institution during this time period. Patients with an operative indication other than nonruptured infrarenal abdominal aortic aneurysm and those without preoperative computed tomography angiography or follow-up data available for review were excluded. The total number of patients included was 389 with an average follow-up of 58.8 months (range, 0-194 months). Follow-up imaging diagnosed 124 patients with TIIEL (32%). Patients with TIIEL were significantly older (P < .0001) and more likely to be hypertensive (P < .05) but less likely to be smokers (P = .01). They had a significantly larger sac size increase than patients without TIIEL (9.50 vs -0.78 mm; P < .0001). Those with TIIEL were significantly more likely to develop a TIAEL than patients who did not have TIIEL (14% vs 5%; P = .004), but the rate of rupture was not significantly different (4% vs 2%; P = .33). In those with a TIIEL, the average sac size increase at which TIAEL developed was 13 mm. Patients in the TIIEL group who underwent intervention for their TIIEL survived significantly longer than patients who did not undergo intervention (140 months vs 100 months; P = .004). CONCLUSIONS Our data suggest that there is an increased incidence of late TIAEL in patients with TIIEL compared with those without a TIIEL. Our study also demonstrates an increased overall survival in TIIEL patients who underwent intervention. Future studies are necessary to better define the association between TIIEL with enlarging sac and the development of TIAEL. However, it is reasonable to conclude that intervention for TIIEL should be undertaken at or before a cumulative sac size increase of 13 mm.

Inspired by recent breakthroughs with Zr(OH)4-type materials that decompose chemical warfare agents (CWAs), we explore aerogel forms of zirconium oxyhydroxide (ZrOxHy) as reactive sorbents for dime...

Background With the limited number of available suitable donor hearts resulting in plateaued numbers of heart transplantations, short- and long-term mechanical circulatory support devices, including the implantation of total artificial hearts (TAHs) are modalities that are increasingly being used as treatment options for patients with end-stage heart failure. The superior vena cava syndrome has been described in this context in various disease processes. We report successful venoplasty for superior vena cava syndrome in a patient with a TAH. Case presentation A 65-year-old man with a history of nonischemic cardiomyopathy had received a left ventricular assist device, and then 2 years later, underwent orthotopic heart transplantation using the bicaval anastomosis technique. The postprocedural course was complicated by primary graft failure, resulting in the need for implantation of a TAH. About 5 months after TAH implantation, he started to develop complications such as volume retention, swelling of the upper extremities, and was diagnosed to have a superior vena cava syndrome. The patient underwent a successful venoplasty of his superior vena cava by interventional radiology with resolution of upper body edema, normalization of renal, and liver function. Conclusion Potential fatal complications caused by catheter or wire entrapment in the right-sided mechanical valve of a TAH have been reported. We describe a safe method for the treatment of superior vena cava syndrome in patients with TAH.

Nanoscale coatings of a solution-processable conducting polymer, 3,4-propylenedioxythiophene–dimethyl-3,4-propylenedioxythiophene (ProDOT-dimethyl-ProDOT, PDMP), are incorporated within a macroscal...

Although there is some evidence to support the use of hypofractionated (HF) radiation therapy (RT) postmastectomy, it is not currently the standard of care. RT noncompletion and delayed completion have been shown to lead to inferior outcomes. This study assesses the association between the choice of an HF versus conventionally fractionated regimen and completion.RT orders placed in 2016 and 2017 for patients with a national health plan, along with the associated claims, were extracted. Each order was assigned a target date for timely completion, as well as a date 30 days after the target, which was used to assess delayed completion. Univariate analyses and logistic regressions were conducted to test for an association between regimen and completion. A Poisson regression was used to examine the association between regimen and length of treatment delay among patients completing RT.Of the 743 orders meeting inclusion criteria, 56 (7.5%) were for HF. Unadjusted analyses found that the timely and delayed completion rates were significantly (P.001) higher for patients receiving HF. The adjusted odds ratios (HF order versus CF order) were 3.96 (95% confidence interval, 2.23-7.01) for timely completion and 2.64 (95% confidence interval, 1.43-5.15) for completion within 30 days of the target. Among completers, an order for HF was significantly (P .001) associated with less delay.When an HF regimen was ordered, patients were more likely to complete therapy without a delay, to complete therapy overall, and, if experiencing a delay, to experience a shorter delay.