Search

Abstract Tailwater recovery (TWR) systems, which divert phosphorus‐rich drainage water from cranberry (Vaccinium macrocarpon Ait.) farms into reservoirs, have the potential to improve water quality of freshwater lakes in Massachusetts. However, residents and environmentalists have questioned the potential water quality benefits of TWR systems. In the southeastern United States, research shows that TWR systems decrease agricultural inputs of phosphorus (P) to surface water by 23%–92%. Additionally, a case study in Massachusetts demonstrated the efficacy of TWR and other best management practices in reducing P concentrations in White Island Pond. Although TWR systems appear effective as part of a P management strategy, more research is needed to quantify their environmental benefits and allay public concerns. We propose filling three critical research gaps to strengthen and broaden support for TWR systems in cranberry agriculture in Massachusetts: (1) quantifying physical properties, (2) quantifying water storage potential, and (3) quantifying P retention capacity. Core Ideas Excess phosphorus (P) from cranberry farms may contribute to the eutrophication of freshwater lakes. Agricultural tailwater recovery (TWR) systems are increasingly used to conserve water and improve water quality. Earth removal, critical to create TWR ponds and provide sand for cranberry farms, has drawn public opposition. Research and case studies indicate TWR systems may decrease P inputs from cranberry farms to surface water. Long‐term monitoring, research, and stakeholder engagement are needed to assess efficacy of cranberry TWR systems.

Background Adoptive cell therapy, such as chimeric antigen receptor (CAR)-T cell therapy, has improved patient outcomes for hematological malignancies. Currently, four of the six FDA-approved CAR-T cell products use the FMC63-based αCD19 single-chain variable fragment, derived from a murine monoclonal antibody, as the extracellular binding domain. Clinical studies demonstrate that patients develop humoral and cellular immune responses to the non-self CAR components of autologous CAR-T cells or donor-specific antigens of allogeneic CAR-T cells, which is thought to potentially limit CAR-T cell persistence and the success of repeated dosing.Methods In this study, we implemented a one-shot approach to prevent rejection of engineered T cells by simultaneously reducing antigen presentation and the surface expression of both Classes of the major histocompatibility complex (MHC) via expression of the viral inhibitors of transporter associated with antigen processing (TAPi) in combination with a transgene coding for shRNA targeting class II MHC transactivator (CIITA). The optimal combination was screened in vitro by flow cytometric analysis and mixed lymphocyte reaction assays and was validated in vivo in mouse models of leukemia and lymphoma. Functionality was assessed in an autologous setting using patient samples and in an allogeneic setting using an allogeneic mouse model.Results The combination of the Epstein-Barr virus TAPi and an shRNA targeting CIITA was efficient and effective at reducing cell surface MHC classes I and II in αCD19 ‘stealth’ CAR-T cells while retaining in vitro and in vivo antitumor functionality. Mixed lymphocyte reaction assays and IFNγ ELISpot assays performed with T cells from patients previously treated with autologous αCD19 CAR-T cells confirm that CAR T cells expressing the stealth transgenes evade allogeneic and autologous anti-CAR responses, which was further validated in vivo. Importantly, we noted anti-CAR-T cell responses in patients who had received multiple CAR-T cell infusions, and this response was reduced on in vitro restimulation with autologous CARs containing the stealth transgenes.Conclusions Together, these data suggest that the proposed stealth transgenes may reduce the immunogenicity of autologous and allogeneic cellular therapeutics. Moreover, patient data indicate that repeated doses of autologous FMC63-based αCD19 CAR-T cells significantly increased the anti-CAR T cell responses in these patients.

FOXG1 syndrome is a neurodevelopmental disorder caused by mutations in the FOXG1 gene. Here, an induced pluripotent stem cell (iPSC) line was generated from human dermal fibroblasts of an individual with the c.490dupG (p.Glu154fs) mutation in the FOXG1 gene. Fibroblasts were reprogrammed using non-integrating episomal plasmids and pluripotency marker expression was confirmed by both immunocytochemistry and quantitative PCR in the resultant iPSC line. There were no karyotypic abnormalities and the cell line successfully differentiated into all three germ layers. This cell line may prove useful in the study of the pathogenic mechanisms that underpin FOXG1 syndrome.

The Pseudomonas syringae acetyltransferase HopZ1a is delivered into host cells by the type III secretion system to promote bacterial growth. However, in the model plant host Arabidopsis thaliana, HopZ1a activity results in an effector-triggered immune response (ETI) that limits bacterial proliferation. HopZ1a-triggered immunity requires the nucleotide-binding, leucine-rich repeat domain (NLR) protein, ZAR1, and the pseudokinase, ZED1. Here we demonstrate that HopZ1a can acetylate members of a family of 'receptor-like cytoplasmic kinases' (RLCK family VII; also known as PBS1-like kinases, or PBLs) and promote their interaction with ZED1 and ZAR1 to form a ZAR1-ZED1-PBL ternary complex. Interactions between ZED1 and PBL kinases are determined by the pseudokinase features of ZED1, and mutants designed to restore ZED1 kinase motifs can (1) bind to PBLs, (2) recruit ZAR1, and (3) trigger ZAR1-dependent immunity in planta, all independently of HopZ1a. A ZED1 mutant that mimics acetylation by HopZ1a also triggers immunity in planta, providing evidence that effector-induced perturbations of ZED1 also activate ZAR1. Overall, our results suggest that interactions between these two RLCK families are promoted by perturbations of structural features that distinguish active from inactive kinase domain conformations. We propose that effector-induced interactions between ZED1/ZRK pseudokinases (RLCK family XII) and PBL kinases (RLCK family VII) provide a sensitive mechanism for detecting perturbations of either kinase family to activate ZAR1-mediated ETI.

Detailed knowledge of the shape of the seafloor is crucial to humankind. Bathymetry data is critical for safety of navigation and is used for many other applications. In an era of ongoing environmental degradation worldwide, bathymetry data (and the knowledge derived from it) play a pivotal role in using and managing the world’s oceans in a way that is in accordance with the United Nations Sustainable Development Goal 14 – conserve and sustainably use the oceans, seas and marine resources for sustainable development. However, the vast majority of our oceans is still virtually unmapped, unobserved, and unexplored. Only a small fraction of the seafloor has been systematically mapped by direct measurement. The remaining bathymetry is predicted from satellite altimeter data, providing only an approximate estimation of the shape of the seafloor. Several global and regional initiatives are underway to change this situation. This paper presents a selection of these initiatives as best practice examples for bathymetry data collection, compilation and open data sharing as well as the Nippon Foundation-GEBCO (The General Bathymetric Chart of the Oceans) Seabed 2030 Project that complements and leverages these initiatives and promotes international collaboration and partnership. Several non-traditional data collection opportunities are looked at that are currently gaining momentum as well as new and innovative technologies that can increase the efficiency of collecting bathymetric data. Finally, recommendations are given toward a possible way forward into the future of seafloor mapping and toward achieving the goal of a truly global ocean bathymetry.

TRIM5α is an interferon inducible restriction factor which contributes to intrinsic defense against HIV infection by targeting the HIV capsid protein CA. Although human TRIM5α (huTRIM5α) does not potently inhibit HIV-1 infection, the ability of huTRIM5α to exhibit some control of HIV-1 infection is evidenced by a single nucleotide polymorphism in huTRIM5α which substitutes aspartic acid to glycine at position 249 (G249D) in the L2 region and is associated with higher susceptibility to HIV-1 infection. To understand the mechanistic basis for the reduced antiviral activity, we employed biophysical and cell biological methods coupled with molecular dynamics simulations to compare WT and the G249D polymorphism of huTRIM5α. We investigated the differences in conformational dynamics of rhesus and huTRIM5α Coiled Coil-Linker 2 (CC-L2) dimers utilizing circular dichroism and single molecule-Fluorescence Energy Transfer (sm-FRET). These methods revealed that the G249D dimer exhibits secondary structure and conformational dynamics similar to WT huTRIM5α. Homology modelling revealed that G249 was present on the hairpin of the antiparallel dimer, in a position which may act to stabilize the adjacent BBox2 domain which mediates the inter-dimeric contacts required for the formation of TRIM5 assemblies. We therefore asked if the G249D mutant forms assemblies in cells with the same efficiency as WT protein by expressing these proteins as YFP fusions and quantifying the number of assemblies in cells. In cells expressing comparable amounts of protein, the G249D mutant formed fewer assemblies than WT protein, in agreement with our homology modeling predictions and molecular dynamics simulations of dimers and higher oligomers of TRIM5α, providing a mechanistic explanation of the reduced antiviral activity of the G249D polymorphism.

Activation of G protein-coupled receptors (GPCRs) by agonist ligands is mediated by a transition from an inactive to active receptor conformation. We describe a novel single-molecule assay that monitors activation-linked conformational transitions in individual GPCR molecules in real-time. The receptor is site-specifically labeled with a Cy3 fluorescence probe at the end of trans-membrane helix 6 and reconstituted in phospholipid nanodiscs tethered to a microscope slide. Individual receptor molecules are then monitored over time by single-molecule total internal reflection fluorescence microscopy, revealing spontaneous transitions between inactive and active-like conformations. The assay provides information on the equilibrium distribution of inactive and active receptor conformations and the rate constants for conformational exchange. The experiments can be performed in the absence of ligands, revealing the spontaneous conformational transitions responsible for basal signaling activity, or in the presence of agonist or inverse agonist ligands, revealing how the ligands alter the dynamics of the receptor to either stimulate or repress signaling activity. The resulting mechanistic information is useful for the design of improved GPCR-targeting drugs. The single-molecule assay is described in the context of the β2 adrenergic receptor, but can be extended to a variety of GPCRs.

OBJECTIVE:To evaluate the efficacy, safety, and maternal satisfaction of a newly established integrative postpartum community-based clinic providing comprehensive support for mothers during the first month after discharge from the hospital. Our primary interests were breastfeeding rates, readmission and patient satisfaction. METHODS:A randomized controlled trial was conducted in Ottawa, Canada, where 472 mothers were randomized via a 1:2 ratio to either receive standard of care (n = 157) or to attend the postpartum breastfeeding clinic (n = 315). Outcome data were captured through questionnaires completed by the participants at 2, 4, 12 and 24 weeks postpartum. Unadjusted and adjusted logistic regression models were conducted to determine the effect of the intervention on exclusive breastfeeding at 12 weeks (primary outcome). Secondary outcomes included breastfeeding rate at 2, 4 and 24 weeks, breastfeeding self-efficacy scale, readmission rate, and satisfaction score. RESULTS:More mothers in the intervention group (n = 195, 66.1%) were exclusively breastfeeding at 12 weeks compared to mothers in the control group (n = 81, 60.5%), however no statistically significant difference was observed (OR = 1.28; 95% CI:0.84-1.95)). The rate of emergency room visits at 2 weeks for the intervention group was 11.4% compared to the standard of care group (15.2%) (OR = 0.69; 95% CI: 0.39-1.23). The intervention group was significantly more satisfied with the overall care they received for breastfeeding compared to the control group (OR = 1.96; 95% CI: 3.50-6.88)). CONCLUSION:This new model of care did not significantly increase exclusive breastfeeding at 12 weeks. However, there were clinically meaningful improvements in the rate of postnatal problems and satisfaction that support this new service delivery model for postpartum care. A community-based multidisciplinary postpartum clinic is feasible to implement and can provide appropriate and highly satisfactory care to mother-baby dyads. This model of care may be more beneficial in a population that is not already predisposed to breastfeed. TRIAL REGISTRATION:ClinicalTrials.gov NCT02043119.

Transmedia storytelling involves telling a story using multiple distinct media. The remit of stories that fall under this broad definition is vast, consequently causing theorists to examine different phenomena using tools that are not suitable for all forms of transmedia storytelling. The lack of critical tools means we are unable to describe, compare and analyse different experiences using common language. In this paper, we present our model that can be used to identify the fundamental structural features of a variety of transmedia storytelling forms. We illustrate its usage with twenty case studies and discuss how three groups of patterns emerge which can be identifiable in all transmedia stories. These patterns can be used to extend transmedia language and help form taxonomies, by identifying common patterns and their usages amongst various forms of transmedia stories.

Commercial sod farms occupy about 1.62 x 10³ km² of the landscape of the United States. Land managers generally consider sod farms on an equal footing with other, sustainable agricultural land uses. We measured soil losses associated with sod harvesting in farms in the northeastern United States. Sod harvest resulted in soil losses ranging from 74 to 114 Mg ha-1 yr-1, considerably higher than the tolerable soil loss of 6.7 Mg ha-1 yr-1. Soil losses were proportional to time under sod production, with soil removal rates of 0.833 cm yr-1. We estimate that sod harvesting in the United States results in the net, permanent loss of 12.0 to 18.7 Tg of agriculturally productive soil from sod farms-and associated ecosystem services-every year. The soil losses reported here have important implications in terms of land use planning, transactions involving the purchase of development rights, and tax deductions for soil depletion. [ABSTRACT FROM AUTHOR]