Regulating the dorsal neural tube expression of Ptf1a through a distal 3' enhancer.

Generating the correct balance of inhibitory and excitatory neurons in a neural network is essential for normal functioning of a nervous system. The neural network in the dorsal spinal cord functions in somatosensation where it modulates and relays sensory information from the periphery. PTF1A is a key transcriptional regulator present in a specific subset of neural progenitor cells in the dorsal spinal cord, cerebellum and retina that functions to specify an inhibitory neuronal fate while suppressing excitatory neuronal fates. Thus, the regulation of Ptf1a expression is critical for determining mechanisms controlling neuronal diversity in these regions of the nervous system. Here we identify a sequence conserved, tissue-specific enhancer located 10.8kb 3' of the Ptf1a coding region that is sufficient to direct expression to dorsal neural tube progenitors that give rise to neurons in the dorsal spinal cord in chick and mouse. DNA binding motifs for Paired homeodomain (Pd-HD) and zinc finger (ZF) transcription factors are required for enhancer activity. Mutations in these sequences implicate the Pd-HD motif for activator function and the ZF motif for repressor function. Although no repressor transcription factor was identified, both PAX6 and SOX3 can increase enhancer activity in reporter assays. Thus, Ptf1a is regulated by active and repressive inputs integrated through multiple sequence elements within a highly conserved sequence downstream of the Ptf1a gene.

Comparing an Annual and a Daily Time-Step Model for Predicting Field-Scale Phosphorus Loss.

A wide range of mathematical models are available for predicting phosphorus (P) losses from agricultural fields, ranging from simple, empirically based annual time-step models to more complex, process-based daily time-step models. In this study, we compare field-scale P-loss predictions between the Annual P Loss Estimator (APLE), an empirically based annual time-step model, and the Texas Best Management Practice Evaluation Tool (TBET), a process-based daily time-step model based on the Soil and Water Assessment Tool. We first compared predictions of field-scale P loss from both models using field and land management data collected from 11 research sites throughout the southern United States. We then compared predictions of P loss from both models with measured P-loss data from these sites. We observed a strong and statistically significant ( < 0.001) correlation in both dissolved (ρ = 0.92) and particulate (ρ = 0.87) P loss between the two models; however, APLE predicted, on average, 44% greater dissolved P loss, whereas TBET predicted, on average, 105% greater particulate P loss for the conditions simulated in our study. When we compared model predictions with measured P-loss data, neither model consistently outperformed the other, indicating that more complex models do not necessarily produce better predictions of field-scale P loss. Our results also highlight limitations with both models and the need for continued efforts to improve their accuracy.

Evaluation of the APEX Model to Simulate Runoff Quality from Agricultural Fields in the Southern Region of the United States.

The Agricultural Policy Environmental eXtender (APEX) model has been widely applied to assess phosphorus (P) loss in runoff water and has been proposed as a model to support practical decisions regarding agricultural P management, as well as a model to evaluate tools such as the P Index. The aim of this study is to evaluate the performance of APEX to simulate P losses from agricultural systems to determine its potential use for refinement or replacement of the P Index in the southern region of the United States. Uncalibrated and calibrated APEX model predictions were compared against measured water quality data from row crop fields in North Carolina and Mississippi and pasture fields in Arkansas and Georgia. Calibrated models satisfactorily predicted event-based surface runoff volumes at all sites (Nash-Sutcliffe efficiency [NSE] > 0.47, |percent bias [PBIAS]| < 34) except Arkansas (NSE < 0.11, |PBIAS| < 50) but did not satisfactory simulate sediment, dissolved P, or total P losses in runoff water. The APEX model tended to underestimate dissolved and total P losses from fields where manure was surface applied. The model also overestimated sediments and total P loads during irrigation events. We conclude that the capability of APEX to predict sediment and P losses is limited, and consequently so is the potential for using APEX to make P management recommendations to improve P Indices in the southern United States.

Southern Phosphorus Indices, Water Quality Data, and Modeling (APEX, APLE, and TBET) Results: A Comparison.

Phosphorus (P) Indices in the southern United States frequently produce different recommendations for similar conditions. We compared risk ratings from 12 southern states (Alabama, Arkansas, Florida, Georgia, Kentucky, Louisiana, Mississippi, North Carolina, Oklahoma, South Carolina, Tennessee, and Texas) using data collected from benchmark sites in the South (Arkansas, Georgia, Mississippi, North Carolina, Oklahoma, and Texas). Phosphorus Index ratings were developed using both measured erosion losses from each benchmark site and Revised Universal Soil Loss Equation 2 predictions; mostly, there was no difference in P Index outcome. The derived loss ratings were then compared with measured P loads at the benchmark sites by using equivalent USDA-NRCS P Index ratings and three water quality models (Annual P Loss Estimator [APLE], Agricultural Policy Environmental eXtender [APEX], and Texas Best Management Practice Evaluation Tool [TBET]). Phosphorus indices were finally compared against each other using USDA-NRCS loss ratings model estimate correspondence with USDA-NRCS loss ratings. Correspondence was 61% for APEX, 48% for APLE, and 52% for TBET, with overall P index correspondence at 55%. Additive P Indices (Alabama and Texas) had the lowest USDA-NRCS loss rating correspondence (31%), while the multiplicative (Arkansas, Florida, Louisiana, Mississippi, South Carolina, and Tennessee) and component (Georgia, Kentucky, and North Carolina) indices had similar USDA-NRCS loss rating correspondence-60 and 64%, respectively. Analysis using Kendall's modified Tau suggested that correlations between measured and calculated P-loss ratings were similar or better for most P Indices than the models.

Applicability of models to predict phosphorus losses in drained fields: a review.

Most phosphorus (P) modeling studies of water quality have focused on surface runoff loses. However, a growing number of experimental studies have shown that P losses can occur in drainage water from artificially drained fields. In this review, we assess the applicability of nine models to predict this type of P loss. A model of P movement in artificially drained systems will likely need to account for the partitioning of water and P into runoff, macropore flow, and matrix flow. Within the soil profile, sorption and desorption of dissolved P and filtering of particulate P will be important. Eight models are reviewed (ADAPT, APEX, DRAINMOD, HSPF, HYDRUS, ICECREAMDB, PLEASE, and SWAT) along with P Indexes. Few of the models are designed to address P loss in drainage waters. Although the SWAT model has been used extensively for modeling P loss in runoff and includes tile drain flow, P losses are not simulated in tile drain flow. ADAPT, HSPF, and most P Indexes do not simulate flow to tiles or drains. DRAINMOD simulates drains but does not simulate P. The ICECREAMDB model from Sweden is an exception in that it is designed specifically for P losses in drainage water. This model seems to be a promising, parsimonious approach in simulating critical processes, but it needs to be tested. Field experiments using a nested, paired research design are needed to improve P models for artificially drained fields. Regardless of the model used, it is imperative that uncertainty in model predictions be assessed.

Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome.

Wiskott-Aldrich syndrome (WAS) is a severe X-linked primary immunodeficiency resulting from a diversity of mutations distributed across all 12 exons of the WAS gene. WAS encodes a hematopoietic-specific and developmentally regulated cytoplasmic protein (WASp). The objective of this study was to develop a gene correction strategy potentially applicable to most WAS patients by employing nuclease-mediated, site-specific integration of a corrective WAS gene sequence into the endogenous WAS chromosomal locus. In this study, we demonstrate the ability to target the integration of WAS2-12-containing constructs into intron 1 of the endogenous WAS gene of primary CD34+ hematopoietic stem and progenitor cells (HSPCs), as well as WASp-deficient B cell lines and WASp-deficient primary T cells. This intron 1 targeted integration (TI) approach proved to be quite efficient and restored WASp expression in treated cells. Furthermore, TI restored WASp-dependent function to WAS patient T cells. Edited CD34+ HSPCs exhibited the capacity for multipotent differentiation to various hematopoietic lineages in vitro and in transplanted immunodeficient mice. This methodology offers a potential editing approach for treatment of WAS using patient's CD34+ cells.

The Wiskott-Aldrich syndrome protein is required for positive selection during T-cell lineage differentiation.

The Wiskott-Aldrich syndrome (WAS) is an X-linked primary immune deficiency caused by a mutation in the WAS gene. This leads to altered or absent WAS protein (WASp) expression and function resulting in thrombocytopenia, eczema, recurrent infections, and autoimmunity. In T cells, WASp is required for immune synapse formation. Patients with WAS show reduced numbers of peripheral blood T lymphocytes and an altered T-cell receptor repertoire. In vitro, their peripheral T cells show decreased proliferation and cytokine production upon aCD3/aCD28 stimulation. It is unclear whether these T-cell defects are acquired during peripheral activation or are, in part, generated during thymic development. Here, we assessed the role of WASp during T-cell differentiation using artificial thymic organoid cultures and in the thymus of humanized mice. Although CRISPR/Cas9 WAS knockout hematopoietic stem and progenitor cells (HSPCs) rearranged the T-cell receptor and differentiated to T-cell receptor (TCR)+ CD4+ CD8+ double-positive (DP) cells similar to wild-type HSPCs, a partial defect in the generation of CD8 single-positive (SP) cells was observed, suggesting that WASp is involved in their positive selection. TCR repertoire analysis of the DP and CD8+ SP population, however, showed a polyclonal repertoire with no bias toward autoreactivity. To our knowledge, this is the first study of the role of WASp in human T-cell differentiation and on TCR repertoire generation.

Development of PLEAD: A Database Containing Event-based Runoff Phosphorus Loadings from Agricultural Fields.

Computer models are commonly used for predicting risks of runoff P loss from agricultural fields by enabling simulation of various management practices and climatic scenarios. For P loss models to be useful tools, however, they must accurately predict P loss for a wide range of climatic, physiographic, and land management conditions. A complicating factor in developing and evaluating P loss models is the relative scarcity of available measured field data that adequately capture P losses before and after implementing management practices in a variety of physiographic settings. Here, we describe the development of the P Loss in runoff Events from Agricultural fields Database (PLEAD)-a compilation of event-based, field-scale dissolved and/or total P loss runoff loadings from agricultural fields collected at various research sites located in the US Heartland and southern United States. The database also includes runoff and erosion rates; soil-test P; tillage practices; planting and harvesting rates and practices; fertilizer application rate, method, and timing; manure application rate, method, and timing; and livestock grazing density and timing. In total, >1800 individual runoff events-ranging in duration from 0.4 to 97 h-have been included in the database. Event runoff P losses ranged from <0.05 to 1.3 and 3.0 kg P ha for dissolved and total P, respectively. The data contained in this database have been used in multiple research studies to address important modeling questions relevant to P management planning. We provide these data to encourage additional studies by other researchers. The PLEAD database is available at .