Epidrug mediated re-expression of miRNA targeting the HMGA transcripts in pituitary cells

Transgenic mice overexpressing the high mobility group A (HMGA) genes, Hmga1 or Hmga2 develop pituitary tumours and their overexpression is also a frequent finding in human pituitary adenomas. In some cases, increased expression of HMGA2 but not that of HMGA1 is consequent to genetic perturbations. However, recent studies show that down-regulation of microRNA (miRNA), that contemporaneously target the HMGA1 and HMGA2 transcripts, are associated with their overexpression. In a cohort of primary pituitary adenoma we determine the impact of epigenetic modifications on the expression of HMGA-targeting miRNA. For these miRNAs, chromatin immunoprecipitations showed that transcript down-regulation is correlated with histone tail modifications associated with condensed silenced genes. The functional impact of epigenetic modification on miRNA expression was determined in the rodent pituitary cell line, GH3. In these cells, histone tail, miRNA-associated, modifications were similar to those apparent in human adenoma and likely account for their repression. Indeed, challenge of GH3 cells with the epidrugs, zebularine and TSA, led to enrichment of the histone modification, H3K9Ac, associated with active genes, and depletion of the modification, H3K27me3, associated with silent genes and re-expression of HMGA-targeting miRNA. Moreover, epidrugs challenges were also associated with a concomitant decrease in hmga1 transcript and protein levels and concurrent increase in bmp-4 expression. These findings show that the inverse relationship between HMGA expression and targeting miRNA is reversible through epidrug interventions. In addition to showing a mechanistic link between epigenetic modifications and miRNA expression these findings underscore their potential as therapeutic targets in this and other diseases

Effects of combined conservation practices on soil and water quality in the Central Mississippi River Basin

Conventional cultivation of claypan soils leads to soil and water quality degradation because of high runoff and associated soil erosion. The Goodwater Creek Experimental Watershed, which is part of the USDA Agricultural Research Service Benchmark Conservation Effects Assessment Project, Watershed Assessment Studies, was established to address these issues. Plot studies have highlighted trade-offs between erosion control and herbicide or nutrient runoff. There is a need for long-term field-scale evaluation of combined practices that reduce sediment, nutrient, and herbicide losses by runoff. A 36 ha field located in Missouri was under a conventional corn (Zea mays L.)-soybean (Glycine max L.) system from 1993 to 2003 with fertilizer application and tillage prior to planting in the spring. A precision agriculture system defined by two main management zones was implemented from 2004 to 2014: Wheat (Triticum aestivum L.) and soybean in 60% of the field, and corn and soybean in the remaining 40%. The system included no-till, cover crops, atrazine split-applications based on weed pressure, variable rates of nitrogen (N), and variable rates of fall-applied phosphorus (P). The objective of this study was to compare runoff water quality from the two management systems, based on flow and load duration curves, cumulative distribution functions, and conclusions from replicated plot studies. The precision agriculture system did not affect annual runoff, but it did increase the frequency of low flows. Sediment losses were reduced by 87% as a result of no-till and cover crops. Atrazine and P losses were lower than expected, despite the lack of incorporation into the soil. Atrazine losses were possibly lower because of the wheat area acting as a buffer, greater atrazine adsorption and retention in the field, and faster decay. Dissolved P losses as a fraction of applied remained the same, likely because of greater adsorption and lower runoff risk when applying P. Finally, nitrate-N (NO3-N) losses decreased and resulted in an overall decrease of N losses despite a slight increase of ammonium-N (NH4-N) losses. Explanations includeincluded a greater soil water content, a different timing of N applications, and N uptake by cover crops. Building on these successes, an aspirational management system is proposed to further improve on the performance and practicality of the precision agriculture system

Sceptical Employees as CSR Ambassadors in Times of Financial Uncertainty

This chapter offers new insights into the understanding of internal (employee) perceptions of organizational corporate social responsibility (CSR) policies and strategies. This study explores the significance of employees’ involvement and scepticism upon CSR initiatives and focuses on the effects it may have upon word of mouth (WOM) and the development of employee–organisation relationships. Desk research introduces the research questions. Data for the research questions were gathered through a self-completion questionnaire distributed in a hardcopy form to the sample. An individual’s level of scepticism and involvement appears to affect the development of a positive effect on employees’ WOM. Involvement with the domain of the investment may be a central factor affecting relationship building within the organization, and upon generation of positive WOM. The chapter offers a conceptual framework to public relations (PR) and corporate communications practitioners, which may enrich their views and understanding of the use and value of CSR for communication strategies and practices. For-profit organisations are major institutions in today’s society. CSR is proffered as presenting advantages for (at macro level) society and (micro level) the organization and its employees. Concepts, such as involvement and scepticism, which have not been rigorously examined in PR and corporate communication literature, are addressed. By examining employee perceptions, managers and academic researchers gain insights into the acceptance, appreciation and effectiveness of CSR policies and activities upon the employee stakeholder group. This will affect current and future CSR communication strategies. The knowledge acquired from this chapter may be transferable outside the for-profit sector

XTE J0111.2-7317 : a nebula-embedded X-ray binary in the SMC

The observed characteristics of the nebulosity surrounding the SMC High Mass X-ray Binary XTE J0111.2-7317 are examined in the context of three possible nebular types: SNR, bowshock and HII region. Observational evidence is presented which appears to support the interpretation that the nebulosity surrounding XTE J0111.2-7317 is an HII region. The source therefore appears to be a normal SMC Be X-ray binary (BeXRB) embedded in a locally enhanced ISM which it has photoionised to create an HII region. This is supported by observations of the X-ray outburst seen with BATSE and RXTE in 1998-1999. It exhibited characteristics typical of a giant or type II outburst in a BeXRB including large spin-up rates, Lx~10E38 erg/sq.cm-s, and a correlation between spin-up rate and pulsed flux. However, the temporal profile of the outburst was unusual, consisting of two similar intensity peaks, with the first peak of shorter duration than the second.Comment: Accepted for publication by MNRA

Updated Three-Stage Model for the Peopling of the Americas

Background: We re-assess support for our three stage model for the peopling of the Americas in light of a recent report that identified nine non-Native American mitochondrial genome sequences that should not have been included in our initial analysis. Removal of these sequences results in the elimination of an early (i.e.,40,000 years ago) expansion signal we had proposed for the proto-Amerind population. Methodology/Findings: Bayesian skyline plot analysis of a new dataset of Native American mitochondrial coding genomes confirms the absence of an early expansion signal for the proto-Amerind population and allows us to reduce the variation around our estimate of the New World founder population size. In addition, genetic variants that define New World founder haplogroups are used to estimate the amount of time required between divergence of proto-Amerinds from the Asian gene pool and expansion into the New World. Conclusions/Significance: The period of population isolation required for the generation of New World mitochondrial founder haplogroup-defining genetic variants makes the existence of three stages of colonization a logical conclusion. Thus, our three stage model remains an important and useful working hypothesis for researchers interested in the peopling of th

Water channel pore size determines exclusion properties but not solute selectivity

Aquaporins (AQPs) are a ubiquitous family of transmembrane water channel proteins. A subgroup of AQP water channels also facilitates transmembrane diffusion of small, polar solutes. A constriction within the pore, the aromatic/arginine (ar/R) selectivity filter, is thought to control solute permeability: previous studies on single representative water channel proteins suggest narrow channels conduct water, whilst wider channels permit passage of solutes. To assess this model of selectivity, we used mutagenesis, permeability measurements and in silico comparisons of water-specific as well as glycerol-permeable human AQPs. Our studies show that single amino acid substitutions in the selectivity filters of AQP1, AQP4 and AQP3 differentially affect glycerol and urea permeability in an AQP-specific manner. Comparison between in silico-calculated channel cross-sectional areas and in vitro permeability measurements suggests that selectivity filter cross-sectional area predicts urea but not glycerol permeability. Our data show that substrate discrimination in water channels depends on a complex interplay between the solute, pore size, and polarity, and that using single water channel proteins as representative models has led to an underestimation of this complexity

Active-Optical Reflectance Sensing Corn Algorithms Evaluated over the United States Midwest Corn Belt

Uncertainty exists with corn (Zea mays L.) N management due to year-to-year variation in crop N need, soil N supply, and N loss from leaching, volatilization, and denitrification. Activeoptical reflectance sensing (AORS) has proven effective in some fields for generating N fertilizer recommendations that improve N use efficiency, but locally derived (e.g., within a US state) AORS algorithms have not been tested simultaneously across a broad region. The objective of this research was to evaluate locally developed AORS algorithms across the US Midwest Corn Belt region for making in-season corn N recommendations. Forty-nine N response trials were conducted across eight states and three growing seasons. Reflectance measurements were collected and sidedress N rates (45–270 kg N ha–1 on 45 kg ha–1 increments) applied at approximately V9 corn development stage. Nitrogen recommendation rates from AORS algorithms were compared with the end-of-season calculated economic optimal N rate (EONR). No algorithm was within 34 kg N ha–1 of EONR \u3e 50% of the time. Average recommendations differed from EONR 81 to 147 kg N ha–1 with no N applied at planting and 74 to 118 kg N ha–1 with 45 kg of N ha–1 at planting, indicating algorithms performed worse with no N applied at planting. With some algorithms, utilizing red edge instead of the red reflectance improved N recommendations. Results demonstrate AORS algorithms developed under a particular set of conditions may not, at least without modification, perform very well in regions outside those within which they were developed

Improving an Active-Optical Reflectance Sensor Algorithm Using Soil and Weather Information

Active-optical reflectance sensors (AORS) use light reflectance characteristics from a crop canopy as an indicator of the plant’s N health. However, studies have shown AORS algorithms used in conjunction with measured reflectance characteristics for corn (Zea mays L.) N fertilizer rate recommendations are not consistently accurate. Our objective was to determine if soil and weather information could be utilized with an AORS algorithm developed at the University of Missouri (ALGMU) to improve in-season (~V9 corn development stage) N fertilizer recommendations. Nitrogen response trials were conducted across eight states over three growing seasons, totaling 49 sites with soils ranging in productivity. Nitrogen fertilizer rates according to the ALGMU were compared to economic optimal nitrogen rate (EONR). Without soil and weather information included, the root mean square error (RMSE) of the difference between ALGMU and EONR (MUDIFF) was 81 and 74 kg N ha–1 for treatments receiving 0 and 45 kg N ha–1 applied at planting, respectively. When ALGMU was adjusted using weather (seasonal precipitation and distribution prior to sidedress) and soil clay content, the RMSE was reduced by 24 to 26 kg N ha–1. Without adjustment, 20 and 29% of sites were within 34 kg N ha–1 of EONR with 0 and 45 kg N ha–1 at planting, respectively. But with adjustment for soil and weather data, 45 and 51% of sites were within 34 kg N ha–1 of EONR. These results show that weather and soil information could be used to improve ALGMU N recommendation performance

ANGPTL6 genetic variants are an underlying cause of familial intracranial aneurysms

BACKGROUND AND PURPOSE: To understand the role of the angiopoietin-like 6 gene (ANGPTL6) in intracranial aneurysms (IA) we investigated its role in a large cohort of familial IAs. METHODS: Inclusion of individuals with family history of IA recruited to the Genetic and Observational Subarachnoid Haemorrhage (GOSH) study. The ANGPTL6 gene was sequenced using Sanger sequencing. Identified genetic variants were compared to a control population. RESULTS: We found six rare ANGPTL6 genetic variants in 9/275 individuals with a family history of IA (3.3%), none of them were present in controls: Five missense and one nonsense mutation leading to a premature stop codon. One of these had been previously reported: c.392A>T (p.Glu131Val) on exon 2, another was very close: c.332G>A (p.Arg111His). Two further genetic variants lie within the fibrinogen-like domain of the ANGPTL6 gene, which may influence function or level of the ANGPTL6 protein. The last two missense mutations lie within the coiled-coil domain of the ANGPTL6 protein. All genetic variants were well conserved across species. CONCLUSION: ANGPTL6 genetic variants are an important cause of IA. Defective or lack of ANGPTL6 protein is therefore an important factor in blood vessel proliferation leading to IA; dysfunction of this protein is likely to cause abnormal proliferation or weakness of vessel walls. With these data, not only do we emphasise the importance of screening familial IA cases for ANGPTL6 and other genes involved in IA, but also highlight the ANGPTL6 pathway as a potential therapeutic target. CLASSIFICATION OF EVIDENCE: This is a Class III study showing some specificity of presence of the ANGPTL6 gene variant as a marker of familial intracranial aneurysms in a small subset of those with familial aneurysms

Validation of the SCID-hu Thy/Liv mouse model with four classes of licensed antiretrovirals.

BackgroundThe SCID-hu Thy/Liv mouse model of HIV-1 infection is a useful platform for the preclinical evaluation of antiviral efficacy in vivo. We performed this study to validate the model with representatives of all four classes of licensed antiretrovirals.Methodology/principal findingsEndpoint analyses for quantification of Thy/Liv implant viral load included ELISA for cell-associated p24, branched DNA assay for HIV-1 RNA, and detection of infected thymocytes by intracellular staining for Gag-p24. Antiviral protection from HIV-1-mediated thymocyte depletion was assessed by multicolor flow cytometric analysis of thymocyte subpopulations based on surface expression of CD3, CD4, and CD8. These mice can be productively infected with molecular clones of HIV-1 (e.g., the X4 clone NL4-3) as well as with primary R5 and R5X4 isolates. To determine whether results in this model are concordant with those found in humans, we performed direct comparisons of two drugs in the same class, each of which has known potency and dosing levels in humans. Here we show that second-generation antiretrovirals were, as expected, more potent than their first-generation predecessors: emtricitabine was more potent than lamivudine, efavirenz was more potent than nevirapine, and atazanavir was more potent than indinavir. After interspecies pharmacodynamic scaling, the dose ranges found to inhibit viral replication in the SCID-hu Thy/Liv mouse were similar to those used in humans. Moreover, HIV-1 replication in these mice was genetically stable; treatment of the mice with lamivudine did not result in the M184V substitution in reverse transcriptase, and the multidrug-resistant NY index case HIV-1 retained its drug-resistance substitutions.ConclusionGiven the fidelity of such comparisons, we conclude that this highly reproducible mouse model is likely to predict clinical antiviral efficacy in humans