Prouroguanylin’s role in the homeostatic response to dietary sodium: hormone precursor or active signaling agent?

High blood pressure affects more than 70 million Americans, putting them at a much greater risk for heart disease or a stroke. Sodium (Na+) is a major determinant of blood osmolarity and volume, and therefore plays a critical role in blood pressure regulation. Prouroguanylin (proUgn) has been implicated as a signal being sent from the small intestine to the kidney, in response to oral Na+ intake, that intestinal Na+ adsorption is eminent and renal Na+ excretion is necessary to maintain homeostasis. This concept, known as post-prandial natriuresis, is based on past observations of a faster natriuretic response from an oral salt load than from an equimolar intravenous infusion of NaCl. This role for proUgn is contentious however, as it has not been clearly shown if the role played by proUgn is that of a primary messenger from the intestine to the kidney in response to salt, if it is a intrarenal paracrine mechanism secondary to some other extrarenal signal in response to oral Na+ or even if it is proUgn itself or one of its' metabolites that are acting in the kidney. My studies address identification and quantification of proUgn and Ugn in relevant tissue compartments and additionally, I look at the effects of dietary Na+ on changes in the expression of proUgn and Ugn in these tissues. A novel adaptation of a binding assay technique allowed for urinary Ugn measurements that were found to directly correlate with dietary Na+ intake on a time scale that the prior techniques were insufficiently sensitive to achieve. Enteric and plasma proUgn do not change in response to dietary Na+, however a renal proUgn expression and Ugn in the urine do. Changes in plasma proUgn elicit a greater natriuretic response with a lower urinary Ugn concentration than urinary Ugn concentrations from infusions of Ugn in the blood. My conclusion is that proUgn is a secondary agent involved in volume and Na+ homeostasis, acting though some as yet unidentified renal metabolite/receptor mechanism. Further studies should focus on if there is another active cleavage product of proUgn and what receptor it (proUgn or Ugn) acts through

Landspotting: Social gaming to collect vast amounts of data for satellite validation

At present there is no single satellite-derived global land cover product that is accurate enough to provide reliable estimates of forest or cropland area to determine, e.g., how much additional land is available to grow biofuels or to tackle problems of food security. The Landspotting Project aims to improve the quality of this land cover information by vastly increasing the amount of in-situ validation data available for calibration and validation of satellite-derived land cover. The Geo-Wiki (Geo-Wiki.org) system currently allows users to compare three satellite derived land cover products and validate them using Google Earth. However, there is presently no incentive for anyone to provide this data so the amount of validation through Geo-Wiki has been limited. However, recent competitions have proven that incentive driven campaigns can rapidly create large amounts of input. The LandSpotting Project is taking a truly innovative approach through the development of the Landspotting game. The game engages users whilst simultaneously collecting a large amount of in-situ land cover information. The development of the game is informed by the current raft of successful social gaming that is available on the internet and as mobile applications, many of which are geo-spatial in nature. Games that are integrated within a social networking site such as Facebook illustrate the power to reach and continually engage a large number of individuals. The number of active Facebook users is estimated to be greater than 400 million, where 100 million are accessing Facebook from mobile devices. The Landspotting Game has similar game mechanics as the famous strategy game "Civilization" (i.e. build, harvest, research, war, diplomacy, etc.). When a player wishes to make a settlement, they must first classify the land cover over the area they wish to settle. As the game is played on the earth surface with Google Maps, we are able to record and store this land cover/land use classification geographically. Every player can play the game for free (i.e. a massive multiplayer online game). Furthermore, it is a social game on Facebook (e.g. invite friends, send friends messages, purchase gifts, help friends, post messages onto the wall, etc). The game is played in a web browser, therefore it runs everywhere (where Flash is supported) without requiring the user to install anything additional. At the same time, the Geo-Wiki system will be modified to use the acquired in-situ validation information to create new outputs: a hybrid land cover map, which takes the best information from each individual product to create a single integrated version; a database of validation points that will be freely available to the land cover user community; and a facility that allows users to create a specific targeted validation area, which will then be provided to the crowdsourcing community for validation. These outputs will turn Geo-Wiki into a valuable system for earth system scientists

An iron-oxygen intermediate formed during the catalytic cycle of cysteine dioxygenase

Cysteine dioxygenase is a key enzyme in the breakdown of cysteine, but its mechanism remains controversial. A combination of spectroscopic and computational studies provides the first evidence of a short-lived intermediate in the catalytic cycle. The intermediate decays within 20 ms and has absorption maxima at 500 and 640 nm

Identification of BPIFA1/SPLUNC1 as an epithelium-derived smooth muscle relaxing factor

Asthma is a chronic airway disease characterized by inflammation, mucus hypersecretion and abnormal airway smooth muscle (ASM) contraction. Bacterial permeability family member A1, BPIFA1, is a secreted innate defence protein. Here we show that BPIFA1 levels are reduced in sputum samples from asthmatic patients and that BPIFA1 is secreted basolaterally from healthy, but not asthmatic human bronchial epithelial cultures (HBECs), where it suppresses ASM contractility by binding to and inhibiting the Ca2+ influx channel Orai1. We have localized this effect to a specific, C-terminal α-helical region of BPIFA1. Furthermore, tracheas from Bpifa1−/− mice are hypercontractile, and this phenotype is reversed by the addition of recombinant BPIFA1. Our data suggest that BPIFA1 deficiency in asthmatic airways promotes Orai1 hyperactivity, increased ASM contraction and airway hyperresponsiveness. Strategies that target Orai1 or the BPIFA1 deficiency in asthma may lead to novel therapies to treat this disease

Robust Henderson III estimators of variance components in the nested error model

Common methods for estimating variance components in Linear Mixed Models include Maximum Likelihood (ML) and Restricted Maximum Likelihood (REML). These methods are based on the strong assumption of multivariate normal distribution and it is well know that they are very sensitive to outlying observations with respect to any of the random components. Several robust altematives of these methods have been proposed (e.g. Fellner 1986, Richardson and Welsh 1995). In this work we present several robust alternatives based on the Henderson method III which do not rely on the normality assumption and provide explicit solutions for the variance components estimators. These estimators can later be used to derive robust estimators of regression coefficients. Finally, we describe an application of this procedure to small area estimation, in which the main target is the estimation of the means of areas or domains when the within-area sample sizes are small

Spurious correlations in simultaneous EEG-fMRI driven by in-scanner movement

Simultaneous EEG-fMRI provides an increasingly attractive research tool to investigate cognitive processes with high temporal and spatial resolution. However, artifacts in EEG data introduced by the MR-scanner still remain a major obstacle. This study employing commonly used artifact correction steps shows that head motion, one overlooked major source of artifacts in EEG-fMRI data, can cause plausible EEG effects and EEG-BOLD correlations. Specifically, low frequency EEG

A novel synonymous KMT2B variant in a patient with dystonia causes aberrant splicing

BACKGROUND: Heterozygous KMT2B variants are a common cause of dystonia. A novel synonymous KMT2B variant, c.5073C>T (p.Gly1691=) was identified in an individual with childhood-onset progressive dystonia. METHODS: The splicing impact of c.5073C>T was assessed using an in vitro exon-trapping assay. The genomic region of KMT2B exons 23-26 was cloned into the pSpliceExpress plasmid between exon 2 and 3 of the rat Ins2 gene. The c.5073C>T variant was then introduced through site-directed mutagenesis. The KMT2B wild-type and c.5073C>T plasmids were transfected separately into HeLa cells and RNA was extracted 48 hours after transfection. The RNA was reverse transcribed to produce cDNA, which was PCR amplified using primers annealing to the flanking rat Ins2 sequences. RESULTS: Sanger sequencing of the PCR products revealed that c.5073C>T caused a novel splice donor site and therefore a 5-bp deletion of KMT2B exon 23 in mature mRNA, leading to a coding frameshift and premature stop codon (p.Lys1692AsnfsTer7). CONCLUSION: To our knowledge, this is the first report of a KMT2B synonymous variant associated with dystonia. Reassessment of synonymous variants may increase diagnostic yield for inherited disorders including monogenic dystonia. This is of clinical importance, given the generally favourable response to deep brain stimulation for KMT2B-related dystonia