MICROBIAL GENOME MINING IN UNDERSTANDING HUMAN BACTERIAL PATHOGENESIS AND CYANOBACTERIAL NATURAL PRODUCTS BIOSYNTHESIS

Recent accessibility of microbial genome sequencing data has enabled broad investigations into the nature of microbial physiology and their consequences on humans and their environment. Through an approach known as genome mining, an in silico technique that enables the identification of uncharacterized gene clusters based on sequence homology, the investigations described herein provide new insight into two important groups of microbes, namely human bacterial pathogens and cyanobacteria. In Part I of this dissertation, genome mining techniques have identified two evolutionarily-conserved cryptic biosynthetic operons in the human pathogens Acinetobacter baumannii and Pseudomonas aeruginosa. Interestingly, the findings demonstrate the significance of these gene clusters and their small molecule products to be important contributors to the pathogenesis of these organisms. As a result, the proteins encoded by these gene clusters are expected to be important targets in the development of next-generation antibiotics. In Part II, a novel platform for the investigation of natural products biosynthesis from cyanobacteria is described. Cyanobacteria have proven to be important yet relatively unexplored sources of bioactive compounds. Recent genome sequencing has indicated the substantial potential of these microbes to synthesize compounds that can be developed into new medicines. The collective findings in this dissertation demonstrate the considerable utility of emerging microbial genome sequencing information and its future impact on human health and disease

Steric antisense inhibition of AMPA receptor Q/R editing reveals tight coupling to intronic editing sites and splicing

Adenosine-to-Inosine (A-to-I) RNA editing is a post-transcriptional mechanism, evolved to diversify the transcriptome in metazoa. In addition to wide-spread editing in non-coding regions protein recoding by RNA editing allows for fine tuning of protein function. Functional consequences are only known for some editing sites and the combinatorial effect between multiple sites (functional epistasis) is currently unclear. Similarly, the interplay between RNA editing and splicing, which impacts on post-transcriptional gene regulation, has not been resolved. Here, we describe a versatile antisense approach, which will aid resolving these open questions. We have developed and characterized morpholino oligos targeting the most efficiently edited site--the AMPA receptor GluA2 Q/R site. We show that inhibition of editing closely correlates with intronic editing efficiency, which is linked to splicing efficiency. In addition to providing a versatile tool our data underscore the unique efficiency of a physiologically pivotal editing site

A mathematical framework for contact detection between quadric and superquadric surfaces

The calculation of the minimum distance between surfaces plays an important role in computational mechanics, namely, in the study of constrained multibody systems where contact forces take part. In this paper, a general rigid contact detection methodology for non-conformal bodies, described by ellipsoidal and superellipsoidal surfaces, is presented. The mathematical framework relies on simple algebraic and differential geometry, vector calculus, and on the C2 continuous implicit representations of the surfaces. The proposed methodology establishes a set of collinear and orthogonal constraints between vectors defining the contacting surfaces that, allied with loci constraints, which are specific to the type of surface being used, formulate the contact problem. This set of non-linear equations is solved numerically with the Newton-Raphson method with Jacobian matrices calculated analytically. The method outputs the coordinates of the pair of points with common normal vector directions and, consequently, the minimum distance between both surfaces. Contrary to other contact detection methodologies, the proposed mathematical framework does not rely on polygonal-based geometries neither on complex non-linear optimization formulations. Furthermore, the methodology is extendable to other surfaces that are (strictly) convex, interact in a non-conformal fashion, present an implicit representation, and that are at least C2 continuous. Two distinct methods for calculating the tangent and binormal vectors to the implicit surfaces are introduced: (i) a method based on the Householder reflection matrix; and (ii) a method based on a square plate rotation mechanism. The first provides a base of three orthogonal vectors, in which one of them is collinear to the surface normal. For the latter, it is shown that, by means of an analogy to the referred mechanism, at least two non-collinear vectors to the normal vector can be determined. Complementarily, several mathematical and computational aspects, regarding the rigid contact detection methodology, are described. The proposed methodology is applied to several case tests involving the contact between different (super)ellipsoidal contact pairs. Numerical results show that the implemented methodology is highly efficient and accurate for ellipsoids and superellipsoids.Fundação para a Ciência e a Tecnologia (FCT

Methylthioadenosine reprograms macrophage activation through adenosine receptor stimulation

Regulation of inflammation is necessary to balance sufficient pathogen clearance with excessive tissue damage. Central to regulating inflammation is the switch from a pro-inflammatory pathway to an anti-inflammatory pathway. Macrophages are well-positioned to initiate this switch, and as such are the target of multiple therapeutics. One such potential therapeutic is methylthioadenosine (MTA), which inhibits TNFα production following LPS stimulation. We found that MTA could block TNFα production by multiple TLR ligands. Further, it prevented surface expression of CD69 and CD86 and reduced NF-KB signaling. We then determined that the mechanism of this action by MTA is signaling through adenosine A2 receptors. A2 receptors and TLR receptors synergized to promote an anti-inflammatory phenotype, as MTA enhanced LPS tolerance. In contrast, IL-1β production and processing was not affected by MTA exposure. Taken together, these data demonstrate that MTA reprograms TLR activation pathways via adenosine receptors to promote resolution of inflammation. © 2014 Keyel et al

Understanding Crowd-Powered Search Groups: A Social Network Perspective

Background: Crowd-powered search is a new form of search and problem solving scheme that involves collaboration among a potentially large number of voluntary Web users. Human flesh search (HFS), a particular form of crowd-powered search originated in China, has seen tremendous growth since its inception in 2001. HFS presents a valuable test-bed for scientists to validate existing and new theories in social computing, sociology, behavioral sciences, and so forth. Methodology: In this research, we construct an aggregated HFS group, consisting of the participants and their relationships in a comprehensive set of identified HFS episodes. We study the topological properties and the evolution of the aggregated network and different sub-groups in the network. We also identify the key HFS participants according to a variety of measures. Conclusions: We found that, as compared with other online social networks, HFS participant network shares the power-law degree distribution and small-world property, but with a looser and more distributed organizational structure, leading to the diversity, decentralization, and independence of HFS participants. In addition, the HFS group has been becoming increasingly decentralized. The comparisons of different HFS sub-groups reveal that HFS participants collaborated more often when they conducted the searches in local platforms or the searches requiring a certain level of professional knowledge background. On the contrary, HFS participants did not collaborate much when they performed the search tas

The Receptor Tyrosine Kinase FGFR4 Negatively Regulates NF-kappaB Signaling

NFκB signaling is of paramount importance in the regulation of apoptosis, proliferation, and inflammatory responses during human development and homeostasis, as well as in many human cancers. Receptor Tyrosine Kinases (RTKs), including the Fibroblast Growth Factor Receptors (FGFRs) are also important in development and disease. However, a direct relationship between growth factor signaling pathways and NFκB activation has not been previously described, although FGFs have been known to antagonize TNFα-induced apoptosis. assays. FGF19 stimulation of endogenous FGFR4 in TNFα-treated DU145 prostate cancer cells also leads to a decrease in IKKβ activity, concomitant reduction in NFκB nuclear localization, and reduced apoptosis. Microarray analysis demonstrates that FGF19 + TNFα treatment of DU145 cells, in comparison with TNFα alone, favors proliferative genes while downregulating genes involved in apoptotic responses and NFκB signaling.These results identify a compelling link between FGFR4 signaling and the NFκB pathway, and reveal that FGFR4 activation leads to a negative effect on NFκB signaling including an inhibitory effect on proapoptotic signaling. We anticipate that this interaction between an RTK and a component of NFκB signaling will not be limited to FGFR4 alone

Genetic Associations of Type 2 Diabetes with Islet Amyloid Polypeptide Processing and Degrading Pathways in Asian Populations

10.1371/journal.pone.0062378PLoS ONE86

Neuroanatomical heterogeneity and homogeneity in individuals at clinical high risk for psychosis.

Individuals at Clinical High Risk for Psychosis (CHR-P) demonstrate heterogeneity in clinical profiles and outcome features. However, the extent of neuroanatomical heterogeneity in the CHR-P state is largely undetermined. We aimed to quantify the neuroanatomical heterogeneity in structural magnetic resonance imaging measures of cortical surface area (SA), cortical thickness (CT), subcortical volume (SV), and intracranial volume (ICV) in CHR-P individuals compared with healthy controls (HC), and in relation to subsequent transition to a first episode of psychosis. The ENIGMA CHR-P consortium applied a harmonised analysis to neuroimaging data across 29 international sites, including 1579 CHR-P individuals and 1243 HC, offering the largest pooled CHR-P neuroimaging dataset to date. Regional heterogeneity was indexed with the Variability Ratio (VR) and Coefficient of Variation (CV) ratio applied at the group level. Personalised estimates of heterogeneity of SA, CT and SV brain profiles were indexed with the novel Person-Based Similarity Index (PBSI), with two complementary applications. First, to assess the extent of within-diagnosis similarity or divergence of neuroanatomical profiles between individuals. Second, using a normative modelling approach, to assess the 'normativeness' of neuroanatomical profiles in individuals at CHR-P. CHR-P individuals demonstrated no greater regional heterogeneity after applying FDR corrections. However, PBSI scores indicated significantly greater neuroanatomical divergence in global SA, CT and SV profiles in CHR-P individuals compared with HC. Normative PBSI analysis identified 11 CHR-P individuals (0.70%) with marked deviation (>1.5 SD) in SA, 118 (7.47%) in CT and 161 (10.20%) in SV. Psychosis transition was not significantly associated with any measure of heterogeneity. Overall, our examination of neuroanatomical heterogeneity within the CHR-P state indicated greater divergence in neuroanatomical profiles at an individual level, irrespective of psychosis conversion. Further large-scale investigations are required of those who demonstrate marked deviation